WO2022011436A1

WO2022011436A1 - Methods of treating coronavirus infection with bovine-hyperimmune colostrum

Info

Publication number: WO2022011436A1
Application number: PCT/AU2021/050772
Authority: WO
Inventors: Jerry Kanellos; Roger Aston; Peter ANASTASIOU
Original assignee: Immuron Limited
Priority date: 2020-07-17
Filing date: 2021-07-17
Publication date: 2022-01-20

Abstract

A method of treating and/or preventing human coronavirus infection in a subject comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

Description

METHODS OF TREATING CORONAVIRUS INFECTION WITH BOVINE-HYPERIMMUNE COLOSTRUM

Technical Field

[001] This invention relates to compositions and methods for the treatment and/or prevention of COVID-19 and COVID-19 associated disease.

Background of Invention

[002] As of June 2020, more than 8 million confirmed cases of COVID-19 caused by the SARS-CoV-2 virus have been reported worldwide, involving all global regions and resulting in over 433,000 deaths. Although the majority of cases are clinically mild or asymptomatic, early reports from China estimated that 20% of all COVID-19 patients progressed to severe disease and required hospitalisation, with 5-16% of these individuals going on to require management within an Intensive Care Unit (ICU). Pulmonary disease leading to respiratory failure has been the major cause of mortality in severe cases. The ability of health systems around the world to cope with increasing case numbers in coming months is of major concern. All levels of the system will be challenged, from primary care, pre-hospital and emergency department (ED) services, to inpatient units and ultimately ICUs. Stresses on clinical care provision will result in increased morbidity and mortality. These consequences have tragically already been observed even in high income countries that provide whole population access to quality medical care. Greater impacts will be observed over coming months in low and middle-income countries where access to high level care is extremely limited. Availability of ICU beds and ventilators has proven critical for the adequate management of severe cases, with overwhelming demand initiating complex ethical discussions about rationing of scarce resources. In readiness for this challenge, Australia has drawn on approaches developed over many years to prepare for influenza pandemics and rapidly produced a national COVID-19 pandemic plan. The plan has reoriented relevant strategies towards this new pathogen, based on emerging understanding of its anticipated transmissibility and severity, which are the determinants of clinical impact (8). Early imposition of stringent border measures, high levels of testing, active case-finding and quarantine of contacts have all bought time to reinforce public health and clinical capacity. However, a recent influx of cases among travellers returning from countries with rapidly growing epidemics has been associated with establishment of community transmission in several Australian states. [003] As of 8 April 2020, the global COVID-19 vaccine R&D landscape includes 115 vaccine candidates, of which 78 are confirmed as active and 37 are unconfirmed (development status cannot be determined from publicly available or proprietary information sources); of these candidates there is a focus on live attenuated viruses, inactivated viruses, non-replicating viral vectors, replicating viral vectors, recombinant proteins, peptide based vaccines, virus like particles, and DNA and RNA vaccines.

[004] The approaches being applied for COVID-19 vaccine development — which involve a new virus target and often novel vaccine technology platforms and novel development paradigms as well — are likely to increase the risks associated with delivering a licensed vaccine and will require careful evaluation of effectiveness and safety at each step.

[005] There is a need for methods and compositions for treating and/or preventing COVID-19 associated disease and transmission of SARS-CoV-2 infection.

Summary of Invention

[006] In one embodiment, the present invention provides a method of treating and/or preventing human coronavirus infection in a subject comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[007] In another embodiment, the present invention provides a method of reducing viral shedding from a subject infected with a human coronavirus comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[008] In a further embodiment, the present invention provides a method of treating and/or preventing human coronavirus-associated diarrhoea in a subject comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[009] In a further embodiment, the present invention provides a method of inhibiting coronavirus mediated cytotoxicity comprising contacting one or more human cells with an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS. [0010] In a further embodiment, the present invention provides a method of improving cell viability comprising contacting one or more human cells with an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0011] In a further embodiment, the present invention provides a method of maintaining gastrointestinal cell viability comprising contacting one or more human cells with an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0012] In a further embodiment, the present invention provides a method as described herein, wherein the vaccine comprising ETEC LPS comprises one or more LPS O serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159.

[0013] In a further embodiment, the present invention provides a method as described herein, wherein the vaccine comprising ETEC LPS comprises two or more LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159.

[0014] In a further embodiment, the present invention provides a method as described herein, wherein the vaccine comprising ETEC LPS comprises LPS 0 serotypes 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159.

[0015] In a further embodiment, the present invention provides a method as described herein, wherein the vaccine comprising ETEC LPS comprises the lipid A core and O-polysaccharide regions of the LPS.

[0016] In a further embodiment, the present invention provides a method as described herein, wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to ETEC LPS.

[0017] In a further embodiment, the present invention provides a method as described herein, wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to one or more LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 071 , 0114, 0115, 0117, 0128, 0148, 0153, and 0159, 0167. [0018] In a further embodiment, the present invention provides a method as described herein, wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to one or more LPS serotypes of a gram negative bacteria selected from the group consisting of Enteropathogenic Escherichia coli strain E2348/69, Klebsiella pneumoniae strain ATCC 26, Pseudomonas aeruginosa strain ATCC 27853, Salmonella typhimurium strain ATCC 14028, Vibrio cholerae strain 6239, Yersinia enterocolitica strain 67R, C. jejuni, Vibrio cholera Ogawa, V. cholera Inaba, and V. cholera NON 01/0139, C. coli, C. jejuni subsp. Doylei, C. jejuni subsp. Jejuni, C. upsaliensis, S. boydii, S. boydii 2, S. boydii 18, S. dysenteriae 2, S. dysenteriae 4, S. dysenteriae 12, S. flexneri, S. flexneri 1b, S. flexneri 2 variant (ll:3,4,7,8), S. flexneri 2a, S. flexneri 3a, S. flexneri 4, S. flexneri 4av, S. flexneri 6, and S. sonnei..

[0019] In a further embodiment, the present invention provides a method as described herein, wherein the composition comprising bovine-hyperimmune colostrum is prepared using a first vaccine comprising ETEC LPS and a second vaccine comprising ETEC LPS.

[0020] In a further embodiment, the present invention provides a method as described herein, wherein the composition comprising bovine-hyperimmune colostrum is prepared using a first vaccine comprising ETEC LPS and a second vaccine comprising a bovine coronavirus antigen and/or a bovine coronavirus.

[0021] In a further embodiment, the present invention provides a method as described herein, wherein the composition comprising bovine-hyperimmune colostrum is prepared using a vaccine comprising ETEC LPS administered to an animal sequentially- or co-administered a vaccine comprising a bovine coronavirus antigen and/or a bovine coronavirus.

[0022] In one embodiment the vaccine comprising a bovine coronavirus is an attenuated bovine coronavirus.

[0023] In a further embodiment, the present invention provides a method as described herein, wherein the composition comprising bovine-hyperimmune colostrum is prepared using a vaccine comprising ETEC LPS administered to an animal producing antibodies to bovine coronavirus. [0024] In a further embodiment, the present invention provides a method as described herein, wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to bovine coronavirus.

[0025] In a further embodiment, the present invention provides a method as described herein, wherein the human coronavirus is SARS-COV-2.

[0026] In a further embodiment, the present invention provides a method as described herein, wherein the hyperimmune colostrum is prepared by administering a bovine animal with a vaccine comprising ETEC LPS and collecting hyperimmune colostrum from the animal.

[0027] In a further embodiment, the present invention provides a method as described herein, wherein the hyperimmune colostrum is prepared by administering one bovine animal with a first vaccine and a second bovine animal with a second vaccine, and subsequently combining the hyperimmune colostrum collected from the animals.

[0028] In a further embodiment, the present invention provides a method as described herein, wherein the hyperimmune colostrum comprises one or more immunoglobulin classes selected from the group of IgG, IgM and IgA.

[0029] In a further embodiment, the present invention provides a method as described herein, wherein the hyperimmune colostrum further comprises a component selected from oligosaccharides, beta lactoglobulin, alpha-lactalbumin, lactoferrin, bovine serum albumin, growth factors, lactoperoxidase, plasm in, lipoprotein lipase, esterase, ribonucleases A, B, C, D and 11-1 , Lysozyme, enzyme inhibitors, cytokines, and lipids.

[0030] In a further embodiment, the present invention provides a method as described herein, wherein the effective amount of a composition comprising bovine- hyperimmune colostrum is administered to the subject orally, nasally or using pulmonary administration.

[0031] In a further embodiment, the present invention provides a method as described herein, wherein the contacting is performed in vivo in a human. [0032] In a further embodiment, the present invention provides a method as described herein, wherein a symptom of coronavirus infection is treated and/or prevented.

[0033] In a further embodiment, the present invention provides a method as described herein, wherein the symptom of coronavirus infection is selected from the group consisting of a secondary bacterial infection, sepsis, bowel pain, nausea, vomiting and diarrhoea.

[0034] In a further embodiment, the present invention provides a method as described herein, wherein the symptom of coronavirus infection is the load of coronavirus in stool.

[0035] In a further embodiment, the present invention provides a method of treating and/or preventing coronavirus infection in one or more human subjects comprising applying to a surface selected from air filters, PPE, room surfaces and respiratory mucosal membranes an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0036] In one embodiment the composition comprising bovine-hyperimmune colostrum is adsorbed on a particulate carrier of a size in the range from 50 to 120 microns.

[0037] In one embodiment the composition comprising bovine-hyperimmune colostrum is adsorbed onto a solid carrier for administration by dry powder inhaler.

[0038] In one embodiment the composition comprising bovine-hyperimmune colostrum is administered as an aerosol.

[0039] In one embodiment the composition comprising bovine-hyperimmune colostrum is in dried form.

[0040] In a further embodiment, the present invention provides a method of decreasing levels of LPS in the blood of a subject, comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0041 ] In a further embodiment, the present invention provides a method of treating and/or preventing sepsis in a subject, comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0042] In a further embodiment, the present invention provides a method of decreasing inflammation in a subject, comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0043] In a further embodiment, the present invention provides a method of treating and/or preventing cytokine storm in a subject comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0044] In a further embodiment, the present invention provides a method of treating and/or preventing a symptom associated with coronavirus infection in a subject comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0045] In one embodiment the symptom associated with coronavirus infection is selected from the group consisting of loss of gastrointestinal flora, increased inflammation, elevated ALT levels, elevated AST levels, pathologic change, LPS in the blood, and cytokine storm, human coronavirus-associated diarrhoea. In one embodiment the symptom associated with coronavirus infection is secondary to SARS-CoV-2 infection. In one embodiment the pathologic change is gastrointestinal tract pathology, or lung pathology.

[0046] In a further embodiment, the present invention provides a method as described herein wherein the increased inflammation is increased inflammation in the lungs, increased inflammation in the liver, and/or systemic inflammation.

[0047] In a further embodiment, the present invention provides a method of reducing viral shedding from a subject infected with a human coronavirus comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0048] In one embodiment the subject is a subject with SARS-CoV-2 infection. Description of Figures

[0049] Figure 1 A shows LPS extracted from ETEC strains in the ETEC vaccine, run on 15% Tris-tricine SDS-PAGE and stained using LPS specific silver stain. 0- polysaccharide chains are shown as ladders and lipid A core region is marked with an arrow. Shown are extracted LPS from 1 - ETEC strain B2C [serotype 06]; 2 - ETEC strain C55 3/3c3 [serotype 08]; 3 - ETEC strain PE 595 [serotype 015]; 4 - ETEC strain E11881A [serotype 025]; 5 - ETEC strain C1064-77 [serotype 027]; 6 - ETEC strain PE 672 [serotype 063]; 7 - ETEC strain E20738/0 [serotype 0114]; 8 - ETEC strain PE 724 [serotype 0115]; 9 - ETEC strain El 37-2 [serotype 0128]; 10 - ETEC strain B7A [serotype 0148]; 11 - ETEC strain E8772/0 [serotype 0153]; and 12 - ETEC strain PE 768 [serotype 0159] Not shown is LPS extracted from ETEC strain H 10407 [078: H11]; cows are also immunised with 078 LPS. Figure 1 B shows western transfer to PVDF membrane and blotted with Bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS, made from a pool of colostrum from over 1000 cows. This figure shows binding of anti-LPS antibodies against both the O-polysaccharide ladders and arrowed lipid A core region of all the serotypes included in the ETEC vaccine (078, data not shown).

[0050] Figure 2A shows a silver stained 15% T ris-tricine gel showing LPS extracted from ETEC serotypes not included in the ETEC vaccine. Shown are LPS extracted from 2 - ETEC strain M452C1 [serotype 020: H-]; 3 - ETEC strain T0225-C4 [075: H4]; 4 - ETEC strain 83-552 [0126: FI-]; 5 - ETEC strain G33 [0126: H 12]; 6 - ETEC strain M145C2 [0128:H(NT)]; 7 - ETEC strain E23477/0/A [0139:H25]; 8 - ETEC strain ND782 [0141 :H4]; 9 - ETEC strain ND748 [O149:H10]; and 10 - ETEC strain

E11881A [025:H42] (+ control). Figure 2B shows western transfer to PVDF membrane and blotted with Bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS, made from a pool of colostrum from over 1000 cows. This figure shows binding of anti-LPS antibodies against both the 0- polysaccharide ladders and arrowed lipid A core region of all the serotypes not included in the ETEC vaccine.

[0051 ] Figure 3A shows a silver stained 15% T ris-tricine gel showing LPS extracted from pathogens not included in the ETEC vaccine. Shown are LPS extracted from 2 - Enterobacter aerogenes strain ATCC 13048; 3 - Enteropathogenic Escherichia coli strain E2348/69; 4 - Klebsiella pneumoniae strain ATCC 26; 5 - Pseudomonas aeruginosa strain ATCC 27853; 6 - Salmonella typhimurium strain ATCC 14028; 7 - Vibrio cholerae strain 6239; 8 - Yersinia enterocolitica strain 67R; 9 - Citrobacter rodentium strain DSB100; and 10 - ETEC strain E11881A [025: H42] (+ control). Figure 3B shows western transfer to PVDF membrane and blotted with Bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS, made from a pool of colostrum from over 1000 cows. This figure shows binding of anti-LPS antibodies against both the O-polysaccharide ladders and arrowed lipid A core region of LPS extracted from pathogens not included in the ETEC vaccine.

[0052] Figure 4 shows hyperimmune colostrum powder Batch # 1710002659 had an EC50 of 44.7 ug/ml (left hand curve, raw data points “+”) and a CC50 of 1085.3 ug/ml (right hand curve, raw data points “°”). This batch of colostrum inhibits viral replication at a dose at which there is no cell toxicity, and at a dose that inhibits 90% of viral replication there is no cell toxicity (an EC90 of 53.15 ug/ml).

[0053] Figure 5 shows hyperimmune colostrum powder Batch # 1710002660 had an EC50 of 91 .9 7 ug/ml (left hand curve, raw data points “+”) and a CC50 of 1790.6 ug/ml (right hand curve, raw data points “°”). This batch of colostrum inhibits viral replication at a dose at which there is no cell toxicity, and at a dose that inhibits 90% of viral replication there is no cell toxicity (an EC90 of 155.39 ug/ml).

[0054] Figure 6 shows hyperimmune colostrum powder Batch # 1710002662 had an EC50 of 40.5 7 ug/ml (bottom curve, raw data points “+”) and a CC50 of 2053.3 ug/ml (top/right hand curve, raw data points “°”). This batch of colostrum inhibits viral replication at a dose at which there is no cell toxicity, and at a dose that inhibits 90% of viral replication there is no cell toxicity (an EC90 of 45.17 ug/ml).

[0055] Figure 7 shows hyperimmune colostrum powder Batch # 1710002859 had an EC50 of 42.6 7 ug/ml (left hand curve, raw data points “+”) and a CC50 of 2506.6 ug/ml (right hand curve, raw data points “°”). This batch of colostrum inhibits viral replication at a dose at which there is no cell toxicity, and at a dose that inhibits 90% of viral replication there is no cell toxicity (an EC90 of 48.69 ug/ml).

[0056] Figure 8 shows pro-milk powder Batch # 6068 N50 (control) had an EC50 of > the CC50 (EC50 - bottom curve, raw data points “+”), and a CC50 of 4818.6 ug/ml (top curve, raw data points “°”). Accordingly, this control milk powder inhibits viral replication at a dose >25000 ug/ml, and importantly does not inhibit viral replication at the doses at which it is cytotoxic to cells.

[0057] Figure 9 shows the positive control, remdesivir, has an EC50 of 1 16pM.

[0058] Figure 10 shows a composition prepared according to Example 1 decreases levels of the liver enzymes ASL and ALT in human subjects when administered orally.

[0059] Figure 11 shows a composition prepared according to Example 1 decreases LPS in the serum of human subjects when administered orally.

Detailed Description

[0060] The present inventors have developed a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS that inhibits SARS-CoV-2 infection of cells. For example, Example 2 shows hyperimmune colostrum powder compositions of the present invention, such as those prepared in Example 1 , inhibit viral replication at a dose at which there is no cell toxicity. The compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS described herein are stable at room temperature, have a desirable safety profile, and/or do not require the induction of a humoral immune response in subjects administered.

[0061] Accordingly, in one embodiment the present invention provides a method of treating and/or preventing human coronavirus infection in a subject comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0062] In one embodiment, the subject is a subject with impaired liver function. In one embodiment, the subject with impaired liver function has decreased AST and/or ALT levels.

[0063] In another embodiment of the methods described herein, the method includes a step of selecting a subject with impaired liver function prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0064] In another embodiment of the methods described herein, the method includes a step of selecting a subject with decreased AST and/or ALT levels prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0065] In one embodiment, the subject is a subject with human coronavirus- associated diarrhoea.

[0066] In another embodiment of the methods described herein, the method includes a step of selecting a subject with human coronavirus-associated diarrhoea prior to administering an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0067] In one embodiment, the subject is a subject with sepsis.

[0068] In another embodiment of the methods described herein, the method includes a step of selecting a subject with sepsis prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0069] In one embodiment, the subject is a subject with increased inflammation. In one embodiment the increased inflammation is increased inflammation in the lungs, increased inflammation in the liver, and/or systemic inflammation.

[0070] In another embodiment of the methods described herein, the method includes a step of selecting a subject with increased inflammation prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0071] In another embodiment of the methods described herein, the method includes a step of selecting a subject with increased inflammation in the lungs prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0072] In another embodiment of the methods described herein, the method includes a step of selecting a subject with increased inflammation in the liver prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0073] In another embodiment of the methods described herein, the method includes a step of selecting a subject with increased systemic inflammation prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0074] In one embodiment, the subject is a subject with LPS-induced inflammation. In one embodiment the subject has elevated levels of LPS in the serum of the subject.

[0075] In another embodiment of the methods described herein, the method includes a step of selecting a subject with LPS-induced inflammation prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0076] In another embodiment of the methods described herein, the method includes a step of selecting a subject with elevated levels of LPS in the serum prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0077] In one embodiment, the subject is a subject with cytokine storm.

[0078] In another embodiment of the methods described herein, the method includes a step of selecting a subject with cytokine storm prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0079] In one embodiment, the subject is a subject with loss of gastrointestinal flora.

[0080] In another embodiment of the methods described herein, the method includes a step of selecting a subject with loss of gastrointestinal flora prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

In one embodiment, the subject is a subject with increased pathologic change. In one embodiment, the pathologic change is gastrointestinal tract pathology, or lung pathology.

[0081] In another embodiment of the methods described herein, the method includes a step of selecting a subject with increased pathologic change prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0082] In another embodiment of the methods described herein, the method includes a step of selecting a subject with increased gastrointestinal tract pathology prior to administering an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0083] In another embodiment of the methods described herein, the method includes a step of selecting a subject with increased lung pathology prior to administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[0084] As used herein the term “human coronavirus” includes severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2): SARS-CoV-2 is the virus strain that causes coronavirus disease 2019 (COVID-19), a respiratory illness. It is colloquially known as the coronavirus and was previously referred to by its provisional name 2019 novel coronavirus (2019-nCoV). SARS-CoV-2 is a positive-sense single- stranded RNA virus. It is contagious in humans, and the World Health Organization (WHO) has designated the ongoing pandemic of COVID-19 a Public Health Emergency of International Concern. Taxonomically, SARS-CoV-2 is a strain of Severe acute respiratory syndrome-related coronavirus (SARS-CoV). It is believed to have zoonotic origins and has close genetic similarity to bat coronaviruses, suggesting it emerged from a bat-borne virus. An intermediate animal reservoir such as a pangolin is also thought to be involved in its introduction to humans. The virus shows little genetic diversity, indicating that the spillover event introducing SARS-CoV-2 to humans is likely to have occurred in late 2019. Epidemiological studies estimate each infection results in 1.4 to 3.9 new ones when no members of the community are immune and no preventive measures taken. The virus is primarily spread between people through close contact and via respiratory droplets produced from coughs or sneezes. It mainly enters human cells by binding to the receptor angiotensin converting enzyme 2 (ACE2).

[0085] In a preferred embodiment, the human coronavirus is SARS-COV-2.

[0086] As used herein the term “subject” is used interchangeably with “patient”, and refers to a human infected with, or at risk of infection with a human coronavirus.

[0087] As used herein, effective amount includes a “therapeutically effective amount” and a "prophylactically effective amount". A “therapeutically effective amount” refers to the amount of the composition comprising bovine-hyperimmune colostrum which when administered alone or in combination to a subject for treating Covid-19, or at least one of the clinical symptoms of Covid-19, is sufficient to affect such treatment of the disease, or symptom. The therapeutically effective amount can vary depending, for example, on the formulation of the composition, the infection, and/or symptoms of the infection, severity of the infection, and/or symptoms of the infection, the age, weight, and/or health of the patient to be treated, and the judgment of the prescribing physician. An appropriate therapeutically effective amount in any given instance may be ascertained by those skilled in the art or capable of determination by routine experimentation. A therapeutically effective amount is also one in which any toxic or detrimental effects of the antibody are outweighed by the beneficial effects. A "prophylactically effective amount" is any amount of the antibody that, when administered alone or in combination to a patient, inhibits or delays the onset or recurrence of Covid-19, or at least one of the clinical symptoms of Covid-19. In some embodiments, the prophylactically effective amount prevents the onset or recurrence of Covid-19 infection entirely. "Inhibiting" the onset means either lessening the likelihood of the infection's onset or preventing the onset entirely. The term includes preventing the onset of the symptoms of the disorder in a subject at risk of developing the disorder.

[0088] Each dose form may, for example, comprise the colostrum equivalent of up to 20g per day but preferably less than 1200 mg (dry weight basis), preferably less than 800 mg, preferably less than 400 mg, more preferably less than 200 mg. Colostrum equivalent refers to the amount of raw colostrum, howsoever purified, which is processed to provide the contents of a dose form, for example an oral, a nasal, or a pulmonary dose form.

[0089] For oral administration, the oral dose form may comprise 5 mg to 800 mg bovine colostrum powder (BCP) (dry weight basis), e.g. 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 70, 80, 90, 100, 125, 150, 175, 200, 225, 250, 500, 550, 600, 650, 700, 750 or 800 mg.

[0090] Suitable dosage ranges are, e.g. from about 30mg to about 20000mg/day, preferably 50mg to about 5000mg/day, more preferably 500mg to about 5000mg/day, or most preferably 1500mg to about 2000mg/day BCP (dry weight basis). In one preferred embodiment, the dose is 1800mg/day BCP (dry weight basis). [0091 ] In one embodiment the composition is formulated for administration at a dose of about 30 mg to about 10000 mg per day, or formulated for administration at a dose of about 1800 mg per day.

[0092] In one embodiment the composition is administered at a dose of about 30 mg to about 10000 mg per day, or administered at a dose of about 1800 mg per day.

[0093] In one embodiment the antibodies are present in the composition for administration in an amount sufficient to provide at least 30% by dry weight of the composition of IgG.

[0094] Accordingly, for oral administration, the oral dose form may comprise 1 .5 mg to 240 mg IgG, e.g. 1.5, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 4045, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 125, 150, 175, 200, 240, 400, 600, 800 or 1000 mg IgG.

[0095] For nasal administration, the nasal dose form may comprise 1.5 mg to 240 mg IgG, e.g. 1.5, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 40 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 125, 150, 175, 200, 240, 400, 600, 800 or 1000 mg IgG.

[0096] For pulmonary administration, the pulmonary dose form may comprise 1.5 mg to 240 mg IgG, e.g. 1.5, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 25, 30, 35, 4045, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 125, 150, 175, 200, 240, 400, 600, 800 or 1000 mg IgG.

[0097] In another embodiment the antibodies are present in the composition for administration in an amount sufficient to provide at least 36% by weight of the composition of IgG.

[0098] Suitable dosage ranges are, e.g. from about 10 to about 3333 mg/day, preferably 20 to about 4000 mg/day, more preferably 200 to about 2000mg/day, or most preferably 600 mg to about 800 mg/day IgG. In one preferred embodiment, the dose is 600 mg/day IgG.

[0099] In one embodiment the antibodies that bind to the antigen are present in the composition for administration in an amount sufficient to provide at least 5% specific IgG of the weight of IgG. [00100] Accordingly, for oral administration, the oral dose form may comprise 0.075 mg to 12 mg specific IgG, e.g. 0.075, 0.1 , 0.125, 0.15, 0.175, 0.2, 0.4, 0.6, 0.8, 1 , 1.2,

1.4, 1.6, 1.8, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 11.0 or 12 mg specific IgG.

[00101] For nasal administration, the nasal dose form may comprise 0.075 mg to 12 mg specific IgG, e.g. 0.075, 0.1 , 0.125, 0.15, 0.175, 0.2, 0.4, 0.6, 0.8, 1 , 1.2, 1.4, 1.6, 1.8, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, 10.0, 11.0 or 12 mg specific IgG.

[00102] Accordingly, for pulmonary administration, the pulmonary dose form may comprise 0.075 mg to 12 mg specific IgG, e.g. 0.075, 0.1 , 0.125, 0.15, 0.175, 0.2, 0.4, 0.6, 0.8, 1 , 1.2, 1.4, 1.6, 1.8, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0,

8.5, 9.0, 9.5, 10.0, 11.0 or 12 mg specific IgG.

[00103] In one embodiment the antibodies that bind to the antigen are present in the composition for administration in an amount sufficient to provide at least 10% specific IgG of the weight of IgG.

[00104] Suitable dosage ranges are, e.g. from about 0.5 to about 167 mg/day, preferably 10 to about 150 mg/day, more preferably 15 to about 100mg/day, or most preferably 30 mg to about 100 mg/day specific IgG. In one preferred embodiment, the dose is 30 mg/day specific IgG.

[00105] The dose form may be administered for about 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30 or 31 days.

[00106] In one embodiment, the composition is administered for 30 days.

[00107] The magnitude of prophylactic or therapeutic dose of the active ingredients can, of course, vary with the nature of the severity of the condition to be treated. It can also vary according to the age, weight and response of the individual patient, and may be administered in subject in single or divided doses. On the other hand, it may be necessary to use dosages outside the ranges provided herein in some cases.

[00108] As used herein, the term “prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS” refers to bovine-hyperimmune colostrum prepared by administering to a bovine animal a vaccine comprising enterotoxigenic E. coli (ETEC) LPS and collecting colostrum from the bovine animal. [00109] Accordingly, in one embodiment, the hyperimmune colostrum is prepared by administering a bovine animal with a vaccine comprising ETEC LPS and collecting hyperimmune colostrum from the animal. In another embodiment, the hyperimmune colostrum is prepared by administering one bovine animal with a first vaccine and a second bovine animal with a second vaccine, and subsequently combining the hyperimmune colostrum collected from the animals.

[00110] It will be understood that if the hyperimmune material is raised by vaccination of bovines it will contain polyclonal antibodies. As used herein, the terms "antibody", "antibodies" and the like include any monospecific molecule comprising a portion of the light chain variable region and/or the heavy chain variable region to effect binding to the epitope to which the antibody has binding specificity. Exemplary antibodies and fragments thereof that may be prepared according to this aspect of the invention include intact immunoglobulin molecules, substantially intact immunoglobulin molecules and fragments that contain a paratope.

[00111] Fragments, as used herein, typically include a portion of an antibody molecule that retains the ability to specifically bind to an antigen (e.g., a SARS-CoV-2 and/or LPS antigen) and include, but are not limited to, Fab, Fab', F(ab')2 and F(v). Antibody fragments may be obtained from antibodies such as described above by methods such as digestion by enzymes, such as pepsin or papain and/or by cleavage of the disulfide bridges by chemical reduction. Single chain antibodies are also intended to be encompassed within the term "fragment”.

[00112] According to one specific embodiment, the bovine hyperimmune colostrum comprises antibodies (e.g. “hyperimmune material-derived antibodies”) that bind to LPS.

[00113] According to another specific embodiment, the bovine hyperimmune colostrum comprises antibodies (e.g. “hyperimmune material-derived antibodies”) that bind to LPS, and antibodies (e.g. “hyperimmune material-derived antibodies”) that bind to a human coronavirus or antigen thereof.

[00114] According to one specific embodiment, the bovine hyperimmune colostrum comprises antibodies (e.g. “hyperimmune material-derived antibodies”) may comprise monomeric, dimeric or multimeric immunoglobulin selected from the group consisting of IgG, IgA and IgM and any fragments thereof. In ruminants, the principal compositional difference between colostrum and mature milk is the very high content of colostral immunoglobulin, of which IgG class makes up 80-90%.

[00115] In a preferred embodiment, the hyperimmune colostrum comprises one or more immunoglobulin classes selected from the group of IgG, IgM and IgA.

[00116] Thus, according to a specific embodiment, the hyperimmune material- derived antibodies mainly comprise IgG, specifically, lgG1 and lgG2.

[00117] The term “Immunoglobulin G” (IgG) as used herein, is a multimeric immunoglobulin, built of two heavy chains and two light chains. Each complex has two antigen binding sites. This is the most abundant immunoglobulin and is approximately equally distributed in blood and in tissue liquids, constituting 75% of serum immunoglobulins in humans. In general, the number of IgG subclasses varied widely between different species, ranging from one subclass in rabbits to seven subclasses in horses, making it difficult to find orthologues. In humans, for example, IgG 1 and lgG3 are the most pro-inflammatory IgG subclasses. In mice, however, lgG2a and lgG2b are the most pro-inflammatory IgG molecules showing a greater activity than mouse IgG 1 and lgG3 in many in vivo model systems.

[00118] Optionally or additionally, hyperimmune material-derived antibodies may comprise a secretory antibody, specifically, slgA.

[00119] Dimeric and multimeric IgA and IgM are secreted by a number of exocrine tissues. IgA is the predominant secretory immunoglobulin present in colostrum, milk, saliva, tears, bronchial secretions, nasal mucosa, prostatic fluid, vaginal secretions, and mucous secretions from the small intestine. IgA output exceeds that of all other immunoglobulins, making it the major antibody produced by the body daily and is the major immunoglobulin found in human milk, whey and colostrum. IgM secretion is less abundant but can increase to compensate for deficiencies in IgA secretion. J chain containing IgA is produced and secreted by plasma B immunocytes located in the lamina propria just beneath the basement membrane of exocrine cells. IgA has a typical immunoglobulin four-chain structure (Mr 160,000) made up of two heavy chains (Mr 55,000) and two light chains (Mr 23,000). In humans, there are two subclasses of IgA. These are IgAI and lgA2 that have one and two heavy chains, respectively. IgA can occur as monomers, dimers, trimers or multimers. In plasma, 10% of the total IgA is polymeric while the remaining 90% is monomeric. The secreted IgA binds to a Mr 100,000 poly-lg receptor positioned in the basolateral surface of most mucosal cells. The receptor-lgA complex is next translocated to the apical surface where IgA is secreted. The binding of dimeric IgA to the poly-lg receptor is completely dependent upon the presence of a J chain. Monomeric IgA will not bind to the receptor.

[00120] The difference in function of IgG and IgA, includes the position where the molecules operate. IgA is found mainly on mucosal surfaces where there is little in the way of tissue fluid to carry immune cells and chemicals. Therefore, IgA (often as a dimer) would be preferably used for physical neutralisation of pathogens and may be too effective at other immune functions. IgGs are present in the tissue fluid and blood where there is the full collection of leukocytes, complement system, macrophages etc. may physically neutralize a pathogen effectively and are also more effective in a communication/presentation role than IgA, i.e. , they tend to induce better opsonisation by phagocytes (e.g., Killer T cells and macrophages) and switch on the complement system better.

[00121] In another embodiment, the hyperimmune material-derived antibodies of the invention may be obtained from any one of colostrum, colostrum fractions hyperimmunised milk or colostrum, colostrum whey (either cheese or casein), cheese or casein whey, directly from skim milk, whole milk, or a reconstituted form of such streams.

[00122] In one embodiment, the present invention provides a method of preparing hyperimmune colostrum that binds to human coronavirus, said method comprising a step of administering a bovine animal with a vaccine comprising ETEC LPS and collecting hyperimmune colostrum from the animal.

[00123] It should be appreciated that hyperimmune material within the composition of the invention may be any fraction of colostrum. Thus, the term colostrum where used herein includes colostral milk, processed colostral-milk such as colostral milk processed to partly or completely removes one or more of fat, cellular debris, lactose and casein.

[00124] The colostrum, or milk, containing antibodies that bind to LPS and optionally, containing the antibodies that bind to a human coronavirus or antigen thereof, the antigen-specific antibodies may be preferably collected by milking the animal colostrum or milk thus collected can either be used directly, may be further processed, for instance to purify anti-antigen antibodies and optionally, antigen-specific antibodies. Methods for the (partial) purification of antibodies from colostrum or milk are present in the art.

[00125] The present inventors have demonstrated in Example 1 the preparation of compositions comprising bovine-hyperimmune colostrum using a vaccine comprising enterotoxigenic LPS from ETEC strain B2C [serotype 06]; ETEC strain C55 3/3c3 [serotype 08]; ETEC strain PE 595 [serotype 015]; ETEC strain E11881 A [serotype 025]; ETEC strain C1064-77 [serotype 027]; ETEC strain PE 672 [serotype 063]; ETEC strain E20738/0 [serotype 0114]; ETEC strain PE 724 [serotype 0115]; ETEC strain El 37-2 [serotype 0128]; ETEC strain B7A [serotype 0148]; ETEC strain E8772/0 [serotype 0153]; and ETEC strain PE 768 [serotype 0159], and LPS extracted from ETEC strain H 10407 [078: H11]

[00126] Accordingly, in one embodiment the vaccine comprising ETEC LPS comprises one or more LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159. In another embodiment, the vaccine comprising ETEC LPS comprises LPS 0 serotypes 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159.

[00127] As used herein the term LPS refers to Lipopolysaccharide, which comprises a lipid and a polysaccharide composed of O-antigen, core oligosaccharide (e.g. outer and inner core) joined by a covalent bond, and lipid A.

[00128] In one embodiment, the vaccine comprising ETEC LPS comprises the lipid A, core and O-polysaccharide regions of the LPS.

[00129] In one embodiment, the compositions of the present invention are prepared using the methods described in PCT/AU2004/000277, hereby incorporated by reference in its entirely.

[00130] The present inventors have demonstrated in Example 1 that compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic LPS comprise bovine immunoglobulin that binds to ETEC LPS.

[00131] In particular, the present inventors have demonstrated in Example 1 that compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic LPS from ETEC strain B2C [serotype 06]; ETEC strain C55 3/3c3 [serotype 08]; ETEC strain PE 595 [serotype 015]; ETEC strain E11881 A [serotype 025]; ETEC strain C1064-77 [serotype 027]; ETEC strain PE 672 [serotype 063]; ETEC strain E20738/0 [serotype 0114]; ETEC strain PE 724 [serotype 0115]; ETEC strain El 37-2 [serotype 0128]; ETEC strain B7A [serotype 0148]; ETEC strain E8772/0 [serotype 0153]; and ETEC strain PE 768 [serotype 0159], and LPS extracted from ETEC strain H10407 [078:H11] comprise bovine immunoglobulin that binds to one or more LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 071, 0114, 0115, 0117, 0128, 0148, 0153, and 0159, 0167.

[00132] Accordingly, in one embodiment the present invention provides a method as described herein wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to ETEC LPS.

[00133] Accordingly, in another embodiment the present invention provides a method as described herein wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to one or more LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 071, 0114, 0115, 0117, 0128, 0148, 0153, and 0159, 0167.

[00134] The present inventors have also demonstrated in Example 1 that compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic LPS from ETEC strain B2C [serotype 06]; ETEC strain C55 3/3c3 [serotype 08]; ETEC strain PE 595 [serotype 015]; ETEC strain E11881 A [serotype 025]; ETEC strain C1064-77 [serotype 027]; ETEC strain PE 672 [serotype 063]; ETEC strain E20738/0 [serotype 0114]; ETEC strain PE 724 [serotype 0115]; ETEC strain El 37-2 [serotype 0128]; ETEC strain B7A [serotype 0148]; ETEC strain E8772/0 [serotype 0153]; and ETEC strain PE 768 [serotype 0159], and LPS extracted from ETEC strain H10407 [078:H11] comprise bovine immunoglobulin that binds to one or more LPS serotypes of a gram negative bacteria selected from the group consisting of Enteropathogenic Escherichia coli strain E2348/69, Klebsiella pneumoniae strain ATCC 26, Pseudomonas aeruginosa strain ATCC 27853, Salmonella typhimurium strain ATCC 14028, Vibrio cholerae strain 6239, Yersinia enterocolitica strain 67R, C. jejuni, Vibrio cholera Ogawa, V. cholera Inaba, and V. cholera NON 01/0139, C. coli, C. jejuni subsp. Doylei, C. jejuni subsp. Jejuni, C. upsaliensis, S. boydii, S. boydii 2, S. boydii 18, S. dysenteriae 2, S. dysenteriae 4, S. dysenteriae 12, S. flexneri, S. flexneri 1b, S. flexneri 2 variant (ll:3,4,7,8), S. flexneri 2a, S. flexneri 3a, S. flexneri 4, S. flexneri 4av, S. flexneri 6, and S. sonnei..

[00135] Accordingly, in another embodiment the present invention provides a method as described herein wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to one or more LPS serotypes of a gram negative bacteria selected from the group consisting of Enteropathogenic Escherichia coli strain E2348/69, Klebsiella pneumoniae strain ATCC 26, Pseudomonas aeruginosa strain ATCC 27853, Salmonella typhimurium strain ATCC 14028, Vibrio cholerae strain 6239, Yersinia enterocolitica strain 67R, C. jejuni, Vibrio cholera Ogawa, V. cholera Inaba, and V. cholera NON 01/0139, C. coli, C. jejuni subsp. Doylei, C. jejuni subsp. Jejuni, C. upsaliensis, S. boydii, S. boydii 2, S. boydii 18, S. dysenteriae 2, S. dysenteriae 4, S. dysenteriae 12, S. flexneri, S. flexneri 1b, S. flexneri 2 variant (ll:3,4,7,8), S. flexneri 2a, S. flexneri 3a, S. flexneri 4, S. flexneri 4av, S. flexneri 6, and S. sonnei..

[00136] In one embodiment, a bovine mammal is co-administered with more than one vaccine, for example a first vaccine comprising one or more LPS from a one or more strains of ETEC and a second vaccine comprising one or more LPS from a one or more strains of ETEC.

[00137] Accordingly, in one embodiment the present invention provides a method as described herein wherein the composition comprising bovine-hyperimmune colostrum is prepared using a first vaccine comprising ETEC LPS and a second vaccine comprising ETEC LPS.

[00138] The present inventors have demonstrated in Example 2 that compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) inhibit SARS-CoV-2 replication, and inhibit SARS-CoV- 2 replication at doses that are not cytotoxic to cells.

[00139] In one embodiment, a bovine mammal infected or previously infected with bovine coronavirus is administered a vaccine comprising one or more LPS from a one or more strains of ETEC. Bovine coronavirus (BCoV) is a widely distributed pathogen, causing disease and economic losses in the cattle industry worldwide.

[00140] Without wishing to be bound by theory, the present inventors propose that administration of vaccine comprising one or more LPS from a one or more strains of ETEC to a bovine animal infected or previously infected with bovine coronavirus results in bovine-hyperimmune colostrum that can inhibit SARS-CoV-2 replication of human cells. Cross-protection against a human enteric coronavirus and a virulent bovine enteric coronavirus in gnotobiotic calves has been demonstrated, and the present inventors propose administration of a vaccine comprising one or more LPS from a one or more strains of ETEC can enhance titres of anti-coronavirus antibodies.

In one embodiment, a bovine mammal is co-administered with more than one vaccine, for example a first vaccine comprising one or more LPS from a one or more strains of ETEC and second vaccine comprising a coronavirus antigen and/or a coronavirus.

[00141] Accordingly, in one embodiment the present invention provides a method as described herein wherein the composition comprising bovine-hyperimmune colostrum is prepared using a first vaccine comprising ETEC LPS and a second vaccine comprising a coronavirus antigen and/or a coronavirus.

[00142] In one embodiment, the composition comprising bovine-hyperimmune colostrum is prepared using a vaccine comprising ETEC LPS administered to an animal sequentially- or co-administered a vaccine comprising a coronavirus antigen and/or a coronavirus.

[00143] In one embodiment, the vaccine comprising a coronavirus is an attenuated coronavirus.

[00144] In another embodiment, the composition comprising bovine-hyperimmune colostrum is prepared using a vaccine comprising ETEC LPS administered to an animal producing antibodies to coronavirus.

[00145] In another embodiment, the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to coronavirus.

[00146] In one embodiment, the coronavirus antigen is a human or bovine coronavirus antigen.

[00147] In another embodiment, the coronavirus is a human or bovine coronavirus.

[00148] It should be further appreciated that any adjuvants may be added to the compositions of the invention. Appropriate adjuvants therefore may be any antigen, antibody, glycosphingolipids, proteins, cytokines, adhesion molecules, and component that can activate or alter the function of antigen presenting cell or of any other cell related to the immune system in a direct and indirect manner.

[00149] Alternatively, antibodies that bind to LPS and/or antibodies that bind to a human coronavirus or antigen thereof may be an affinity purified antibody or any fragment thereof. The term "antibody" is meant to include both intact molecules as well as fragments thereof, such as, for example, Fab and F(ab')2, which are capable of binding antigen. Fab and F(ab')2 fragments lack the Fc fragment of intact antibody, clear more rapidly from the circulation, and may have less non-specific tissue binding than an intact antibody. It will be appreciated that Fab and F(ab')2 and other fragments of the antibodies useful in the present invention may be used for immuno-modulation, according to the methods disclosed herein for intact antibody molecules. Such fragments are typically produced by proteolytic cleavage, using enzymes such as papain (to produce Fab fragments) or pepsin (to produce F(ab')2 fragments).

[00150] According to another embodiment, the composition of the invention may optionally further comprise colostrum component/s such as for example, alarmins, defensins, colostrinin, and any other colostrum or milk derived carbohydrates, glycolipids or any other molecules or components that may further enhance or inhibit modulation of an immune response, or any preparations, mixtures or combinations thereof. Moreover, the composition of the invention may comprise any additional adjuvant. Appropriate adjuvants therefore may be any antigen, antibody, glycosphingolipids, proteins, cytokines, adhesion molecules, and component that can activate or alter the function of antigen presenting cell or of any other cell related to the immune system in a direct and indirect manner.

[00151] In one embodiment, the hyperimmune colostrum further comprises a component selected from oligosaccharides, beta lactoglobulin, alpha-lactalbumin, lactoferrin, bovine serum albumin, growth factors, lactoperoxidase, plasm in, lipoprotein lipase, esterase, ribonucleases A, B, C, D and 11-1 , Lysozyme, enzyme inhibitors, cytokines, and lipids.

[00152] In another embodiment, the hyperimmune colostrum further comprises an effective amount of a therapeutic agent, for example, an antiviral agent. [00153] In one embodiment, the effective amount of a composition comprising bovine-hyperimmune colostrum is administered to the subject orally, nasally or using pulmonary administration.

[00154] In a preferred embodiment, the composition is formulated for oral administration.

[00155] Orally administrated antibodies would be expected to be degraded in the gastrointestinal tract, given the low gastric pH and the presence of gastric and intestinal proteases. However, bovine colostral IgG (BCIg) has been cited as particularly resistant to Gl destruction, relative to other immunoglobulins. Early studies of BCIg cited remarkable "resistance to proteolytic digestion in the intestine of a heterologous host". There is also evidence that bovine lgG1 is somewhat more resistant to proteolysis by trypsin, chymotrypsin and pepsin than other Igs. These results drove much of the early development of oral antibody therapy. More specifically, the composition of the invention may be suitable for mucosal administration, for example, pulmonary, buccal, nasal, intranasal, sublingual, rectal, vaginal administration and any combination thereof.

[00156] As indicated above, although oral administration is preferred, it should be appreciated that any other route of administration may be applicable, for example, intravenous, intravenous, intramuscular, subcutaneous, intraperitoneal, parenteral, intravaginal, intranasal, mucosal, sublingual, topical, rectal or subcutaneous administration, or any combination thereof.

[00157] According to one preferred embodiment, any of the compositions of the invention may be administered orally or by inhalation as an aerosol or by intravenous, intramuscular, subcutaneous, intraperitoneal, parenteral, transdermal, intravaginal, intranasal, mucosal, sublingual, topical, rectal or subcutaneous administration, or any combination thereof.

[00158] The compositions described herein may be administered in dosage formulations containing conventional non-toxic acceptable carriers and may also include one or more acceptable additives, including acceptable salts, polymers, solvents, buffers, excipients, bulking agents, diluents, excipients, suspending agents, lubricating agents, adjuvants, vehicles, deliver systems, emulsifiers, disintegrants, absorbents, preservatives, surfactants, colorants, flavourants or sweeteners. An optional dosage form of the present invention may be a powder for incorporation into beverages, pills, syrup, capsules, tablets, granules, beads, chewable lozenges or food additives, using techniques known in the art. Thus, immuno-modulating composition of the invention may be administered in a form selected from the group consisting of orally-active powders, pills, capsules, teas, extracts, dried extracts, sublinguals, sprays, dispersions, solutions, suspensions, emulsions, foams, syrups, lotions, ointments, gels, pastes, dermal patches, injectables, creams and suppositories. Therapeutic formulations may be administered in any conventional dosage formulation. Formulations typically comprise at least one active ingredient, as defined above, together with one or more acceptable carriers thereof.

[00159] Each carrier should be both pharmaceutically and physiologically acceptable in the sense of being compatible with the other ingredients and not injurious to the patient. Formulations include those suitable for oral, rectal, nasal, or parenteral (including subcutaneous, intramuscular, intravenous and intradermal or by inhalation) administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The nature, availability and sources, and the administration of all such compounds including the effective amounts necessary to produce desirable effects in a subject are well known in the art and need not be further described herein.

[00160] The preparation of pharmaceutical compositions is well known in the art and has been described in many articles and textbooks, see e.g., Remington's Pharmaceutical Sciences, Gennaro A. R. ed., Mack Publishing Co., Easton, PA, 1990, and especially pp. 1521-1712 therein, fully incorporated herein by reference.

[00161] The compositions of the invention generally comprise a buffering agent, an agent that adjusts the osmolarity thereof, and optionally, one or more pharmaceutically acceptable carriers, excipients and/or additives as known in the art. Supplementary active ingredients can also be incorporated into the compositions. The carrier can be solvent or dispersion medium containing, for example, water, ethanol, polyol (for example, glycerol, propylene glycol, and liquid polyethylene glycol, and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating, such as lecithin, by the maintenance of the required particle size in the case of dispersion and by the use of surfactants. [00162] As used herein "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, coatings, antibacterial and antifungal agents and the like. The use of such media and agents for pharmaceutical active substances is well known in the art. Except as any conventional media or agent is incompatible with the active ingredient, its use in the therapeutic composition is contemplated.

[00163] In instances in which oral administration is in the form of a tablet or capsule, the active components can be combined with a non-toxic pharmaceutically acceptable inert carrier such as lactose, starch, sucrose, glucose, modified sugars, modified starches, methylcellulose and its derivatives, dicalcium phosphate, calcium sulfate, mannitol, sorbitol, and other reducing and non-reducing sugars, magnesium stearate, stearic acid, sodium stearyl fumarate, glyceryl behenate, calcium stearate and the like. For oral administration in liquid form, the active drug components can be combined with non-toxic pharmaceutically acceptable inert carriers such as ethanol, glycerol, water and the like. When desired or required, suitable binders, lubricants, disintegrating agents and colouring and flavouring agents can also be incorporated into the mixture. Stabilizing agents such as antioxidants, propyl gallate, sodium ascorbate, citric acid, calcium metabisulphite, hydroquinone, and 7-hydroxycoumarin can also be added to stabilize the dosage forms. Other suitable compounds can include gelatin, sweeteners, natural and synthetic gums such as acacia, tragacanth, or alginates, carboxymethylcellulose, polyethylene, glycol, waxes and the like.

[00164] In one embodiment the present invention provides a method of treating and/or preventing coronavirus infection in one or more human subjects comprising applying to a surface selected from air filters, PPE, room surfaces and respiratory mucosal membranes an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00165] In another embodiment, the surface is a filter on which the immune material is adsorbed. The filter may be used to inhibit infection of people with SARS-CoV-2 which may be present in the airstream.

[00166] In a further embodiment, the antibodies or fragments are dissolved or dispersed in a liquid which is then used in a nebulising device or to provide an antibody aerosol which is released into the atmosphere of a hospital ward or sick room or other atmosphere containing SARS-CoV-2 virus.

[00167] In a further embodiment, the antibody aerosol is directed into an air stream comprising polluted air which is then passed over or through a surface, for example a porous surface or a hairy surface or some other surface having an extended surface area. In this case the role of the surface is to capture both SARS-CoV-2 viruses or virions and also antibody moieties, leading to at least partial neutralisation of the polluted air stream.

[00168] In another embodiment, the antibodies or fragments are deposited on the surface of a face mask, or within the fabric of a face mask.

[00169] The personal protective equipment and air filters include marks or personal air filters such as may be worn over the nose and mouth of the subjects or the filters may be building or air conditioner filters in buildings inhabited by the subjects.

[00170] In another embodiment, the surface is any surface in a room frequented by subjects with Covid-19, for example a hospital room.

[00171] In another embodiment, the composition comprising bovine-hyperimmune colostrum is adsorbed on a particulate carrier of a size in the range from 50 to 120 microns

[00172] In another embodiment, the composition comprising bovine-hyperimmune colostrum is adsorbed onto a solid carrier for administration by dry powder inhaler.

[00173] In one embodiment, the antibodies or fragments thereof are provided in the form of an inhalational dose.

[00174] In one embodiment, the composition comprising bovine-hyperimmune colostrum is administered as an aerosol.

[00175] In one embodiment, the composition comprising bovine-hyperimmune colostrum in dried form.

[00176] The compositions may be provided in milled particulate form of the immune material wherein the average particle size is less than 20 microns, preferably less than 10 microns, more preferably less than 5 microns. Once small particles have been produced, the micronized substance may be blended with an excipient. Examples of suitable carriers may include one or more carbohydrate, such as fructose, glucose, galactose, sucrose, lactose, trehalose, raffinose, melezitose; alditols, such as mannitol and xylitol; maltodextrins, dextrans, cyclodextrins, amino acids, such as glycine, arginine, lysine, aspartic acid, glutamic acid and polypeptides, such as human serum albumin and gelatin. To mask the unpleasant taste of some inhaled drug compounds, flavouring particles containing maltodextrin and peppermint oil may be incorporated into dry powder formulations. Large sized particles increase mouth deposition and reduce lung deposition. Lactose is a particularly preferred carrier.

[00177] The carrier particles are typically relatively large such as on the order of 50 to 120 microns, or approximately 50 times bigger than the milled particles containing the bovine hyperimmune material. These carrier particles help to facilitate the dispersion of the small particles and allow precise filling into the dry powder inhaler (DPI) powder storage system in a reproducible manner. Milled bovine hyperimmune colostrum is blended with lactose at concentrations ranging from <1 to 50% by weight. Finally, the blend is filled into the powder storage systems of an inhaler at weights ranging from approximately 3-25 mg. Preferred powder storage systems are shown in U.S. Pat. Nos. 5,492,112; 5,645,051 ; 5,622,166; and 5,921 ,237, incorporated herein by reference. The inhaler and storage systems shown in U.S. Pat. Nos. 5,921 ,237 and 5,622,166, incorporated herein by reference, are appropriate for use with vaccines. In these systems, a dry powder formulation is sealed into a foil blister that protects the powder from exposure to high humidity, reduces the risk of contamination, and can prevent inactivation of the vaccine by sunlight. The process of preparing dry powder blends for aerosol delivery involves three basic steps.

[00178] Size reduction may be accomplished by a variety of techniques including spray drying, precipitation from supercritical fluids, and jet milling or micronization. Preferably, jet milling is used. This technique uses high pressure, high velocity gas to cause particle to particle attrition to generate small particles at high efficiencies. Multidose DPIs may be used with disposable cassettes or foil blister disks, strips or unit dose blisters to deliver many doses, contributing to the cost effectiveness of this approach as compared with syringes, particularly single use syringes. These DPI's may be provided with disposable mouthpieces that can be used in mass dosing campaigns. Alternatively, unit dose DPIs with vaccine sealed in the aerosolization chamber can be used. [00179] The inhaler may have a body, a mouthpiece and an airflow passage. A restrictor plate may be used having flow control openings in the airflow passage opposite from the mouthpiece. A dose of dry powder vaccine sealed by a foil strip or capsule is received within the body of the inhaler. In use, the foil strip or capsule is pulled out or back, peeling or breaking open blister formed around the dose or a capsule is received in a chamber provided with means such as a sharp object for piercing the capsule to release the powder which may occur in response to actuation by the user to urge the sharp object into the capsule or blister by means of a trigger lever, button or the like. The subject inhales on the mouthpiece, and the dose is drawn into the lungs.

[00180] Alternatively, or in addition, the inhaler may be provided with means for actively generating an air flow in response to actuation by manual operation of the user or commencement of the inhalation process by the user.

[00181] In this embodiment the composition is administered so as to come in contact with an airway surface of a human subject in the upper respiratory tract, preferably an airway surface within 3 or 4 branch points of the trachea.

[00182] In yet another embodiment, the composition optionally further comprises colostrum, milk or milk products component/s and any adjuvant/s, preferably, alarmins, defensins, colostrinin and any preparation, mixture or combination thereof. It should be further appreciated that the composition of the invention may comprise any additional adjuvant. Appropriate adjuvants therefore may be any antigen, antibody, glycosphingolipids, proteins, cytokines, adhesion molecules, and component that can activate or alter the function of antigen presenting cell or of any other cell related to the immune system in a direct and indirect manner. It should be noted that according to certain embodiments the present invention further provides the use of colostrum or any colostrum-derived preparations in the combined compositions of the invention for enhancing an immunomodulatory effect of an immunomodulatory therapeutic agent.

[00183] The term alarmin denotes an array of structurally diverse multifunctional host proteins that are rapidly released during infection or tissue damage, and that have mobilizing and activating effects on receptor-expressing cells engaged in host defence and tissue repair. Innate-immune mediators that have alarmin function include defensins, eosinophil-derived neurotoxin, cathelicidins and HMGB1. [00184] Defensins are small (15-20 residue) cysteine-rich cationic proteins found in both vertebrates and invertebrates. They are active against bacteria, fungi and enveloped viruses. They consist of 15-20 amino acids including six to eight conserved cysteine residues. Cells of the immune system contain these peptides to assist in killing phagocytized bacteria, for example in neutrophil granulocytes and almost all epithelial cells. Most defensins function by penetrating the microbes cell membrane by way of electrical attraction, and once embedded, forming a pore in the membrane which allows efflux.

[00185] The term "Colostrinin", as use herein refers to a polypeptide which, in its natural form, is obtained from mammalian colostrum. Colostrinin is sometimes known as "colostrinine", and has a molecular weight in the range 16,000 to 26,000 Daltons. Colostrinin may form a dimer or trimer of sub-units (each having a molecular weight in the range 5,000 to 10,000 Daltons, preferably 6,000 Daltons), and contains mostly proline (the amount of proline is greater than the amount of any other single amino acid). Colostrinin is characterized in that it stimulates the production of cytokines, especially gamma interferon (IFN-y), tumor necrosis factor TNF-a), interleukins (e.g. IL-6 and IL-10) and various growth factors.

[00186] During the 2003 outbreak of severe acute respiratory syndrome (SARS), “viral detection in [gastrointestinal] biopsy specimens and stool even in discharged patients ... partially provide[d] explanations for the gastrointestinal symptoms, potential recurrence and transmission of SARS from persistently shedding human[s].” Importantly, it has been demonstrated that more than 50% of people infected with SARS-CoV-2 have the virus in their stool; some patients have vomiting and diarrhea; and some test positive for virus in stool even after respiratory samples test negative for the pathogen. The first patient in the United States diagnosed with SARS-CoV-2 infection reported two days of nausea and vomiting before presenting to the hospital, and had loose bowel movements while in the hospital. Both stool and respiratory specimens from the patient tested positive for SARS-CoV-2. A growing body of clinical evidence indicates the digestive system may serve as an alternative route of SARS- CoV-2 infection in addition to the respiratory tract, suggesting fecal-oral transmission. Furthermore, a retrospective cohort study of the clinical course and risk factors for mortality of adult inpatients with COVID-19 in Wuhan demonstrated that while no bacterial pathogens were detected in COVID-19 patients on admission, more than half of patients developed bacterial sepsis.

[00187] Importantly, many patients with severe COVID-19 show signs of a cytokine storm.

[00188] Since the first reports on COVID-19 disease, it appeared clear that Acute respiratory distress syndrome (ARDS) accounted for a significant number of deaths among infected patients and that ARDS should be regarded as the hallmark immune- mediated clinical consequence in SARS-CoV-2, similarly to what described for SARS- CoV and MERS-CoV infections. Acute respiratory distress syndrome (ARDS) is a devastating event, with an estimated mortality of approximately 40 %, defined as the presence of bilateral lung infiltrates and severe hypoxemia. ARDS can occur in a variety of clinical situations, including pneumonia, sepsis, pancreatitis, blood transfusion. ARDS pathogenesis involves inflammatory injury to the alveolo-capillary membrane, which results in increased lung permeability and the exudation of protein- rich pulmonary edema fluid into the airspaces, leading in the end to respiratory insufficiency. As shown by previous data in the literature, increased circulating levels of pro-inflammatory cytokines (eg, Interferon y, interleukin (IL-) 1 B, IL-6, IL-12) and chemokines (CXCL10, and CCL2) are associated with pulmonary inflammation and extensive lung involvement in SARS patients, similarly to what happens in MERS-CoV infection. As far as COVID 19 infection is concerned, Huang et al. recently reported that infected patients also show high levels of pro-inflammatory cytokines and chemokines. The demonstration of increased levels of IL-1 B, IFNy, CXCL10, and CCL2 strongly pointed toward an activation of T-helper-1 (Th1 ) cell function. More importantly, the so called “cytokine storm” emerged as a main factor driving a more severe clinical course. This concept originated from the observation that COVID-19 patients requiring ICU admission displayed higher concentrations of CXCL10, CCL2 and TNFa as compared to those in which the infection was less severe and did not require an ICU admission. To further complicate the issue, it should be highlighted that, in patients with SARS-Cov-2 infection, at difference from SARS-CoV infection, there is also an increased secretion Th2-immune-oriented cytokines such as IL-4 and IL-10, whose main effect is to suppress inflammation.

[00189] Taken together, these data clearly indicate that, in SARS-CoV infection, ARDS is the ultimate result of a cytokine storm. In this scenario, the release by immune effector cells of large amounts of pro-inflammatory cytokines (IFNa, IFNy, IL-1 b, IL-6, IL-12, IL-18, IL-33, TNFa, TGFP) and chemokines (CXCL10, CXCL8, CXCL9, CCL2, CCL3, CCL5) precipitates and sustains the aberrant systemic inflammatory response. The cytokine storm is readily followed by the immune system “attacking” the body, which in turn will cause ARDS and multiple organ failure, the final result being death, at least in the most severe cases of SARS-CoV-2 infection.

[00190] The cytokine storm, and the consequent ARDS, results from the effects of a combination of many immune-active molecules.

[00191] In other infectious and non-infectious diseases in which the immune system is affected, such as HIV-AIDS and inflammatory bowel disease, damage to gut- associated lymphocyte tissues occurs, enabling luminal bacteria to enter into the circulation. Lipopolysaccharide (LPS) is a bacterial product that stimulates macrophages, leading to the production of pro-inflammatory cytokines and other soluble factors such as MIF, which in turn activate lymphocytes. Continuous and exaggerated stimulation causes exhaustion of the T-cell compartment, contributing to immunosuppression. Importantly, release of lipopolysaccharide (LPS) into the bloodstream, which triggers an uncontrolled systemic inflammatory response (cytokine storm) leading to multiorgan failure and death.

[00192] The present inventors have demonstrated in Example 4 that the bovine hyperimmune colostrum compositions described herein can decrease levels of LPS in the serum of patients with inflammation. Without wishing to be bound by theory, the present inventors propose that the bovine hyperimmune colostrum compositions described herein can be used to treat and/or prevent one or more symptoms associated with Covid-19, including cytokine storm.

[00193] In one embodiment the present invention provides methods of decreasing levels of LPS in the blood of a subject comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00194] In another embodiment the present invention provides methods of treating and/or preventing sepsis in a subject comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS. [00195] In another embodiment the present invention provides methods of decreasing inflammation in a subject comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00196] In another embodiment the present invention provides methods of treating and/or preventing cytokine storm in a subject comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00197] In one embodiment the cytokine storm is LPS-associated cytokine storm.

[00198] In another embodiment the subject is a patient infected with a coronavirus.

[00199] In a further embodiment, the coronavirus is SARS-Cov-2.

[00200] The skilled artisan would appreciate that decreasing toxic cytokine release or toxic cytokine levels comprises decreasing or inhibiting production of toxic cytokine levels in a subject, or inhibiting or reducing the incidence of cytokine release syndrome or a cytokine storm in a subject by decreasing LPS in the serum of the subject. In another embodiment toxic cytokine levels are reduced during cytokine storm. In another embodiment, decreasing or inhibiting the production of toxic cytokine levels comprises treating cytokine storm. In another embodiment, decreasing or inhibiting the production of toxic cytokine levels comprises preventing cytokine storm. In another embodiment, decreasing or inhibiting the production of toxic cytokine levels comprises alleviating a cytokine storm. In another embodiment, decreasing or inhibiting the production of toxic cytokine levels comprises ameliorating a cytokine storm. In another embodiment, the toxic cytokines comprise pro-inflammatory cytokines. In another embodiment, pro-inflammatory cytokines comprise IL-6. In another embodiment, pro inflammatory cytokines comprise IL-1. In another embodiment, pro-inflammatory cytokines comprise TNF-a. In another embodiment, pro-inflammatory cytokines comprise IL-6, IL- 13, or TNF-a, or any combination thereof.

[00201] In one embodiment, cytokine storm is characterized by elevated levels of several inflammatory cytokines and adverse physical reactions in a subject such as low blood pressure, high fever and shivering. In another embodiment, inflammatory cytokines comprise IL-6, IL-1 , and TNF-a. In another embodiment, cytokine storm is characterized by elevated levels of IL-6, IL- 13, or TNF-a, or any combination thereof. [00202] In another embodiment, cytokine storm is characterized by elevated levels of IL-8, or IL-13, or any combination thereof. In another embodiment, a cytokine storm is characterized by increases in TNF-alpha, IFN-gamma, IL-lbeta, IL-2, IL-6, IL-8, IL- 10, IL-13, GM CSF, IL-5, CX3CL1 , or a combination thereof or a subset thereof. In yet another embodiment, IL-6 comprises a marker of cytokine storm. In another embodiment, IFN-y comprises a marker of cytokine storm. In another embodiment, patients with coronavirus infection have higher incidence and severity of cytokine storm.

[00203] The present inventors have demonstrated in Examples 3 and 4 that the bovine hyperimmune colostrum compositions described herein can decrease markers of inflammation in human subjects. Without wishing to be bound by theory, the present inventors propose that the bovine hyperimmune colostrum compositions described herein can be used to treat and/or prevent one or more symptoms associated with inflammation, such as immune pathology associated with CARS-CoV-2 infection.

[00204] In another embodiment the present invention provides methods of treating and/or preventing a symptom associated with coronavirus infection. in a subject comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00205] In one embodiment the symptom associated with coronavirus infection is a cytokine storm, LPS-associated cytokine storm, inflammation, pathologic change, LPS in the blood, elevated ALT, ASL levels.

[00206] The present inventors have demonstrated that the bovine hyperimmune colostrum compositions described herein can decrease AST and ALT levels in patents with inflammation. As shown, by the Example 3, the compositions of the invention, significantly decreased the serum levels of ALT and AST. Therefore, according to one embodiment, the composition of the invention leads to at least one of a decrease in the serum levels of ALT and AST in a subject with inflammation. Decrease, reduction, inhibition, as used herein refers to a reduction of about 5% to 99% of the serum level of ALT or AST in a subject. More specifically, such reduction may be a reduction of about, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% and over 99%, as compared to the levels prior to the treatment, or the levels of untreated control.

[00207] In one embodiment, the pathologic change is gastrointestinal tract pathology, or lung pathology.

[00208] In another embodiment, the inflammation is systemic inflammation, inflammation in the lungs or inflammation in the liver.

[00209] In another embodiment the subject is a patient infected with a coronavirus.

[00210] In a further embodiment, the coronavirus is SARS-Cov-2.

[00211 ] In a further embodiment, the subject is an elderly subject.

[00212] As used herein the term "cytokine" includes cytokines (e.g., interferon gamma, granulocyte macrophage colony stimulating factor, tumor necrosis factor alpha), chemokines (e.g., MIP 1 alpha, MIP 1 beta, RANTES), and other soluble mediators of inflammation, such as reactive oxygen species and nitric oxide.

[00213] As used herein, cytokine storm is characterized by any or all of the following symptoms: fever with or without rigors, malaise, fatigue, anorexia, myalgias, arthalgias, nausea, vomiting, headache, skin rash, nausea, vomiting, diarrhea, Tachypnea, hypoxemia Cardiovascular Tachycardia, widened pulse pressure, hypotension, increased cardiac output (early), potentially diminished cardiac output (late), Elevated D-dimer, hypofibrinogenemia with or without bleeding, Azotemia Hepatic Transaminitis, hyperbilirubinemia, mental status changes, confusion, delirium, word finding difficulty or frank aphasia, hallucinations, tremor, dysmetria, altered gait, seizures. In another embodiment, a cytokine storm is characterized by IL-2 release and lymphoproliferation.

[00214] In another embodiment, cytokine storm leads to potentially life-threatening complications including cardiac dysfunction, adult respiratory distress syndrome, neurologic toxicity, renal and/or hepatic failure, and disseminated intravascular coagulation.

[00215] In one embodiment, measurement of cytokine levels or concentration, as an indicator of cytokine storm, may be expressed as -fold increase, percent (%) increase, net increase or rate of change in cytokine levels or concentration. In another embodiment, absolute cytokine levels or concentrations above a certain level or concentration may be an indication of a subject undergoing or about to experience a cytokine storm. In another embodiment, absolute cytokine levels or concentration at a certain level or concentration, for example a level or concentration normally found in a control subject not undergoing CAR-T cell therapy, may be an indication of a method for inhibiting or reducing the incidence of a cytokine storm in a subject undergoing CAR T-cell.

[00216] A skilled artisan would appreciate that the term "cytokine level" may encompass a measure of concentration, a measure of fold change, a measure of percent (%) change, or a measure of rate change. Further, the methods for measuring cytokines in blood, saliva, serum, urine, and plasma are well known in the art.

[00217] In one embodiment, despite the recognition that cytokine storm is associated with elevation of several inflammatory cytokines, IL-6 levels may be used as a common measure of cytokine storm and/or as a common measure of the effectiveness of a treatment for cytokine storms. A skilled artisan would appreciate that other cytokines may be used as markers of a cytokine storm, for example any of TNF-a, IL- 6, IL-8, IL-13, or INF-y, or any combination above may be used as a marker a cytokine storm. Further, that assay methods for measuring cytokines are well known in the art.

[00218] As used herein, "treatment" refers to the reduction or elimination of the severity of a symptom of the disease, the frequency with which such a symptom is exhibited, or both. The term includes an action that occurs while a patient is suffering from Covid-19 or associated disorder that reduces the severity of one or more symptoms or effects of Covid-19 or a Covid-19 associated disease or symptom.

[00219] As used herein, "prophylaxis" or “prevention” refers to completely or partially preventing or inhibiting a symptom of the disease or the frequency with which such a symptom is exhibited, or a reduction of the risk of acquiring a given symptom, of Covid- 19. Prophylaxis includes inhibiting Covid-19 associated diarrhea, preventing Covid- 19 associated diarrhea, decreasing the severity of Covid-19 associated diarrhea or improving signs and symptoms related to having Covid-19 associated diarrhea. Prophylaxis includes inhibiting, preventing, or decreasing the severity a symptom of Covid-19 as described herein. The term includes action that occurs before a patient begins to suffer from Covid-19 or associated disorder, such as but not limited to bowel or gastrointestinal disorder, that delays the onset of, and/or inhibits or reduces the severity of Covid-19 or Covid-19 associated disease or symptom.

[00220] In one embodiment the present invention provides a method as described herein wherein a symptom of coronavirus infection is treated and/or prevented.

[00221] Although SARS-Cov-2 primarily causes lung infection through binding of ACE2 receptors present on the alveolar epithelial cells, yet it was recently reported that SARS-CoV-2 RNA was found in the faeces of infected patients. Interestingly, the intestinal epithelial cells particularly the enterocytes of the small intestine also express ACE2 receptors. The role of the gut microbiota in influencing lung disease is well known.

[00222] SARS-CoV-2 is shed in the faeces of infected individuals. Reducing SARS- CoV-2 shedding includes decreasing the number of SARS-CoV-2 virions in the faeces of an infected individual.

[00223] Accordingly, in another embodiment, the at least one symptom of Covid-19 is selected from the group consisting of diarrhea, abdominal pain, bowel pain, fever, loss of appetite, SARS-CoV-2 viral shedding, nausea, vomiting, and Covid-19 associated mortality.

[00224] As discussed above, ACE2 mainly enters human cells by binding to the receptor angiotensin converting enzyme 2 (ACE2) also expressed by gastrointestinal epithelial cells. Accordingly, in another embodiment, the at least one symptom of Covid-19 is SARS-CoV-2 infection in the gastrointestinal tract.

[00225] The present inventors have demonstrated in Example 2 that compositions comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-2 replication, improve cell viability. These data suggest that compositions comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-2 replication improve cell viability of ACE2 expressing cells, such as gastrointestinal epithelial cells.

[00226] Accordingly, in one embodiment the present invention provides a method of improving cell viability comprising contacting one or more human cells with an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS. [00227] In another embodiment the present invention provides a method of maintaining gastrointestinal cell viability comprising contacting one or more human cells with an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00228] In one embodiment, the contacting is performed in vivo.

[00229] In one embodiment the present invention provides a method of improving cell viability in a subject comprising to a subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00230] In another embodiment the present invention provides a method of maintaining gastrointestinal cell viability in a subject comprising to a subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00231] In one embodiment the subject is a subject with a SARS-CoV-2 infection.

[00232] Without wishing to be bound by theory, the present inventors propose that infection of gastrointestinal epithelial cells with SARS-CoV-2 and the resultant pathology allows secondary bacterial infections in the gastrointestinal tract to flourish. Furthermore, the extensive use of antibiotics for acute respiratory infection can cause a loss of gastrointestinal flora resulting in an increased susceptibility to secondary bacterial infections in the gastrointestinal tract.

[00233] Accordingly, in one embodiment the at least one symptom of Covid-19 is loss of gastrointestinal flora secondary to SARS-CoV-2 infection.

[00234] In another embodiment the at least one symptom of Covid-19 is bacterial sepsis associated with SARS-CoV-2 infection or bacterial sepsis secondary to SARS- CoV-2 infection.

[00235] Without wishing to be bound by theory, Gl epithelial cell infection by SARS- CoV-2 and associated pathology would cause a series of secondary effects including cytokine storm and increased severity of other gastrointestinal conditions. Accordingly, in one embodiment the at least one symptom of Covid-19 is cytokine storm associated with SARS-CoV-2 infection or increased severity of a gastrointestinal disease associated with to SARS-CoV-2 infection. [00236] In one embodiment the present invention provides a method of reducing viral shedding from a subject infected with a human coronavirus comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00237] In one embodiment the present invention provides a method of treating and/or preventing human coronavirus-associated diarrhoea in a subject comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

[00238] In one embodiment the present invention provides a method of inhibiting coronavirus mediated cytotoxicity comprising contacting one or more human cells with an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS. In one embodiment the contacting is performed in vivo in a human.

[00239] In one embodiment the symptom of coronavirus infection is selected from the group consisting of nausea, vomiting and diarrhoea.

[00240] In one embodiment the symptom of coronavirus infection is the load of coronavirus in stool.

[00241] In one embodiment the symptom of coronavirus infection is bowel pain. [00242] The present disclosure is illustrated by the following non-limiting Examples.

Examples

[00243] Example 1: Bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS binds to other pathogens.

[00244] Bovine-hyperimmune colostrum is prepared using the methods described in PCT/AU2004/000277, hereby incorporated by reference in its entirely. In brief, a composition comprising bovine-hyperimmune colostrum is prepared by immunising cows with a vaccine comprising ETEC LPS of LPS O serotypes 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159, and a vaccine comprising 078, wherein the LPS comprises the lipid A, core and O-polysaccharide regions of the LPS.

[00245] Cytokines / hormones / growth factors detected in the composition comprising bovine-hyperimmune colostrum is prepared by immunising cows with a vaccine comprising ETEC LPS of LPS 0 serotypes 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159, and a vaccine comprising 078, wherein the LPS comprises the lipid A, core and O-polysaccharide regions of the LPS by ELISA included in batches tested TNF-alpha, IGF-1 , Lactoferrin, and Bovine Growth Hormone, with lgG1 making up between 65% and 90% of total immunoglobulins, TGFP2 in the range of 0.3 - 0.7 pg/gl, and lactoferrin in the range of 1.4 - 3.8 mg/g.

Figure 1A shows LPS extracted from ETEC strains in the ETEC vaccine, run on 15% Tris-tricine SDS-PAGE and stained using LPS specific silver stain. O-polysaccharide chains are shown as ladders and lipid A core region is marked with an arrow. Shown are extracted LPS from 1 - ETEC strain B2C [serotype 06]; 2 - ETEC strain C553/3c3 [serotype 08]; 3 - ETEC strain PE 595 [serotype 015]; 4 - ETEC strain E11881A [serotype 025]; 5 - ETEC strain C1064-77 [serotype 027]; 6 - ETEC strain PE 672 [serotype 063]; 7 - ETEC strain E20738/0 [serotype 0114]; 8 - ETEC strain PE 724 [serotype 0115]; 9 - ETEC strain El 37-2 [serotype 0128]; 10 - ETEC strain B7A [serotype 0148]; 11 - ETEC strain E8772/0 [serotype 0153]; and 12 - ETEC strain PE 768 [serotype 0159] Not shown is LPS extracted from ETEC strain H10407 [078:H11]; cows are also immunised with 078 LPS.

[00246] Figure 1 B shows western transfer to PVDF membrane and blotted with Bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS, made from a pool of colostrum from over 1000 cows. This figure shows binding of anti-LPS antibodies against both the O-polysaccharide ladders and arrowed lipid A core region of all the serotypes included in the ETEC vaccine (078, data not shown).

[00247] These data indicate that hyperimmune colostrum from cattle immunised with LPS 0 serotypes 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159 comprises bovine immunoglobulin that binds LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159, and lipid A cores of 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159 LPS.

[00248] The ability of bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS to bind to LPS of other ETEC pathogens was examined. [00249] Figure 2A shows a silver stained 15% Tris-tricine gel showing LPS extracted from ETEC serotypes not included in the ETEC vaccine. Shown are LPS extracted from 2 - ETEC strain M452C1 [serotype 020: H-]; 3 - ETEC strain T0225-C4 [075: H4]; 4 - ETEC strain 83-552 [0126:H-]; 5 - ETEC strain G33 [0126: H 12]; 6 - ETEC strain M145C2 [0128:H(NT)]; 7 - ETEC strain E23477/0/A [0139:H25]; 8 - ETEC strain ND782 [0141 :H4]; 9 - ETEC strain ND748 [O149:H10]; and 10 - ETEC strain

E11881 A [025: H42] (+ control).

[00250] Figure 2B shows western transfer to PVDF membrane and blotted with Bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS, made from a pool of colostrum from over 1000 cows. This figure shows binding of anti-LPS antibodies against both the O-polysaccharide ladders and arrowed lipid A core region of all the serotypes not included in the ETEC vaccine.

[00251] The ability of bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS to bind to LPS of other pathogens was examined.

[00252] Figure 3A shows a silver stained 15% Tris-tricine gel showing LPS extracted from pathogens not included in the ETEC vaccine. Shown are LPS extracted from 2 - Enterobacter aerogenes strain ATCC 13048; 3 - Enteropathogenic Escherichia coli strain E2348/69; 4 - Klebsiella pneumoniae strain ATCC 26; 5 - Pseudomonas aeruginosa strain ATCC 27853; 6 - Salmonella typhimurium strain ATCC 14028; 7 - Vibrio cholerae strain 6239; 8 - Yersinia enterocolitica strain 67R; 9 - Citrobacter rodentium strain DSB100; and 10 - ETEC strain E11881A [025: H42] (+ control)

[00253] Figure 3B shows western transfer to PVDF membrane and blotted with Bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS, made from a pool of colostrum from over 1000 cows. This figure shows binding of anti-LPS antibodies against both the O-polysaccharide ladders and arrowed lipid A core region of LPS extracted from pathogens not included in the ETEC vaccine.

[00254] The ability of the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS to inhibit bovine mannose resistant hemagglutination (MRHA) by ETEC strains representing common CF types (below) was also examined. [00255] Table 1 : Further ETEC strains tested

[00256] In brief, Hemagglutination Inhibition (HAI) Assay with ETEC of the colonization factor (CF) types above was performed. In brief, in the HAI assay, the IgG adheres to the CF of the ETEC strain preventing bovine MRHA. The bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS exhibited the strongest inhibitory effect of MRHA by ETEC expressing CFA/I, CS2, and CS3 as shown in the Table 2. The HAI titer is the dilution of the the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS stock solution (140 mg/ml) which inhibited MRHA by the ETEC strain. The 140 mg/ml stock concentration corresponds to the concentration of 50 mg/ml IgG in the the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS colostrum. The inhibitory concentration of the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS IgG refers to the amount of IgG needed to bind to the CF to prevent adherence of the bacteria to the bovine red blood cells resulting in no agglutination. Consequently, the lower the concentration of IgG, the stronger the inhibitory effect. Promilk 85, a product containing powdered milk proteins, was used as a negative control at the 140 mg/ml concentration. Promilk did not inhibit hemagglutination by any of the ETEC strains tested.

[00257] Table 2. Hemagglutination titers and inhibitory concentrations resulting from the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS combined with ETEC expressing common CF types.

*ETEC strain 8786-1 which expresses CF type CS15 was not tested in the HAI assay due to its low MHT in the MRHA.

**Average of 4 titers resulting from two assays done in duplicate on separate days. The value of 0.5 was assigned when no inhibition was observed. The value of 1 was assigned for inhibition at the undiluted (140 mg/ml) concentration.

[00258] These data demonstrate that the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS binds to the ETEC strains H 10407, WS1974AC91f, WS2010A, BANG10-SP, ETEC8, DS02-2, WS3294A, LSN02-013966/A, WS0115A, and WS2173A, of serotypes 078:H11, 08:H9, 06:H16, 08:H9. 025:H42, 0167:H5, 0114:H49, 078:H18. 0117:H4, 0115: H51 , 0114:H- and 071:H4. [00259] The ETEC bacterial suspensions used in the HAI were electrophoresed on SDS PAGE and immunoblotted. The blots were immunodetected with the bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS also or Promilk 85. Promilk is a negative control containing powdered milk proteins. The bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS colostrum reacted with many whole cell lysate proteins in each of the HAI strains tested. Interestingly, some of the strongest reacting proteins appear around 15 kDa for H 10407 (CFA/I), C91f (CS2), and WS2010A (CS3). These bands most likely correspond to the major subunits of the CFA/I (CfaB), CS2 (CotA), and CS3 (CstG and CstH) fimbrial proteins. The bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS also detected all of the fimbrial proteins with varying degrees. None of these proteins reacted with the Promilk (data not shown).

[00260] The ability of the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS to cross react with Campylobacter jejuni antigens was also examined by immunoblotting. In brief, whole cell lysates of C. jejuni strains were immunodetected with the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS and Promilk 85. C. jejuni strains were selected from among the most prevalent Penner (capsule) types worldwide.

[00261] Table 3: Campylobacter jejuni strains tested

[00262] Cross reactivity of the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS with the polysaccharide surface antigens of C. jejuni was determined by immunoblots of crude proteinase K digestions of whole cells. There was no reactivity seen with the high molecular weight capsule of any C. jejuni strain tested when samples were run on either 12.5% gels or on 16% gels (data not shown). However, the LOS cores of all strains tested reacted with bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS , but not with Promilk 85. Since several of the strains used in this study contained ganglioside mimics in their LOS cores (see Table 1 ), we also examined reactivity with a mutant of one strain, 3208, in which the LOS core of strain 81-176 was genetically truncated such that it lost the ganglioside mimicry of the outer core, but retained the inner core. The LOS core of this mutant still reacted with the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS, indicating that the reactivity was with the inner core and not with the gangliosides found on the outer core.

[00263] These data demonstrate that the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS binds to C. jejuni strains, for example, MSC 57360, NCTC 11168, BH-01-0142, GC8486, RM3409, GC8421 , 81-176, and 3208.

[00264] The ability of the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS to cross react with Vibrio cholera strains was also examined by immunoblotting. In brief, whole cell lysates of 71 strains of Vibrio cholera Ogawa, V. cholera Inaba, and V. cholera NON 01/0139 were immunodetected with the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS and Promilk 85. Western blot analyses revealed all 71 isolates of V. cholera Ogawa, V. cholera Inaba, and V. cholera NON 01/0139 showed similar pattern of immunoreactive protein bands, there are only three immune reactive bands, which are between 37 to 75 kD (data not shown).

[00265] Table 4: List of Vibrio isolates tested

[00266] These data demonstrate that the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS binds to Vibrio cholera strains, for example, Vibrio cholera Ogawa, V. cholera Inaba, and V. cholera NON 01/0139

[00267] The ability of the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS to cross react with Campylobacter (n=60) and Shigella strains (n=60) strains was also examined by immunoblotting. In brief, whole cell lysates of ETEC, Campylobacter and Shigella were immunodetected with the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS and Promilk 85.

[00268] For the further Campylobacter strains, Western blot analyses revealed only single immune reactive band between 50 to 75 kD in all strains, however some isolated also showed another immune reactive band between 100 to 150 kD. Four isolates of C. jejuni subsp. jejuni (# BRH14-007, JH13-225, BR11-143, and CW12-420) and one isolate of C. jejuni subsp. doylei (# SPH14-009) have an extra prominent immune- reactive protein band close to 100 kD. Campylobacter isolates from Bhutan, Cambodia, Nepal and Thailand showed similar pattern of western blot results.

[00269] These data demonstrate that the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS binds to C. coli strains, C. jejuni subsp. Doylei, C. jejuni subsp. Jejuni and C. upsaliensis strains.

[00270] For ETEC, while the immune-reactive bands are different among 14 ETEC- LT strains, the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS reacted with all ETEC strains tested (data not shown). There are five prominent immune-reactive protein bands, which are observed among 60 ETEC strains from Bhutan, Cambodia, Nepal and Thailand. Those bands are of approx. 14 KD, approx. 50 KD, approx. 75 KD approx. 150 KD and approx. 200 KD. However, The ~14 kD band was found only in ETEC strains with colonization factor antigen I (CFA/I). Only two ETEC isolates showed the presence of CFA/I band, however, the ETEC isolates with CS antigens (CS1 , CS2, CS3, CS6, CS7, CS12, CS13, CS17, CS20, CS21 , PCF071 ) did not show that specific bands at low MW.

[00271] These data demonstrate that the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS binds to ETEC strains CHC03-194-3-1 , SR08-098-5-1 , CR08-603-3, PN08-462-2, CG-2010#3-4-1 , BR11- 033-5, BR11-097-5, BR11-104-1 , BR11-151-1 , BR11 -161 -1-3, ENT-CG-15-109-1 , KR08-377, PN08-462-2-2, ENT-CG-16-1002-1 , ENT-CG04-S-550, CW16-530-2-1 , CW1 6-503-1 , CW16-088-8, CW16-533-1 , CW16-017-3, CW16-055-7, CW16-532-8, CW1 6-010-4, CW16-021-2, CW16-514-6, CW16-068-2, CW16-514-1. CW16-020-5, CW1 6-030-1 , CW16-009-1 , MR514-001-2, SPH14-023-4, BRH14-109-5, MR5C14- 060-3, SPHC14-022-4, SPH14-048-1 , BRH14-123-2, SPH14-060-1 , MR5C14-062-1 , BRHC1 4-094-1 , BRH14-109-1 , BRH14-115-3, BRH14-178-3, SPH14-139-2, MR514- 288-5, JH 13-465-5. JH13-551-2, JHC13-541-5, JM13-242-1 , JMC13-240-1 , JG13- 563-1 , JH13-463-2, JH13-475-1 , JH13-492-3, JHC13-556-2, JH13-592-3, JH13-600- 3, JHC13-383-6, JH13-594-1 and JH13-597-1 .

[00272] For Shigella, the immune-reactive protein bands are similar among the 4 isolates of S. boydii; two prominent immune-reactive bands are uniquely shown between 37 - 75 kD. Unlike others, the isolate# JH13-575 (S. boydii 18) has an additional prominent band between 37 - 50 kD. The western blot immune-reactive bands of S. boydii isolates from Thailand and Bhutan are similar. For S. dysenteriae, the immune-reactive protein bands are slightly different among the 3 isolates of S. dysenteriae; one prominent immune-reactive band between 50 - 75 kD is uniquely shown in all isolates and the prominent bands between 37 - 50 kD have different immune-reactive pattern among the 3 isolates, and two bands are detected in the isolates# KRC-131 (S. dysenteriae-4) and JPC13-737 (S. dysenteriae-12). For S. flexneri, The immune-reactive protein bands are similar among the 15 isolates of S. flexneri. Two prominent immune-reactive bands are shown between 50 - 75 kD and 37 - 50 kD. Unlike others, the isolate# TBFI-01-0001 (S. flexneri 2a) has an additional prominent immune-reactive band at 37 kD. The western blot immune-reactive bands of 15 S. flexneri isolates from Bhutan, Cambodia, Nepal and Thailand are similar. All isolates of S. flexneri 3a, S. flexneri 4, S. flexneri 4av, and S. flexneri 6 have two prominent immune-reactive bands shown between 50 - 75 kD. For S. sonnei, all S. sonnei isolates showed similar pattern of immunoreactive bands. There is one prominent band between 37 to 50 kD, and several bands between 50 to 75 with one prominent band.

[00273] These data demonstrate that the bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS binds to Shigella strains, including S. boydii, S. boydii 2, S. boydii 18, S. dysenteriae 2, S. dysenteriae 4, S. dysenteriae 12, S. flexneri, S. flexneri 1b, S. flexneri 2 variant (ll:3,4,7,8), S. flexneri 2a, S. flexneri 3a, S. flexneri 4, S. flexneri 4av, S. flexneri 6, and S. sonnei.

[00274] Example 2: Bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS inhibits SARS-CoV-2 viral replication.

[00275] In-vitro susceptibility of viruses to an antiviral agent is usually assessed using a quantitative assay to measure virus replication in the presence of increasing concentrations of the product compared to replication in the absence of the product. The effective concentration is the concentration of product at which virus replication is inhibited by 50 percent (EC50 for cell-based assays). Assays that evaluate antiviral activity include, but are not limited to, virus inactivation assays, plaque reduction assays, cytopathic effect inhibition assays, peripheral blood mononuclear cell (PBMC) assays, and binding and fusion assays.

[00276] The ability of bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS to inhibit SARS-CoV-2 replication was examined. In brief, five batches of colostrum powder were prepared as per Example 1 , and these bovine-hyperimmune colostrum preparations (“Test Compounds”) were tested in a cytopathic effect inhibition assay against SARS-CoV-2 hCoV- 19/AustraliaA/IC01/2020 according to the following protocol.

[00277] Table 5: Bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS

[00278] Table 6: Key Reagents and Materials

[00279] Virus

[00280] SARS-CoV-2 hCoV-19/Australia/VIC01/2020 was a gift from Melbourne’s Peter Doherty Institute for Infection and Immunity (Melbourne, Australia). Documentation received with the parent stock indicated that prior to receipt, the virus had been passaged as follows: two passage in Vero cells. A working stock was generated at 360biolabs by two further passages in Vero cells in virus growth media, which comprised Minimal Essential Medium without L-glutamine supplemented with 1 % (w/v) L-glutamine 1.0pg/mL of TPCK-Trypsin, 0.2% BSA, 1x Pen/Strep, and 1 % Insulin Transferrin Selenium (ITS).

[00281] Cells

[00282] African Green Monkey Kidney (Vero E6) cells (ATCC-CRL1586) were sub cultured to generate cell bank stocks in cell growth medium, which comprised Minimal Essential Medium without L-glutamine supplemented with 10% (v/v) heat-inactivated Fetal Bovine Serum and 1 % (w/v) L-glutamine. Cell stocks were frozen at -80°C overnight and then transferred to liquid nitrogen.

[00283] Vero E6 cells were passaged for a maximum of 13 passages, after which a new working cell bank stock was retrieved from liquid nitrogen for further use.

[00284] Experimental Methods

[00285] Preparation of cells for Assay

[00286] Vero cells were seeded into 96-well plates at 2x104 cells/well in 100pL seeding media (Minimal Essential Medium supplemented with 1 % (w/v) L-glutamine, 1 % ITS, 0.2% BSA). Plates were incubated overnight at 37°C, 5% C02.

[00287] Test and Control Compound Preparation

[00288] Products were prepared on the day of assay. Powders were dissolved in PBS (pH 7.4), with shaking at room temperature for 2 hours. Supernatants were subsequently collected after centrifugation at 4,000 X g for 10 minutes and used in the assay.

[00289] The positive control compound Remdesivir was prepared as a 10mM stock in DMSO and stored at -20°C.

[00290] Addition of Test and Control Articles to Assay Plate

[00291 ] Compound dilutions were prepared on the day of experimentation. A dilution series of each product was performed by addition of 800pL (100mg/mL) to row A, B, C, D, E, column 2 of a deep well plate. A volume of 500pL of virus growth media (Minimal Essential Medium supplemented with 1 % (w/v) L-glutamine, 1 % ITS, 0.2% BSA, 1 pg/mL TPCK-Trypsin) was added to row A-E, columns 3-11. Products were serially diluted 1 :3 by transfer of 250pL from column 2 to column 3, column 3 to column 4 and continued to column 10.

[00292] A 50pL volume from each product dilution series was added to rows B-G of a fresh assay plate. One assay plate / compound was generated.

[00293] A DMSO dilution series of Remdesivir was generated by addition of 15pL (1 OmM) compound to column 3 and 10pL DMSO to columns 2, 4-11 . Remdesivir was serially diluted 1 :3 by transfer of 5pL compound from column 3 to column 4, column 4 to column 5 and continued to column 10 and then discarded. An intermediate dilution series was prepared by addition of 4pL from columns 2-11 to 496pL virus growth media. A 50pL volume from the intermediate dilution series was added to rows B-G of the assay control plate containing pre-seeded cells in 100pL seeding media. Virus was then added as outlined below.

[00294] Addition of Virus

[00295] A 50pL volume of SARS-CoV-2 (B2) diluted in virus growth media to generate a multiplicity of infection (MOI) of 0.05, was added to rows B, C and D of the 96-well plates containing the product (test compound) dilution series . This MOI was previously determined to provide 100% CPE in 4 days. Virus growth media minus the virus was added to rows E, F and G. Plates were incubated for 1 hour at 37°C, 5% C02.

[00296] Product / virus mixture was then added to rows B, C and D of a 96-well plate containing pre-seeded Vero E6 cells to assess antiviral activity and Product / virus growth media without virus was added to rows E, F and G to assess cytotoxicity. Plates were incubated at 37°C, 5% C02 for 4 days prior to staining with MTT.

[00297] Cvtopathic Effect (CPE) Determination

[00298] After incubation for four days, viable cells were determined by staining with MTT. A 100pL volume of a 3mg/ml_ solution of MTT was added to plates and incubated for 4 hours at 37°C in a 5% C02 incubator. Wells were aspirated to dryness using a multichannel manifold attached to a vacuum chamber and formazan crystals solubilised by the addition of 200pL 100% 2-Propanol at room temperature for 30 minutes. Absorbance was measured at 540 - 650nm on a plate reader.

[00299] Determination of Effective Concentration 50 (EC50)

[00300] The percent cell protection achieved by the positive control and test articles in virus-infected cells was calculated by the formula of Pauwels et al. as shown below:

Percent cell protection = ([ODt]virus - [ODc]virus / [ODc]mock - [ODc]virus) x 100

Where:

[ODt]virus = the optical density measured in a well examining the effect of a given concentration of test article or positive control on virus-infected cells. [ODc]virus = the optical density measured in a well examining the effect of the negative control on virus-infected cells.

[ODc]mock = the optical density measured in a well examining the effect of the negative control on mock-infected cells

EC50 values were calculated from the percent cell protection results by non-linear regression analysis using the Hill (sigmoid Emax) formula as shown below:

Abbreviations: x, test or control article concentration; y, percent cell protection; Min, minimum; Max, maximum; D, slope coefficient.

[00301] The 50% cytotoxic concentration (CC50) is defined as the concentration of the test compound that reduces the absorbance of the mock infected cells by 50% of the control value. The CC50 value was calculated as the ratio of (ODt)mock/(ODc)mock.

[00302] IDBS XLFit4 Excel Add-in (ID Business Solutions Inc., Alameda, CA) was used to perform the above calculations.

[00303] Results

[00304] Figure 4 shows hyperimmune colostrum powder Batch # 1710002659 had an EC50 of 44.7 ug/ml and a CC50 of 1085.3 ug/ml. This batch of colostrum inhibits viral replication at a dose at which there is no cell toxicity, and at a dose that inhibits 90% of viral replication there is no cell toxicity (an EC90 of 53.15 ug/ml).

[00305] Figure 5 shows hyperimmune colostrum powder Batch # 1710002660had an EC50 of 91.9 7 ug/ml and a CC50 of 1790.6 ug/ml. This batch of colostrum inhibits viral replication at a dose at which there is no cell toxicity, and at a dose that inhibits 90% of viral replication there is no cell toxicity (an EC90 of 155.39 ug/ml).

[00306] Figure 6 shows hyperimmune colostrum powder Batch # 1710002662 had an EC50 of 40.5 7 ug/ml and a CC50 of 2053.3 ug/ml. This batch of colostrum inhibits viral replication at a dose at which there is no cell toxicity, and at a dose that inhibits 90% of viral replication there is no cell toxicity (an EC90 of 45.17 ug/ml). [00307] Figure 7 shows hyperimmune colostrum powder Batch # 1710002859 had an EC50 of 42.6 7 ug/ml and a CC50 of 2506.6 ug/ml. This batch of colostrum inhibits viral replication at a dose at which there is no cell toxicity, and at a dose that inhibits 90% of viral replication there is no cell toxicity (an EC90 of 48.69 ug/ml).

[00308] Figure 8 shows pro-milk powder Batch # 6068 N50 (control) had an EC50 of > the CC50, and a CC50 of 4818.6 ug/ml. Accordingly, this control milk powder inhibits viral replication at a dose >25000 ug/ml, and importantly does not inhibit viral replication at the doses at which it is cytotoxic to cells.

[00309] Figure 9 shows the positive control, remdesivir, has an EC50 of 1 16pM.

[00310] These results indicate that compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) inhibit SARS-CoV-2 replication, and inhibit SARS-CoV-2 replication at doses that are not cytotoxic to cells.

[00311] Importantly, cell toxicity was diminished by treatment with compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-2 replication.

[00312] Figure 4 shows hyperimmune colostrum powder Batch # 1710002659 improved cell viability by 260% relative to control, at 102.8 ug/ml.

[00313] Figure 5 shows hyperimmune colostrum powder Batch # 1710002660 improved cell viability by 212% relative to control, at 102.8 ug/ml.

[00314] Figure 6 shows hyperimmune colostrum powder Batch # 1710002662 improved cell viability by 186% relative to control, at 102.8 ug/ml.

[00315] Figure 7 shows hyperimmune colostrum powder Batch # 1710002859 improved cell viability by 218% relative to control, at 102.8 ug/ml.

[00316] Figure 8 shows pro-milk powder Batch # 6068 N50 (control) did not improve cell relative to control.

[00317] Figure 9 shows the positive control, remdesivir, did not improve cell relative to control. [00318] These results indicate that compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-2 replication, improve cell viability.

[00319] Example 3: Compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-replication improve liver inflammation.

[00320] Mao et al. demonstrated that the pooled prevalence of abnormal liver functions from 35 studies including 6686 patients was 19%, and subgroup analysis showed patients with severe COVID-19 had higher rates of abdominal pain (odds ratio [OR] 7-10 [95% Cl 1 -93-26 07]; p=0 003; l²=0%) and abnormal liver function including increased ALT (1 -89 [1 -30-2-76]; p=0-0009; l²=10%) and increased AST (3-08 [2-14- 4-42]; p<0-00001 ; l²=0%) compared with those with non-severe disease.

[00321] The ability of compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS- CoV-replication to alter liver function were examined.

[00322] To examine the effect of compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-replication to alter liver function, 133 biopsy proven NASH patients were administered colostrum powder prepared as per Example 1. In brief, a multinational, randomized, double-blind study comparing 2 doses of colostrum powder prepared as per Example 1 were compared to placebo in adults with any stage biopsy- proven NASH. The trial enrolled 133 patients across 25 clinical sites in Australia (6), Israel (2) and the USA (17). The trial had 12 scheduled visits over a 28-week study duration, with 24 weeks of treatment and four weeks of follow-up and screened a total of 237 patients. The patients were randomized into three arms: placebo, high dose (1200mg), and low dose (600mg). Endpoints included changes in ASL and ALT as well as other liver enzymes and metabolic markers of liver inflammation.

[00323] Figure 10 shows that patients administered with colostrum powder prepared as per Example 1 have a statistically significant improvement in both AST and ALT levels (e.g. a 15% decrease in AST and ALT or a 30% decrease in AST and ALT) relative to placebo. [00324] These results indicate that compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-2 replication, improve liver function.

[00325] Example 4: Compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-replication decrease levels of LPS in the serum of patients with inflammatory disease.

[00326] The pathogenesis of COVID-19 is believed to involve viral-induced suppression of innate pathogen surveillance systems. Under normal conditions, pathogen pattern recognition receptors (PPR) on resident innate immune cells sense viral RNA molecules that trigger anti-viral, interferon (IFN) expression which prevents replication and promotes the removal of infected cells. However, genomic studies conducted on the original severe acute respiratory syndrome coronavirus (SARS- CoV) demonstrate that the virus encodes for proteins that serve as innate immune antagonists by suppressing the expression of IFN and promoting evasion of viral RNA from host defense mechanisms, independent of pro-inflammatory cytokine release. As a result, early in infection, innate toll-like receptor (TLR) and PRR signaling pathways continue to potentiate the release of pro-inflammatory mediators, such as cytokines (i.e. , TNFa, IL-6, IP10 or CXCL10, etc.) while viral replication remains unchecked. Hence, it has been theorized, coronaviruses pathogenesis involves the delayed release of IFN and an accumulation of monocyte/macrophages together with an inappropriate T-cell response. Adding support to this etiology, severe cases of COVID- 19 appear to present with dysregulated T-cell counts and elevated inflammatory cytokine levels. In the case of severe COVID-19, this disruption leads to a condition described as a “cytokine storm”, in which excessive amounts of pro-inflammatory cytokines are produced and may contribute to morbidity and mortality in these patients.

[00327] Similarly, sepsis caused by Gram-negative bacteria is the consequence of an unrestrained infection that continuously releases lipopolysaccharide (LPS) into the bloodstream, which triggers an uncontrolled systemic inflammatory response leading to multiorgan failure and death. In particular, TLR4 is expressed on a wide range of immune cells, which specifically recognize bacterial lipopolysaccharide, and activation of TLR4 leads to the synthesis of proinflammatory cytokines and chemokine with the ultimate goal of identifying and destroying the pathogen. However, activation of the innate immune system occasionally leads to host tissue collateral damage, resulting in organ dysfunction and death.

[00328] The ability of compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS- CoV-replication to decrease serum levels of LPS, a cytokine storm inducing molecule, was examined.

[00329] To examine the effect of compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-replication to alter serum LPS levels, 133 biopsy proven NASH patients were administered colostrum powder prepared as per Example 1. In brief, a multinational, randomized, double-blind study comparing 2 doses of colostrum powder prepared as per Example 1 were compared to placebo in adults with any stage biopsy- proven NASH. The trial enrolled 133 patients across 25 clinical sites in Australia (6), Israel (2) and the USA (17). The trial had 12 scheduled visits over a 28-week study duration, with 24 weeks of treatment and four weeks of follow-up and screened a total of 237 patients. The patients were randomized into three arms: placebo, high dose (1200mg), and low dose (600mg). Endpoints included serum LPS levels.

[00330] Figure 11 shows that patients administered with colostrum powder prepared as per Example 1 have an improvement in serum LPS levels relative to placebo, at all cut-offs, and statistically significantly improved. These differences are significant for cut-offs of at least a 15% decrease, 10%, 5% or any decrease. Furthermore, these data demonstrate that there is less deterioration in the 1200mg arm than placebo, significant at all cut-offs.

[00331] These results indicate that compositions comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) that inhibit SARS-CoV-2 replication, decrease serum LPS.

Claims

1 . A method of treating and/or preventing human coronavirus infection in a subject comprising administering an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

2. A method of reducing viral shedding from a subject infected with a human coronavirus comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

3. A method of treating and/or preventing human coronavirus-associated diarrhoea in a subject comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

4. A method of inhibiting coronavirus mediated cytotoxicity comprising contacting one or more human cells with an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

5. A method of improving cell viability comprising contacting one or more human cells with an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

6. A method of maintaining gastrointestinal cell viability contacting one or more human cells with an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

7. A method according to any one of claims 1 to 6 wherein the vaccine comprising ETEC LPS comprises one or more LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159.

8. A method according to any one of claims 1 to 5 wherein the vaccine comprising ETEC LPS comprises two or more LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159.

9. A method according to any one of claims 1 to 7 wherein the vaccine comprising ETEC LPS comprises LPS 0 serotypes 078, 06, 08, 015, 025, 027, 063, 0114, 0115, 0128, 0148, 0153 and 0159.

10. A method according to any one of claims 1 to 8 wherein the vaccine comprising ETEC LPS comprises the lipid A core and O-polysaccharide regions of the LPS.

11. A method according to any one of claims 1 to 10 wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to ETEC LPS.

12. A method according to any one of claims 1 to 11 wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to one or more LPS 0 serotypes selected from the group consisting of 078, 06, 08, 015, 025, 027, 063, 071, 0114, 0115, 0117, 0128, 0148, 0153, and 0159, 0167.

13. A method according to any one of claims 1 to 12 wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to a gram negative bacteria selected from the group consisting of Enteropathogenic Escherichia coli strain E2348/69, Klebsiella pneumoniae strain ATCC 26, Pseudomonas aeruginosa strain ATCC 27853, Salmonella typhimurium strain ATCC 14028, Vibrio cholerae strain 6239, Yersinia enterocolitica strain 67R, C. jejuni, Vibrio cholera Ogawa, V. cholera Inaba, and V. cholera NON 01/0139, C. coli, C. jejuni subsp. Doylei, C. jejuni subsp. Jejuni, C. upsaliensis, S. boydii, S. boydii 2, S. boydii 18, S. dysenteriae 2, S. dysenteriae 4, S. dysenteriae 12, S. flexneri, S. flexneri 1b, S. flexneri 2 variant (ll:3,4,7,8), S. flexneri 2a, S. flexneri 3a, S. flexneri 4, S. flexneri 4av, S. flexneri 6, and S. sonnei.

14. A method according to any one of claims 1 to 13 wherein the composition comprising bovine-hyperimmune colostrum is prepared using a first vaccine comprising ETEC LPS and a second vaccine comprising ETEC LPS.

15. A method according to any one of claims 1 to 14 wherein the composition comprising bovine-hyperimmune colostrum is prepared using a first vaccine comprising ETEC LPS and a second vaccine comprising a bovine coronavirus antigen and/or a bovine coronavirus.

16. A method according to any one of claims 1 to 14 wherein the composition comprising bovine-hyperimmune colostrum is prepared using a vaccine comprising ETEC LPS administered to an animal sequentially- or co-administered a vaccine comprising a bovine coronavirus antigen and/or a bovine coronavirus.

17. A method according to claim 16 wherein the vaccine comprising a bovine coronavirus is an attenuated bovine coronavirus.

18. A method according to any one of claims 1 to 17 wherein the composition comprising bovine-hyperimmune colostrum is prepared using a vaccine comprising ETEC LPS administered to an animal producing antibodies to bovine coronavirus.

19. A method according to any one of claims 1 to 18 wherein the composition comprising bovine-hyperimmune colostrum comprises bovine immunoglobulin that binds to bovine coronavirus.

20. A method according to any one of claims 1 to 18 wherein the human coronavirus is SARS-COV-2.

21. A method according to anyone of claims 7 to 20 wherein the hyperimmune colostrum is prepared by administering a bovine animal with a vaccine comprising ETEC LPS and collecting hyperimmune colostrum from the animal.

22. A method according to claim 21 wherein the hyperimmune colostrum is prepared by administering one bovine animal with a first vaccine and a second bovine animal with a second vaccine, and subsequently combining the hyperimmune colostrum collected from the animals.

23. A method according to any one of claims 1 to 22 wherein the hyperimmune colostrum comprises one or more immunoglobulin classes selected from the group of IgG, IgM and IgA.

24. A method according to any one of claims 1 to 23 wherein the hyperimmune colostrum further comprises a component selected from oligosaccharides, beta lactoglobulin, alpha-lactalbumin, lactoferrin, bovine serum albumin, growth factors, lactoperoxidase, plasmin, lipoprotein lipase, esterase, ribonucleases A, B, C, D and II- 1 , Lysozyme, enzyme inhibitors, cytokines, and lipids.

25. A method according to any one of claims 1 to 3 and 7 to 24, wherein the effective amount of a composition comprising bovine-hyperimmune colostrum is administered to the subject orally, nasally or using pulmonary administration.

26. A method according to any one of claims 4 to 6 wherein the contacting is performed in vivo in a human.

27. A method according to any one of claims 1 to 26, wherein a symptom of coronavirus infection is treated and/or prevented.

28. A method according to claim 27 wherein the symptom of coronavirus infection is selected from the group consisting of a secondary bacterial infection, sepsis, bowel pain, nausea, vomiting and diarrhoea.

29. A method according to claim 27 wherein the symptom of coronavirus infection is the load of coronavirus in stool.

30. A method of treating and/or preventing coronavirus infection in one or more human subjects comprising applying to a surface selected from air filters, PPE, room surfaces and respiratory mucosal membranes an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

31. A method according to claim 30 wherein the composition comprising bovine- hyperimmune colostrum is adsorbed on a particulate carrier of a size in the range from 50 to 120 microns

32. A method according to claim 30 wherein the composition comprising bovine- hyperimmune colostrum is adsorbed onto a solid carrier for administration by dry powder inhaler

33. A method according to claim 30 wherein the composition comprising bovine- hyperimmune colostrum is administered as an aerosol.

34. A method according to claim 30 wherein the composition comprising bovine- hyperimmune colostrum in dried form.

35. A method of decreasing levels of LPS in the blood of a subject, comprising administering to the subject an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

36. A method of treating and/or preventing sepsis in a subject, comprising administering to the subject an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

37. A method of decreasing inflammation in a subject, comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

38. A method of treating and/or preventing cytokine storm in a subject comprising administering to the subject an effective amount of a composition comprising bovine- hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

39. A method of treating and/or preventing a symptom associated with coronavirus infection. in a subject comprising administering to the subject an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

40. A method according to claim 39 wherein the symptom associated with coronavirus infection is selected from the group consisting of loss of gastrointestinal flora, increased inflammation, elevated ALT levels, elevated AST levels, pathologic change, LPS in the blood, and cytokine storm, human coronavirus-associated diarrhoea.

41. A method according to claim 39 or claim 40 wherein the symptom associated with coronavirus infection is secondary to SARS-CoV-2 infection.

42. A method according to claim 40 or 41 wherein the pathologic change is gastrointestinal tract pathology, or lung pathology.

43. A method according to claim 40 or 41 wherein the increased inflammation is increased inflammation in the lungs, increased inflammation in the liver, and/or systemic inflammation.

44. A method of reducing viral shedding from a subject infected with a human coronavirus comprising administering an effective amount of a composition comprising bovine-hyperimmune colostrum prepared using a vaccine comprising enterotoxigenic E. coli (ETEC) LPS.

45. A method according to any one of claims 30 to 44 wherein the subject is a subject with SARS-CoV-2 infection.