GB2594032A - Methods and compositions comprising carboxylic acids - Google Patents

Abstract

A composition comprising one or more carboxylic acid or derivatives thereof for the prevention and/or treatment of a porcine viral infection is provided. Preferably, the carboxylic acid is selected from formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, lauric acid, myristic acid, palmitic acid, stearic acid and oleic acid. The porcine viral infection is preferably selected from African Swine Fever Virus (ASFV), Porcine Reproductive and Respiratory Syndrome (PRRSV), Senecavirus A (SVA, also known as Seneca Valley Virus), Porcine Epidemic Diarrhoea Virus (PEDv), ORF virus (OV), pseudorabies virus (PRV) and classical swine fever (CSF). A composition comprising formic acid and propionic acid (pHorce®) in treating PEDv in pigs is exemplified.

Description

Methods and Compositions Comprising Carboxylic Acids The invention relates to a composition comprising one or more carboxylic acids or derivatives thereof for the prevention and/or treatment of viral infection in swine. The invention also relates to a method of preventing and/or treating porcine viral infection. In certain embodiments, the invention relates to a composition comprising one or more carboxylic acids or derivatives for inhibiting viral pathogens in a viral infected feed.

Over the last decade, the number of serious diseases affecting porcine populations with significant socio-economic consequences has increased. Controlling the spread of these diseases -many of which are caused by viral infection -has been extremely challenging, with wide scale culling of infected animals.

ASFV (African Swine Fever Virus) causes African Swine Fever in pigs. It is a large, enveloped DNA virus belonging to the Asfivirus genus having a double stranded DNA genome of approx 190kb pairs. It comprises a capsid having several concentric lipid and protein layers below an external lipid membrane. ASFV in pigs is characterised by progressive decrease in cellular protein synthesis (and increase in viral protein synthesis) leading to death of -90% of the herd. Notification of the disease and whole herd culling are the only current mitigating factors for sero-positive herds as there is no cure.

Porcine Reproductive and Respiratory Syndrome virus (PRRSV) infects domestic pigs. PRRSV is an enveloped virus belonging to the family Arteriviridae, genus Porartevirus having a capsid that encases a single stranded RNA genome. PPRSv causes massive losses for the pig industry each year through reproductive failure, increased mortality and reduced growth. There is currently no cure for PRRSv.

Senecavirus A (SVA) (formerly known as Seneca Valley virus) causes a vesicular disease in pigs characterized by lethargy, anorexia, fever, lameness, and vesicles on the snout and coronary bands. SVA in classified as the genus, Senecavirus, family Picornaviridae. It is a single-stranded positive-sense, nonenveloped RNA virus with an RNA genome of 7.2kb long.

Porcine Epidemic Diarrhoea virus or PEDv is caused by a virus from the Coronaviridae family of the genus Coronavirus. The structure of these viruses includes proteins of a spike (S), envelope (E), membrane (M) and nucleocapsid (N). When viewed using electron microscopy, coronaviruses have a characteristic "halo" or "crown" appearance, corresponding to the viral spike (5) proteins on their surface. PEDv comprises a single-stranded, positive-sense RNA genome of about 30kb in size. The RNA is enveloped in virus nucleocapsid protein, surrounded by a lipid envelope containing membrane glycoprotein, envelope protein and a homotrimer complex of virus spike (5) glycoprotein, in which three molecules constitute a spike on the virion that mediates cell entry.

PEDv in pigs is characterised by watery diarrhoea, weight loss and vomiting and it closely resembles transmissible gastroenteritis. Transmission can occur via the faecal-oral route. During the infection, viral particles are shed in the faeces. Infection of healthy animals occurs via the ingestion of contaminated faeces.

Other common porcine viral infections include ORF virus (OV), pseudorabies virus (PRV) and classical swine fever (C5F).

Furthermore, it has been shown that secondary bacterial infections such as E. Coli and Salmonella can further complicate the picture.

These viral infections are notoriously difficult to prevent and treat. Spread of the viruses has caused significant economic loss in recent years. Many current treatments concentrate on treating only the secondary bacterial infections. For some of these viruses there are a small number of licensed vaccines available. For others, no successful vaccines have been developed. Further many vaccines are restricted to certain geographical areas and commercially available vaccines do not exist elsewhere and are not likely to be cost-effective.

immunoprophylactic agents have also been suggested as a way to treat PEDv. One report (2014) has also suggested the use of formaldehyde in animal feed. Many control initiatives rely heavily on biosecurity programmes and rigorous disinfection using agents such as formaldehyde, quaternary ammonium compounds and calcium hydroxide (lime), which may have harmful effects on the animals and humans working with these chemicals.

It would be desirable to provide an improved composition for the prevention and/or treatment of porcine viral infection.

Summary of Invention

One aspect of the invention provides a composition comprising one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.

One aspect of the invention provides a composition comprising carvacrol and thymol and/or one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.

One aspect of the invention provides the use of a composition comprising one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.

One aspect of the invention provides a composition comprising one or more carboxylic acids or derivative thereof for inhibiting viral pathogens in a pig feed.

One aspect of the invention provides a composition comprising one or more carboxylic acids or derivative thereof for prevention transmission of viral pathogens from an animal feed to an animal.

One aspect of the invention provides the use of a composition comprising one or more carboxylic acids or derivative thereof for prevention transmission of viral pathogens from an animal feed to an animal.

The animal may be a porcine animal and the feed may be a pig feed.

Advantageously, the composition may be provided as a dietary supplement and/or a disinfectant in the feed of pigs.

In certain embodiments, the porcine viral infection comprises one or more of porcine Coronavirus infection; porcine Asfi virus infection; porcine Seneca virus infection or porcine Porartevirus infection.

In certain embodiments, the porcine viral infection or viral pathogens comprises one or more of ASVF (African Swine Fever Virus), Porcine Epidemic Diarrhoea virus or PED, Senecavirus (SVA).

In certain embodiments, the porcine viral infection or viral pathogens comprises one or more of ORE virus (OV), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV) and classical swine fever (CSF).

The porcine viral infection may comprise a plurality of viruses.

In one embodiment the porcine Coronavirus infection comprises porcine epidemic diarrhoea virus. Optionally, the composition comprises a carvacrol and thymol composition comprising oregano oil.

In certain embodiments, the composition comprises a carboxylic acid composition comprising one or more carboxylic acids as a free acid.

Optionally, the one or more derivative comprises an ammonium salt.

Optionally, the one or more derivative comprises an ester. The ester may be a glyceride ester.

In a preferred embodiment, the one or more carboxylic acids comprises a mid chain carboxylic acid.

The carboxylic acid may be any carboxylic acid selected from Cl to C12.

The carboxylic acid may be selected from: formic acid, propionic acid, acetic acid, butyric acid.

In certain embodiment, the carboxylic acid comprises a lipophilic carboxylic acid.

In certain embodiments, the one or more carboxylic acids comprises formic acid and propionic acid.

Optionally, the carboxylic acid composition comprises formic acid as an active ingredient at around 25 to 40 wt%.

Preferably, the carboxylic acid composition comprises formic acid as an active ingredient at around 25 to 35 wt%.

More preferably, the carboxylic acid composition comprises formic acid as an active ingredient at around 30 to 35 wt%.

Optionally, the carboxylic acid composition comprises propionic acid as an active ingredient at around 5 to 25 wt%.

Preferably, the carboxylic acid composition comprises propionic acid as an active ingredient at around 15 to 25 wt% More preferably, the carboxylic acid composition comprises propionic acid as an active ingredient at around 15 to 20 wt%.

In one embodiment, the carvacrol and thymol composition comprises oregano oil at around 1 to 15 wt%.

In one embodiment, the carvacrol and thymol composition comprises oregano oil at around 0.5 to 15 wt%.

In one embodiment, the carvacrol and thymol composition comprises oregano oil at around 0.03 to 15 wt%.

Preferably, the carvacrol and thymol composition comprises oregano oil at around 1 to 10 wt%.

More preferably, the carvacrol and thymol composition comprises oregano oil at around 3 to 6 wt%.

More preferably, the carvacrol and thymol composition comprises oregano oil at around 5 wt%.

In one embodiment, the oregano oil is natural oregano oil comprising carvacrol at around 76-84 wt% and thymol at around 0.5-4.0 wt%.

In certain embodiments, the carvacrol and thymol composition comprises carvacrol as an active ingredient at around 2 to 5 wt%.

More preferably, the carvacrol and thymol composition comprises carvacrol as an active ingredient at around 3 to 4.5 wt%.

In certain embodiments, the carvacrol and thymol composition comprises thymol as an active ingredient at around 0.01 to 0.5 wt%.

More preferably, the carvacrol and thymol composition comprises thymol as an active ingredient at around 0.02 to 0.3 wt%.

The composition may comprise a liquid or a substantially dry solid.

The composition may further comprise a carrier.

Optionally, the carrier is a purified silicic acid and is free from respirable silicas and fibrous materials.

The carrier may be a micropearl carrier.

The carrier may be a soluble silica carrier.

Advantageously, the composition may comprise carvacrol and thymol and one or more carboxylic acids or derivatives thereof on the same carrier.

One aspect of the invention comprises an animal feed comprising a composition comprising carvacrol and thymol and/or one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.

In a preferred embodiment the animal feed comprises the composition at around 0.5 to 10kg per 1000kg of feed, more preferably around 1 to 5kg per 1000kg of feed and even more preferably 2 to 4kg per 1000kg of feed.

In a preferred embodiment the animal feed comprises the composition at around 0.5 to 10kg per 1000kg of feed, more preferably around 1 to 6kg per 1000kg of feed and even more preferably 2 to 4kg per 1000kg of feed.

One aspect of the invention comprises an animal feed additive comprising a composition comprising carvacrol and thymol and/or one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.

Another aspect of the invention comprises the use of a composition comprising carvacrol and thymol and/or one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.

Another aspect of the invention comprises the use of a composition comprising one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.

Another aspect of the invention comprises the use of a composition comprising carvacrol and thymol and/or one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.

In one embodiment the porcine viral infection comprises porcine epidemic diarrhoea virus.

Another aspect of the invention comprises a method of preventing and/or treating porcine viral infection comprising administering a composition comprising carvacrol and thymol and/or one or more carboxylic acids or derivative thereof to a subject.

Another aspect of the invention comprises a method of preventing and/or treating porcine viral infection comprising administering a composition comprising carvacrol and thymol and/or one or more carboxylic acids or derivative thereof to a feed and/or local environment of a subject.

In one embodiment the composition comprises one or more carboxylic acids and their ammonium salts.

Preferably, the subject is a porcine subject.

Optionally, the subject is a lactating sow.

In certain embodiments, the subject is a gestating sow. Optionally, the subject is a piglet.

Preferably, the composition is administered orally. The composition may be administered as a feed. Preferably, the composition comprises a gestation feed. The composition may be administered by oral drenching.

In certain embodiments, the composition comprises Orego-stim ® Meriden Animal Health) and/or pHorce® (Kiotechagil) The porcine coronavirus infection may comprise Type I and/or Type II porcine epidemic diarrhoea virus.

In a preferred embodiment, the method comprises the step of providing a carvacrol and thymol composition and/or carboxylic acid composition in a feed at around 2-4kg per 1000kg.

In one embodiment the animal feed comprises a carvacrol and thymol composition at around lkg per metric ton of feed.

In one embodiment the animal feed comprises a carboxylic acid composition at around 2kg per metric ton of feed.

In one embodiment the animal feed comprises a carboxylic acid composition at around 4kg per metric ton of feed.

In one embodiment the animal feed comprises a carvacrol and thymol composition at around lkg per metric ton and a carboxylic acid composition at around 2kg per metric ton of feed.

In one embodiment the animal feed comprises a carvacrol and thymol composition at around 2kg per metric ton of feed.

In one embodiment the animal feed comprises a carvacrol and thymol composition at around 2kg per metric ton and a carboxylic acid composition at around 4kg per metric ton of feed.

More preferably, the method comprises the step of providing a carvacrol and thymol composition and/or carboxylic acid composition in a feed at around 4kg per 1000kg.

Preferably, the method comprises the step of feeding the carvacrol and thymol composition and/or carboxylic acid composition in a feed to a pig.

The composition may inhibit the virus in a viral-infected feed.

The carboxylic acid composition may comprise one or more carboxylic acids or derivate thereof. Advantageously, the method provides a reduction in the pre-weaning mortality rate of piglets. Advantageously, the method provides a higher weaning weight of piglets.

Yet another aspect of the invention comprises a method of reducing the pre-weaning mortality rate of piglets and providing a higher weaning weight of piglets preventing and/or treating porcine coronavirus infection comprising administering a composition comprising carvacrol and thymol and/or one or more carboxylic acids or derivative thereof to a subject.

In one embodiment, the subject comprises a lactating sow. In one embodiment the subject comprises a piglet.

Brief Description of the Figures

Figure 1 shows pre-weaning Mortality Rate and Sow Culling Rate before and after the inclusion of the carvacrol and thymol composition in the diet.

Figure 2 shows Litter weight at Weaning.

Detailed Description

It has been surprisingly found that a composition comprising carboxylic acid is effective in preventing and/or treating porcine viral infection.

In this document, the term porcine is intended to mean any animal of the genus Sus and any related animals including domestic pigs, wild boar and warthogs.

When added to the feed of pigs, the carboxylic acid composition is believed to accelerate the speed of disinfection, providing a powerful preventative anti-viral in feed and the local environment. The blend of carboxylic acids provides improved efficacy over a single acid. A particular blend of propionic and formic acid provides powerful anti-viral properties. The addition of carvacrol and thymol may further impede transmission of viruses from feed and the environment, while also helping to combat any secondary infections in infected animals. A silica carrier system improves efficacy of the anti-viral.

Advantageously, the composition may be provided as a dietary supplement and a disinfectant in the feed of pigs.

During a localised outbreak of PEDv, a carvacrol and thymol composition added to feed in neighbouring sow barns in lactation and gestation prevented PEDv outbreak in animals receiving the supplement.

In addition to the preventative effects, it has also been demonstrated in a number of trials including PEDv infected animals that the composition is effective as a direct treatment of the virus itself, as well as its secondary symptoms.

Trials utilising oregano oil-based Orego-stim® (Meriden Animal Health) as a composition comprising carvacrol and thymol and pHorce® (Kiotechagil) as a carboxylic acid composition showed effective treatment of PEDv pigs when included in feed at inclusion rates of around 1kg per metric ton or 2lbs per short ton of carvacrol and thymol composition and around 2kg per metric ton or 4lbs per short ton of carboxylic acid composition.

A combined treatment comprising combining compositions or a single composition containing carvacrol, thymol and a carboxylic acid (or carboxylic acid derivative) would be effective at lower inclusion rates in each case, such as from around 0.5 to 1kg per metric ton carvacrol and thymol composition and from around 1 to 2kg per metric ton carboxylic acid composition.

Natural products, such as Orego-Stim®, made using oregano oil containing the natural phenols of carvacol and thymol, have been shown to support intestinal health. The indirect action of those phenols prevents the replication of the virus in the intestinal epithelial cells by speeding up the shedding process of the enterocytes. A quick replenishment of these cells creates a hostile environment for the virus disrupting its multiplication, thus controlling PEDy in piglets.

The combined action of carboxylic acids together with phenols carvacrol and thymol is believed to provide a particularly effective treatment for the prevention and/or treatment of PEEN due to a synergistic effect involving a combined attack of short chain lipids in the virion spike by lipophilic propionic acid together with the phenols, allowing phenols, propionic and formic acid into the virus.

Furthermore, treatment with carvacrol and thymol formulations increases the intestinal dimensions and the villus height-crypt ratio in pigs. The larger villi ensure a more absorptive surface for nutrients and electrolytes, combating villus atrophy and preventing maldigestion and malabsorption.

Carvacrol and thymol in oregano oil can further help with the treatment of secondary gastro-intestinal infections. Oregano oil has known antibacterial properties and can kill bacteria within the gastrointestinal tract, preventing further complications. It has also been shown to significantly increase intestinal enzyme activity such as alkaline phosphate, leucine, aminopeptidase, maltase, sucrase and lactase within the gut, further aiding digestion and electrolyte nutrient absorption. Carboxylic acids combined with oregano oil in the animal feed may also act as a feed inhibitor, inhibiting the virus and other pathogens in the feed.

When included in the ration of lactating sows, oregano oil preserves the lactation production rate. The milk yield of the sow is crucial for suckling piglet survivability during the first couple of weeks, as it contains all the essential antibodies needed by the piglet to combat infections. It has further been shown that the inclusion of carvacrol and thymol in lactating sow diets results in the more efficient utilisation of the feed offered, which helps the sow to preserve lactation production.

In a trial where 2000 sows had a PEDy outbreak, it has been found that the inclusion of an oregano oil based product comprising carvacrol and thymol effectively improved the PEDv cases.

The inclusion of Orego-Stim® for the sows started one week before farrowing until 25 days after farrowing. The piglets were supplemented with Orego-Stim® Liquid through oral drenching.

From Figure land Figure 2, it can be seen that this inclusion offered a reduction in the pre-weaning mortality rate of the piglets as well as a higher weaning weight.

In conclusion, oregano oil, thanks to its phenolic constituents that have proven very valuable in combating intestinal diseases, can prove a valuable tool in the fight against PEDv. It can help reduce the load of secondary bacterial infections on the sow prior to farrowing and thereafter on the piglets. By minimising the risk for those secondary infections, and by creating an environment that is hostile to the life cycle of the virus, the inclusion of this natural product can substantially improve the chances of a successful weaned healthy piglet.

Acidifiers may be added to animal feeds to acidify the feed, which improves palatability and digestibility. Acidification may also help to reduce bacterial numbers in raw materials and protect against moulds and mycotoxins. One example of a commercially available carboxylic acid based animal feed acidifier is pHorce (Kiotechagil), which is added to animal feed at 0.5-1kg per 1000g. pHorce comprises formic and propionic acid and ammonium salts.

Carboxylic acids include but are not limited to formic acid, acetic acid, propionic acid, butyric acid, valeric acid, caproic acid, lauric acid, myristic acid, palmitic acid, stearic acid, oleic acid, and are well known to those in the art.

It has been surprisingly found that a carboxylic acid composition is effective for the prevention and/or treatment of porcine coronavirus infection and in particular, porcine epidemic diarrhoea virus.

In a trial 4 PEDv and PRRS negative/naive pigs around 21 days old were fed an infected diet, which also contained a carboxylic acid composition (pHorce6). The trial included providing the carboxylic acid composition in the feed at around 2kg per 1000kg of feed. Results indicated that the composition was more effective against PEDv at higher levels and was particularly effective at around 4kg per 1000kg of feed.

Pigs were tested for PEDv via daily rectal swabs and PCR testing for 5 days post feeding. Pigs showed no clinical symptoms, firm stools and healthy appetites. Results indicated that the carboxylic acid composition inhibited PEDv.

A further trial design to test efficacy of treatment with a carvacrol and thymol composition alone, a carboxylic acid treatment alone, and carvacrol and thymol composition and carboxylic acid treatment in combination included a two part study consisting of an initial study to prove the infectivity of mash feed product that is to be contaminated with PEDv fed to a number of pigs and a second part (approximately 48 hours after the completion of the Feed Infectivity portion) and conducted per the below protocol.

Protocol (PEDv) To show the effectiveness of essential oil of Oregano (provider of carvarcol and thymol phenols) and organic acid (providing propionic and formic acid) to combat PEDv contaminated feed, either alone or in combination and to demonstrate the effectiveness of carvacrol and thymol and carboxylic acids in the presence of PEDv in swine.

Trial Design (PEDv): Trial Animals: Seventy (72) weaned piglets of approximate 21 days of age. Housing: USDA approved BL2 facility. The single room with Seventy-Five (75) single pig pens with individual feed and water. Feeders are: Kane PNF 18 feeders, Water provided by Trojan 45 Nipple waterers. Each pen is approximately 3 square feet including feeder and water system. Feed:

Base feed -ground feed adequate to meet the nutritional needs of the pigs for the duration of the trial. Feed contaminated with PEDv virus at an agreed upon ct. Contaminated feed fed Adlib to the pigs for the duration of the trial to treatments 2-8.

Treatments: 1 Negative Control PEDv -Naïve herd, no exposure to PEDv 2 Positive Control PEDv-Feed Infectivity 3 Orego-Stim at 1Kg per metric ton 4 pHorce at 2Kg per metric ton Orego-Stim at 1Kg and pHorce at 2Kg per metric ton 6 Orego Stim at 2 Kg per metric Ton 7. pHorce at 4 Kg per metric Ton S. Orego Stim at 2 Kg and pHorce at 4 Kg per metric ton Replication: Treatment 1 is the Naive PEDv herd pigs are sourced from.

Treatment 2 pigs (Positive Control), -tested for feed infectivity prior to Treatment 3-8 pigs going on test. Virus harvested and utilized for Treatment 3-8 contaminated feed.

For Treatments 3 through 8 -Twelve (12) Replicates per treatment for a total of 72 pigs and treatments.

Treatments 3-8 pigs -randomly assigned to a treatment and pen. Animal Care: Proper care of all trial animals based on IACUC standards. Animal care -includes Euthanasia of animals by a licensed Veterinarian or trained personnel, in the event of extreme distress to the research subject.

Data Collection: 1 Feed Intake over the seven day period.

2 ADG over the seven day period.

3 Stool Scores -Day 0,1,3,5,7 4 Feed PCR Pre-contamination Post-contamination End of Trial 5. Fecal PCR via swabs -Day 0, 3, 7 Trial Design (PRRSV 174, PEDV and SVA): Trial protocol: An evaluation the efficacy of feed additive candidates at reducing the risk of infection following consumption of feed contaminated with PRRSV 174, PEDV and SVA.

Introduction: In 2014, feed was first identified as a risk factor for porcine epidemic diarrhea viral infection of pigs. This observation was expanded across multiple viruses at both the domestic and transboundary levels. In an effort to reduce this risk, feed additives were tested to determine their ability to negatively affect viral survival and viral load. These testing platforms, while informative, have been primarily bench-top level and bioassay-based. Now the need for further evaluation of candidate mitigants under more rigorous, field-based models is needed to better assess their potential efficacy.

Objective: to evaluate the efficacy of select feed additives at reducing the risk of infection following consumption of feed contaminated with PRRSV 174, PEDV and SVA.

Materials and Methods Experimental design and viral challenge: The experimental design consisted of 6 groups, including: Group 1-5: complete feed mixed with feed additive mitigant candidates (one candidate per room) Group 6: complete feed positive control, non-mitigated feed A 14-day period study period employed. Room as the experimental unit with 100 pigs/room (6 pens/room) with a designated feed bin/room. Pigs (15 kg) from a source herd documented to be free of all 3 viral pathogens by monthly testing and clinical history.

Challenge procedure Viral challenge of feed involves a 454g "Ice Block Model", consisting of 100 mL SVA (5 logs TCID50/mL, Ct = 20.72), 100 mL PRRSV 174(5 logs TCID50/mL, Ct = 21.38), 100 mL PEDV (5 logs TCID50/mL, Ct = 24.25) and balanced with 154 mL MEM. Blocks are frozen at -80C and dropped into each feed bin on days 0 and 6. The blocks are allowed to melt, with liquid permeating the feed, which is then augured into room for pigs to consume via natural feeding behaviour.

Metrics Ante-mortem measurements Ante-mortem samples, including oral fluids and Swiffer samples of feeders, collected across the 6 pens from each of the rooms at 0 and 6 days post-inoculation (DPI) of feed. Feed samples alone collected at 14 DPI. For collection of feed samples, individual Swiffer cloths drawn across the feed trough, contacting feed particles present. Cloths immersed in sterile saline with a 3 mL aliquot decanted for testing.

Post-mortem measurements Post-mortem samples collected from 30 pigs from each room at 14 DPI. Samples collected include tonsil for SVA, serum for PRRSV and rectal swabs for PEDV. Samples evaluated for the presence of viral nucleic acid by PCR and nucleic acid sequencing of the ORF 5 performed on select samples, as needed.

Performance measurements Start and end weights collected from all pigs (0 and 14 DPI). Data described using descriptive statistics and differences in Ct value (cycle threshold) between groups will be analyzed for significance (p <0.05) using ANOVA or T test.

Clinical measurements Pigs scored daily for the presence of the following clinical signs: 1. Lameness/vesicles (WA) 2. Dyspnea/weight loss/rough hair coat (PRRW) 3. Diarrhea (PEDV) A novel approach to the study includes the use of controlled field conditions involving population size, and the use of natural feeding behavior. In addition, a robust 3-way viral challenge model is employed with multiple metrics, such as diagnostics and performance measures collected.

Claims (1)

1. Claims Claim 1 A composition comprising one or more carboxylic acids or derivative thereof for the prevention and/or treatment of porcine viral infection.