AU2019201630A1 - Formulation for Transferring Passive Immunity to New Born Domestic Animals - Google Patents

Abstract

A colostrum supplement formulation for transferring passive immunity to new born domestic animals, the formulation comprising freeze dried plasma, wherein the plasma has been separated from mammalian blood. Abattoir 101 Freeze Drying Facility 106 Unclotted Horse Blood 102 + citrate 1,000L into IBC 4 ~50Kg dried 107 plasma powder Chiller facility for 2 days 4 plasma settles 103 Final Filln15g per bottle 107A Decant plasma off 104 Sterilization of booted batch by 4500L plasma Gamma Irradiation QC Testing Dispose of packed blood 108 cells 104A Initial STABILITY STUDIES Storage at room temperature Administer recommended dosage

Description

FORMULATION FOR TRANSFERRING PASSIVE IMMUNITY TO NEW BORN

DOMESTIC ANIMALS

BACKGROUND

Any references to methods, apparatus or documents of the prior art are not to be taken as constituting any evidence or admission that they formed, or form part of the common general knowledge.

Colostrum is a lactation product produced by the mother after birth (parturition) of mammals such as humans, cows or horses. Medical researchers have been attracted to colostrum because of the immunological aspects. Colostrum is known to contain a number of leukocytes, immunoglobulins and lysozymes. The typical immunoglobulins include IgA, IgD, IgE, IgG and IgM, with IgG being the most abundant, and the one of most interest in this colostrum supplement.

By way of further background, there are two types of immunity - active and passive. Passive immunity is where antibodies are produced outside the newborn and “passively” transferred to it - e.g. by plasma transfusion, or colostrum ingestion. The antibodies received by colostrum or transfusion help fight infection in the newborn, but there is a finite quantity of antibodies transferred. Typically, antibodies have a half-life in the animal’s plasma, circulating in its blood stream, of 21 days. They will deplete much faster if they are needed to fight an infection, such as in an environment where there is a high load of pathogens - e.g. an outbreak of a very contagious disease.

Active immunity is where the animal’s body is “conditioned” to produce its own antibodies to an infection - either by exposure to vaccines (vaccination), or by sublethal exposure to pathogens in the animal’s environment. An animal typically needs

2019201630 08 Mar 2019 exposures/vaccinations to a naturally occurring infection before it can be considered “immune” to that infection. This immunity is conferred by the animal producing its own antibodies to fight the infection. In a disease outbreak where there is a high environmental pathogen load, actively acquired immunity will see unlimited quantities of antibody produced by the infected animal, as it has had the required previous exposures, either naturally acquired or by vaccination, to give it the “immunological memory” to continue producing antibodies for as long as is needed to combat the constantly invading pathogen.

As is common with most domestic animals, equine foals are born without immunity. Passive immunity is transferred on a postpartum basis from a dam (mother) to the newborn foal through the colostrum. It is reported and recognized that the nursing newborn receives antibodies from the mother via her colostrum (or first milk), which is present in colostrum during early feeding after birth. Typically, the term colostrum is used to designate the mammalian milk produced immediately before birth, and present in the udder for about 1 - 2 days after parturition. Colostrum is yellow in colour since the concentration of carotenoids is high, as much as ten times that of mature milk (that produced about 21-22 days after parturition). It is also known that colostrum has higher protein and mineral content as compared to mature milk: it is also lower in fat and lactose content than mature milk. In order for adequate transfer of antibodies from the mare to the foal to take place, the foal must suckle normally from its mother 3-5 times during its first 24 hours of life. A mare produces colostrum only once per pregnancy. A foal is able to absorb the immunoglobulins from its intestine into its blood stream for its first 24 hours of life. Antibodies (immunoglobulins) are protein molecules of high molecular weight, and as such require a special transport mechanism by the cells lining the intestinal villi, to transport them from the ingested colostrum in the intestinal

2019201630 08 Mar 2019 lumen into the blood stream. This transport mechanism is known as “Pinocytosis”.

Pinocytosis only exists in the intestinal lining cells for a period of 24 hours after its birth.

Thus, three windows need to align in order for ingestion and absorption of adequate quantities colostral immunoglobulins by the foal, such that failure of passive transfer will not occur - (1) the mare must have sufficient quantities of good-quality colostrum (= colostrum of normal or sufficient immunoglobulin concentration) in her udder when the foal is born - (2) the foal must be given sufficient opportunity to ingest sufficient quantities of this colostrum to avoid failure of passive transfer, and (3) the foal must ingest this quantity of good-quality colostrum in its first 24 hours of life.

The initial colostrum secretion contains rapidly diminishing (due to suckling) levels of immunologically active, large molecular weight proteins known as immunoglobulins (abbreviated below as Ig). These Ig molecules are antibodies and are actively produced by mature animals, and provide the foal with immunity to infection by bacteria, viruses or parasites. At birth, a foal lacks Ig in its blood serum. Only as a direct response to ingestion and absorption of a quantity and quality of Ig from maternal colostrum shortly after birth, can a foal’s immune system function efficiently in the neonatal period, until when immune-competency is reached at about 2 months of age.

A first essential element of the natural passive immunity transfer mechanism relates to the characteristics of the maternal colostrum. To achieve ideal passive immunity, the maternal colostrum should contain an adequate concentration of Ig, having an appropriate distribution of pathogen-specific antibodies and an appropriate concentration of each pathogen specific antibody. If the maternal colostrum contains

2019201630 08 Mar 2019 an insufficient concentration of important pathogen specific antibodies, the foal will absorb an insufficient quantity of these antibodies and will develop a deficient level of immunity to the diseases which such antibodies attack. This situation is known as “Failure of Passive Transfer” can typically occur if the mare leaks colostrum from her udder before the foal is born. As stated previously, mammals only produce colostrum once per pregnancy, and if colostrum is leaked from the udder before birth, much if not all of the antibody will be lost and therefore not available to the foal, once born.

The second important element of the natural passive immunity transfer mechanism is foal-oriented and relates to the quantity and time of colostrum ingestion. As to quantity of ingestion, previous studies have indicated that there is a limit to the volume of colostrum that can be ingested to maximize the level of Ig absorbed into the circulatory system of the foal. A significant issue with newborn foals is that they are unable to ingest large quantities of liquid within their initial feeding period. It is also important to note that the permeability of the newborn mammal’s gut to the large molecular weight Ig molecules diminishes very rapidly after birth as a result of intestinal cell maturation. This well-known natural intestinal mechanism is called Pinocytosis, and the period in which pinocytosis occurs may be referred to as the critical period of absorption, which defines the short postpartum interval during which the foal must consume and absorb the optimum quantity of ideal potency colostrum to achieve an ideal level of passive immunity. The first 24 hours of the foal’s life is typically regarded as the duration of this “critical period of absorption”. Although colostrum consumption as late as twenty-four hours postpartum may achieve some immune transfer, subsequent colostrum consumption will have very little effect on blood antibody levels. Ideally, colostrum ingestion should occur within the first eighteen hours postpartum. The optimum level of immunoglobulins in a normal foal’s bloodstream at 24 hours old is 8g/L or above.

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Foals with less than 2g/L are known to be suffering from Failure of Passive Transfer, and foals with 2-8g/L are classified as “Partial Failure of Passive Transfer” cases.

In practice, a high percentage of foals either consume far less than an ideal quantity and quality of colostrum or fail to consume colostrum within the critical absorption period. The resulting adverse effects due to the lack of passive immune transfer are demonstrated by high death rates, increased susceptibility to disease and incidence of disease and reduced growth rates.

Accordingly, there is a need to provide an improved method and formulation to address some of the shortcomings of the prior art.

SUMMARY OF INVENTION

In one aspect, the invention provides a colostrum supplement formulation for transferring passive immunity to new born domestic animals, the formulation comprising freeze dried plasma, said plasma being separated from mammalian blood (equine blood in this case).

Preferably, the formulation comprises immunologically active immunoglobulin such that ratio of the immunoglobulin to animal weight is at least 0.09wt%. More preferably the ratio of the immunoglobulin to animal weight of the new born animal is greater than 0.1 wt%. More preferably, the ratio of the immunoglobulin to animal weight is in the range of 0.1 wt% to 0.25wt%.

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In another aspect, the invention provides a method for transferring passive immunity to a neonate domestic animal during a critical absorption period and for enhancing initiation of active immunity in said animal, the method comprising the steps of:

providing plasma separated from mammalian blood wherein the separated plasma comprises a measurable quantity of immunologically active immunoglobulin;

freeze drying said separated plasma; and administering a predetermined quantity of said freeze dried plasma to said domestic animal during the critical absorption period.

In an embodiment, the step of administering comprises oral administration of the freeze dried plasma. Administration may not be confined to this particular route (referring to the possibility that, with further treatment steps, administration by injection may also be a possibility.

In an embodiment, the administering step involves administering immunoglobulin such that the weight of the immunoglobulin in said freeze dried plasma is at least 0.09wt% and more preferably greater than 0.15wt% of the body weight of the animal.

In an embodiment, the method comprises the steps of collecting mammalian blood comprises the additional step of:

mixing an anticoagulant with the blood for preventing clotting of the blood,

2019201630 08 Mar 2019 allowing the mixture of the anticoagulant and blood to mix in a container such that a supernatant liquid comprising plasma containing immunoglobulin is separated from all of the other components of the blood and anticoagulant mixture, collecting the supernatant liquid and freezing the supernatant liquid; and freeze drying the supernatant liquid to form said freeze dried plasma.

In another aspect, the invention provides the use of frozen plasma separated from mammalian blood for the manufacture a colostrum supplement formulation for transferring passive immunity to new born domestic animals.

DETAILED DESCRIPTION

In order to better illustrate the advantages of the invention and its contributions to the art, a preferred embodiment of the formulation and process for transferring passive immunity to new born domestic animals will be described in further detail.

The following embodiment focuses on new born horses or foals. However, it must be understood that the invention is in no way limited to horses and encompasses use of the present invention for other animals.

Embodiments of the present invention focuses on extracting plasma from horses for producing a colostrum supplement formulation for transferring passive immunity to new born domestic animals.

Figure 1 illustrates a step wise method of producing the colostrum supplement formulation. The initial step involves collecting blood from horses at an abattoir. Typically, blood may be collected from the horse by way of exsanguination after being stunned for slaughter, as shown in step 101. The blood may be collected from the

2019201630 08 Mar 2019 horse by any known means and placed in a container under cold conditions. In the preferred method an anticoagulant is introduced into the collection container before the addition of blood into the container (step 102) to prevent clotting of the blood when the blood from step 101 is introduced into the collection container. Anticoagulants, commonly referred to as blood thinners, are chemical substances that prevent or reduce coagulation of blood, prolonging the clotting time. By way of example, sodium citrate may be used as an anti-coagulant to stabilise the blood and prevent clotting. Other additional or alternative anti-coagulants may also be used without departing from the spirit and scope ofthe invention.

Once the blood has been added to the anticoagulant and the two mixed gently and thoroughly, the blood-anticoagulant mixture is placed in a collection container at temperatures in the range of 1 -6°C as shown in step 103 for a stipulated period of time (for example, 12 hours). Placing the mixture for an extended period of time allows the heavier red blood cells (erythrocytes) plus the white blood cells and platelets, to settle at the bottom of the collection container which may be disposed in an optional step 104A. On the other hand, the supernatant liquid comprising blood plasma can be easily decanted or separated by using any other known means as shown in step 104 and collected for freezing in step 105. The frozen plasma is then subjected to a freeze drying in step 106.

Freeze drying ofthe collected plasma in step 106 potentially offers a long term storage solution without requiring refrigeration. The freeze drying step 106 may optionally involve pre-treating the plasma before undertaking the freezing step 105. Alternatively, or additionally the frozen plasma may be pre-treated with one or more substances to

2019201630 08 Mar 2019 promote granulation in the plasma. The freeze drying step 106 may involve rapid cooling of the plasma into a frozen product of a second predetermined size followed by drying said frozen product by means of sublimation of a water content in the plasma. By way of example, the freeze dried formulation may be formed from clusters and/or porous substance possessing a size of not greater than 1 mm and comprising granules possessing a size of several pm to several 100 pm. The next step 107 involves collection of the freeze dried plasma sample. The collected sample may be packaged in pre-determined quantities (say 150gms) in a packaging step 107A followed by a post packaging sterilization step 107B. In one embodiment, a gamma radiation using Cobalt 60 may be used to expose the packaged sample to the radiation. Gamma radiation can easily penetrate packaging and quickly sterilize the freeze dried plasma in the packaging. A further qualitative analysis/control step 108 is undertaken to ensure that the freeze dried plasma possesses sufficient antibody potency. It is important to note that the level of antibody concentration or potency determines the dosage rate of the colostrum supplement. Therefore, quantity of the freeze dried formulation administered to the newborn horse is dependent upon the antibody potency of the freeze dried plasma in the formulation. The freeze dried plasma containing formulation may be packaged in predetermined quantities for sale as shown in step 109. Another quality control test may also be carried out to analyse bacterial, fungal and protozoal contamination of the freeze-dried plasma powder. This test is likely to be performed by an outside commercial laboratory, suitably qualified and accredited for the performance of such tests, and is the same test that the United Nations applies to ensure safety of all food products for human use by ingestion.

Example-In one example, a standard foal with an average body weight of 35kgs may be administered a dose of approximately 170gms of the freeze dried colostrum

2019201630 08 Mar 2019 supplement formulation in accordance with an exemplary embodiment of the present invention. Qualitative analysis of the formulation of example indicates that 170gms of the formulation contains 68gms of pure immunologically active antibodies. This is the amount of Immunoglobulin G required to give a 35Kg foal a blood antibody level of 8g/L.

The inventor has realised that providing the immunologically active immunoglobulin (or antibodies) as a freeze dried colostrum supplement allows a larger dosage of the immunoglobulin to be administered. Previous methods of providing liquid supplements have required large quantities of liquids to be fed to the newly born foals. However, as discussed previously, newly born foals have a limited capacity to ingest liquids. Using a formulation comprising freeze dried plasma provides the formulation in a powdered form and alleviates the need for using large quantities of liquids. As a result, larger quantities of antibodies or immunoglobulin can be administered by adopting the freeze dried colostrum supplement in accordance with embodiments of the present invention.

In compliance with the statute, the invention has been described in language more or less specific to structural or methodical features. The term “comprises” and its variations, such as “comprising” and “comprised of” is used throughout in an inclusive sense and not to the exclusion of any additional features.

It is to be understood that the invention is not limited to specific features shown or described since the means herein described comprises preferred forms of putting the invention into effect.

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The invention is, therefore, claimed in any of its forms or modifications within the proper scope of the appended claims appropriately interpreted by those skilled in the art.

Claims (11)

1. A colostrum supplement formulation for transferring passive immunity to new born domestic animals, the formulation comprising freeze dried plasma, wherein the plasma has been separated from mammalian blood.

2. A colostrum supplement formulation in accordance with claim 1 wherein the formulation comprises immunologically active immunoglobulin such that ratio of the immunoglobulin to animal weight is at least 0.09wt%.

3. A colostrum supplement formulation in accordance with claim 1 or claim 2 wherein the ratio of the immunoglobulin to animal weight of the new born animal is greater than 0.1 wt%.

4. A colostrum supplement formulation in accordance with any one of the preceding claims wherein the ratio of the immunoglobulin to animal weight is in the range of 0.1 wt% to 0.25wt%.

5. A method for transferring passive immunity to a neonate domestic animal during a critical absorption period and for enhancing initiation of active immunity in said animal, the method comprising the steps of:

2019201630 08 Mar 2019 providing plasma separated from mammalian blood wherein the separated plasma comprises a measurable quantity of immunologically active immunoglobulin;

freeze drying said separated plasma; and administering a predetermined quantity of said freeze dried plasma to said domestic animal during the critical absorption period.

6. A method for transferring passive immunity to a neonate domestic animal during a critical absorption period and for enhancing initiation of active immunity in said animal, the method comprising the steps of:

providing plasma separated from mammalian blood wherein the separated plasma comprises a measurable quantity of immunologically active immunoglobulin;

freeze drying said separated plasma; and administering an effective amount of said freeze dried plasma to said domestic animal during the critical absorption period.

7. A method in accordance with claims 5 or 6 wherein the step of administering comprises oral administration of the freeze dried plasma.

8. A method in accordance with claims 5 or 6 wherein the step of administering comprises injection of the freeze dried plasma to the neonate domestic animal.

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9. A method in accordance with any one of claims 5 to 8 wherein the administering step involves administering immunoglobulin such that the weight of the immunoglobulin in said freeze dried plasma is at least 0.09wt% and more preferably greater than 0.15wt% of the body weight of the animal.

10. A method in accordance with any one of claims 5 to 10 comprising an initial step of collecting and processing mammalian blood which further comprises the steps of:

mixing an anticoagulant with the blood for preventing clotting of the blood, allowing the mixture of the anticoagulant and blood to mix in a container such that a supernatant liquid comprising plasma containing immunoglobulin is separated from all of the other components of the blood and anticoagulant mixture, collecting the supernatant liquid and freezing the supernatant liquid; and freeze drying the supernatant liquid to form said freeze dried plasma.

11. The use of frozen plasma separated from mammalian blood for the manufacture a colostrum supplement formulation for transferring passive immunity to new born domestic animals.