The Equine Neonate Failure of Passive Transfer and Use of Plasma





FAILURE OF PASSIVE TRANSFER

All foals are born, because of their epitheliochorial placentation, with a deficiency of humoral immunity.1 They must rely on the adequate intake of good quality colostrum within a few hours of birth to provide essential antibodies in order to withstand the challenges of infectious agents during the following weeks. Unfortunately, this transfer of antibodies from dam to of offspring does not always occur successfully and "failure of passive transfer" (FPT) is said to exist, as depicted by the foal's serum gammaglobulin (usually specifically IgG) level being less than 400-800mg/dl (see page 3). If this deficiency is confirmed before the gut's ability to absorb antibodies ceases (12 -18 hours of age), oral supplementation of good quality colostrum (or possibly plasma) may well rectify the situation. If, on the other hand, the foal is over 24 hours old when this deficiency is diagnosed, the foal must be considered at risk to infection, especially in the "high challenge" (see page 3) environment. In such circumstances an intravenous transfusion of plasma should be considered, the benefits of which are well documented.2,3 POLYMUNE and POLYMUNE-PLUS are two forms of commercially available high quality equine plasma, specifically produced for this purpose.

THE DIAGNOSIS OF FPT

Accurately assessing the immunoglobulin status of the equine neonate on a clinical basis cannot be accomplished. Certain factors relative to the foaling process, such as prior leakage of "liquid" from the udder, failure to nurse soon after birth and the presence of agalactia will warn of the probability of low IgG levels. Veterinary Dynamics has recognized this problem and through its research and development program has devised the GAMMA-CHECK-C and GAMMA-CHECK-E tests to provide a simple cost effective means of semi-quantitatively assessing colostrum and blood gammaglobulin levels. In addition, an accurate and standardized IgG test is available in the form of the EQUINE-RID kit. All these tests can be easily performed by either the veterinarian or technical staff.


TREATMENT OF FPT

Foals under 18 hours old:

High quality colostrum (Spg. >1.060 or + Gamma-Check-C) is the method of choice. Research has shown that 60g of good quality colostrum are needed to achieve a significant increase in the circulating IgG level. Presently in vogue, are various non colostrum oral forms of IgG and their efficacy awaits to be seen. What they do not contain are all the other essential components of colostrum, such as epithelial growth factor, transferrin, various cytokines, etc.

Foals over 24 hours old:

Plasma is the treatment of choice and has been for many years. Commercial plasma is well proven for efficacy and safety. Reactions are rare. With sophisticated vaccine strategies, antibodies to the common neonatal pathogens are present in the plasma, even though it might be produced 2000 miles from where the mare and foal are.

Other sources of IgG are now being made available. Serum products suffer from the frequent problem of being contaminated with endotoxin and prostaglandin metabolites,5-8 which may often cause severe reactions. Because of USDA requirements, these pasteurized serum products often contain damaged proteins and have abnormal electrophoretic patterns.8 For these reasons they have to be diluted and administered very slowly.9,10 Many of the essential cytokines present in plasma are not present in serum.

Lyophilized IgG is now available, but again does not have the broad spectrum of components that plasma has. Whether it will provide adequate protection awaits to be seen. It is distinctly possible that we have become preoccupied with IgG and we are forgetting there are many other essential components to the healthy immune system. It could well be that it is not sufficient just to increase the IgG level to cause the foal to stay healthy. Time will tell!

One leading equine practitioner in Kentucky11 aptly described plasma as "life in a bag."

EQUINE PLASMA (POLYMUNE and POLYMUNE-PLUS)

Indications

Equine plasma is indicated where a foal that is 24 hours old or more has been diagnosed as having inadequate circulating immunoglobulin G levels. The definition of "inadequate" is open to some degree of interpretation and dependent on several factors.12 In a recent research project, 85% of foals totally deprived of colostrum became ill within a very short period of time.13 Other studies have shown that in the right environment foals can survive with very small amounts of maternally derived antibodies.14 The "right" environment might, for example, be created by one foal being in a well tended 20 acre field, with only its dam to share the land. Conversely, a "high challenge" situation could be produced if 50 mares and foals shared the same field. The "right" environment is, unfortunately, not present in most situations and the majority of foals with FPT do succumb to some form of infection during the early weeks of life.2-13

Generally accepted veterinary medical standards (and often insurance company requirements) now dictate that foals be checked for IgG within the first day or so of life and those showing a deficiency given some form of supplemental antibodies.

Dosage

A. Oral:

Research has shown that the amount absorbed is directly dependent on the time of administration. Close to 100% could be absorbed if the plasma is given within the first hour or two. This rapidly diminishes, so that only 50% is absorbed at 10 hours and less than 20% at 15 hours. The dose required to correct complete colostrum lack is over 60g and this will be supplied by about two liters of POLYMUNE-PLUS (see Treatment of FPT, page 3).

B. Intravenous:

A 50 Kg foal has a blood volume of approximately 5 liters (or 3 liters of plasma) and therefore any transfused antibodies will be immediately distributed within this volume. The administration of one liter of plasma containing 25 grams IgG will initially raise the recipient's whole blood level by 5g (25 divided by 5). However, within the 24 hours following transfusion there is some movement out of the circulation and only about 50% remains in the vascular spaces after 24 hours. (This is why it is important to wait at least 24 hours before measuring the IgG level after transfusion). Therefore, after the transfusion of the liter of plasma with g IgG, the foal's circulating whole blood level will be increased by about 2.5g/L (250mg/dl whole blood). If we wish to express the concentration in the plasma, then the level will be 25 divided by 2 = 1 2.5g in the blood vessels which is now distributed within 3 liters of plasma, i.e. 4.17g per liter or 417mg/dl plasma.

When having a test done it is important to know whether the results are mg/dl of whole blood or plasma. As shown in the above example, there is a 65% difference. This has been well documented in the past.15 By knowing the initial level and the desired final level, the amount of plasma to be administered can be calculated.

IT SHOULD BE POINTED OUT THAT ANY IgG ADMINISTERED TO A FOAL WILL BE OF SOME BENEFIT. It is also important to realize that passive immunoglobulin transfer is not a simple "yes" or "no" equation and there is no absolute level of circulating IgG that will ensure the foal's health. To afford maximum protection against disease, the immunoglobulin should be available at the place where an organism is invading. It must also be of the appropriate immunoglobulin type to attach to the specific invading organism and be present in sufficient quality to neutralize the agent.16

In the presence of sepsis, immunoglobulins are rapidly consumed having, therefore, a very short half-life. (Plasma proteins provided by transfusion normally have a half-life similar to autologous proteins, which in the case of immunoglobulins is about 21 days). In the presence of infection the half-life might be as low as a few hours and a plasma transfusion might not appear to give the expected increase in IgG when given to a sick foal. It needs to be emphasized that the sick foal would have had a lower IgG if plasma had not been given. To keep IgG levels up in the face of rapid consumption, severely compromised foals can require many liters over a few days.

Non-antibody factors in Polymune

As well as specifically increasing resistance to infection by providing antibodies, plasma also provides non-specific protection against diseases. The protection is provided by cytokines, Iymphokines and other bioactive peptides.17 These compounds increase the activity of neutrophils and enhance phagocytosis. Plasma also enhances the benefits of fluid therapy, because of its albumin content. It is indicated for intravenous use in horses of any age with hypovolemia and/or hypoproteinemia, and as supportive therapy in major equine surgery.

Production

Veterinary Dynamics Inc. produces normal equine plasma in California from a closed herd of donor horses, under the regulations of the USDA and the California Department of Agriculture. Prior to acceptance into the herd, the animals are screened by special arrangement with the Department of Serology, University of California, Davis, the USDA and the California State Diagnostic Laboratory. The tests ensure that these horses are essentially free from diseases and that they are cleared as plasma donors.

To ensure the presence of high levels of immunoglobulin (specifically IgG), a special vaccination protocol is utilized. Accurate IgG measurements are done on each batch using a standardized radial immunodiffusion test. The level of IgG in the plasma ranges from 1600 >3500mg/dl. In addition, the total protein is measured during the production stage by refractometer and ranges from 50 g/L to 68 g/L.

Collection of plasma is by the process of plasmapheresis, using equipment initially devised for human donors, but with modifications to facilitate their use in large animals. This completely closed process ensures sterility and freedom from endotoxins. Nevertheless, each batch is also subjected to sterility tests using thioglycollate broth. Plasma is collected directly into one liter bags which are immediately labeled, batch numbered, packaged and placed in the freezer at-18° C.

Specific Antibodies

As well as providing plasma with a range of antibodies against common neonatal pathogens, Veterinary Dynamics has the ability to produce plasma with high levels of antibodies to specific organisms. These antibody specific plasmas are not presently licensed by the USDA but are available in California and Europe though our UK facility.

For many years rotavirus has been implicated as a cause of foal diarrhea.18,19. Consequently, Veterinary Dynamics (UK) has had a vaccine prepared by Moredun Animal Health Ltd. incorporating the equine H2 strain of the virus. The vaccine, utilizing ISCOM technology has been used in their donors to produce high titer rotavirus plasma. These titers are confirmed, using the serum-virus neutralization test, by the Moredun Research Institute. This plasma can be provided (in Europe) in pre-filled syringes for oral use or in liter bags for intravenous use in the severely ill and dehydrated foal. We are looking at the possibility of producing a similar product in California.

More recently, Clostridium perfringens has been implicated in foal diarrhea20 and in 1995 donors were vaccinated with this antigen. In another situation isolates of Rhodococcus equi, have been made into an autogenous vaccine which is used to immunize some of our donors. The plasma subsequently produced by these donors has then been used prophylactically. This is similar to established protocols employed successfully in California, Texas, Kentucky, and Brazil.21,22

Endotoxemia is a serious problem in horses of all ages and, whilst its treatment is a controversial topic, we produce plasma containing gram negative core antibodies by using a vaccine produced by Hygiea Biological Laboratories, CA. In human medicine it has been documented that antiendotoxin plasma is most beneficial when there is a concomitant bacteremia.23 In horses good results have been obtained in a double blind study24 at U.C. Davis, California.

Veterinary Dynamics, Inc. has been developing hyperimmune serum to Clostridium botulinum types B and C. Type B and Type C are available on a limited basis. There is a possibility that we could supply the type B product for use in horses when the veterinarian client relationship is established. Consultation is available with Dr. Denise Jones, our Director of Research. An application for conditional use is pending with the USDA.

Storage

Plasma must be handled carefully when frozen, as the blood transfer bags are somewhat brittle in this state and might crack if knocked or dropped. It is advisable, therefore, to keep it packaged (as supplied) until required. The shelf-life of the frozen product is three years.

Administration

Plasma is best thawed immediately prior to administration. The most convenient means is to place the bag into warm water with a temperature of about 40°C (this will feel like a warm shower - if you cannot keep your hand in it, it is too hot. Thawing in water too hot will denature certain proteins and cause excessive fibrin precipitation). Keep adding more warm water as the plasma thaws. At the correct temperature the whole process will take about 20 minutes. Plasma that has been thawed slowly, but not heated will have a large amount of precipitate in it. This is called cryoprecipate and contains several important clotting factors. By allowing plasma to warm to about body temperature this precipitate will dissolve and the liquid should be relatively clear before administering. Occasionally some fibrin strands are present and these are filtered out by the administration set.

Plasma should be given USING A BLOOD ADMINISTRATION KIT WITH APPROPRIATE FILTER (40-180 microns). It should be used straight from the bag, with nothing added. Foals may be mildly sedated if necessary and the jugular site prepared aseptically. It is also recommended that the skin be anaesthetized and a 16 G x 2" catheter or similar be used. One liter of plasma can be safely administered to a 50 Kg foal in 15-20 minutes.

Plasma which has been thawed and not brought to body temperature can be stored in the refrigerator for up to two weeks or refrozen. We do not recommend that you keep plasma or refreeze it, if it has been warmed to body temperature.

Adverse Reactions

As with any biological (or pharmaceutical) product, reactions can occur . Fortunately, the incidence with commercially produced plasma are very rare . Over the ten years we have been selling this product (amounting to thousands of liters) we have had only a hand full of reactions reported. Some of those, such as reactions caused by not using a filter are easily avoided.

There are potentially several types of adverse reactions:

1. Volume overload - as the new born foal has a total blood volume of about 5 liters, it is easy to understand how volume overload might occur. A healthy foal is able to handle the transfusion of 1 liter of plasma in about 15-20 minutes, but 2 liters should be given over a minimum of two hours. A sick and compromised foal will require much longer for the administration of the same volume. Signs of volume overload include hyperventilation, tachycardia and sweating. The rate of administration must be immediately slowed down if these signs appear and stopped completely if these signs do not quickly abate.

2. Anaphylaxis - there are two types:

A -this usually occurs when the foal is in some way sensitized to the plasma being transfused. In the neonate the likely cause of this is by receiving sensitizing antibodies through the colostrum.25 We are aware of several instances where this appears to have occurred in mares with previous histories of severe dystocia. It is theorized that the dystocia leads to the mare being exposed to foreign (foal) antigens and then producing antibodies to these. Signs includes hyperventilation, frothing at the mouth and gurgling sounds from the lungs. The transfusion should be stopped immediately and epinephrine and anti-inflammatory drugs administered. Forced ventilation, thoracic massage and oxygen might be needed. Mares producing such foals do so year after year and if a transfusion is essential in a subsequent foal, intradermal skin testing could be performed with the plasma prior to transfusion.

B -if using plasma from a donor not screened for red cell antibodies, then agglutination and/or hemolysis might occur. Signs include hyperventilation, but not profound bubbling. This is not a problem with commercial plasma from screened donors.

3. Fibrin entering the system - by not using a filter. Symptoms include rapid dilation of the pupil, apnoea and sudden death. It is likely that a fibrin clump enters the cardiac circulation causing myocardial ischemia.

All these problems are easily avoided by using properly harvested plasma from screened donors. The only occasion which might cause an unexpected problem is when a foal is transfused born to a mare with unknown hypersensitivity from a previous foaling.

Delivery

Plasma is shipped frozen in insulated containers by express courier to reach destinations in most parts of the USA by noon the following day. (Orders must be placed by 3 p.m. to ensure next day delivery). Special arrangements can be made for weekend delivery and when emergency situations arise.

 

LEGEND

a. Gamma Check-E Veterinary Dynamics, Inc. San Luis Obispo, CA

b. Gamma Check-C Veterinary Dynamics, Inc. San Luis Obispo, CA

c. Colostrometer Juergensen Loveland, CO

d. Equine RID Veterinary Dynamics, Inc. San Luis Obispo, CA

e. POLYMUNE and

POLYMUNE-PLUS Veterinary Dynamics, Inc. San Luis Obispo, CA

 

REFERENCES

1. Jeffcott LB: Some practical aspects of the transfer of passive immunity to newborn foals. Equine Vet J, 1974; 6:109-115.

2. Perryman LE, and Crawford TB: Diagnosis and management of immune system failures in foals. Am Assoc Equine Pract, 1979; 235-244.

3. Rumbaugh GE, et al: Identification and treatment of colostrum deficient foals. J Am Vet Med Assoc, 1979; 174:273-275.

4. Jones D, and Brook D: Investigation of the GAMMA-CHECK-C test as a means of evaluating IgG levels in Equine Colostrum. J of Eq Vet Sc, 1995; 15: 269-273.

5. Durando MM, et al.: Effects of polymyxin B and Salmonella typhimurium antiserum on horses given endotoxin intravenously. Am J Vet Res, 1994; 55 (7):921-926.

6. Mouchawar AM, et al.: Serum but not Plasma Produces Injury in the Perfused Rabbit Lung. Anesth Analg, 1994; 79:40-45.

7. Cullor J: Dept of Pathology, UC Davis, Davis, CA. Personal Communication, 1994.

8. Brown CM: Uncertainties in the significance, diagnosis and treatment of FPT in foals. ISVP Newsletter, Ed. Shauna Spurlock, Veterinary Learning Systems, New Jersey, 1991.

9. Immvac, Inc., Columbia, Missouri. Instructions for Using Endoserum

10. Sera, Inc., Kansas. Instructions for using Seramune

11. Byars TD: Personal communication, Lexington, KY, 1995

12. Koterba AM, Brewer B, and Drummond WH: Prevention and control of infection. Vet Clin North Am Equine Pract, 1985; 1:41-50.

13. Robinson JA et al.: A prospective study of septicemia in colostrum-deprived foals. Equine Vet J, 1993; 25:214-219.

14. Baldwin JL et al.: Immunoglobuiin G and early survival of foals: a three year field study. Proc Am Assoc Equine Pract, 1989; 35:179-185.

15. White SL: Exogenous IgG in the treatment of foals with failure of passive transfer and/or sepsis. In: Sixth Annu ACVIM Proc, 1988; 145.

16. Garry F, and Wells S: Colostrum Management: How Good is Your Program. Second Western Large Herd Dairy Management Conference, Las Vegas, NV, 1995.

17. Harris JA, et al.: Effects of ant-R.equi hyperimmune serum on equine neonatal alveolar macrophage function. In: Third Int Conf of ISVP, 1993; 16-17.

18. Dugdale D: Outbreak of rotavirus diarrhoea in two successive years on a stud farm. Equine Vet Education, 1992; 4:233-236.

19. Dwyer RM, et al.: Infectious foal diarrhea: A three year study of epidemiology and prevention. Proc Am Coll Vet Int Med, 1990; 8:569-57.

20. Anon; Foal Diarrhoea, Animal Health trust, Newmarket. England. Scientific Report, 1992-1993.

21. Veterinary Dynamics Newsletter, 1994. San Luis Obispo, California.

22. Madigan JE and Muller N: Acquisition of passive immunity against Rhodococcus equi by administration of hyperimmune plasma. Proceedings 35th MEP Convention, 1989; 35:521-523.

23. Buamgartner JD: Anti-endotoxin antibodies as treatment for sepsis - lessons to be learnt. Reviews in Med Microbiol, 1994; S3:183-190.

24. Spier SJ, et al.: Protection against clinical endotoxemia in horse by using plasma containing antibodies to an Rc mutant E.coli (J5). Circular. Shock 1989; 28:235-248.

25. Tizard I: Personal communication, Texas A and M University, Texas, USA, 1994.

 VETERINARY DYNAMICS INC. 1535 TEMPLETON ROAD TEMPLETON, CALIFORNIA 93465

Telephone 805 434-0321


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