Animal plasma for use in a treatment of enteropathy caused by a pathogen

Abstract

The present invention pertains to animal plasma for use in the treatment and / or prevention of enteropathy, preferably chronic enteropathy, caused by a pathogen, wherein the animal plasma is obtained from an animal selected from the group consisting of porcine, bovine and ovine. The invention further pertains to compositions comprising the animal plasma for use according to the invention. Preferably the treatment and / or prevention of enteropathy is in an animal, such as a dog or cat.

Description

[0001] Title: ANIMAL PLASMA FOR USE IN A TREATMENT OF ENTEROPATHY CAUSED BY A PATHOGEN

[0002] TECHNICAL FIELD

[0003] The present application relates to animal plasma for use in the treatment and / or prevention of enteropathy caused by a pathogen.

[0004] BACKGROUND OF THE INVENTION

[0005] Dogs frequently experience gastrointestinal issues, with up to 20% of visits to the veterinarian attributed to gut-related problems. One of the most severe manifestations is chronic enteropathy (CE), characterized by persistent inflammation of the gut and often accompanied by symptoms such as anorexia, vomiting, and diarrhoea. The inflammation may be caused by various factors, including the presence of food allergens in the diet, an imbalance in the gut microbiome, or chronic or recurrent infections with pathogens. Common pathogens implicated in chronic enteropathy in dogs include canine parvovirus (CPV), Clostridium spp., Enterococcus faecalis and Escherichia coli.

[0006] Enteropathy caused by pathogens in companion animals refers to a spectrum of gastrointestinal disorders stemming from infections by various microorganisms, including bacteria, viruses, parasites, and fungi. This condition manifests through inflammation, damage, or dysfunction of the intestinal tract, leading to a range of clinical signs such as diarrhoea, vomiting, abdominal discomfort, and dehydration.

[0007] Common pathogens implicated in enteropathy include Escherichia coli, Salmonella spp., Campylobacter spp., parvoviruses, and various parasitic agents. Enteropathy can affect dogs, cats, and other companion animals, with severity varying based on factors such as the type of pathogen, the age and health status of the animal, and the presence of predisposing factors.

[0008] Parvoviruses are a group of small, non-enveloped viruses known to cause hostspecific diseases in various animal species.

[0009] Canine parvovirus (CPV) is commonly encountered in dogs and causes a highly contagious and potentially life-threatening disease known as canine parvoviral enteritis. CPV primarily affects puppies and young dogs, causing symptoms such as severe diarrhoea, vomiting, dehydration, lethargy, and loss of appetite. The virus targets rapidly dividing cells in the intestinal lining and bone marrow, leading to severe gastrointestinal and immune system dysfunction. CPV can lead to acute enteropathy, but also increases the incidence of gut problems, including later in life. CPV is mainly prevalent in India, North Africa and parts of South America, but its spread is also seen in Europe. Although vaccination levels are high, the incidence of the virus is common and re-infection with lighter symptoms can occur.

[0010] In cats, feline parvovirus, also known as feline panleukopenia virus (FPV) or feline distemper virus, is a highly contagious viral infection that primarily affects young kittens and unvaccinated cats. Symptoms of feline parvovirus infection include fever, vomiting, diarrhea, lethargy, and loss of appetite. The virus can cause severe damage to the intestinal lining and bone marrow, leading to a decrease in white blood cells (leukopenia) and anemia.

[0011] Currently, the approach to treating chronic enteropathy, including CPV-induced chronic enteropathy, involves dietary changes, antibiotic therapy, or, if these interventions prove ineffective, the use of immunomodulatory medications such as immunosuppressants. These modulate the immune response and reduce inflammation associated with chronic enteropathy.

[0012] However, both antibiotics and immunosuppressants are associated with side effects. Antibiotics can reduce the population of infective bacteria, but they also diminish beneficial bacteria in the gut. Consequently, this alteration in the gut microbiome can influence sensitivity to allergens, necessitating dietary adjustments post-treatment. On the other hand, immunosuppressants compromise overall health and may pose toxic effects for dogs. The long-term use of immunosuppressants has not undergone extensive study but is frequently deemed inadequate.

[0013] Potential alternatives to antibiotics and immunosuppressants for treating gastrointestinal disorders may be pre- and probiotics and dietary modifications to support gastrointestinal health. This may involve feeding a highly digestible diet, eliminating potential food allergens or irritants, or providing specialized diets formulated to support gut health. These treatments are not specific for the treatment and / or prevention of enteropathy caused by a pathogen.

[0014] Hence, there is a need for improved treatments of enteropathy, especially chronic enteropathy, caused by a pathogen that has reduced side effects associated with treating enteropathy. Preferably such treatment is readily available and may be conveniently administered without requiring the assistance of a medically trained professional.

[0015] In addition, there is a need for treatments that reduce the risk of acquiring enteropathy, especially chronic enteropathy, caused by a pathogen. Such treatments need to be preferably convenient and cost-effective, so that they can be consistently applied for prolonged periods of time.

[0016] The above needs are particularly felt in the treatment and / or prevention of enteropathy caused by parvovirus in dogs and cats. Parvovirus is an important cause of (severe) chronic enteropathy in these animals, in particular canine parvovirus (CPV) in dogs. While current treatments, such as supportive care, intravenous fluids, anti-emetics, antibiotics, and plasma transfusions, can help manage the symptoms and improve survival rates, parvovirus infections remain a significant cause of morbidity and mortality in affected animals.

[0017] Considering the above, it is an object of the present invention amongst other objects, to provide improved treatment and / or prevention of enteropathy caused by a pathogen, in particular chronic enteropathy caused by a pathogen.

[0018] The present invention meets the above object, amongst other objects.

[0019] SUMMARY OF THE INVENTION

[0020] In a first aspect, the present invention achieves the above object, amongst other objects, by providing an animal plasma for use in the treatment and / or prevention of enteropathy caused by a pathogen. Preferably the animal plasma is obtained from an animal selected from the group consisting of porcine, bovine and ovine.

[0021] Animal plasma for use in treating or preventing enteropathy is advantageous as feeding spray-dried animal plasma has no side effects in dogs. The present invention may therefore provide an improved therapy for treating and / or preventing enteropathy caused by pathogens with reduced side effects.

[0022] Another advantage of the present invention is that animal plasma for use in the treatment and / or prevention of enteropathy provides a more user- and animal-friendly manner of treating and / or preventing enteropathy caused by a pathogen. Unlike presently known treatments or prevention regimes, animal plasma is appealing to cats and dogs due to its pleasant taste. A further advantage is that animal plasma may facilitate a more consistent dosing adherence. This is particularly important for prevention and / or continuation of treatment when symptoms of enteropathy are subsiding or have dissipated. When the animal plasma is included as a component of an animal’s feed the (preventative) treatment will continue to be administered routinely as part of the animal’s feeding schedule.

[0023] In preferred embodiments, the animal plasma for use in the treatment and / or prevention of enteropathy caused by a pathogen is animal plasma obtained from porcine. Porcine immunoglobulin, in particular the IgG in the porcine plasma, may activate immune effector responses, such as phagocytosis, via Fey receptor binding in dogs and cats. An interaction of porcine IgG with Fc-receptors of canine or feline immune cells may therefore enable improved immune responses against targeted pathogens resulting in an improved treatment and / or prevention of enterophathy caused by a pathogen. Cross-mammalian immunoactivity was previously shown in the use of cow’s milk and its effect on gut health in humans, pigs and dogs. However such cross-mammalian immunoactivity was not shown in the context of treating and / or preventing enteropathy caused by a pathogen, particularly not in cats and / or dogs.

[0024] The present inventors found that multiple immunoglobulins (at least IgG, IgA, IgM) can have direct neutralizing effect on intestinal pathogens commonly found in dogs or cats - in particular parvovirus. The combined neutralizing effects may provide for a broader immune activity and a synergistic effect in ameliorating enteropathy.

[0025] A particularly advantageous embodiment according to the present invention is porcine plasma for use in the treatment and / or prevention of enteropathy (preferably chronic enteropathy) caused by parvovirus, wherein the subject to be treated is a cat or a dog.

[0026] Parvoviruses are host-specific, meaning they typically infect and cause disease in specific animal species. Canine parvovirus (CPV) is a virus infecting dogs, feline parvovirus (FPV) infects cats while porcine parvovirus (PPV) infects pigs. Each parvovirus species has evolved to efficiently infect its specific host species. Parvovirus infection in these species also causes different symptoms. Severe enteropathy is the dominant symptom of parvovirus in dogs and cats, while porcine parvovirus primarily affects the reproductive system of pigs, causing infertility, embryonic death, mummification of foetuses, stillbirths, and the birth of weak or runty piglets. In the case of parvoviruses, immunity generated against one species of parvovirus, such as CPV or PPV, is not expected to provide significant protection against infection with a different species of parvovirus. The inventors have, contrary to widely held believe, surprisingly found that porcine plasma comprises porcine immunoglobulins capable of binding canine and / or feline parvovirus. As a result of this finding, the invention may provide an improved treatment and prevention of parvovirus in dogs and cats. Unlike treatments presently available, porcine plasma for use in treating and / or preventing parvovirus in a dog or cat may specifically target the virus, while simultaneously enhancing the immune response.

[0027] The invention further provides animal plasma for the treatment of enteropathy caused by parvovirus and a bacterium. This dual anti-viral and anti-bacterial action of animal plasma, preferably porcine plasma, may be particularly effective in treating and / or preventing parvovirus in a cat or a dog as secondary bacterial infections occur regularly in parvovirus- infected animals. Such secondary infections lead to prolonged and / or worsening gastrointestinal symptoms.

[0028] In a second aspect, the present invention provides a composition for use in the treatment and / or prevention of enteropathy caused by a pathogen, wherein the composition comprises animal plasma for use as disclosed herein. BRIEF DESCRIPTION OF THE FIGURES

[0029] Figure 1 : Binding of immunoglobulin G (IgG) in spray-dried porcine plasma (SDPP) to canine parvovirus (CPV): (top, left) Binding of IgG by SDPP (PP70 and PP80) and purified IgG to CPV; (top, right) Zoomed-in portion of previous graph showing binding of IgG in SDPP (PP70 and PP80) to CPV; (bottom) Competitive ELISA using purified porcine IgG.

[0030] Figure 2: Binding of canine parvovirus by IgA and IgM in PP70 and PP80. As PP70 and PP80 differ in protein and immunoglobulin concentration, potential of IgA and IgM in both samples to bind CPV was measured using competitive ELISA. A) Presence of CPV-specific IgA in PP70, B) Presence of CPV-specific IgM in PP70, C) Presence of CPV-specific IgA in PP80, D) Presence of CPV-specific IgM in PP80. Data are represented as mean ± SD and are representative of three independent experiments (n = 3). No statistics carried out as the aim of the experiment was to research whether porcine immunoglobulins IgA and IgM from different sources could bind CPV.

[0031] Figure 3: Binding of IgG present in SDPP and purified IgG to bacterial species associated with canine chronic enteropathy (CE). Two strains per bacterial species were tested.

[0032] DETAILED DESCRIPTION OF THE INVENTION

[0033] In a first aspect, the invention relates to an animal plasma for use in the treatment and / or prevention of enteropathy caused by a pathogen, wherein the animal plasma is preferably obtained from an animal selected from the group consisting of porcine, bovine and ovine.

[0034] Source of animal plasma

[0035] Animal plasma is the liquid component of blood in animals, excluding the cells. Animal plasma comprises proteins from animal blood or a fraction thereof. It may further comprise electrolytes, hormones, gases, waste products, and nutrients. As animal plasma is a by-product of the meat industry, its use has the advantage that it reduces waste and maximizes resource utilization.

[0036] Animal plasma may be obtained by methods known by the skilled person. Preferably, animal plasma is obtained by allowing blood to clot naturally, or by centrifugation after clotting has occurred. The clotting process removes the cells (red blood cells and white blood cells), so that a liquid component of blood is obtained, excluding the cells. Such animal plasma lacks clotting factors and may also be referred to as serum. More preferably, animal plasma is obtained by removing cellular material from whole blood by centrifuging whole blood in the presence of an anticoagulant. The remaining liquid (i.e., the extracellular matrix of whole blood) is animal plasma. Animal plasma obtained in such a manner is not serum as it comprises clotting factors.

[0037] In one embodiment, animal plasma is processed animal plasma. Processed animal plasma may be obtained by various methods known to the person skilled in the art, such as concentration or fractionation.

[0038] In one embodiment, animal plasma is enriched animal plasma. Enriched animal plasma may be obtained by processing animal plasma so that the concentration of one or more of its components, in particular the amount of immunoglobulins, is increased beyond its natural range. Preferably, the enriched animal plasma comprises at least 45 wt.% immunoglobulins, more preferably at least 60 wt.%, most preferably at least 75 wt.%, wt.% based on the total protein content of the animal plasma. The immunoglobulin may be a combination of IgG, IgM and IgA. Preferably, the immunoglobulin is IgG.

[0039] In one embodiment, the animal plasma in the context of the present invention is obtained, obtainable or is from an animal selected from the group consisting of porcine, bovine and ovine. The animal plasma may comprise a combination of porcine, bovine and ovine. Preferably, the animal plasma in the context of the present invention is obtained, obtainable or is from porcine.

[0040] Porcine means pig, swine or hog or any animal belonging to genus Sus. Preferably porcine is a domestic pig (Sus scrofa domesticus). Bovine means cattle or any animal belonging to the genus Bos. Preferably bovine is domestic cattle (Bos taurus). Ovine means sheep or any animal belonging to the genus Ovis. Preferably ovine is domestic sheep (Ovis aries).

[0041] In one embodiment, animal plasma is obtained, obtainable or is from an animal selected from the group consisting of porcine, bovine and ovine, wherein the animal has immunity against a pathogen.

[0042] Immunity in mammals like pigs, cows, and sheep refers to their ability to resist or defend against pathogens, which are disease-causing organisms such as bacteria, viruses, parasites, and fungi. Preferably the immunity is adaptive immunity, which is a type of immunity that develops over time as an animal is exposed to specific pathogens or antigens. Adaptive immunity involves the production of specialized proteins called immunoglobulins, also known as antibodies. Such immunity may be acquired through vaccination. The term “pathogen” in the context of the present invention encompasses viruses, bacteria, fungi and parasites. The pathogen in the context of the present invention is most preferably a virus or bacterium.

[0043] In one embodiment, the virus as disclosed herein is preferably one or more selected from the group consisting from parvovirus, coronavirus, rotavirus, distemper virus, adenovirus type 1, more preferably parvovirus. In one embodiment, the virus is selected from the group consisting of canine parvovirus, canine coronavirus, canine rotavirus, canine distemper virus and canine Adenovirus Type 1 , more preferably canine parvovirus. In one embodiment, the virus is selected from the group consisting of feline parvovirus (i.e. feline panleukopenia virus), feline coronavirus, feline rotavirus, feline calicivirus and feline viral diarrhoea complex, more preferably feline parvovirus.

[0044] In one embodiment, the pathogen as disclosed herein is a bacterium selected from the group consisting of Salmonella spp., Campylobacter spp., Clostridium spp., Streptococcus spp. and Enterococcus spp.. In a preferred embodiment, the pathogen may be a bacterium selectedfrom the group consisting of Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli.

[0045] Parvovirus refers to a group of viruses belonging to the Parvoviridae family. Different parvoviruses affect different host species. Each virus has evolved to infect their respective host species. Bovine parvovirus (BPV) affects cattle, causing reproductive disorders such as infertility, embryonic death, and abortion. Porcine parvovirus (PPV) targets pigs, leading to reproductive failure in sows. Porcine parvovirus may have a genomic sequence comprising a nucleic acid sequence with at least 97% sequence identity, preferably at least 99% sequence identity, most preferably 100% sequence identity with GenBank accession number OR046036.1 (Genbank Release 259.0).

[0046] If the animal plasma is obtained, obtainable or is from a porcine, preferably the porcine comprises immunity against a pathogen selected from the group consisting of parvovirus, Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coir, more preferably, the porcine comprises immunity against parvovirus, most preferably porcine parvovirus (PPV).

[0047] In some embodiments, the porcine may have immunity to a plurality of pathogens, preferably at least parvovirus. More preferably Escherichia coli and parvovirus. Most preferably Clostridium perfringens, Escherichia coli and parvovirus.

[0048] In one embodiment, animal plasma is obtained, obtainable or is from a porcine, wherein the porcine is a domestic pig immunized against parvovirus, preferably porcine parvovirus (PPV). Porcine Parvovirus (PPV) is relatively common in pig populations worldwide. It is considered one of the major viral pathogens affecting swine production, causing reproductive problems such as infertility, stillbirths, mummified fetuses, and runt pigs. PPV infections are endemic in many pig-producing regions and can occur in both intensive and extensive production systems. Vaccination against PPV is commonly practiced in many swine-producing regions to control the spread of the virus and minimize its impact on reproductive performance in breeding herds.

[0049] Vaccination, also known as immunization, is a medical intervention that stimulates the immune system to produce a protective response against specific pathogens, such as bacteria or viruses. Vaccine in the context of the present invention encompasses inactivated and (modified) live vaccines. The inactivated vaccine may comprise one or more inactivated virus particles derived from parvovirus disclosed herein. The live vaccine may comprise attenuated (weakened) strains of the parvovirus disclosed herein.

[0050] In one embodiment, the animal plasma is obtained, obtainable or is from a porcine, wherein the porcine is a domestic pig vaccinated against parvovirus, preferably porcine parvovirus (PPV).

[0051] In one embodiment, the animal plasma is obtained, obtainable or is from a porcine, wherein the porcine is a domestic pig vaccinated against Escherichia coli.

[0052] In one embodiment, the animal plasma is obtained, obtainable or is from a porcine, wherein the porcine is a domestic pig vaccinated against Clostridium spp..

[0053] In some embodiments, the animal plasma is obtained, obtainable or is from a porcine, wherein the porcine is a domestic pig vaccinated against porcine parvovirus, Clostridium spp. and Escherichia coir, preferably at least porcine parvovirus and Escherichia coli.

[0054] In one embodiment, the animal plasma as disclosed herein is not obtained from an animal that is a canine or feline.

[0055] Physical form of animal plasma

[0056] Animal plasma in the context of the present invention may be used in any suitable form, including both dried and liquid forms. The term 'liquid form' also encompasses reconstituted forms of dried plasma. Preferably, the animal plasma is in a dried form. Dried refers to plasma that has had its water content removed through a drying process. In addition or alternatively, the terms “dry” or “dried” can mean a water content of no more than 10 wt.%, 7.5 wt.%, 5 wt.%, 2.5 wt.% or 1 wt.%. The powder may have a water content of typically 1-10 wt.%, preferably 2-9 wt.%, more preferably 3-8 wt.%, even more preferably 4-7 wt.%. Plasma in dried form has the advantage of a longer shelf life and stability at ambient temperature compared to liquid forms. In addition or alternatively, a dried plasma in the context of the present invention may have a moisture content of less than 100 g / kg, preferably less than 90 g / kg and more preferably in an amount of between 20 g / kg to 80 g / kg, wherein the moisture content is expressed as a percentage of the weight of the dried plasma powder. The skilled person knows how to determine moisture content of dried animal plasma. Moisture content may be determined by placing a sample of dried animal plasma at a temperature of at least 100 degrees Celsius. When the weight of the sample remains constant, all moisture has evaporated. By comparing the weight of the sample before and after before all moisture evaporated, the moisture content can be determined.

[0057] If the animal plasma is in a dried form it may be obtained by spray drying, freeze drying, fluid bed drying, rotary drum drying and spray freeze drying. Freeze drying is also known as lyophilization. Preferably, the dried animal plasma is obtained or obtainable from freeze-drying or spray-drying. It was found that spray-drying and freeze-drying preserve the functional properties of the plasma. More preferably, the dried animal plasma is obtained by spray-drying.

[0058] In one embodiment animal plasma is spray-dried animal plasma powder. Spray drying is advantageous in terms of efficiency, speed, and cost-effectiveness. Spray-drying leads to uniform particle size distribution, improved solubility, enhanced stability, and extended shelf life. These characteristics make spray-dried immunoglobulin products convenient to handle, store, and incorporate into various end products. In the context of the present invention, it was found that spray-drying is preferred as it maintains the therapeutic efficacy of the animal plasma. Immunoglobulins present in the animal plasma retain their biological function. In particular, spray-drying was advantageous as it is effective in maintaining biological activity of immunoglobulins capable of binding parvovirus or a bacterium selected from the group consisting of Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli.

[0059] In one embodiment the dried animal plasma is in a powder form. Plasma powder refers to a fine, dry powder composed of dried animal plasma. This form may be obtained by spray drying or freeze drying. Preferably, the animal plasma powder is obtained by spraydrying.

[0060] Plasma powder produced by spray drying tends to have a relatively uniform particle size distribution, with particles typically ranging from a few micrometers to several tens of micrometers in diameter. The skilled person knows how to modify the exact particle size distribution by controlling parameters such as the atomization rate, inlet temperature, and airflow rate during spray drying. Plasma powder may have a particle size of 1 pm to 100 pm, preferably 2 pm to 50 pm.

[0061] Composition of animal plasma Animal plasma is primarily composed of proteins, lipids (fats), carbohydrates, vitamins, minerals, and water if in liquid form. The composition of animal plasma may be determined by Weende analysis.

[0062] In one embodiment, the animal plasma comprises at least 50 wt.% protein on a dry matter basis. On a dried matter basis means based on dry weight of the animal plasma. Dry weight of animal plasma is the weight of the solids present in animal plasma, excluding the weight of water. Preferably, the animal plasma comprises 65-95 wt.% protein on a dry matter basis. Proteins may include albumin, globulins, fibrinogen, immunoglobulins (antibodies) and enzymes.

[0063] Immunoglobulins (Ig), also known as antibodies, are proteins produced by the immune system in response to the presence of foreign substances, known as antigens. There are several classes or isotypes of immunoglobulins, each with distinct functions and properties. Main classes of immunoglobulin include immunoglobulin G (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin D (IgD) and imunoglobulin E (IgE).

[0064] In one embodiment, an immunoglobulin is an immunoglobulin selected from the group consisting of IG (IgG), immunoglobulin A (IgA), immunoglobulin M (IgM), immunoglobulin D (IgD) and imunoglobulin E (IgE). Preferably, the immunoglobulin is selected from the group consisting of immunoglobulin G (IgG), immunoglobulin A (IgA), and immunoglobulin M (IgM).

[0065] Immunoglobulin G (IgG) is a class of immunoglobulins that plays a crucial role in an animal’s immune system's defense against pathogens. Immunoglobulin G (IgG) is the most abundant type of antibody in the bloodstream and is found in extracellular fluids such as blood. IgG molecules are large Y-shaped proteins composed of four polypeptide chains: two identical heavy chains and two identical light chains. Each IgG molecule has two antigenbinding sites located at the tips of the Y-shaped arms. IgG antibodies exhibit high specificity for antigens. The variable regions of the antibody molecules determine their antigen-binding specificity, enabling the immune system to mount targeted responses to different pathogens.

[0066] Immunoglobulin M (IgM) has a pentameric structure composed of five identical monomeric subunits, each consisting of two identical heavy chains and two identical light chains. IgM molecules ate the largest type of antibodies in mammals and have ten antigenbinding sites. IgM is primarily located in the bloodstream and lymphatic system.

[0067] Immunoglobulin A (IgA) has two subclasses (lgA1 and lgA2) and can be produced as a monomeric as well as a dimeric form. The IgA dimeric form is the most prevalent and, when it has bound the Secretory component, is also called secretory IgA (slgA). slgA is the main immunoglobulin found in mucous secretions, including tears, saliva, sweat, colostrum and secretions from the genitourinary tract, gastrointestinal tract, prostate and respiratory epithelium. It is also found in small amounts in blood. In one embodiment, the immunoglobulin is immunoglobulin G (IgG).

[0068] In one embodiment, the animal plasma comprises at least 10 wt.% immunoglobulin (Ig) on a dry matter basis. Preferably, the animal plasma comprises from 10 wt.% to 90 wt.% immunoglobulin (Ig) on a dry matter basis. More preferably, the animal plasma comprises from 15 wt.% to 80 wt.%, from 18 wt.% to 70 wt.%, from 20 wt.% to about 65 wt.%, immunoglobulin (Ig) on a dry matter basis. Most preferably, the animal plasma comprises from 20 wt.% to 60 wt.% immunoglobulin (Ig) on a dry matter basis. The immunoglobulin content of animal plasma can be measured using various analytical techniques known to the skilled person, such as enzyme-linked immunosorbent assay (ELISA) or immunoblotting.

[0069] In one embodiment, the animal plasma comprises at least 10 wt.% IgG based on the total protein content of the animal plasma. Preferably, the animal plasma comprises from 10 wt.% to 90 wt.% IgG based on the total protein content of the animal plasma. More preferably, the animal plasma contains from 10% to 85 wt.%, IgG based on the total protein content of the animal plasma; most preferably from 10 wt.% to 40 wt.% IgG based on the total protein content of the animal plasma.

[0070] In one embodiment, the animal plasma comprises at least 0.5 wt.% IgA based on the total protein content of the animal plasma. Preferably, the animal plasma comprises from 1 wt.% to 25 wt.% IgA based on the total protein content of the animal plasma. More preferably, the animal plasma comprises 2 wt.% to 10 wt.% IgA based on the total protein content of the animal plasma.

[0071] In one embodiment, the animal plasma comprises at least 1 wt.% IgM based on the total protein content of the animal plasma. Preferably, the animal plasma comprises from 4 wt.% to 15 wt.% IgM based on the total protein content of the animal plasma. More preferably, the animal plasma comprises 6 wt.% to 12 wt.% IgM based on the total protein content of the animal plasma.

[0072] In one embodiment, immunoglobulins are porcine immunoglobulins.

[0073] In certain embodiments, the protein component of the animal plasma may comprise: at least 10 wt.% IgG; preferably 10-90 wt.%, most preferably 20-60 wt.% IgG, based on the total protein content of the animal plasma; at least 0.5 wt.% IgA; preferably 1-25 wt.%, most preferably 2.0-10 wt.% IgA, based on the total protein content of the animal plasma; and at least 1 wt.% IgM; preferably 4-20 wt.%, most preferably 6.0-12.0 wt.% IgM, based on the total protein content of the animal plasma.

[0074] In a preferred embodiment, the protein component comprises 20-30 wt.% IgG, 0.5- 10 wt.% IgA and 5-15 wt.% IgM, based on the total protein content of the animal plasma.

[0075] In one embodiment, the animal plasma comprises an immunoglobin capable of binding to a pathogen selected from the group consisting of parvovirus, Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli. The immunoglobulin may be selected from the group consisting of IgM, IgG and IgA. Preferably the immunoglobulin is IgG.

[0076] An immunoglobulin capable of binding to a pathogen means that the immunoglobulin has the ability to attach to the surface or components of the pathogen. This binding interaction may occur between molecules on the surface of the immunogen (e.g., antigens) and complementary molecules on the surface of the pathogen (e.g., receptors).

[0077] The strength of binding between an immunoglobulin (antibody) and a pathogen can be expressed using various methods, including qualitative assessments, quantitative measurements, and functional assays. The skilled person is familiar with these methods and can therefore determine whether an immunoglobulin is capable of binding a pathogen in a routine manner. In particular enzyme-linked immunosorbent assay (ELISA) may be used, and is a preferred method, to determine the binding capability of an immunoglobulin to a pathogen. In this assay, the pathogen or its components are immobilized on a solid surface. The immunoglobulin is then added, and if binding occurs, it forms antigen-antibody complexes. Detection is typically achieved by adding a secondary antibody conjugated to an enzyme, followed by a substrate that produces a measurable signal. Positive binding is indicated by a signal above background levels. Controls and validation steps ensure specificity and reliability. ELISA provides quantitative data on the strength of the interaction, aiding in the assessment of the antibody's effectiveness in recognizing and neutralizing the pathogen.

[0078] A preferred method to determine binding of immunoglobulin to lipopolysaccharide (LPS) or a pathogen as disclosed herein comprises the steps of: a) Coating a first well of a NUNC 96-well coat, flat bottom, medium binding microplate with a coating buffer by adding 100 pl of the coating buffer to each well, wherein the coating buffer comprises the lipopolysaccharide (LPS) or the pathogen (preferably CPV or FPV) at a concentration of 10 pg / mL protein; b) Coating a second well with purified porcine IgG (Merck; positive control) and a third well with ultrapure water (Milli-Q; negative control); c) Covering the plate with a plate cover and incubating the wells overnight at 4 °C; d) Providing a sample by providing animal plasma in a liquid form or by weighing dried animal plasma on a scale, dissolving the animal plasma at 10 mg / mL in ultrapure water and mixing the ultrapure water comprising the dissolved plasma using a vortex and a roller bench; e) Determining protein amount in the sample with a spectrophotometer (Nanodrop machine; Implen) by calibrating the spectrophotometer using ultrapure water (blanking) and subsequently measuring per sample 2 to 4 times absorption at 280 nm. If the standard deviation between measurements is less than 5%, an average absorption is calculated and based on this average, a stock solution is made of 1 mg / mL protein content by diluting the sample in ultrapure water; f) Diluting the stock solution in a blocking buffer to 100 pg / mL protein, wherein the blocking buffer comprises 1 wt.% gelatin hydrolysate in ultrapure water (Roche); g) Washing the first, second and third well of the microplate three times using the blocking buffer; h) Adding to each well the diluted stock solution and incubating for 1 hr at 37 °C and subsequently washing each well 3 times using the blocking buffer; i) Adding 100 pL of a blocking buffer comprising biotin-labelled goat IgG diluted 1:20000 to each well and incubating for 1 hour at room temperature; the IgG is selected from the group consisting of anti-pig IgG (Bethyl, #A100-113), anti- bovine IgG (Sanbio, ARG81168096) and anti-ovine IgG (Sanbio, 41-IGGOV-E01) j) Washing each well 3 times with the blocking buffer; k) Adding 100 pL of a blocking buffer comprising horse-radish peroxidase (HRP; Biolegend, #405210) diluted 1:1000 and incubating for 30 minutes at room temperature; l) Removing the blocking buffer comprising horse-radish peroxidase from each well and subsequently washing each well 3 times with the blocking buffer; m) Adding 100 pL of 3,3',5,5'-Tetramethylbenzidine (TMB; Biolegend, 421101) to each well and incubating for 5 to 10 minutes to form a blue-colored product and then stopping the colour formation by adding 100 pL of 1 M HCI per well; n) Measuring the absorption at 450 nm for each well using a microplate reader (FilterMax5, Multi-mode microplate reader) o) Optionally, repeating steps (a) to (n) with the stock solution diluted in the blocking buffer to 33 pg / mL and 10 pg / mL protein;

[0079] The strength of binding between antibodies and pathogens may be expressed as a titer. This involves determining the highest dilution of the antibody sample that still produces a detectable binding signal in an assay.

[0080] In one embodiment, animal plasma is obtained, obtainable or is from a porcine, and comprises a porcine immunoglobin capable of binding to a parvovirus. As porcine parvovirus (PPV) is relatively common in pig populations worldwide, porcine plasma comprising porcine immunoglobulins capable of binding to a parvovirus is readily available.

[0081] In one embodiment, animal plasma comprises an immunoglobin capable of binding to parvovirus and a immunoglobin capable of binding of a further pathogen selected from the group consisting of Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli. The immunoglobulins may be selected from the group consisting of IgM, IgG and IgA. Preferably the immunoglobulins are IgG.

[0082] The binding of an immunoglobulin to a selected pathogen can be determined experimentally using immunological assays known to the skilled person. Examples are enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), immunoprecipitation (IP) and flow cytometry. In particular, ELISA may be used to determine immunoglobulin concentrations in animal plasma. The skilled person will thus know how to determine whether the animal plasma comprises immunoglobulins capable to binding to a pathogen.

[0083] The skilled person also knows how to obtain animal plasma, in particular porcine plasma, comprising immunoglobulins capable of binding a pathogen selected from the group consisting of parvovirus, Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli. In particular, commercial vaccines exist for porcine for Clostridium, E. coli and porcinevirus. These vaccines may for example be used to vaccinate a porcine to obtain animal plasma comprising an immunoglobin capable of binding to a pathogen selected from the group consisting of parvovirus, Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli. In particular, the skilled person is aware that commercial vaccines for porcine parvovirus (PPV) are available and widely used in the swine industry to prevent infection and reduce the economic impact of PPV-associated reproductive failure in pigs. These vaccines are typically administered to breeding females (sows and gilts) to prevent fetal death, mummification, and other reproductive problems caused by PPV infection during pregnancy.

[0084] In one embodiment, the animal plasma has a pathogen-specific immunoglobulin titer of at least 1:50 as determined by direct enzyme-linked immunosorbent assay (direct ELISA), wherein the pathogen is selected from the group consisting of parvovirus, Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli. Preferably the titer is at least 1:100; more preferably at least 1:200; even more preferably at least 1:400; most preferably at least 1:1000. The immunoglobulin may be selected from the group consisting of IgM, IgG and IgA. Preferably the immunoglobulin is IgG.

[0085] In one embodiment, immunoglobulin is porcine immunoglobulin.

[0086] In preferred embodiments, the immunoglobin is capable of binding to a parvovirus. Preferably, the parvovirus is selected from the group consisting of canine parvovirus, feline parvovirus and mink parvovirus. More preferably, the parvovirus is a canine parvovirus and / or feline parvovirus. Most preferably, the parvovirus is a canine parvovirus. The canine parvovirus may be a CPV2-type canine parvovirus. There are two types of canine parvovirus called canine minute virus (CPV1) and CPV2. CPV2 is the most pathogenic in dogs. The CPV2-type canine parvovirus may be a CPV-2a, CPV-2b or CPV-2c variant. Preferably, the CPV2-type canine parvovirus is a CPV-2a variant.

[0087] Preferably, the animal plasma has a parvovirus-specific immunoglobulin titer of at least 1 :50 as determined by direct enzyme-linked immunosorbent assay (direct ELISA). Preferably the titer is 1 :100 to 1:2000; more preferably 1 :200 to 1:2000. The immunoglobulin may be selected from the group consisting of IgM, IgG and IgA. Preferably the immunoglobulin is IgG.

[0088] Use of animal plasma

[0089] The present invention relates to animal plasma for use in the treatment and / or prevention of enteropathy caused by a pathogen. Enteropathy caused by a pathogen refers to a condition where the intestines become inflamed or damaged due to infection by a pathogen. The pathogen may be a bacterium, a virus, a parasite, or a fungus. These infections may lead to gastrointestinal symptoms such as diarrhea, abdominal pain, nausea, and vomiting.

[0090] In one embodiment, animal plasma may be for use in the treatment of enteropathy preferably chronic enteropathy, caused by a pathogen. Treatment goals may include symptom relief, disease management, prevention of complications, and improvement of overall quality of life.

[0091] Enteropathy caused by a pathogen refers to a condition characterized by inflammation or damage to the intestines (enteropathy) resulting from infection with a disease-causing microorganism. This condition can manifest as various gastrointestinal symptoms, including diarrhoea, abdominal pain, vomiting, and sometimes fever. Pathogens that can cause enteropathy include bacteria, viruses, parasites, and fungi. Enteropathy caused by a pathogen is typically diagnosed based on a combination of clinical symptoms, laboratory tests (such as stool analysis), and sometimes imaging studies (such as abdominal ultrasound or computed tomography).

[0092] Chronic enteropathy caused by a pathogen is a long-lasting inflammation or disease of the intestines (enteropathy) that is attributed to the presence or infection of a specific pathogen. Preferably the inflammation or disease of the intestines (enteropathy) is present for at least 3 weeks. Unlike acute or short-term gastrointestinal infections, chronic enteropathy caused by a pathogen describes persistent or recurrent gastrointestinal disorders resulting from prolonged or repeated exposure to a pathogenic microorganism. In one embodiment, animal plasma may be for use in the prevention of enteropathy, preferably chronic enteropathy, caused by a pathogen. Prevention means reducing the risk of enteropathy occurrence, while treatment addresses existing health conditions to improve outcomes and quality of life. In an embodiment, the enteropathy is infectious enteropathy, meaning enteropathy caused by one or more infectious agents such as bacteria, viruses, parasites, or fungi. The cause of enteropathy can be diagnosed by clinical evaluation, fecal examination laboratory tests, blood tests and and / or imaging studies.

[0093] In one embodiment of the invention, the enteropathy is chronic enteropathy, more preferably chronic enteropathy caused by a pathogen.

[0094] In one embodiment of the invention, the enteropathy caused by a pathogen is chronic enteropathy. Chronic enteropathy is a prolonged or persistent form of enteropathy meaning that it persists over an extended period, typically lasting weeks, months, or even years. These conditions often involve repeated episodes of symptoms interspersed with periods of remission or relative symptom relief. Chronic enteropathies may require long-term management and monitoring to control symptoms, prevent complications, and maintain quality of life. Chronic enteropathy caused by a pathogen may result from a persistent infection, an inadequate immune response, or a complication of the initial infection.

[0095] In one embodiment of the invention, the pathogen causing the enteropathy is a virus and / or a bacterium.

[0096] In one embodiment the enteropathy is caused by a parvovirus. The parvovirus may be a canine parvovirus, feline parvovirus or mink parvovirus. Preferably, the parvovirus is a canine parvovirus or feline parvovirus. More preferably, the parvovirus is a canine parvovirus. The canine parvovirus may be a CPV2-type canine parvovirus. In particular, the CPV2-type canine parvovirus may be a CPV-2a, CPV-2b or CPV-2c variant.

[0097] The canine parvovirus may have a genomic sequence comprising a nucleic sequence selected from the group consisting of GenBank accession numbers M 19296.1, MH545963.1, JQ268284.1 and KY073269.1 (Genbank Release 259.0). In particular, CPV2 comprising a nucleic sequence selected from the group consisting of GenBank accession numbers M19296.1, MH545963.1 (CPV-2a), JQ268284.1 (CPV-2b) and KY073269.1 (CPV- 2c) (Genbank Release 259.0).

[0098] The feline parvovirus may have a genomic sequence comprising a nucleic sequence selected from the group consisting of GenBank accession numbers MT614366.1 (Genbank Release 259.0).

[0099] In one embodiment, the enteropathy is caused by a bacterium selected from the group consisting of Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli.

[0100] In one embodiment the enteropathy is caused by a parvovirus and a second pathogen, preferably a bacterium. Enteropathy caused by parvovirus may lead to secondary bacterial infections, which can occur due to damage to the intestinal lining and a weakened immune system. These infections may lead to complications such as sepsis. One advantage of the animal plasma for use according to the present invention is that it can treat enteropathy caused by the parvovirus, while simultaneously treating and / or preventing enteropathy caused by a second pathogen. Preferably, the second pathogen is selected from the group consisting of Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli. More preferably, the second pathogen is Clostridium perfringens or Escherichia coli.

[0101] The skilled person may obtain porcine plasma comprising a porcine immunoglobulin capable of binding a parvovirus and a further porcine immunoglobulin capable of binding a second pathogen, preferably a bacterium, without undue burden as pigs are generally immunized for various bacteria, e.g., by routine vaccination programs.

[0102] In one embodiment of the invention, the subject to be treated is a dog, a cat or a mink. Preferably, the treatment and / or prevention of enteropathy is in a dog or cat; most preferably a dog.

[0103] In one embodiment of the invention, the subject to be treated is a dog. Preferably, the pathogen causing the chronic enteropathy is canine parvovirus. In dogs, canine parvovirus (CPV) infection can be severe, especially in puppies and young dogs. CPV infection may result in severe enteropathy, including gastrointestinal symptoms such as (bloody) diarrhoea, loss of appetite and vomiting resulting in dehydration and weakness. In some cases, CPV infection may be fatal, especially if left untreated or if the affected dog's immune system is compromised. Dogs with canine parvovirus (CPV) infection may develop secondary bacterial infections due to damage to the intestinal lining and suppression of the immune system. The most common secondary bacterial infections associated with CPV include Clostridium infection and Escherichia coli (E. coli) infection. Enteropathy caused by a second pathogen may contribute to the persistence of gastrointestinal symptoms in a parvovirus-infected dog.

[0104] In one embodiment of the invention, the subject to be treated is a cat. Preferably, the pathogen causing the chronic enteropathy is feline parvovirus. In cats, feline parvovirus (FPV) infection (also known as feline panleukopenia virus or feline distemper) may cause severe illness, particularly in kittens and young cats. Symptoms of FPV infection may be enteropathy, including gastrointestinal symptoms such as vomiting, diarrhea, and loss of appetite. Similar to canine parvovirus infection in dogs, feline parvovirus infection can damage the intestinal lining and suppress the immune system, increasing the risk of a secondary bacterial infection. Common secondary bacterial infections that may occur in cats with feline parvovirus infection include: Clostridium infection and Escherichia coli (E. coli) infection. Enteropathy caused by a second pathogen may contribute to the persistence of gastrointestinal symptoms in a parvovirus-infected cat.

[0105] In one embodiment of the invention, the animal plasma is administered orally, preferably in feed. The terms food and feed have the same meaning in the context of the present application and may be used interchangeably. Oral administration, including administration in feed, is challenging compared to administration through injection, e.g., intravenously or subcutaneously. When animal plasma is administered orally, it must pass through the digestive system of the subject, which is generally expected to degrade the ingested products, for example by degrading proteins into smaller peptides, such as peptide fragments, dipeptides, tripeptides, or even singular amino acids (or some combination of these smaller peptides). Thus, a therapeutic product that can be effectively administered through an injectable route (e.g., intravenously or subcutaneously) often cannot be effectively administered orally due to the degradation of the therapeutic product in the subject’s digestive system. Yet the animal plasma as described herein may be effectively and therapeutically delivered orally.

[0106] In one embodiment, the animal plasma is administered at a dose of 5 mg to 1 g per kg of body weight of the animal to be treated per day.

[0107] A “therapeutically effective amount” can be, but is not limited, to an amount of animal plasma composition that is sufficient to provide an improvement of at least one symptom of enteropathy in the subject. The dosage and number of doses (e.g., single or multiple dose) administered to the subject will vary depending upon a variety of factors, including the route of administration, conditions and characteristics of subject (sex, age, body weight, health, size), extent of symptoms, concurrent treatments, frequency of treatment and the effect desired.

[0108] In one embodiment, the animal plasma is administered for at least 3 months.

[0109] In one embodiment, the animal plasma for use according to the present invention may be administered in the form of a tablet, a capsule, an ampule for oral use, a powder or a cream. In certain embodiments, the animal plasma may be administered in liquid form or suspended or dissolved in a suitable liquid, such as water, saline, or milk.

[0110] In one embodiment, the animal plasma is not administered to an animal through intravenous administration.

[0111] In a second aspect, the present invention relates to a composition for use in the treatment and / or prevention of enteropathy caused by a pathogen, wherein the composition comprises animal plasma as disclosed herein. The animal plasma may be combined with a pharmaceutically acceptable carrier such as a suitable liquid vehicle or excipient and an optional auxiliary additive or additives. The liquid vehicles and excipients are conventional and commercially available. Illustrative thereof are distilled water, physiological saline, aqueous solutions of dextrose, and the like. In general, in addition to the active compounds, the compositions of this invention may contain suitable excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used pharmaceutically. In certain embodiments, the plasma proteins may also be microencapsulated, thereby protecting and stabilizing them.

[0112] A composition for use according to the present invention may be for the same use as described herein for the animal plasma.

[0113] In one embodiment, the composition for use according to the present invention may be in the form of a tablet, a capsule, an ampule for oral use, a powder or a cream.

[0114] The compositions for use in the present invention are manufactured in a manner which is known in the art. For example, the preparations may be made by means of conventional mixing, granulating, dragee-production, dissolving, and / or lyophilizing processes. The processes to be used will depend ultimately on the physical properties of the ingredients used and the desired form of the end product.

[0115] In one embodiment, the composition is a feed, preferably an animal feed.

[0116] Method of treating and / or preventing enteropathy

[0117] In a third aspect, the present invention relates to a method for treating and / or preventing enteropathy caused by a pathogen, the method comprising the steps of:

[0118] Providing an animal plasma as disclosed herein

[0119] Orally administrating the animal plasma to a cat, dog or mink; preferably a cat or dog; most preferably a dog.

[0120] In one embodiment, the animal plasma may be in the form of a composition as disclosed herein, preferably this composition is a feed, preferably an animal feed, such as dog kibbles.

[0121] In one embodiment of the invention, the enteropathy caused by a pathogen is chronic enteropathy.

[0122] In one embodiment of the invention, the pathogen causing the enteropathy is a virus and / or a bacterium.

[0123] In one embodiment the enteropathy is caused by a parvovirus. The parvovirus may be a canine parvovirus, feline parvovirus or mink parvovirus. Preferably, the parvovirus is a canine parvovirus or feline parvovirus. More preferably, the parvovirus is a canine parvovirus. The canine parvovirus may be a CPV2-type canine parvovirus. In particular, the CPV2-type canine parvovirus may be a CPV-2a, CPV-2b or CPV-2c variant. In one embodiment, the enteropathy is caused by a bacterium selected from the group consisting of Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli.

[0124] In one embodiment the enteropathy is caused by a parvovirus and a second pathogen, preferably a bacterium.

[0125] In one embodiment, the animal plasma may be administered orally at a dose of 5 mg to 1 g per kg of body weight of the animal to be treated per day.

[0126] In one embodiment, the animal plasma may be administered for at least 3 months.

[0127] In one embodiment, the animal plasma may be administered in the form of a tablet, a capsule, an ampule for oral use, a powder or a cream. In certain embodiments, the animal plasma may be administered in liquid form or suspended or dissolved in a suitable liquid, such as water, saline, or milk.

[0128] In a fourth aspect, the present invention relates to the use of animal plasma as disclosed herein for the treatment and / or prevention of enteropathy caused by a pathogen in any manner, wherein the use is any use as described herein for animal plasma.

[0129] In a fifth aspect, the present invention relates to a method for treating and / or preventing enteropathy caused by a pathogen, the method comprising the steps of: Providing an animal plasma as disclosed herein

[0130] Orally administrating the animal plasma to a cat, dog or mink; preferably a cat or dog; most preferably a dog.

[0131] In a sixth aspect, the present invention relates to the use of animal plasma as disclosed herein for the treatment and / or prevention of enteropathy, preferably chronic enteropathy, caused by a pathogen as disclosed herein.

[0132] In a seventh aspect, the present invention relates to the use of animal plasma as disclosed herein for the manufacture of a medicament for the treatment and / or prevention of enteropathy, preferably chronic enteropathy, caused by a pathogen as disclosed herein.

[0133] Without being bound by any theory, it appears that the amelioration of enteropathy is drastically further improved when the plasma and / or immunoglobulin content when the amount of immunoglobulin administered to dogs or cats is above a certain threshold amount. The following dosages are particularly effective in ameliorating enteropathy such as caused by canine or feline parvovirus. In particular, it is considered particularly advantageous to administer at least 10 mg, even more preferably at least 100 mg, total immunoglobulin per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight. Hence, in preferred embodiments, the amount of plasma administered is chosen such that at least 50 mg, or at least 100 mg, or at least 250 mg, or at least 500 mg, or at least 1000 mg of total immunoglobulin is administered per kg animal per day, and in addition or alternatively, no more than 1000 mg, or no more than 750 mg, or no more than 500 mg, or no more than 250 mg total immunoglobulin is administered per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight. In preferred embodiments, the amount of plasma administered is chosen such that 50-1000 mg, preferably 100-500 mg total immunoglobulin is administered per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight.

[0134] In preferred embodiments, the amount of plasma administered is chosen such that at least 10 mg, or at least 25 mg, or at least 50 mg, or at least 100 mg, or at least 200 mg of IgG is administered per kg animal per day, and in addition or alternatively, no more than 500 mg, or no more than 250 mg, or no more than 200 mg, or no more than 100 mg IgG is administered per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight. In preferred embodiments, the amount of plasma administered is chosen such that 50-1000 mg, preferably 100-500 mg IgG is administered per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight.

[0135] In preferred embodiments, the amount of plasma administered is chosen such that at least 1 mg, or at least 5 mg, or at least 10 mg, or at least 25 mg, or at least 50 mg of IgA is administered per kg animal per day, and in addition or alternatively, no more than 100 mg, or no more than 50 mg, or no more than 25 mg, or no more than 10 mg IgA is administered per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight. In preferred embodiments, the amount of plasma administered is chosen such that 5-200 mg, preferably 10-100 mg IgA is administered per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight.

[0136] In preferred embodiments, the amount of plasma administered is chosen such that at least 1 mg, or at least 10 mg, or at least 25 mg, or at least 50 mg, or at least 100 mg of IgM is administered per kg animal per day, and in addition or alternatively, no more than 500 mg, or no more than 250 mg, or no more than 200 mg, or no more than 100 mg IgM is administered per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight. In preferred embodiments, the amount of plasma administered is chosen such that 25-500 mg, preferably 50-250 mg IgM is administered per kg animal per day, wherein the weight (in kg) preferably is the metabolic weight.

[0137] Without being bound by theory, the present inventors consider that appropriate dosage of the plasma in animal feed, in particular kibbles, can drastically further improve the effect in ameliorating enteropathy, such as caused by canine or feline parvovirus. The clinical effects of plasma are surprisingly high at a clinical dose of about 6% based on the caloric intake as calculated using the metabolic weight. In preferred embodiments, the plasma is provided in feed, preferably kibbles in an amount of at least 1%, or at least 2%, or at least 3%, or at least 4%, or at least 3%, or at least 5%, or at least 6%, or at least 7%, or at least 8%, or at least 9%, or at least 10% based on the caloric intake as calculated using the metabolic weight, and in addition or alternatively, the plasma is provided in feed, preferably kibbles in an amount no more than 20%, or no more than 15%, or no more than 12%, or no more than 10%, or no more than 9, or no more than 8%, or no more than 7%, or no more than 6%, or no more than 5% based on the caloric intake as calculated using the metabolic weight. The given amounts of plasma in the feed (e.g. kibbles) may provide for the effective amelioration of enteropathy, without further negative influences such as on digestive tolerance, palatability or overstimulation of the immune system.

[0138] The “weight” or “body weight” in the context of the present invention can be the absolute body weight, but is more preferably the metabolic weight. The metabolic weight (also referred to interchangeably as metabolic body weight) in the context of the present invention is calculated with the formula:

[0139] Metabolic weight = Absolute body weight0 75

[0140] For example, the metabolic weight for a 20 kg dog is 9.46 kg (20°75). The metabolic weight takes into consideration that the many of the animals’ physiological processes generally don't scale directly with an animal’s total body weight.

[0141] EXAMPLES

[0142] Example 1 : Material & Methods

[0143] Materials

[0144] Two spray dried plasma samples were used: PP70 and PP80. PP70 contains 70% protein and PP80 contains 80% protein. All plasma samples were derived from Darling Ingredients International, Loenen. To compare the effect of plasma with the effect of purified IgG, commercially available purified porcine IgG was used as control in experiments.

[0145] Immunoglobulin and lactoferrin measurement

[0146] The concentration of immunoglobulins (IgG, IgA and IgM) in SDPP was tested by coating a medium binding, flat bottom 96-well plate with 100 pl / well of 10 pg / mL of capture antibody. Plates were coated overnight at 4°C. In between steps, wells were washed thrice using PBS containing 0.05% Tween20. Wells were blocked using 1% gelatin hydrolysates in PBS-Tween for 1 hr at 37 °C. 100 pl / well of samples and controls, diluted in blocking buffer, were added to the wells and incubated for 1h at RT. Binding was detected by adding 100 pl / well of biotinylated goat anti-porcine IgA or IgM (1 :50,000 and 1:20,000, respectively), or 100 pl / well of HRP-conjugated goat anti-porcine IgG (1:80,000) in blocking buffer, followed by incubation for 1h at RT. For IgA and IgM measurement, 100 pl of horseradish peroxidase (HRP; 1:1000) was added to every well and incubated 30 min at room temperature (RT). 100 pl / well TMB (3,3',5,5'-Tetramethylbenzidine) substrate was incubated at RT for 2-4 min. The reaction was stopped by adding 100 pl / well of 1M HCI. Absorbance was measured at 450 nm (background: 580 nm) on a microplate reader.

[0147] Bacterial binding of IgG from SDPP

[0148] Medium binding 96-well plates were coated overnight at 4°C with 100 pl / well of a bacterial solution in coating buffer. Bacterial solution was made by diluting samples to 0.5 McFarland in PBS, then further diluting the sample in coating buffer. The ELISA was then carried out as described above using HRP-conjugated goat anti-porcine IgG (1 :80,000).

[0149] Detection canine parvovirus specific IgG, IgA or IgM

[0150] To detect canine parvovirus binding IgG, IgA or IgM in SDPP, 96-well plates were coated overnight at 4°C with 100 pl / well of 10 pg / mL CPV vaccine solution in coating buffer. The ELISA was carried out as described above, using the goat-anti porcine IgG-bioinylated antibody (1:20,000), goat-anti porcine IgA-bioinylated antibody (AAI40B, Bio-rad, 1:50,000) or goat-anti porcine IgM-bioinylated antibody (AAI48B, Bio-rad, 1:20,000) as detection antibodies. To prove that binding of canine parvovirus by porcine IgG was specific, competitive ELISAs were carried out. Here, samples consisting of porcine IgG, IgA, IgM or SDPP and CPV were pre-incubated for 1 h, before being added and incubated for 1h at RT. The ELISA was then carried out as described above.

[0151] Neutralization of canine parvovirus To test whether SDPP was able to neutralize CPV, both a plaque formation and a microneutralization assay were carried out. For the plaque neutralization assay, CrfK cells (ATCC) were cultured in 6-well tissue cell culture plates. 100 pl of diluted virus was added and samples were incubated for 60 min at 37°C. An agarose plug was formed and plates were kept for 72h at 37°C. Wells were then washed and fixed, after which methylene blue was used for staining. Plaque forming units were calculated according to standard formula. The microneutralization assay was carried out as follows: SDPP and CPV were diluted and added to respective wells of a 96-well plate. 1.5 x 104cells were added to every well and allowed to incubate at 37°C for 18-20h. Wells were washed and fixed, after which the anticanine parvovirus antibody was added to each well and incubated for 1h at RT. After washing, the HRP-conjugated goat-anti mouse lgG2a was added to each well and incubated for 1h at RT. After washing, TMB and stop solution were used to visualize. Plates were measured at 450 nm with a background reading at 580 nm using a microplate reader.

[0152] Statistical analysis

[0153] All data are reported as the mean ± standard deviation. Statistical significant differences between treatment groups were determined using one-way ANOVA and defined as P<0.05.

[0154] Example 2: Binding of SDPP to parvovirus

[0155] The capacity of SDPP to bind parvovirus (PV) was tested. A direct ELISA showed the potential of IgG present in SDPP to bind CPV (Figure 1 , top left and right panel). To determine whether binding was specific, a competitive ELISA was set up using purified porcine IgG. Here, pre-incubating purified porcine IgG with CPV led to decreased binding to CPV coated plates, indicating porcine IgG is able to bind CPV specifically (Figure 1, bottom). The competitive ELISA was also set up for PP70 and PP80, showing the presence of CPV specific antibodies in both types of SDPP. Similar results are obtained for feline parvovirus (FPV).

[0156] A similar competitive ELISA as aforementioned was set up using purified porcine IgA and IgM. As PP70 and PP80 differ in protein and immunoglobulin concentration, potential of IgA and IgM in both samples to bind CPV was measured using competitive ELISA. It was found that pre- incubating purified porcine IgA or IgM with CPV led to decreased binding to CPV coated plates, indicating porcine IgA and IgM are similarly able to bind CPV specifically (Figure 2). Example 3: Binding of bacterial strains by SDPP

[0157] Binding of SDPP to 4 bacterial species associated with canine chronic enteropathy was tested using ELISA. All 4 bacterial species (2 strains per species) were bound by both purified IgG and the IgG present in SDPP (Figure 3).

[0158] Example 4: treatment and / or prevention of enteropathy

[0159] Based on the results shown above, present inventors consider that oral administration of animal plasma for use as disclosed herein yields therapeutic effects as shown in Table 1 , wherein:

[0160] (-) indicates no to little improvement and (+) indicates significant improvement of symptoms with further plus signs (+) indicating a stronger improvement.

[0161] Table 1 : Considered effect of animal plasma on enteropathy, particularly chronic enteropathy.

[0162] Animal plasma’s (ovine, bovine, porcine) have reduced / no to little improvement in treating and / or preventing enteropathy in other animals (e.g. ferret, rodents).

[0163] CLAUSES

[0164] 1. Animal plasma for use in the treatment and / or prevention of enteropathy caused by a pathogen, wherein the animal plasma is obtained from an animal selected from the group consisting of porcine, bovine and ovine.

[0165] 2. Animal plasma for use according to clause 1 , wherein the animal is porcine.

[0166] 3. Animal plasma for use according to any of the preceding clauses, wherein the animal plasma is in the form of a powder.

[0167] 4. Animal plasma for use according to clause 3, wherein the plasma powder is obtained by spray drying. 5. Animal plasma for use according to any of the preceding clauses, wherein the enteropathy is a chronic enteropathy.

[0168] 6. Animal plasma for use according to any of the preceding clauses, wherein the use is in a dog or cat, preferably dog.

[0169] 7. Animal plasma for use according to any of the preceding clauses, wherein the animal plasma is orally administered.

[0170] 8. Animal plasma for use according to any of the preceding clauses, wherein the pathogen is a virus or a bacterium.

[0171] 9. Animal plasma for use according to any of the preceding clauses, wherein the pathogen is a parvovirus, preferably a canine parvovirus (CPV) or feline parvovirus (FPV).

[0172] 10. Animal plasma for use according to any of the preceding clauses, wherein the pathogen is a bacterium selected from the group consisting of Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli.

[0173] 11. Animal plasma for use according to any of the preceding clauses, wherein the animal plasma comprises at least 5 wt.% immunoglobin based on total protein content, preferably 10-90 wt.%.

[0174] 12. Animal plasma for use according to any one of the previous clauses, wherein the animal plasma comprises an immunoglobin capable of binding to a pathogen selected from the group consisting of parvovirus, Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli.

[0175] 13. Animal plasma for use according to clause 11 or 12, wherein the immunoglobin is capable of binding to canine parvovirus (CPV) and / or feline parvovirus (FPV), preferably canine parvovirus (CPV).

[0176] 14. Animal plasma for use according to any one of clauses 11 to 13, wherein the immunoglobin is one or more selected from the group consisting of IgM, IgG and IgA. 15. Animal plasma for use according to any one of clauses 11 to 14, wherein the animal plasma has a pathogen-specific immunoglobulin titer of at least 1 :100 for the selected pathogen as determined by direct enzyme-linked immunosorbent assay (ELISA); preferably at least 1:200.

[0177] 16. Composition for use in the treatment and / or prevention of enteropathy caused by a pathogen, wherein the composition comprises animal plasma according to any of the preceding clauses.

[0178] 17. Composition for use according to clause 16, wherein the use is as defined in any one of clauses 1 to 14.

[0179] 18. Composition for use according to clause 16 or 17, wherein the composition is in the form of a tablet, capsule, ampoule for oral use, granulate powder or cream.

[0180] 19. Composition according to clause 16 or 17, wherein the composition is a feed, preferably an animal feed.

Claims

CLAIMS1. Animal plasma for use in the treatment and / or prevention of enteropathy caused by a pathogen, wherein the animal plasma is obtained from an animal selected from the group consisting of porcine, bovine and ovine, wherein the use is in a dog or cat, preferably dog.

2. Animal plasma for use according to claim 1, wherein the animal plasma is porcine plasma.

3. Animal plasma for use according to any of the preceding claims, wherein the animal plasma is in the form of a powder.

4. Animal plasma for use according to claim 3, wherein the plasma powder is obtained by spray drying.

5. Animal plasma for use according to any of the preceding claims, wherein the enteropathy is a chronic enteropathy.

6. Animal plasma for use according to any of the preceding claims, wherein the animal plasma is orally administered.

7. Animal plasma for use according to any of the preceding claims, wherein the pathogen is a virus or a bacterium.

8. Animal plasma for use according to any of the preceding claims, wherein the pathogen is a parvovirus, preferably a canine parvovirus (CPV) or feline parvovirus (FPV).

9. Animal plasma for use according to any of the preceding claims, wherein the pathogen is a bacterium selected from the group consisting of Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli.

10. Animal plasma for use according to any of the preceding claims, wherein the animal plasma comprises at least 5 wt.% immunoglobin based on total protein content, preferably 10-90 wt.%.

11. Animal plasma for use according to any one of the previous claims, wherein the animal plasma comprises an immunoglobin capable of binding to a pathogen selected from the group consisting of parvovirus, Streptococcus canis, Clostridium perfringens, Enterococcus faecalis and Escherichia coli.

12. Animal plasma for use according to claim 10 or 11, wherein the immunoglobin is capable of binding to canine parvovirus (CPV) and / or feline parvovirus (FPV), preferably canine parvovirus (CPV).

13. Animal plasma for use according to any one of claims 10 to 12, wherein the immunoglobin is one or more selected from the group consisting of IgM, IgG and IgA.

14. Animal plasma for use according to any one of claims 10 to 13, wherein the animal plasma has a pathogen-specific immunoglobulin titer of at least 1:100 for the selected pathogen as determined by direct enzyme-linked immunosorbent assay (ELISA); preferably at least 1:200.

15. Animal plasma for use according to any one of the previous claims, wherein the animal plasma is provided in a composition in the form of a tablet, capsule, ampoule for oral use, granulate powder or cream.

16. Animal plasma for use according to any one of the previous claims, wherein the animal plasma is provided in a feed composition, preferably an animal feed composition.

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