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FIRST MILKING BOVINE COLOSTRUM Colost rum is a natura l secret ion produced by the mammary g lands of a l l mammals that is in tended for ingest ion by the newborn dur ing the f i rs t hours of l i f e. In most mammals, such as humans, many of the b io log ical ly act ive substances in colost rum that are essent ia l for development and survival of the fetus cross the p lacenta and reach the newborn pr ior to b i r th. However , bovines (cows) are very d i f ferent and essent ia l ly none of the required b io log ical ly act ive substances cr oss the p lacenta. Instead, they remain in the mammary g land (udder) and must be conveyed to the calf by suck l ing dur ing the ear ly hours of i ts l i f e . As such, complete bovine colost rum col lected in the f i rst hours af ter the calf is born is a unique, conce nt rated resource for a wide var iety of h ighly benef ic ia l b io log ical ly act ive substances. The format ion and del ivery of co lost rum in cows involves three processes that occur in associat ion wi th the animal 's mammary t issue. The f irst of these is colostrogenesis , which inc ludes the events that in f luence the format ion of co lost rum. The second is lactogenesis or the process by which mammary t issue changes f rom the stage where i t does not secrete f lu id (non - lactat ing) to the stage where i t does ( lactat ing). The th i rd process is cal led galactopoesis and i t inc ludes the steps through which the mammary g land shif ts f rom the del ivery of co lost rum to the product ion of mi lk . Each of these processes is cont ro l led by a speci f ic set of hormones, but they are a lso inf l uenced by several physical factors associated wi th the mammary g land. In the pregnant cow, development of mammary t issue wi th the abi l i t y to create substances and produce secret ions is promoted main ly by growth hormone and associated insul in - l ike growth f actors. The ent i re process is regulated by a ser ies of hormones, the most impor tant being progesterone, which at taches to specia l s i tes on the cel ls that l ine the inter ior of the mammary g land and prevents the cel ls f rom secret ing any f lu ids in to the g land unt i l a few weeks before b ir th. Colostrogenesis - the formation of colostrum. The format ion of co lost rum in the pregnant cow starts about 3 -4 weeks before b i r th of the cal f when a very smal l amount of f lu id is re leased into the developing mammary t issu es. This f lu id conta ins t iny amounts of growth hormone, insul in - l ike growth factors and other t issue t ransforming substances. The presence of these substances in the mammary g land st imulates the appearance of specia l act ive s i tes on the surface of the ce l ls in the inter ior of the mammary g land. These s i tes wi l l be used to t ransfer the substances necessary for the development and survival of the calf f rom the mother 's b loodst ream into the mammary g land. About two weeks before b i r th, more f lu id enters the udder and some of the s i tes on the surface of the cel ls become fu l ly act ivated. Immunoglobul in molecules ( IgG) f rom the mother 's b lood at tach to the act ive s i tes and are t ranspor ted by specia l ized prote ins through the cel ls in to the f lu id in the mammary g land. At the same t ime, specia l ized whi te b lood cel ls f rom the mother at tach to d i f ferent act ive s i tes and are a lso t ransferred into the

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mammary g land. These cel ls immediate ly beg in to re lease other immunoglobul in molecules ( IgM and IgA) in to the mammar y f lu id. Addi t ional s i tes become act ivated dur ing the 3 -5 day per iod before b i r th and serve to t ranspor t the other b io log ical ly act ive substances f rom the mother 's b loodst ream into the mammary g land f lu id. About two days before b i r th, the hormo nal balance and cont ro l of the mammary g land begins to change rapid ly in the mother . The quant i t ies of a hormone known as pro lact in and, to a lesser extent , other hormones increase sharply in the mother 's b loodstream, removing some of the inhibi tory ef fec ts of progesterone. This s tar ts the product ion of large volumes of f lu id that f i l ls the mammary g land and a lso turns on the abi l i ty of ce l ls in the mammary g land to make cer ta in substances, l ike lactose (mi lk sugar) . W hen bir th occurs and the p lacenta is e l iminated, the amount of progesterone in the b lood of the mother drops dramat ical ly and i ts cont ro l over the secret ions in the mammary g land is complete ly removed. At the same t ime, a unique prote in develops ins ide the cel ls l in ing the mammary g land. T his prote in complete ly b locks the process by wh ich b io log ical ly act ive substances were t ransferred f rom the mother 's b loodst ream into the mammary g land. Therefore, at th is point , the composit ion of co lostrum in the mammary g land represents t rue and comple te colost rum that is maximal ly enr iched wi th the b io log ical ly act ive substances required to suppor t the development and wel l -being of the calf . In addi t ion to being a th ick , golden-colored l iqu id, th is mater ia l has the fo l lowing character is t ics:

a h igh prote in concent rat ion, a large por t ion of which is IgG;

the h ighest concent rat ion of g rowth promot ing substances, other hormones and addi t ional b io log ical ly act ive substances;

r ich in mi lk fat ; and

low in lactose. Lactogenesis - gett ing ready to secrete colostrum. The process by which the cow beg ins to prepare to secrete the f lu ids developed in i ts mammary g land, or lactogenesis , star ts before colost rogenesis f in ishes. W hen the leve l of progesterone in the mother 's b loodstream beg ins to fa l l a few days before b i r th, s ignif icant volumes of f lu id beg in to enter the mammary g land, a l though the volumes that enter are not as great as wi l l occur af ter b i r th. At th is point in t ime, i t is physical ly possib le to remove some colost rum f rom the mammary g land, however , the colost rum usual ly is not fu l ly formed and the removal has been shown to d imin ish the quant i ty and qual i ty of f lu id col lected af ter b i r th. Ful ly funct ional lactat ion in the cow beg ins wi th in moments af ter b i r th when the p lacenta is e l iminated an d a l l inh ib i t ion of secret ions by progesterone is removed. Galactopoeisis - the production of milk. The potent ia l to produce mi lk wi th in the mammary g land a lso occurs a lmost s imul taneously wi th e l iminat ion of the p lacenta and removal of the cont ro l l ing ef fects of progesterone. Af ter b i r th, one of the most

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in f luent ia l factors on the composi t ion of what wi l l be secreted f rom the mammary g land is physical remova l of the f lu id wi th in the g land. Dur ing the hours af ter b i r th, removal of even a smal l amount of f lu id f rom the g land resul ts in the product ion of very large amounts of f lu id by cel ls in the g land i tse l f . Since the t ransfer of b io logical ly act ive substances f rom the mother 's b loodst ream was b locked at b i r th, a l l of the substances found in the adde d f lu id wi l l have been made by cel ls in the g land i tse lf and the composi t ion of the remain ing f lu id wi l l change. This secondary f lu id that is produced whol ly by the mammary g land af ter b i r th, but may conta in remnants of the or ig inal colost rum, is cal led " t ransi t ional mi lk " . Thus, the larger the quant i ty of or ig inal complete colostrum removed at the f i rs t mi lk ing af ter b i r th, the more adul terated the remain ing colostrum becomes wi th t ransi t ional mi lk . Factors that affect colostrum composit ion. Another impor tant character is t ic that in f luences the composi t ion of co lost rum is the fact that , i f the colost rum is not removed f rom the mammary g land wi th in 6 hours af ter b i r th, the mother 's system wi l l beg in to reabsorb many of the b io log ical ly act ive substances, inc luding the hormones and immunoglobul ins, back into her b loodst ream f rom the mammary g land. Therefore, the composi t ion of the secret ions f rom the mammary g land change rapid ly dur ing the hours and days af ter b i r th so that there is a cont inuous t ransi t ion f rom complete colost rum to whole mi lk . These changes are evident f rom the values shown in the fo l lowing char t taken f rom Fundamentals in Dairy Chemist ry, a reference book for the dairy indust ry publ ished in 1978.

COMPOSIT ION OF BOVINE COLOSTRUM & T RANSIT IONAL MILK

Hours % % A f te r To ta l % % Tota l

Ca lv ing Pro te in Fa t Lac tose So l ids 0 17 .57 5 .10 2 .19 26 .99 6 10 .00 6 .85 2 .71 20 .46 12 6 .05 3 .80 3 .71 14 .53 24 4 .52 3 .40 3 .98 12 .77 48 3 .74 2 .80 3 .97 11 .46 72 3 .86 3 .10 4 .37 11 .86 96 3 .76 2 .80 4 .72 11 .85

As can be seen f rom th is char t , complete colostrum is made up of about 27% sol id mater ia l and 73% wate r at the t ime of bi r th. About 65% of the sol ids are prote ins and approximately 40% of these prote ins are immunoglobul ins, most ly IgG. I f no f lu id is physical ly removed f rom the udder wi th in 6 hours af ter b i r th, the amount of so l ids drops by about 25%; the prote in content is reduced by more than 40% and the tota l prote ins now are only 50% of the tota l so l ids. Six hours la ter , at 12 hours af ter b i r th, the tota l so l ids are only about 55% of what they were at the t ime of b i r th and only 40% of the sol ids are prote ins. This change is even more dramat ic by 24 hours af ter b i r th and cont inues as the mother reabsorbs unut i l ized b io log ical ly act ive substances f rom her mammary g land. In addi t ion to the loss of mater ia l due to

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reabsorpt ion, the f lu id in the mammary g land ref lects other changes that are ongoing af ter b i r th, inc luding the product ion of lactose by cel ls that l ine the mammary g land and the s lowing of fat product ion. The average amount of co lost rum that can be col lected f rom a mature Holste in dairy cow in the f i rst mi lk ing taken wi th in the f i rs t 6 -8 hours af ter b i r th is approximately 10 l i ters (9.5 quar ts) . Since the inner surface of the udder has fo lds and wr ink les in i t , not more than 80% of the f lu id in the udder can be removed in a s ing le mi lk ing. The remain ing 20% is t rapped in the fo lds and wr ink les and is d i lu ted wi th the t ransi t ional mi lk that is generated by the cel ls ins ide the mammary g land to replace the f lu id that was removed. Therefore, the second mi lk ing af ter b i r th, even i f i t is taken wi th in the f i rst 6 hours, only conta ins 20% of the or ig inal complete colost rum that was present in the mammary g land at b ir th. The same 80/20 ru le appl ies every t ime the udder is empt ied and, thus, each succeeding mi lk ing proport ionate ly d i lu tes the res idual or ig inal complete colost rum lef t in the mammary g land unt i l the resul t ing f lu id has the composi t ion of who le mi lk . When the facts are put into perspect ive, i t is easy to understand how important i t is to col lect the maximum amount of colostr um in one milking as soon after birth as possible and def ini tely not more than 6 hours after the calf is born. This is the mater ial that contains the maximum amount of biologically act ive substances and is known as complete f i rst milking bovine colostrum. Colostrum and the cal f . Everyone who uses colost rum worr ies that the dairy farmer is going to col lect as much colost rum as he can and then sel l i t wi thout g iv ing any to the calf . F i rst , we have to understand that the calf in quest ion is valuab le to the dairy farmer as a replacement or expansion animal in h is herd, so he is going to do everyth ing that he can to help that calf survive and develop into a hea l thy, producing animal . In fact , modern dairy farmers in the Uni ted States recognize that a l lowing the calf to suck le i ts mother usual ly resul ts in the cal f receiving less colost rum than i t should. Numerous sc ient i f ic studies have shown that a calf must receive about two quar ts of co lost rum wi th in 6 hours af ter i t is born to min imize i ts r isk of d isea se and support i ts development as a heal thy and product ive animal . I f al lowed to suck le, the cal f wi l l usual ly do so for about 10 -15 minutes af ter b i r th and then wi l l s leep for several hours; ar ise, suck le for 10 -15 minutes and then s leep again for severa l hours; repeat ing th is process for a few days unt i l i t ga ins s t rength. The f i rs t suck l ing a lone removes only a small amount of co lost rum, but causes the generat ion of a substant ia l amount of t ransi t ional mi lk , s ignif icant ly d i lu t ing the res idual colostru m. This d i lu t ion ef fect is magnif ied dur ing each successive suck l ing by the calf , l imi t ing the amount of benef ic ia l b io log ical ly act ive substances i t can receive. Most dairy farms in the Uni ted States mainta in a separate matern i ty ward and nursery where they moni tor pregnant cows 24 hours a day

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dur ing the week just pr ior to del ivery. W hen the calf is born, i t is immediate ly removed f rom i ts mother and not a l lowed to suck le. The colost rum is col lected in a s ing le uni t wi th in a few hours af ter b i r th and the calf is hand-fed two quar ts in a nurs ing bott le before i t is 6 hours o ld. This approach not only assures that the calf got enough colost rum when i t needed to in order to maximize the benef i ts, but i t a lso keeps the calf f rom mak ing contact wi th the s oi led udder and fecal mater ia l f rom i ts mother and other cows, wh ich are recognized resources for the t ransfer of infect ious microorganisms. There is a considerable body of sc ient i f ic evidence that a cal f that fa i ls to receive an adequate quant i ty of h ig h qual i ty complete colost rum shor t ly af ter b i r th wi l l have a reduced capabi l i ty to protect i tse lf against pathogenic microorganisms in i ts environment . In addi t ion, i f the animal survives, i t wi l l have a h igher inc idence of i l lness and i t wi l l develop less body mass than normal . The impact of inadequate colost rum intake dur ing the f i rs t hours of l i f e on the survival and heal th of ca lves was studied in more than 2,200 animals over a f ive year per iod by the Uni ted Kingdom Nat ional Agr icu l tura l Center Cal f Un i t . As shown in the table, they found that calves that received only a smal l amount of co lost rum were s ix t imes more l ike ly to d ie than those that received the required two quar ts. I f the animals that received a smal l amount of co lost rum survived, they we re i l l a lmost three t imes as of ten as the calves that got enough colost rum. Get t ing enough high qual i ty colost rum is essent ia l to the heal th and wel l -being of the cal f and fa i lure to do so wi l l fo l low the animal for the rest of i ts l i f e. Dairy farmers say "how a cal f star ts determines how the cow f in ishes" .

EFFECT OF COLOSTRUM INTAKE ON CALF HEALTH

Co los t rum % % In take D ied I l l L i t t le 7 .9 42 .2 Som e 3 .0 24 .2 Enough 1 .3 15 .4

The rapid ly chang ing composi t ion of colost rum in the mammary g land of the mother f i ts together very wel l wi th events that happen in the body of the newborn cal f . Dur ing the f i rs t s ix hours of l i fe , the calf 's s tomach l in ing does not make any ac id and there are very few, i f any, enzymes present that can break down ingested prote ins. Complete f i rs t mi lk ing colostrum also conta ins substances that inhib i t the act ion of some enzymes. Therefore, these condi t ions work in favor of having the b io log ical ly act ive substances in complete colost rum pass through the calf 's stomach into the upper por t ion of the smal l in test ine wi thout being broken down. Dur ing the f i rs t 6 -8 hours of l i f e, th is area of the smal l in test ine has specia l ized s i tes where the b io log ical ly act ive substances can be absorbed and t ranspor ted d i rect ly in to the calf 's b loodst ream. Af ter th is per iod, the stomach beg ins to ac idi f y, enzymes appear and the specia l ized absorpt ion area in the smal l in test ine changes dramat ical ly so that most of the b io log ical ly act ive substances

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in colost rum are no longer absorbed. This process is a ided by the fact that calves are born wi th a wel l -developed system of lymphoid t issue under their tongue and at the back of their throat that pers ists throughout their ent i re l i fe . Many b io log ical ly act ive substances are absorbed through these t issues when the calf suck les i ts mother or a nurs ing bott le . In adul ts , th is is a major route for absorpt ion of benef ic ia l substances f rom food, wi thout subject ing them to degradat ion in the digest ive t ract , as the animal "chews i ts cud " . Comparison of bovine and human colostrums. Colost rum has evolved in nature as a nut r i t ional supplement that is fed to the newborn by i ts mother dur ing a shor t per iod af ter b i r th. I ts natura l evolut ion can be shown to fo l low a path tha t meets the immediate demands of the newborn and, s ince most species of mammals develop at d i f ferent rates and have d i f ferent nut r i t ional requirements, the colost rum and fo l low -on mi lk of each species has a d i f ferent composi t ion. Some colostrums are very s imi lar and some are ext remely d i f ferent . The composi t ion of bovine colost rum is qui te d i f ferent f rom the colost rum produced by humans. The most substant ia l d i f ference is found in the immunoglobul in (ant ibodies) content of these colost rums. In humans, t he IgG immunoglobul ins are conveyed f rom the mother 's b loodst ream through the p lacenta whi le the fetus is s t i l l in the mother 's uterus and are not found in the colost rum. This provides some degree of temporary passive immuni ty to the newborn against possib le infect ious agents af ter b i r th. In contrast , none of the immunoglobul ins in the pregnant cow's b loodst ream are t ransferred across the p lacenta and, thus, the cal f is essent ia l ly defenseless when i t is born unless i t gets colostrum. The major immunoglobu l in found in bovine colost rum is , therefore, IgG, but , a l though there is only a smal l amount of immunoglobul ins in human colostrum, the major one is IgA. The immunoglobul in ( IgA) in human colostrum and mi lk is unique in that i t has a specia l prote in f ragment at tached to i t that is bel ieved to protect i t f rom the ef fects of s tomach ac id and d igest ive enzymes. I t must a lso be recognized that s ince the immunoglobul ins are t ransferred across the p lacenta in humans, many of the essent ia l g rowth factors are a lso t ransferred. Therefore, human colost rum and the mi lk that fo l lows become supplementary resources for these b io log ical ly act ive substances and the newborn can survive and develop wi thout receiv ing them by suckl ing . In cont rast , none of these factors are t ransfer red across the p lacenta in the pregnant cow and, as d iscussed ear l ier , ca lves that do not receive enough colost rum suf fer ser ious def ic ienc ies in the development of their immune system and their body mass. Bovine and human colost rum also d i f fe r in other ways that ref lect the d i f ferent needs of the respect ive newborns. For example, human colost rum and mi lk conta in the h ighest amount of lactose (mi lk sugar) of any species. The lactose provides about 40% of a l l of the calor ies avai lable to the suck l ing infant . The h igh lactose content is bel ieved to

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serve two purposes: (1) the infant bra in is very large, much larger than any other species, and requires a lo t of g lucose to develop rapid ly; lactose is easi ly broken down into g lucose and galactose before i t is absorbed by the intest ines; and (2) for physical chemical reasons, the h igh lactose content causes the secret ion of a large amount of water in to the mammary g land, assur ing that the infant 's needs to support f lu id loss through sweat ing and ur i ne format ion are met. Bovine colost rum and mi lk conta in substant ia l ly more casein than is found in human colost rum or mi lk . Casein is a complex prote in that is acted upon by enzymes in the stomach to form a curd that has the consis tency of cot tage cheese. In the cal f , th is curd is much harder than the curd that forms in babies that are nursed. The curd a l lows t ime for addi t ional enzymes to break the prote in down into smal l pept ides and amino ac ids. These breakdown products are absorbed into the body and serve as the bui ld ing b locks for new prote ins to bui ld muscle. The ext ra casein in bovine colost rum and mi lk ref lects the need for a ready source for more muscle -bui ld ing capaci ty s ince the body mass of a calf develops much more rapid ly than that of a newborn baby. Colost rum is an amazing mater ia l that , l ike many other th ings in nature, ref lects the evo lut ionary development of a unique composi t ion that wi l l serve the needs of the of fspr ing for which i t is in tended. The most unique of the colostrums from mammalian species occurs in bovine species where everything required for the development of a heal thy, product ive offspring is provided in the colostrum. As such, i t represents a special ized resource that offers the broadest possible spectrum of biol ogically act ive substances that can promote the development of a sound body mass and support the act ivat ion and maintenance of a ful ly funct ional immune system capable of ef f icient ly and ef fect ively combating potent ial insults f rom microorganisms and other deleterious sources. Bovine colostrum is also compatible with almost any species and can readi ly convey i ts ful l benef its to humans by rout ine dietary supplementation .

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THE COMPOSITION OF BOVINE COLOSTRUM The previous sect ion provides an ins ight in to how bovine colost rum is formed and descr ibes what complete f i rs t mi lk ing colost rum real ly is . This sect ion d iscusses in some deta i l what i t is composed of and the nature of the major b io log ical ly act ive components. The basic composit ion of colostrum. As indicated in the ear l ier sect ion, complete f i rs t mi lk ing bovine colost rum is a th ick, golden -colored f lu id that conta ins a lo t of prote ins, many of which are b io log ical ly act ive; is r ich in mi lk fat ; and is low in lactose (mi lk sugar) . T he best way to understand what colost rum should be composed of is to look at how the re lat ionship of these three major components changes wi th t ime af ter b i r th of the calf . In th is case, the basic values shown for l iqu id colostrum in the last sect ion have been conver ted to what would be found in the same colost rum if i t was dr ied in powder form, the way most people use colost rum as a d ietary supplement.

COMPOSIT ION OF DRIED BOVINE COLSTRUM & TRANSIT IONAL MILK

Hours %

A f te r To ta l % % Ca lv ing Pro te in Fa t Lac tose

0 65 .10 18 .90 8 .11 6 48 .90 33 .48 13 .25 12 41 .64 26 .15 25 .53 24 35 .40 26 .62 31 .17 48 32 .64 24 .43 34 .64 72 32 .55 26 .14 36 .85 96 31 .73 29 .60 39 .83

I t is real ly obvious how fast the re lat ionship of these components changes af ter b i r th of the calf . This chang ing re lat ionship is ext remely impor tant in establ ish ing that the best colost rum is used and in assur ing that i t conta ins the maximum amount of b io log ical ly act ive substances. Protein. Most of the b io log ical ly act ive substances in complete bovine colost rum that can convey s igni f icant heal th be nef i ts are prote ins. As you can see, the protein composit ion of colostrum changes rapidly af ter bir th. Since almost al l of the benef icial proteins are conveyed from the mother 's bloodstream into the colostrum before bir th and the mother then begins to re absorb them after bir th, i t is important to use colostrum that has been col lected during a t ime period that wil l minimize the effect of the reabsorption process . The f i rst sect ion descr ibed the 80/20 ru le and the d i lu t ing ef fects of t ransi t ional mi lk when colost rum is removed, so i t is a lso very

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impor tant to assure that only complete f i rs t mi lk ing colost rum was used. The whole concept for def ining the value of f i rs t mi lk ing colost rum taken wi th in 6 hours af ter bi r th has been establ ished by a number of sc ient i f ic s tudies. Of real s ignif icance is the f inding that by 24 hours af ter b i r th most of the prote ins in the udder f lu id can be accounted for by two indiv idual prote ins that are pr imar i ly only of nut r i t ional va lue. They are casein, which was d iscussed i n the f ist sect ion, and a lbumin, which is s imi lar to the a lbumin found in egg whi tes. Since we cannot col lect co lost rum at the t ime of or wi th in minutes af ter b i r th, good qual i ty complete f i rst mi lk ing colost rum wi l l conta in 45-60% prote in, of which about 40% wi l l be immunoglobul ins. Colostral Fat . The mi lk fat in complete f i rst mi lk ing colost rum is one the most under -rated and misunderstood components by many companies that promote bovine colost rum for human consumpt ion. There are al l k inds of s tor ies, none of wh ich are ever substant iated wi th any sc ient i f ic evidence. One of these says that the fat in colostrum doesn' t serve any purpose and/or that having i t there leads to faster deter iorat ion of the product. Noth ing could be fur ther f rom the t ruth. I n fact , one of the companies that removes the fat f rom what they cal l "co lost rum" adds a component of the fat back to their dr ied products. They c la im that th is makes their "colost rum" more d igest ib le, which was one of the funct ions of the fat in complete colost rum in the f i rst p lace. Remember f rom the last sect ion how casein is broken down in the stomach to smal l pept ides and amino ac ids so that they can be absorbed and used to bui ld new muscle prote in by forming a cot tage cheese - l ike curd in the stomach. This occurs enzymat ical ly in the newborn and the adul t . The basis for the curd that forms is the fat in the colost rum. So wi thout i t , in addi t ion to los ing some s ignif icant b io log ical ly act ive substances that are associated wi th the fat , most of the n ut r i t ional va lue of the casein is a lso lost . That is par t of the reason why the fat content of co lost rum increases wi th t ime af ter b ir th as the amount of casein increases in the secreted f lu id. Mother Nature doesn't waste much and has organized the components of colostrum and their changing pattern in an ef f icient way to maximize the benef its to the offspring that is going to receive i t . High qual i ty f i rs t milk ing bovine colost rum wi l l conta in 20 -30% mi lk fat . The mi lk fat in colost rum is a lso a very im por tant means to del iver some of i ts benef ic ia l b io log ical ly act ive substances. Dissolved in or associated wi th the fat in colostrum are the fo l lowing substances.

Vi tamins A, D, E and K

Stero id hormones

Cor t icostero ids

Some growth factors

Insul in

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Lactose (mi lk sugar) . As can be seen f rom the chart above, in h igh qual i ty complete f i rst mi lk ing colostrum 10-15% of a l l of the sol id mater ia l in the f lu id wi l l be lactose. W e discussed ear l ier how impor tant the lactose is to the calf as an immediate energy source when i t is broken down to g lucose and galactose by an enzyme ( lactase) in the sal iva and the stomach. Therefore, i t makes good sense that the amount of lactose in t ransi t ional mi lk and mature mi lk increases as the body mass of the cal f develops rapid ly dur ing the ear ly days of i ts l i f e . Since most people have the same enzyme ( lactase) in their sa l iva and their d igest ive system, the lactose in the colost rum that they use as a d ietary supplement can provide the same ready source of energy. However , there are indiv iduals who have problems digest ing lactose because their body produces too l i t t le or none of the lactase enzyme. They are said to be " lactose into lerant" . This problem occurs more f requent ly in indiv iduals of As ian and Lat in Amer ican de scent , but is found in people of a l l e thnic backgrounds. As shown in the char t , the amount of lactose in f i rs t mi lk ing colost rum col lected wi th in 6 hours af ter b i r th is about one -half of what i t is at 12 hours af ter b i r th and one- th i rd of what i t becomes by 24 hours. Therefore, a h igh qual i ty complete f i rs t mi lk ing colostrum col lected wi th in 6 hours af ter b i r th can be used as a d ietary supplement by more people wi thout potent ia l ly having them suf fer the d iscomforts associated wi th lactose into lerance. Other composit ional changes. The fo l lowing comparat ive facts about colost rum and mi lk fur ther st ress the need to use a complete f i rs t mi lk ing colost rum in order to maximize the benef i ts that i t can provide.

Colost rum conta ins 10 t imes more vi tamin A than m i lk .

Colost rum conta ins 3 t imes more vi tamin D than mi lk .

Colost rum conta ins at least 10 t imes more i ron than mi lk .

Colost rum conta ins more calc ium, phosphorous and magnesium than mi lk .

The biological ly act ive components. The bio log ical ly act ive compone nts in complete f i rs t mi lk ing colostrum can be d ivided into categor ies based upon the heal th aspect where they exer t their g reatest inf luence. As we go through the discussion of what these substances do, i t can be seen that , in some cases, the funct ions of these components can be c lear ly separated into such categor ies, whi le, in many cases, the d iv id ing l ine is c louded. The major categor ies are the Immune Factors, the Growth Factors and the Metabol ic Factors. In th is sect ion, we intend to make you fami l ia r wi th the substances in these categor ies. However , before we beg in th is d iscussion, i t is very impor tant to recognize that most of the very broad c la ims about what these substances do that are made by many other suppl iers of co lostrum for human consumpt i on are based upon very

11

specia l ized studies in exper imental animals, l ike mice or rats , and represent the company's interpretat ion of the results and not necessari ly that of the or iginal scienti f ic invest igator. The Immune Factors. To comprehend what the Immune Factors are in h igh qual i ty f i rst mi lk ing colostrum and what they do, i t is important to recognize that some of these components have one or more ef fects on the overal l regulat ion and funct ioning of the immune system ( immuno -regulat ing substances) , whi le others are very restr ic ted in what they can do and their benef i ts are usual ly very loca l ized in the body, ord inar i ly exer t ing their ef fects pr imar i ly in the gut (gut protect ive substances). Immuno-regulating substances.

Thymosin (alpha & beta chai ns). A hormone composed of two prote in-based chains that are separate ly present in bovine colost rum. The chains act on the thymus g land independent ly or in concer t wi th each other to s t imulate act ivat ion, development and maintenance of the immune system.

Prol ine-rich pept ide (PRP). A hormone- l ike smal l prote in that acts upon the thymus and other organs associated wi th the immune system to keep i t f rom over -react ing to an insul t .

Cytokines. Smal l prote ins produced by var ious cel ls in the body that induce the generat ion of specia l ized types of whi te b lood cel ls , s ignal them to come to the s i te of an insul t and he lp in their passage through t issues.

Lymphokines. Prote ins of varying s izes that are produced by d i f ferent types of wh i te b lood cel ls that te l l re lated cel ls to t ransform themselves into more funct ional ce l l types that can re lease substances capable of dest roying an invad ing microorganism.

Gut protective substances.

Immunoglobul ins ( IgG, IgM, IgA) . Complex prote ins, bet ter known as ant ibodies, that make up a s ignif icant port ion of the prote ins found in complete f i rs t mi lk ing colostrum. These ant ibodies were produced by the mother 's immune system in response to her exposure to many d i f ferent microorganisms dur ing her l i fe t ime and then transfer red into the colostrum pr ior to b i r th of the cal f . There is no ev idence that any of these ant ibodies are found intact in the b lood of individua ls who ingest colost rum by mouth. However , many of these ant ibodies are react ive against bacter ia, v i ruses a nd fungi that infect the gast ro intest inal t ract of humans and there is sc ient i f ic evidence that some of them can survive passage through the d igest ive system.

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Transfer factors. Smal l prote ins produced in response to the body's exposure to cer ta in type s of microorganisms, par t icu lar ly those that res ide in deep t issues for a long per iod of t ime, l ike the bacter ium that causes tuberculos is . They are specif ic for a par t icu lar microorganism and are carr ied ins ide of certa in types of specia l ized wh i te b lood cel ls . Transfer factors have l imi ted ef fect iveness a lone in defending the body against infect ion by such microorganisms, but , rather , act in concer t wi th var ious whi te b lood cel ls and other factors in an at tempt to keep the microorganisms under cont ro l .

Lactoferr in. A mineral -b inding carr ier prote in that at taches to avai lable i ron. Cer ta in aerobic (grow in the presence of oxygen) bacter ia, l ike E. col i , require i ron to reproduce and, therefore, lactoferr in is an ef fect ive substance, when operat ing in th e presence of a speci f ic ant ibody, to impede the growth of some microorganisms in the gut . A broad number of addit ional c la ims have been made by some providers of co lost rum for human consumpt ion regarding the appl icat ion of lactofer r in as an immuno -regulat ing substance wi th ant iv i ra l , ant ibacter ia l and ant i - tumor propert ies. To date, none of these c la ims have been adequately substant iated through proper ly cont ro l led s tudies.

Transferr in. Another mineral -b inding carr ier prote in that at taches to avai lable i ron and can act independent ly or in concer t wi th lactoferr in to impede the growth of certa in aerobic bacter ia, par t icu lar ly in the gut .

Lysozyme. A very power fu l enzyme that is capable of at taching i tse lf to the cel l wal l o f certa in pathogenic bacter ia and degrading select components, leav ing holes in the wa l l o f the bacter ia.

Lactoperoxidase. A mi ld ly ef fect ive enzyme that can a lso at tach to the wal l o f certa in bacter ia, degrade selected prote ins and inter fere wi th the abi l i ty of the bacter ia to repl i cate themselves.

Xanthine Oxidase. Another mi ld ly ef fect ive enzyme that can a lso at tach to the wal l of cer ta in bacter ia, degrade d if ferent prote ins than those af fected by lactoperoxidase and, therefore, a lso inter fere wi th the abi l i ty of the bacter ia to repl icate themselves.

White blood cells ( leukocytes) . Pr imar i ly, three types of funct ional whi te b lood cel ls are present in colostrum, inc luding neut rophi ls , macrophages and polymorphonuclear cel ls . Each has the abi l i ty to phagocyt ize (engulf ) microo rganisms and other fore ign bodies and apply substances carr ied internal ly to the dest ruct ion of the microorganisms. Their funct ions are dramat ical ly enhanced when ant ibodies f i rst at tach to the microorganisms.

Oligosaccharides and glycoconjugates. Complex carbohydrates (sugars) that can adhere to specif ic s i tes on the inner surface of the gast ro intest inal t ract and prevent the at tachment of microorganisms.

13

The Growth Factors.

Growth hormone. Very smal l quant i t ies of growth hormone are found in complete f i rs t mi lk ing colost rum, but that is a l l that is required s ince th is hormone is ext remely potent . I t has a d i rect ef fect on a lmost every cel l type and s ignif icant ly in f luences the pro l i ferat ion of new cel ls , par t icu lar ly their rate of generat ion. Scient i f ic s tudies have shown that cont inued ingest ion of smal l amounts of g rowth hormone are benef ic ia l in l imi t ing the ongoing deter iorat ion of ce l ls associated wi th the ag ing process.

Insulin-l ike growth factors ( IGFs) . Insul in- l ike growth factor -1 ( IGF-1) and i ts c losely re lated counterpar t insul in - l ike growth factor -2 ( IGF-2) are potent hormones that are found in associat ion wi th a lmost a l l ce l ls in the body. They are par t of a group of more than 90 d i f ferent prote ins, cal led the " IGF Binding Prote in ( IGFB P) Superfami ly" , that is responsib le for the processes by which cel ls g row and reproduce. These substances are a lso responsib le for maintenance of the metabol ic pathways in the body by which cel ls conver t g lucose to g lycogen, a pr imary energy resource, and use amino ac ids to create prote ins. The key event that t r iggers the funct ions of the var ious prote ins in the IGFBP Superfamily is the at tachment of IGF-1 to a specif ic s i te on the surface of a cel l . Many of the growth factors found in colost rum and pre viously def ined by their funct ions are now considered par t of the IGFBP Superfami ly. This inc ludes the fo l lowing substances, among others.

Transforming growth factors A & B. Induces the

t ransformat ion of ce l ls f rom an immature form to a mature, funct ional status.

Epithelial growth factor. Involved in the generat ion and

maintenance of cel ls in the epi thel ia l (outer) layers of the sk in.

Fibroblast growth factor. Associated wi th the regenerat ion

of var ious types of t issue, inc luding sk in and other organ s.

Platelet -derived growth factor. Responsib le for the generat ion of ce l ls and funct ions associated wi th b lood c lot t ing .

The Metabolic Factors.

Lept in. A smal l hormone- l ike prote in that can suppress appet i te and lead to body we ight reduct ion. Mature f at ce l ls (adipocytes) re lease lept in in the presence of insul in , wh ich is a lso found in colost rum. Insul in -producing pancreat ic beta -cel ls have b inding s i tes for lept in and i t is bel ieved that the s ize of fat ce l ls may be a major factor in determining the amount of lept in re leased. Therefore, lept in def ic iency may be associated wi th obesi ty, par t icu lar ly in d iabet ic individuals .

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Insulin. A hormone required for the ef fect ive ut i l izat ion of g lucose (b lood sugar) in the body. Insul in b inds to specif ic s i tes on cel ls , fac i l i ta t ing their in teract ion wi th IGF -1 and, thus, in i t ia t ing the convers ion of g lucose to g lycogen, a h igh energy source carbohydrate.

Vitamin-binding proteins. Smal ler prote ins that act as carr iers to del iver B-complex vi tamins to the body . Carr ier prote ins and the associated vi tamins fo late (B6), B12 and orot ic ac id are found in colost rum.

Fat-associated vi tamins. Signif icant quant i t ies of v i tamins A, D, E and K are d issolved in or associated wi th the fat in colost rum.

Mineral-binding proteins. The i ron-binding prote ins, lactofer r in and t ransfer r in , have a l ready been d iscussed above. In addi t ion to in ter fer ing wi th the repl icat ion of cer ta in microorganisms, they a lso serve to capture i ron f rom the ingested food and present i t in a form that can readi ly be absorbed by the body. Lactoferr in can a lso b ind copper and del iver i t in a form sui table for absorpt ion by the body. In addi t ion, there are two carr ier prote ins in colost rum that ass is t in the absorpt ion of ca lc ium. They are casein, wh ich is a lso an abundant source of amino ac ids to bui ld new prote in molecules, and a lpha- lacta lbumin, wh ich is present in colost rum very soon af ter b i r th.

Cycl ic adenosine monophosphate (cAMP). A phosphorylated nucleot ide in a very specia l ized form tha t t ransfers the chemical energy necessary to dr ive metabol ic react ions to form new prote in, carbohydrate and fat molecules.

Enzyme inhibi tors. These have been cal led "permeabi l i t y factors" by other manufacturers, but are actual ly smal l prote ins that s low down or inhib i t the breakdown of prote ins by cer ta in enzymes. They provide l imi ted protect ion to the immune, growth and metabol ic factors as they pass through the d igest ive t ract .

There are many other substances present in colost rum, but very few of them are of any s igni f icance s ince they are present only in very minute quant i t ies and they provide l i t t le on no benef i t to humans who ingest colost rum rout inely as a d ietary supplement. However , there are two substances, in addi t ion, to those descr ibed abo ve that do provide s ignif icant benef i ts. They are the hormone melatonin , which has a d i rect ef fect on the establ ishment of b io log ical rhythms and proper s leep pat terns; and relaxin , a hormone known to d i rect ly af fect cont racted muscles.

15

CHARACTERIZATION OF COLOSTRUM PRODUCTS

IMMUNO-DYNAMICS

To assure that bovine colost rum conta ins the maximum amount of the avai lable b io log ical ly act ive substances, i t must be col lected only at the f i rst mi lk ing wi th in s ix hours af ter the b i r th of the calf . In addi t ion, the f inal product must be of the same re lat ive composi t ion as the or ig inal co lost rum and not conta in any addi t ives or supplements in order to assure the intactness and funct ional i ty of the b io log ical ly act ive components. Only colost rum products t hat meet these cr i ter ia are capable of del iver ing the maximum benef i ts avai lable through rout ine use. The fo l lowing sect ions examine the character is t ics of the colost rum powder produced by Immuno-Dynamics and some of i ts compet i tors in terms of their chemical composit ion and the content of specif ic components. The values shown in the tables and graphs were der ived f rom assays conducted independent ly and wi thout b ias by a h ighly regarded dairy test ing laboratory and the d iagnost ic laborator ies of a major col lege of veter inary medic ine. Basic Chemical Composit ion As discussed ear l ier , the best way to understand what colost rum should be composed of is to look at how the re lat ionship o f three major components, prote in, fat and lactose, changes wi th t ime af t er b i r th of the calf . In the table below, the basic values shown in ear l ier tables have been conver ted to the t rue uni t of mass that would be found in each gram of colostrum powder .

Composi t ion of Bov ine Colost rum (m g/gm of to ta l so l ids )

Hours

A f te r To ta l B i r th Pro te in Fat Lac tose

0 651.0 189.0 81 .1 6 489.0 334.8 132.5 12 416.4 261.5 255.3 24 354.0 266.2 311.7

The comparat ive chemical composit ions of ten (10) ser ia ls of co lostrum powder manufactured f rom large pools by Immuno -Dynamics are deta i led in the fo l lowing table. The same data is a lso graphical ly d isplayed. Compar ison of these data wi th the values g iven in the table above c lear ly indicates that the ser ia ls are a l l composed of colost rum col lected wi th in s ix (6) hours af ter b i r th of the calf in a s ing le mi lk ing. In addi t ion, the consis tency of the values, ref lect ing the lot - to- lot reproducib i l i ty of the product , is immediate ly apparent.

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CHEM ICAL COM POSIT ION (m g/gm of powder )

Serial Total

No. Protein Fat Lactose

A 516.8 243.8 117

B 559.7 211.7 117

C 545.4 211.0 109

D 557.7 233.5 105

E 549.3 246.0 106

F 549.0 233.8 109 G 547.7 211.8 118

H 546.6 221.0 109

I 568.0 220.1 110

J 556.7 212.1 113 Mean 549.7 224.5 111.3 S.D. 13.371 13.773 4.692 95% Interval 26.742 27.546 9.384 Range 522.96 - 576.44 196.95 - 252.05 101.92 - 120.68

Biological ly Act ive Components A ser ies of e ight (8) of the b io log ical ly act ive components of bovine colost rum were selected as the marker components based upon a) the avai labi l i ty of an immunoassay for the determinat ion of concent rat ion; b) the abi l i ty of the assay or an adaptat ion thereof to perform re l iably wi th colost rum; an c) the importance of the r o le of the component in the b io log ical ef fects observed wi th colost rum.

0

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A B C D E F G H I J

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CHEMICAL COMPOSITION

Protein

Fat

Lactose

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Insulin–Like Growth Factor–1 ( IGF-1) Insul in- l ike growth factor -1 ( IGF-1) and i ts c losely re lated counterpart insul in- l ike growth factor -2 ( IGF-2) are potent hormones that are found in associat ion wi th a lmost every cel l in the body. IGF -1 is the most potent and best descr ibed of th is pair . These molecules are present in a l l mammals and, in every case, have a very s imi lar chemical s t ructure regardless of the species. IGF -1 is absolu te ly necessary for normal cel l g rowth and for the development of the fetus in the uterus. Both IGF-1 and growth hormone are a lso required for normal development outs ide of the uterus and that is why they are both present in colost rum. The chemical s t ruct ure of the IGFs is very s imi lar to insul in and that is where their name comes f rom. Sc ient i f ic knowledge about the IGFs, what they do and how they act on cel ls in the body has developed very quick ly dur ing the past few years. I t is now known that there a re speci f ic s i tes, ca l led receptors, on a lmost a l l ce l ls in the body capable of in teract ing wi th IGF -1. These s i tes have a s t ructure that f i ts perfect ly wi th par t of the IGF molecule and th is in teract ion t r iggers a ser ies of chemical events wi th in the cel l . There are a lso 6 d i f ferent prote ins present ins ide the cel l and on the surface of the cel l that react to the at tachment of IGF -1 to i ts receptor. These are cal led insul in - l ike growth factor b inding prote ins ( IGFBPs) and they cont ro l the act ions of IGF-1 on the cel l . In addi t ion, ins ide the cel l there are at least 87 other re lated prote ins e i ther capable of b inding to IGF-1, a l ter ing i ts act ions, or in f luencing the ef fects of the IGFBPs. These are cal led insu l in - l ike growth factor b inding prote in -re lated prote ins ( IGFBP-rPs). The ent i re col lect ion of these prote ins is refer red to as the Insul in - l ike Growth Factor Binding Prote in ( IGFBP) Superfami ly. The key event that t r iggers the ef fects of any of these prote ins appears to be the interact ion of IGF -1 wi th i ts speci f ic cel l -surface receptor , an event that some of these prote ins regulate. The mul t i tude of avai lable IGF -1binding prote ins and re lated prote ins avai lable in the cel l is ind icat ive of the many potent ia l ef fects that the b inding of IGF-1 to i ts specif ic cel l -surface receptor can have on cel ls . To keep these many ef fects under cont ro l , some of the b inding prote ins act as checks and balances, a l lowing the secondary chemical swi tches in a cel l to be turned on and then turn ing them of f when i t is appropr iate. Therefore, IGF -1 is l ike the capta in of a ship. W hen i t b inds to i ts specif ic receptor , the ship can move forward, but there are a l l k inds of systems in p lace to keep i t moving at the r ight speed and in the r ight d i rect ion. The main t r iggered events inc lude act ivat ion of the process by wh ich the cel l g rows and reproduces i tse lf and maintenance of the metabol ic pathways by which the cel l conver ts g lucose into g lycogen and uses amino ac ids to create prote ins. The actual pathway by which the cel l uses g lucose and conver ts i t to g lycogen is f i rst swi tched on by the b inding of insul in to i ts speci f ic cel l surface receptors. As indicated ear l ier , g lycogen is s tored in the l iver and muscles and is the reserve source of readi ly avai lable energy wh en the muscles are exerc ised.

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The IGFBP Superfami ly a lso has a d i rect ro le in how the cel l uses amino ac ids to bui ld prote ins. As the body of any mammal ages, i ts abi l i ty to create an adequate supply of IGF -1 is d imin ished. Thus, eat ing a wel l -balanced d iet and mainta in ing a constant supply of IGF -1 in the body, keeps the ship moving at the r ight speed and in the r ight d i rect ion. And when the body is exerc ised, th is becomes even more cr i t ica l s ince there is an increased demand for g lycogen to provide energy to the muscles and the preference is to bui ld more muscle prote in. Even more impor tant ly, as the cel ls in the body age, they do not reproduce themselves as we l l and, s ince IGF -1 is a pr imary factor, a long wi th growth hormone, in the abi l i ty of ce l ls to grow and reproduce, i t is h ighly des irable to have an appropr iate level of IGF -1 in the c i rculat ion through d ietary supplementat ion to l imi t the ever increasing rate of ce l l death.

Biological ly Act ive Components

IGF-1 , Lept in , Insul in (m ass /gm of powder )

IGF-1 Leptin Insulin

Serial (ug/gm) (ng/gm) (mIU/gm)

A 2.277 91.82 1.294

B 2.366 138.00 1.508

C 2.526 114.18 1.728

D 2.940 133.18 1.417

E 2.801 145.82 1.517

F 2.665 110.27 1.195

G 2.440 120.64 1.264

H 2.453 93.91 1.076

Mean 2.723 118.45 1.375

S.D. 0.226 19.819 0.209

95% Int 0.452 39.638 0.418

Range 2.271 - 3.175 78.82 - 158.09 0.957 - 1.793

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1

1.5

2

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3

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A B C D E F G H

Serial

INSULIN-LIKE GROWTH FACTOR (IGF-1)

A h igh qual i ty bovine colost rum must conta in at least 2. 25 micrograms of insul in - l ike growth factor -1 per gram of powder to provide maximum benef i ts to the end-user . The colostrum powder produced by Immuno -Dynamics a lways conta ins more than the minimum level of th is impor tant const i tuent . In addi t ion, these ass ays conf i rmed the lot - to- lot consis tency of the concent rat ion of th is component.

Lept in Lept in is a smal l , hormone- l ike prote in that is produced pr imar i ly by adipocytes ( fat ce l ls) and in smal ler amounts in some other per ipheral organs. Smal l adipocytes produce more lept in than large adipocytes, wh ich are predominant in obese individua ls . Lept in operates in the presence of insul in by l ink ing to cel l surface receptors, wh ich are b inding s i tes speci f ic for lept in on the outs ide of the cel l . This in teract ion results in t ransmiss ion of a chemical s ignal to the hypothalamus, causing a suppression of appet i te wi th a s imul taneous st imulat ion of metabol ism. Lept in and lept in receptors are a lso present in the cel ls of the l in ing of the human smal l in test ine where the ef fects on appet i te suppression are local rather than systemic. Ingest ion of smal l amounts of a lcohol appears to temporar i ly impede the funct ioning of the receptors, causing a shor t - term increase in appet i te. Lept in is important in the regulat ion of f at mass and body weight . Adipose t issue ( fat ce l ls) not only secretes lept in, but a lso serves as a s i te of act ion for lept in through the cel l surface receptors. Lept in receptors exis t in at least four forms on the surface of cel ls , a s ing le major isoform and three smal ler isoforms. Recent s tudies indicate that the smal ler lept in receptor isoforms play the most impor tant ro le in body we ight cont ro l. Lept in p lays a s ignif icant ro le in mammal ian physio logy and, thus, may be found in associat ion wi th patho -physio log ical condi t ions. Because of the involvement of lept in in the regulat ion of body weight , the level of lept in in the serum is considered to be one of the best b io log ical markers of body fat in both humans and animals. Obesi ty in humans is

20

accompanied by an increased level of lept in in the c i rculat ion. There are a lso gender d if ferences in the observed levels of lept in in the b lood associated wi th body mass. Lept in levels are corre lated wi th those of a number of endocr ine substances such as insul in , g luc ocor t ico ids, thyro id hormones and testosterone. Recent s tudies a lso indicate that i t may be involved in mediat ing some endocr ine - re lated mechanisms, l ike the onset of puber ty, and impaired or modif ied receptor funct ions may be associated wi th cer ta in medic al condi t ions, such as obesi ty, polycyst ic ovary syndrome, s leep apnea, card iovascular d isease and cer ta in types of cancer and the wast ing associated wi th the d isease. Research f indings also indicate that lept in can act as a growth factor in the fetus and in the neonate. Obesi ty resul ts f rom an imbalance between calor ic energy and energy expendi ture. Genet ic factors p lay an impor tant ro le in i ts pathogenesis . Several s ing le -gene defects responsib le for obesi ty have been ident i f ied. Lept in is t he most notable example of such a gene defect , but a number of other prote ins and neuropept ides have a lso been shown to par t ic ipate in a complex network that regulates food intake and energy expendi ture. In obesi ty, one of the most s ignif icant changes is the shif t f rom smal l adipocytes to large adipocytes wi th a concomitant reduct ion in lept in product ion per cel l . Further, i t has been shown that the lept in receptor populat ion on the large fat ce l ls shi f ts pr imar i ly to the larger isoform and that their funct ioning is impaired when compared to the receptors found on adipocytes in non -obese individuals . The resul t is more lept in backed -up in the c i rculat ion wi th a fa i lure to send the s ignals that would suppress appet i te and increase metabol ism. This is fur ther compl icated by d ietary factors. For example, i t has been shown that exposure of lept in receptors to a var iety of fa t ty ac ids resul ts in s ignif icant impairment of their abi l i ty to b ind lept in.

0

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A B C D E F G H

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LEPTIN

The colostrum powder produced by Immuno-Dynamics evidences a consis tent ly h igh level of lept in in each manufactured ser ial .

21

Insulin Insul in is a smal l pept ide hormone produced in the Is lets of Langerhans of the pancreas. I t regulates the metabol ism of carbohydrates, fats and starches i n the body. The body normal ly conver ts g lucose to g lycogen to der ive the metabol ic energy needed for the maintenance of v i ta l funct ions. This process is in i t ia ted by the b inding of insul in to cel l surface receptors that are very speci f ic for the molecular conf igurat ion of that hormone. This t r iggers a cascade of events that inc ludes the b inding of IGF -1 and lept in to their specif ic receptors. Thus, there is a s t rong metabol ic in ter -dependence between the funct ions of insul in , IGF-1 and lept in that inf luences the metabol ism of carbohydrates and fats and the der ivat ion of the metabol ic energy necessary to support bodi ly funct ions. I f the pancreas produces too l i t t le insul in and/or the interact ion of insul in wi th i ts cel l surface receptors is impaired, the am ount of sugar in the b lood increases, producing a condi t ion known as hyperg lycemia. This condi t ion, in i tse l f , is not le thal , but is a symptom of a ser ious d isease, d iabetes mel l i tus. Diabetes mel l i tus is usual ly c lass i f ied into two types. Type I is cal led insul in-dependent d iabetes mel l i tus ( IDDM) and was previously known as juveni le -onset d iabetes s ince i t occurs pr imar i ly in chi ldren and young adul ts . I t accounts for about 10 -15 percent of a l l cases of d iabetes and can progress very rapid ly. Type I I is cal led non- insul in -dependent d iabetes mel l i tus (NIDDM) and was former ly known as adul t -onset d iabetes s ince i t usual ly occurs in persons over 40 years o ld. This form of d iabetes progresses very s lowly and af fected indiv iduals of ten have no outward s igns of c l in ica l i l lness, but , rather , are detected by the presence of e levated b lood and/or ur ine g lucose levels . In the Type I d iabet ic , the problem is almost a lways associated wi th a severe reduct ion in the amount of insul in produced, whi le, in the Type I I d iabet ic , the pancreas makes suf f ic ient insul in , but the hormone cannot promote the ent ry of g lucose into cel ls . The h igh level of b lood sugar then inact ivates the receptors for insul in that would normal ly be funct ional on the surface of the cel ls . In some in dividua ls , the development of Type I I d iabetes seems to be associated wi th pro longed obesi ty. Both Type I and I I d iabetes have an associated genet ic component through which individuals appear to be predisposed and they a lso have low levels of IGF -1 in thei r c i rcu lat ion. Insul in ord inar i ly is inef fect ive when adminis tered by mouth s ince i t is rapid ly degraded by proteolyt ic enzymes in the stomach. However , at least two factors associated wi th the del ivery of insul in in complete bovine colost rum l ike ly suppo r t i ts absorpt ion into the body in a funct ional form. Firs t , co lost rum conta ins t ryps in inhib i tors and i t has been shown that t ryps in, a proteolyt ic enzyme, is pr imar i ly responsib le for the degradat ion of insul in in the stomach and that the addi t ion of t ryps in inhib i tors to ora l ly adminis tered insul in enhances i ts absorpt ion into the body. In addi t ion, when co lost rum is ingested, the casein and fat are acted upon by another enzyme in the stomach, rennin, forming

22

a cot tage cheese- l ike curd that serves to pro tect the b io log ical ly act ive components f rom the ef fects of s tomach ac id and enzymes, a l lowing them to pass intact in to the smal l in test ine, where they are absorbed.

0

0.3

0.6

0.9

1.2

1.5

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A B C D E F G H

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INSULIN

As wi th the associated components, IGF -1 and lept in, the colost rum powder produced by Immuno-Dynamics evidences a consis tent ly h igh leve l of insul in in each manufactured ser ia l .

Lactoferr in Lactofer r in is a s ing le -chain metal-b inding g lycoprote in wi th a molecular weight of about 78,000 dal tons. I t is present in l arge quant i t ies in both human and bovine co lost rum and, to a lesser extent , in both human and cow’s mi lk , as we l l as in other exocr ine secret ions. A great deal of st ructura l informat ion has been developed about the lactoferr in molecule and i t has been poss ib le to re late th is in format ion to three s ignif icant propert ies of lactofer r in ; a) i ts abi l i ty to b ind a wide var iety of metal ions wi th ext remely h igh af f in i ty; b) i ts abi l i ty to b ind cat ions; and c) i ts abi l i ty to b ind to a number of d i f ferent cel l types . Lactofer r in has the capaci ty to revers ib ly b ind two i ron (Fe) ions concomitant ly wi th two carbonate (CO3=) or b icarbonate (HCO3 -) anions. The features of metal b inding by lactofer r in that are par t icu lar ly s igni f icant inc lude the synerg is t ic re lat ionship between cat ion and anion b inding, the ext remely t ight b inding of i ron, and the exis tence of mechanisms for the re lease of t ight ly bound i ron. Other metals of s imi lar s ize and charge can be subst i tuted for i ron in the two specif ic b inding s i tes. In colost rum, as in other secret ions, lactofer r in is most ly i ron -f ree, wi th a saturat ion level of about 8 -10%. In th is i ron -f ree form, i t has very pronounced bacter iostat ic propert ies, probably dependent upon i ts abi l i ty to b ind advent i t ious i ron very t ight ly, thus depr iv ing cer ta in bacter ia, l ike E. col i , o f i ron that is essent ia l for their g rowth. In addi t ion, sequest rat ion of i ron by lactofer r in rest r ic ts i ron -cata lyzed f ree radical damage to cel ls .

23

The abi l i ty of lactofer r in to b ind to a var iety of normal and le ukemic b lood cel ls has led to the suggest ion that the lactoferr in re leased by neut rophi ls p lays a ro le in modulat ing immune and inf lammatory responses. Lactofer r in promotes the aggregat ion of neut rophi ls and their adhesion to epi thel ia l ce l ls , suggest ing t hat lactofer r in may be the agent that causes neut rophi ls to accumulate at the s i te of in f lammat ion. The lactofer r in molecules der ived f rom di f ferent t issues and secret ions appear to be ident ica l in s t ructure and funct ion, eg the DNA sequence of lactofer r in f rom leukocyte granules matches the amino ac id sequence of the lactofer r in found in colost rum and mi lk . Lactofer r in iso lated f rom var ious mammal ian species has a lso been found to be essent ia l ly ident ica l in both st ructure and funct ion.

Biological ly Act ive Components

Beta-Lactoglobul in , Lactoferr in (m ass /gm of powder )

Beta-

Lactoferrin Lactoglobulin

Serial (mg/gm) (mg/gm)

A 2.927 17.936

B 2.950 19.545

C 3.211 19.555

D 2.840 19.164

E 2.379 21.064

F 2.318 23.545

G 2.740 24.836

H 3.859 22.891

Mean 2.903 21.064

S.D. 0.487 2.441

95% Int 0.974 4.882

Range 1.929 - 3.877 16.182 -25.946

24

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LACTOFERRIN

The colost rum powder produced by Immuno-Dynamics consis tent ly evidences leve ls of lactofer r in above 2.5mg per gram in each manufactured ser ia l .

Beta-Lactoglobulin Beta- lactog lobul in compromises about 10% of mi lk prote in and is the most prevalent prote in in whey, represent ing about 50 -60% of the prote in content. I t has a molecular weight of about 18,300 dal tons and conta ins 162 amino ac ids. The molecular s tructure of beta -lactog lobul in has been extensive ly s tudied and i t has been shown that there are two genet ic var iants, A and B, that d i f fer in the subst i tu t ion of a g lyc ine in Var iant B for an aspart ic ac id in Var iant A. Below pH 3 and above pH 8, beta - lactog lobul in ex is ts as a monomer ic s t ructure, wh i le between pH 3.1 and 5.1 at low temperatures and h igh prote in contents, i t se lf -associates to form an octamer. At other pH va lues, inc luding the pH of mi lk , beta - lactog lobul in tends to be found as a spher ical d imer, which has a solubi l i ty of about 50 mg/ml in mi lk . I t has been shown that the heat t reatment of mi lk can result in the b inding of up to four lactose molecules to each beta - lactog lobul in molecule. Beta- lactog lobul in is manufactured in the mammary g land and is expressed in the mammary g land dur ing pregnancy and lactat ion. Al l co lost rum and mi lk f rom ruminants conta ins beta - lactog lobul in whi le the mi lk f rom most non-ruminants does no t . However , i t is found to a lesser extent in the mi lk f rom horses, dogs, dolphins and whales. The pr imary ro le of beta - lactog lobul in is as a t ransporter of smal l l inear fat ty ac id - l ike hydrophobic molecules, which is a la rge group of substances that b inds molecules such as ret inol (v i tamin A) , t r ig lycer ides and fat ty ac id -synthesis and -breakdown intermediates. The molecular st ructure of beta - lactoglobul in is unique, consis t ing of e ight non-paral le l s t rands of amino ac ids wi th a channel in the center that coi ls around on i tse lf to form a hydrophobic uni t where t ranspor ted molecules are inserted.

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BETA-LACTOGLOBULIN

The colost rum powder produced by Immuno-Dynamics evidences consis tent and substant ia l levels of beta - lactog lobul in as wou ld be character is t ic of a high qual i ty f i rs t mi lk ing bovine colostrum.

Thymosin alpha-1; Thymosin beta-4 The thymus g land, located over the sternum, is an endocr ine organ for wh ich a uni f ied, physio log ical concept of humoral regulat ion of the immune response has emerged in recent years. The thymus g land is the major s i te for maturat ion of immuno-competent T lymphocytes f rom their hematopoet ic s tem cel ls . This very complex process requires d i rect cel l - to-cel l receptor -based interact ions, as wel l as specia l ized chemical s ignals via the numerous cytok ines and thymic hormones produced by cel ls in the thymus. Thymic hormones are polypept ides that have been shown to local ize in the ret icu lo -endothel ia l ce l ls of the thymus. The complex maturat ion sequence of T lymphocytes and the der ivat ion of numerous unique subpopulat ions of these cel ls invo lves the orchestra l in teract ion of var ious thymic -specif ic factors, inc luding the hormone thymosin, and other molecules dur ing the d if ferent iat ion process. Thymosin f ract ion 5 and i ts const i tuent hormone- l ike pept ides, thymosin a lpha-1 and thymosin beta-4, in f luence several propert ies of lymphocytes inc lud ing cyc l ic nucleot ide leve ls , migrat ion inhib i tory factor product ion, T -cel l dependent ant ibody product ion, as wel l as express ion of var ious cel l surface maturat ion and d if ferent iat ion markers. In var ious c l in ica l t r ia ls , these thymic hormones have been shown to s t rengthen the ef fects of immuno -modulators in immuno-def ic ienc ies, auto immune diseases and neoplast ic mal ignancies. Thymosin a lpha-1 is a smal l pept ide wi th a molecular we ight of approximately 5,000 dal tons. I t has been shown to enhance the maturat ion of undif ferent iated stem cel ls in to d isease -f ight t ing T-cel ls and to enhance the product ion of ce l l messengers that coordinat e the complex response of the immune system to d isease. More recent s tudies have a lso shown that thymosin a lpha -1 d i rect ly increases the

26

immune system’s abi l i ty to recognize and target cancerous and vi rus -in fected cel ls among heal thy cel ls . Thymosin beta-4 is par t of a fami ly of beta thymosin pept ides each having a molecular we ight of about 5,000 dal tons. The beta thymosins are present in varying concent rat ions in a lmost every cel l in the body. They b ind monomer ic act in in a 1:1 complex and act as buf fers , prevent ing polymer izat ion into act in f i laments, but supplying a pool of act in monomers when a cel l needs f i laments for d i f ferent iat ion. Thymosin beta-4 is a lso present outs ide of cel ls in b lood p lasma and in wound f lu id. I t has been shown to in f luence t he induct ion of cer ta in enzymes, the migrat ion of whi te b lood cel ls through t issues and the rate of ce l l d i f ferent iat ion. Thymosin beta -4 has a lso been shown to inhib i t in f lammat ion and to l imi t stem cel l d i f ferent iat ion f rom bone marrow.

Biological ly Act ive Components

Thymosin a lpha-1 , Thymosin beta -4 (m ass /gm of powder )

Thymosin Thymosin

alpha-1 beta-4

Serial (ug/gm) (ug/gm)

A 335.45 611.82

B 332.82 617.82

C 329.64 652.73

D 333.55 742.27

E 344.00 591.91

F 338.27 585.45

G 310.73 681.64

H 353.64 673.64

Mean 334.73 644.636

S.D. 12.326 53.301

95% Int 24.652 106.602

Range 310.075 - 359.379 538.034 - 751.238

27

120

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THYMOSIN ALPHA-1

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THYMOSIN BETA-4

The colostrum powder manufactured by Immuno-Dynamics consis tent ly conta ins h igh levels of both thymosin a lpha -1 and thymosin beta -4.

Immunoglobul ins The format ion of co lost rum starts 3 -4 weeks before the b i r th of the cal f , when hormonal changes t r igger the act ivat ion of s i tes wi th in the mother ’s mammary g lands. W hen fu l ly act ivated, these s i tes beg in to t ransfer b io log ical ly -act ive substances into the mammary g land where colost rum is formed. I t is through th is mechanism alone that ant ibodies (specif ica l ly immunoglobul ins o f the IgG c lass), f rom the mother 's b lood enter in to the mammary g land. Specia l ized whi te b lood cel ls f rom the mother are also t ransfer red in the same way. These cel ls immediate ly beg in to re lease other immunoglobul in molecules ( IgM and IgA) into the developing colost rum. Ant ibodies have some specia l ized funct ions, part ly due to the “Y” shape of these long -chain prote ins. The arms of the Y can at tach to an

28

ant igen and, once th is happens, the tai l o f the Y unfo lds. This ta i l reg ion d if fers among the f ive main c lasses of immunoglobul ins ( IgM, IgG, IgA, IgD and IgE). For example, IgM molecules have very shor t ta i l p ieces which are used to connect f ive Y molecules together creat ing a b ig molecule wi th ten possib le s i tes where i t can at tach i tse lf to a mic roorganism. Al l of the other c lasses represent molecules that are about one-th i rd as b ig as IgM, but are much more ef fect ive. IgM and IgG c lasses of ant ibodies funct ion best when they are in the b loodst ream. In cont rast , immunoglobul ins of the IgA c las s can operate in the b loodst ream or in other bodi ly f lu ids l ike the tears and sal iva. IgA molecules can a lso be found in the lungs and intest ines. Here, the ta i ls at tach to mucous membranes so the arms of the molecule can “catch” and immobi l ize invading o rganisms. Since most in fect ious agents enter the body through the respiratory or d igest ive pathways, IgA is very impor tant in res is t ing many infect ions. The ant ibodies in ora l ly ingested bovine colost rum do not enter the b loodst ream, except through a s pecia l ized mechanism in newborn calves. However , these ant ibodies have been shown to be ef fect ive in enter ic infect ions in var ious species. This is because dairy cows are repeatedly exposed to most of the same bacter ia and vi ruses that cause enter ic infect ions in other mammals, inc luding humans, (Escher ichia col i , Staphylococcus species, Salmonella species, rotavi rus, etc. ) . Thus, in addi t ion to assur ing that the colost rum used is only f rom a f i rs t mi lk ing taken dur ing the f i rst s ix hours af ter b i r th, i t is important to ut i l i ze a colost rum product wi th a h igh degree of "ant ibody d ivers i ty. " This means that i t has been col lected f rom many cows on a lo t of d i f ferent dairy farms and potent ia l ly prov ides protect ion against a broad number of microorganisms.

Biological ly Act ive Components

Immunoglobul in G ( IgG) (m ass /gm of powder )

Total

IgG1 IgG2 IgG

Serial (mg/gm) (mg/gm) (mg/gm)

A 232.40 8.79 241.19

B 239.20 10.81 250.01

C 248.40 10.92 260.32

D 248.40 10.92 260.32

E 230.00 10.35 240.35

F 220.80 10.35 231.15

G 230.00 10.47 240.47

H 271.40 10.81 282.21

Mean 240.075 10.428 250.753

S.D. 15.830 0.704 16.313

95% Int 31.660 1.408 32.626

Range 208.415 - 271.735 9.02 - 11.836 218.127 - 283.379

29

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TOTAL IgG

The colostrum powder produced by Immuno-Dynamics evidences a uni form, consis tent ly h igh leve l of immunoglobul in G ( IgG) in each ser ia l manufactured.

30

CHARACTERIZATION OF COLOSTRUM PRODUCTS

COMPETITORS

Basic Chemical Composit ion As d iscussed ear l ier , the best way to determine the basic qual i ty of a co lost rum product is to establ ish the re lat ionship of three major components, prote in, fa t and lactose, a re lat io nship that changes wi th t ime af ter b i r th of the cal f . The basic chemical composi t ion e ight (8) colost rum powder products f rom dif ferent manufacturers is compared wi th the mean values for ten (10) ser ia ls manufactured f rom large pools by Immuno-Dynamics in the fo l lowing table. The same data is a lso graphical ly d isplayed. Compar ison of these data wi th the values g iven in the table f rom the Handbook of Dairy Chemist ry shown ear l ier and again below indicates a) whether the products employ colost rum col lected wi th in s ix (6) hours af ter b i r th of the calf in a s ing le mi lk ing; and whether the colost rum has been al tered, potent ia l ly reducing i ts ef fect iveness.

Composi t ion of Bov ine Colost rum (m g/gm of to ta l so l ids )

Hours

A f te r To ta l B i r th Pro te in Fa t Lac tose

0 651.0 189.0 811.0 6 489.0 334.8 132.5 12 416.4 261.5 255.3 24 354.0 266.2 311.7

Since the data shown ear l ier substant iated that the colostrum powder products manufactured by Immuno -Dynamics meet the cr i ter ia for a h igh qual i ty f i rs t milk ing colostrum col lected wi th in s ix (6) hours af ter b i r th, the mean values for these products wi l l be used as a benchmark for compar ison to establ ish the re lat ive qual i ty of the tested compet i tor products. In a l l cases, a le t ter designat ion for an individual product represents the same product in a l l tables and graphs.

31

CHEM ICAL COM POSIT ION

(m g/gm of powder )

Total Total Total

Product Protein Fat Lactose

I-D 549.7 224.5 111.3

A 295.8 53.9 74.0

B 518.6 131.1 144.0

C 545.1 133.0 322.0

D 413.0 19.2 48.0

E 557.1 170.9 106.0

F 508.4 140.9 134.0

G 740.0 12.4 97.0

H 547.5 53.5 100.0

I 730.1 11.8 108.0

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I-D A B C

Product

CHEMICAL COMPOSITION

Protein

Fat

Lactose

Compar ison of the composi t ion of Product A wi th that of the Immuno -Dynamics ’ ( I -D) products indicates that i t is def ic ient in prote in content wi th the value being indicat ive of co lost rum col lected more than 24 hours af ter b i r th. The very low fat content of th i s product indicates that i t have been adul terated by part ia l removal of the fat dur ing processing. Since lactose is produced by cel ls in the mammary g land and r ises wi th t ime af ter b i r th, the exceeding ly low level of lactose in Product A is indicat ive of f ur ther adul terat ion wi th substant ia l removal of the lactose. The leve l of fat found in Product B is below that expected for colostrum col lected 6 hours af ter b i r th, ind icat ing that some of i t has been removed and the lactose level is h igher than normal ind icat ing that the colost rum used was co l lected more than 6 hours af ter b i r th. The fat content of Product C indicates that some of th is mater ia l has been removed, which ar t i f ic ia l ly causes an increase in

32

observed prote in and lactose values s ince the determin at ions are based upon tota l so l ids. Adjusted values for th is adulterated product indicate that the colost rum used was col lected between 6 -12 hours af ter b i r th.

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I-D D E F

Product

CHEMICAL COMPOSITION

Protein

Fat

Lactose

The composi t ional analys is for Product D a lso indicates that t he fat and a major port ion of the lactose were removed dur ing processing. Adjustment of the prote in content for th is adul terat ion indicates that the colostrum used to produce th is product was col lected more than 24 hours af ter b i r th. The basic chemical co mposi t ion of Products E and F is s imi lar to that of Immuno-Dynamics ’ co lost rum powder a l though the fa t content is somewhat lower , a character is t ic that can occur in colost rum col lected wi th in 6 hours af ter b i r th.

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I-D G H I

Product

CHEMICAL COMPOSITION

Protein

Fat

Lactose

Products G, H and I have been adul terated by removal of the fat wi th that component being a lmost complete ly removed f rom Products G and I . This would ar t i f ic ia l ly in f la te both the prote in and lactose levels and,

33

since the lactose leve l in these products is lower tha n would be expected, i t ind icates that some of that component has been removed f rom each of the products. The adjusted prote in values for Products G and I ind icate that they employed colost rum col lected more than 24 hours af ter b i r th, wh i le the adjusted pr ote in value for Product H indicates the use of colostrum col lected up to 24 hours af ter b i r th.

Biological ly Act ive Components As for the test ing conducted wi th the ser ia ls produced by Immuno -Dynamics, a ser ies of e ight (8) of the b io log ical ly act ive comp onents of bovine colost rum were selected as the marker components based upon a) the avai labi l i ty of an immunoassay for the determinat ion of concent rat ion; b) the abi l i t y of the assay or an adaptat ion thereof to perform re l iably wi th colost rum; an c) the importance of the role of the component in the biological effects observed with colostrum . The assays were conducted by the d iagnost ic laboratory at a major col lege of veter inary medic ine. In th is case, colost rum powder products f rom nine (9) d i f ferent manufacturers were evaluated. Again, in a l l cases, a le t ter designat ion for an individual product represents the same product in a l l tables and graphs.

BIOLOGICALLY ACTIVE COM PONENTS

IGF-1 , Lept in , Insul in

IGF-1 Leptin Insulin

Product (ug/gm) (ng/gm) (mIU/gm)

I-D 2.723 118.45 1.375

A 0.827 47.72 0.931

B 2.455 96.21 1.239

C 1.357 200.41 4.713

D 2.688 54.91 0.060

E 1.147 61.39 0.513

F 2.046 85.89 0.915

G 1.004 54.25 0.397

H 2.943 139.62 1.412

I 1.461 120.75 1.991

34

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I-D A B C D E F G H I

Product

INSULIN-LIKE GROWTH FACTOR

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LEPTIN

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INSULIN

35

Compar ison of the IGF-1, lept in and insul in content of Product A wi th that of the products f rom Immuno -Dynamics indicates that th is product is s igni f icant ly def ic ient in those components, which is not surpr is ing s ince the composit ional data indicated that the colost rum used to manufacture the tested product was a) col lected more than 24 hours af ter b i r th; and b) had been adul terated by removal of both fat and lactose. Product B had normal levels of tota l prote in that would ord inar i ly be found in a h igh qual i ty f i rs t mi lk ing bovine colost rum col lected wi th in 6 hours af ter b i r th, but had a reduced fat content and e levated lactose. However , i ts IGF-1, lept in and insul in contents were consis tent wi th a colost rum col lected wi th in 6 hours af ter b i r th. The composi t ional analys is ind icated that a por t ion of the fat had been removed f rom Product C, ar t i f ic ia l ly inf la t ing the prote in value. In addi t ion, the data showed that the colost rum used to produce the powder had been co l lected between 6 -12 hours af ter b i r th. According ly, the product evidences a reduced content of IGF -1, which is par t ia l ly fat so luble, and substant ia l ly increased lept in and insul in v alues, a phenomenon associated wi th the art i f ic ia l increase in prote in content . Product D was a lso shown to have a markedly reduced fat content in the composit ional analys is and the adjusted values indica ted that the colost rum used was col lected more than 24 hours af ter b i r th. The product evidences a markedly reduced lept in level and i t was essent ia l ly devoid of any insul in . In contrast , the product had essent ia l ly normal IGF-1 levels in compar ison to the powder produced by Immuno-Dynamics. Insul in is a lmost complete ly removed by the format ion of a curd in the product ion of whey, leaving the res idual IGF -1 and lept in essent ia l ly in tact , but ar t i f ic ia l ly in f la t ing their content by the removal of other major components. Thus, the assayed values der ived wi th Product D are character ist ic of dr ied whey. Al though Product E was thought to have a chemical composi t ion essent ia l ly l ike that seen wi th the powder products manufactured by Immuno-Dynamics, i ts IGF-1, lept in and insul in content are considerably lower . As wi th Product B, th is d iscrepancy may be the resul t of us ing h ighly d iverse col lect ion schedules and, in some cases, pools f rom mult ip le mi lk ings in assembl ing the large pool of “co lost rum” used to manufacture the powder tested. Product F a lso evidenced a chemical composi t ion s imi lar to that seen wi th the powder products manufactured by Immuno -Dynamics. This product demonst rated considerably h igher levels of IGF -1, lept in and insul in than Product E, but s t i l l somewhat lower than those found wi th Immuno-Dynamics ’ products. These lower va lues indicate that the colost rum pool used to produce the powder may have ut i l i zed some mater ia ls col lected af ter the 6 -hour per iod and combined them with h igher qual i ty mater ia l . The composit ional analys is for Product G i ndicated that i t was adul terated by removal of fat and lactose and that i t employed

36

colost rum col lected more than 24 hours af ter b i r th. These observat ions were ver i f ied by the very low leve ls of IGF -1, lept in and insul in found in the tested powder . The composi t ional analys is for Product H indicated that a lmost a l l o f the fat and most of the lactose had been removed resul t ing in an ar t i f ic ia l ly e levated prote in content, which a lso resul ted in e levated leve ls of IGF-1, lept in and insul in in the assayed powde r . In cont rast to the values obta ined wi th Product C, wh ich was apparent ly adul terated in a s imi lar fashion, the content of a l l three components was uni formly e levated. Therefore, i t would appear that a method was used for fat and lactose removal that d id not a l ter the IGF-1 content of the colost rum. Product I evidences a reduced IGF -1 content, a normal lept in concent rat ion and a somewhat e levated amount of insul in . The resul ts of these assays are character ist ic of a product that employed colost rum col lected up to 24 hours af ter b i r th, perhaps f rom mult ip le mi lk ings, and adul terated by removal of a l l or some of the fat us ing a method that removed the fat -soluble por t ion of the IGF-1. This would resul t in an ar t i f ic ia l increase in the va lues for the remain ing components.

B IOLOGICALLY ACTIVE COM PONENTS

Lactoferr in , Beta -Lactoglobul in

Beta-

Lactoferrin Lactoglobulin

Product (mg/gm) mg/ml

I-D 2.903 21.064

A 0.646 17.680

B 2.689 26.529

C 1.053 25.454

D 3.069 24.233

E 1.268 36.200

F 2.837 14.295

G 1.471 23.136

H 1.517 18.724

I 1.955 32.711

37

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LACTOFERRIN

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BETA-LACTOGLOBULIN

Compar ison of the lactofer r in and beta - lactog lobul in content of Product A wi th that of the products f rom Immuno -Dynamics indicates that th is product conta ins very l i t t le lactoferr in and a substant ia l ly reduced quant i ty of beta- lactog lobul in . The propor t ional ly h igher quant i ty of beta- lactog lobul in is l ike ly due to the fact that the colostrum used was col lected more than 24 hours af ter b ir th and bovine mi lk conta ins a substant ia l quant i ty of th is component. Product B evidenced essent ia l ly the same levels of lactofer r in and beta- lactog lobul in observed wi th the powder produced by Immuno -Dynamics. However , approximately hal f o f the fat has been removed f rom the colost rums, which would ar t i f ic ia l ly in f la te these values. Product C assay va lues indicated a reduced lactoferr in concent rat ion wi th a normal quant i ty of beta - lactog lobul in . The reduced lactofer r in concent rat ion is cons is tent wi th use of colost rum col lected 6 -12 hours af ter b i r th s ince lactofer r in is t ransferred f rom the mother ’ c i rcu lat ion

38

dur ing late gestat ion and is not produced by cel ls in the mammary g land. In cont rast , beta - lactog lobul in is only produced by cel ls i n the mammary g land and i ts content increases wi th t ime af ter b i r th, which could resul t in a normal -appear ing value wi th la te col lect ion mater ia ls . Ear l ier data indicated that Product D is dr ied whey wi th normal IGF -1 leve ls , but low lept in content and essent ia l ly no insul in . The normal lactoferr in and beta- lactog lobul in levels , in compar ison to the Immuno -Dynamics ’ products, are a lso consis tent wi th a dr ied whey -based product. Consis tent wi th observed low levels of IGF -1, lept in and insul in in Product E, low levels of lactofer r in were a lso seen. However , the product evidenced an ext remely h igh content of beta - lactog lobul in . These character ist ics are a l l ind icat ive of a product that employs colost rum col lected at least 24 hours af ter b i r th f rom mult ip le mi lk ings. In th is case, the pr imary b io log ical ly act ive components wou ld be s ignif icant ly d i lu ted by t ransi t ional mi lk wi th a concomitant increase in beta- lactog lobul in , wh ich is an inherent component of bovine mi lk . Product F evidenced a normal level of la ctofer r in and a somewhat reduced concentrat ion of beta - lactog lobul in in compar ison to the values found wi th the powder produced by Immuno -Dynamics. The observed values are consis tent wi th a product that employs colost rum pools that employ d iverse mater ia ls , much of which was col lected dur ing the f i rst 6 hours af ter b i r th, as was concluded for the IGF -1, lept in and insul in determinat ions. The reduced level of lactofer r in and seeming ly normal level of beta -lactog lobul in observed wi th Product G are consis tent wi th a product that used colost rum col lected more than 24 hours af ter b i r th and was adul terated by removal of the fat and lactose. This fur ther ver i f ies the same conclus ion drawn f rom the composi t ional analys is and the resul ts of assays for IGF-1, lept in and insul in content. Al though Product H evidenced e levated IGF -1, lept in and insul in leve ls , despi te having the fat removed, i ts lactofer r in level was below that seen wi th the Immuno-Dynamics ’ products. In addi t ion, the beta -lactog lobul in content was s l ig ht ly lower than normal . The combined character is t ics for th is adul terated product are indicat ive of one that has been select ive ly manipulated. Removal of the fat should have resul ted in ar t i f ic ia l e levat ion of the content for a l l measured prote ins, but th is occurred only for some selected components. Product I evidences a reduced quant i ty of lactofer r in wi th an increased leve l of beta- lactofer r in . This is consis tent wi th the use of colost rum col lected at least 24 hours af ter b i r th and fur ther supports the same conclus ion der ived wi th the IGF -1, lept in and insul in values.

39

BIOLOGICALLY ACTIVE COM PONENTS

Thymosin a lpha-1 , Thymosin beta -4

Thymosin Thymosin

alpha-1 beta-4

Product (ug/gm) (ug/gm)

I-D 334.727 644.636

A 254.795 295.000

B 326.491 565.909

C 214.382 586.818

D 138.750 BDL

E 223.486 586.364

F 274.218 569.545

G 272.959 678.182

H 262.641 566.818

I 339.836 622.273

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THYMOSIN ALPHA-1

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THYMOSIN BETA-4

Compar ison of the thymosin a lpha-1 and thymosin beta -4 content of Product A wi th the powder products f rom Immuno -Dynamics indicates that th is product conta ins a s ignif icant ly lesser amount of thymosin a lpha-1 and a markedly lower quant i ty of thymosin beta -4. As wi th the previous f indings, these values are consis tent wi th the use of colost rum that was col lected more than 24 hours af ter b ir th. Product B evidenced thymosin a lpha -1 and thymosin beta -4 levels consis tent wi th the range of values observed wi th the powder produced by Immuno-Dynamics, which is the same conclus ion reached for the values obta ined wi th each of the other tested const i tuents. The combined f indings are indicat ive of the use of colost rum col lected at the f i rs t mi lk ing wi th in 6 hours af ter b i r th. The composi t iona l analys is ind icated that a por t ion of the fat had been removed f rom Product C, ar t i f ic ia l ly inf la t ing the prote in value. In addi t ion, the data showed that the colost rum used to produce the powder had been co l lected between 6 -12 hours af ter b i r th. Accordi ng ly, the product evidences a reduced value for thymosin a lpha -1 and a normal level of thymosin beta -4. However , these values are l ike ly ar t i f ic ia l ly in f la ted, due to removal of the fat , as was observed for lept in and insul in . Product D evidences a very l ow level of thymosin a lpha -1 and no detectable thymosin beta -4. The reduced levels are consis tent wi th the use of colostrum col lected more than 24 hours af ter b i r th. However , previous data indicates that the product is dr ied whey and i t is not known what e f fect the generat ion of whey f rom colost rum would have on the concent rat ion of these const i tuents a l though the resul ts imply that most of the thymosin beta -4 is removed. Again consistent wi th observed lower leve ls of IGF -1, lept in, insul in and lactofer r in , Product E exhib i ted low leve ls of thymosin a lpha -1. However , as wi th beta - lactog lobin, the product a lso exhib i ted thymosin beta-4 levels wi th in the normal range. Al though i t is uncer ta in as to why the thymosin beta -4 value wou ld appear normal , the rest o f the

41

character is t ics are consis tent wi th a product that employs colost rum col lected at least 24 hours af ter b i r th. Product F evidenced both thymosin a lpha -1 and thymosin beta -4 values that were just outside of the range of va lues seen wi th the powder products produced by Immuno-Dynamics. These f indings are consis tent wi th previous observat ions for the other const i tuents and are a lso indicat ive of the use of colost rum pools that employ d iverse mater ia ls, much of which was col lected dur ing the f i rst 6 hou rs af ter bi r th. Product G evidenced thymosin a lpha -1 at a level just below the normal range and a normal level of thymosin beta -4. Data for the other const i tuents was consis tent wi th the use of colost rum obta ined more than 24 hours af ter b i r th and the composi t ional analys is demonstrated that the product was adul terated by removal of fa t and lactose. The seeming ly normal levels of thymosin a lpha -1 and thymosin beta -4 may be a manifestat ion of fat removal and the associated ar t i f ic ia l increase in prote in content . The thymosin a lpha-1 and thymosin beta -4 values for Product H were just below the range of values seen wi th the powder products produced by Immuno-Dynamics. This product evidenced a unique prof i le for the b io log ical ly act ive components, e levated IG F-1, lept in, insul in levels wi th lower than normal values for lactoferr in , beta - lactog lobul in , thymosin a lpha-1 and thymosin beta -4. In addi t ion, the composi t ional analys is showed that the fat had been substant ia l ly removed. This product may have been uniq uely manipulated dur ing processing and/or i t used colost rum col lected more than 24 hours af ter b i r th, which or ig inal ly had a low content of act ive components that were substant ia l ly e levated by removal of a lmost a l l of the fat . Product I conta ined essent i a l ly normal leve ls of thymosin a lpha -1 and thymosin beta-4. Resul ts f rom the assays for other const i tuents evidenced reduced concentrat ions of some components and were consis tent wi th the use of colostrum col lected up to 24 hours af ter b i r th, which was not the case for e i ther of the thymosins. However , the resul ts indicated a low leve l of IGF -1 in th is product . Since IGF -1 is par t ia l ly fat so luble and i ts concent rat ion is usual ly lowered when the fat is removed f rom colost rum, i t is possib le that the fat wa s tota l ly or par t ia l ly removed f rom the colost rum used to manufacture the powder . This would ar t i f ic ia l ly e levate the levels of the remain ing prote in const i tuents and could expla in the resul ts observed here. Unfor tunate ly, the chemical composi t ion of th is product was not determined.

42

BIOLOGICALLY ACTIVE COM PONENTS

Immunoglobul in G ( IgG)

Total

IgG1 IgG2 IgG

Product (mg/gm) (mg/gm) (mg/gm)

I-D 240.075 10.428 250.753

A 92.00 4.72 96.72

B 165.60 7.71 173.31

C 87.40 3.45 90.85

D 271.40 12.40 283.80

E 119.60 5.17 124.77

F 188.00 9.20 197.20

G 105.8 5.06 110.86

H 115.00 4.37 119.37

I 131.10 4.60 135.70

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Total IgG

Compar ison of the Immunoglobul in G ( IgG) content of Product A wi th that of the products f rom Immuno -Dynamics indicates that th is product is s ignif icant ly def ic ient in th is component , which is not surpr is ing s ince the composit ional data indicated that the colost rum used to manufacture the tested product a) was col lected more than 24 hours af ter b i r th and the t ransfer of IgG into colost rum ceases at b i r th; and b) had been adul terated by removal of both fat and lactose. Product B had a level of tota l prote in that would normal ly be found in a h igh qual i ty f i rst mi lk ing bovine colost rum col lected wi th in 6 hours af ter b i r th, but some of the fat had been removed, which wou ld ar t i f ic ia l ly in f la te the other values. Despi te th is ef fect , i ts IgG content was lower

43

than that observed wi th Immuno -Dynamics ’ products, ind icat ing that the colost rums was col lect beyond the desirable 6 -hour per iod. The composi t ional analys is ind icated that a por t ion of the fat had been removed f rom Product C, ar t i f ic ia l ly inf la t ing the prote in value. In addi t ion, the data showed that the colost rum used to produce the powder had been co l lected between 6 -12 hours af ter b i r th. According ly, the product evidences a substant ia l ly reduced content of IgG. Product D was a lso shown to have a markedly reduced fat content in the composit ional analys is and the adjusted values indica ted that the colost rum used was col lected more than 24 hours af ter b i r th. The assayed va lues der ived wi th Product D for the var ious b io log ical ly -act ive components are character is t ic of dr ied whey and the product evidences an ar t i f ic ia l ly in f la ted amount of IgG. Al though Product E was thought to have a chemical composi t ion essent ia l ly l ike that seen wi th the powder products manufactured by Immuno-Dynamics, i ts IgG content is considerably lower , as were the values for IGF-1, lept in and insul in . These d iscrepancies may be the resul t of manipulat ion dur ing the manufactur ing process or i t may be due to the use of colost rum wi th h ighly d iverse col lect ion schedules and, in some cases, pools f rom mult ip le mi lk ings in assembl in g the large pool of “co lostrum” used to manufacture the powder tested. Product F a lso evidenced a chemical composi t ion s imi lar to that seen wi th the powder products manufactured by Immuno -Dynamics. This product demonstrated a considerably h igher level of IgG than Product E, but st i l l somewhat lower than that found wi th Immuno -Dynamics ’ products. The lower value indicates that the colost rum pool used to produce the powder may have ut i l ized some mater ia ls col lected af ter the 6-hour per iod and combined them with h igher qual i ty mater ia l . The composit ional analys is for Product G indicated that i t was adul terated by removal of fat and lactose and that i t employed colost rum col lected more than 24 hours af ter b i r th. These observat ions were ver i f ied by the lower l eve l of IgG found in the tested powder . The composi t ional analys is for Product H indicated that a lmost a l l o f the fat and most of the lactose had been removed resul t ing in an ar t i f ic ia l ly e levated prote in content, which a lso resul ted in e levated leve ls of IGF-1, lept in and insul in in the assayed powder . However , th is d id not ar t i f ic ia l ly increase the low leve l of IgG in the product ind icat ing that the powder was produced f rom “colost rum” obta ined in mult ip le mi lk ings taken up to 48 hours af ter b i r th. Product I a lso evidences a reduced IGF-1 content . The chemical composi t ion of th is product indicated that the i t was adul terated by removal of most of the fat . The resul ts of the other assays are character is t ic of a product that employed colost rum col lected up to 24 hours af ter b i r th, perhaps f rom mult ip le mi lk ings.

44

Summary of Observat ions The fo l lowing table provides a d i rect compar ison of a l l compet i tor products tested wi th the mean values for the chemical composi t ion and b io log ical ly act ive components observed wi th ten (10) ser ia ls of powder manufactured by Immuno-Dynamics. In a l l cases, the mean values for the Immuno-Dynamics ’ products are used as the basel ine and, for comparat ive purposes, the values for each product are expressed as a percent (%) of the mean observed value for the Immuno -Dynamics ’ products. The most s ignif icant observat ion for each product is a lso indicated in summary form.

45

COMPARIS ON OF COM PETITOR PRODUCTS ( a s a % o f I m m un o - D yn a m i c s ’ p ro d u c t v a l u e s )

Total Beta- Thymosin Thymosin

Product Protein Fat Lactose IGF-1 Leptin Insulin Lactoferrin Lactglobulin alpha-1 beta-4 IgG Summary of Observations

A 53.81 24.01 66.49 30.37 40.29 67.71 22.25 83.93 76.12 45.76 38.57 Colostrum collected 24+ hrs. after birth, fat removed

B 94.34 58.40 129.38 90.16 81.22 90.11 92.63 125.94 97.54 87.79 69.12 Colostrum collected late, some fat removed

C 99.16 59.24 289.57 49.83 169.19 342.76 36.27 120.84 64.05 91.03 36.23 Colostrum collected late; some fat removed

D 75.13 8.55 43.13 98.71 46.36 4.36 105.72 115.04 41.45 BDL 113.18 Colostrum collected late; fat and lactose removed

E 101.35 76.12 95.24 42.12 51.83 37.31 43.68 343.71 66.77 90.96 49.76 Colostrum collected late; some fat removed

F 92.49 62.76 120.40 75.14 72.51 66.55 97.73 67.86 81.92 88.35 78.64 Colostrum collected 6-8 hrs. after birth, some fat removed

G 134.62 5.52 87.15 36.87 45.80 28.87 50.67 109.84 81.55 105.20 54.12 Colostrum collected late; fat removed

H 99.60 23.83 89.85 108.08 117.87 102.69 52.26 88.89 78.46 87.93 44.21 Colostrum collected late; most of fat removed

I 132.82 5.26 97.04 53.65 101.94 144.80 67.34 155.29 101.53 96.53 47.61 Colostrum collected late; most of fat removed

BDL = Below Detectable Limits of the Assay