Bovine Colostral Whey Research Articles

Production and properties of health-promoting proteins and peptides from bovine colostrum and milk.

The high nutritive value and diverse functional properties of milk proteins are well known. Beyond these qualities, milk proteins have attracted growing scientific and commercial interest as a source of biologically active molecules. Such proteins are found in abundance in colostrum which is the initial milk secreted by mammalian species during late pregnancy and the first few days after birth of the offspring. The best characterized colostrum-based bioactive proteins include alpha-lactalbumin, beta-lactoglobulin, immunoglobulins, lactoferrin, lactoperoxidase and growth factors. All of them can nowadays be enriched and purified on an industrial scale from bovine colostral whey or cheese whey. These native proteins exhibit a wide range of biological activities that are known to affect the digestive function, metabolic responses to absorbed nutrients, growth and development of organs and disease resistance. Also, some of these proteins may prove beneficial in reduction of the risks of chronic human diseases reflected by the metabolic syndrome. It is speculated that such potentially beneficial effects are partially attributed to bioactive peptides derived from intact proteins. These peptides can be liberated during gastrointestinal digestion or fermentation of milk by starter cultures. The efficacy of a few peptides has been established in animal and human studies and the number of commercial products supplemented with specific milk peptides is envisaged to increase on global markets. Bovine colostrum appears as a highly potential source of biologically active native proteins and peptide fractions for inclusion as health-promoting ingredients in various food applications.

임실지역 젖소 초유로부터 분리한 TGF-β 함유 유청 단백질의 면역활성

분만 후 3일 이내에 분비되는 임실 지역의 젖소 초유에서 유청 단백질을 효율적으로 분리하고 RAW 264.7 세포의 증식 및 면역활성에 미치는 영향을 조사하였다. 실험에 사용한 초유의 유청 단백질 g당 TGF-<TEX>${\beta}1$</TEX>은 875 pg/mL,TGF-<TEX>${\beta}2$</TEX>는 6,600 pg/mL이었으며 실험에 사용한 초유 유청 단백질 내 총 TGF-<TEX>${\beta}$</TEX>의 양은 7,475 pg/mL이었다. RAW264.7 세포에 대한 초유 유청 단백질 첨가에 따른 세포증식 정도를 알아본 결과 10 mg/mL의 농도까지는 세포성장을 유도하였으나 20 mg/mL 이상의 농도에서는 세포성장을 억제하였다. 이에 RAW 264.7 세포에서의 초유 유청 단백질의 면역관련 실험에 사용할 농도는 최대 10 mg/mL까지로 결정하였다. RAW 264.7 세포에 초유 유청 단백질(0-10 mg/mL)과 LPS(1 <TEX>${\mu}g/mL$</TEX>)를 차례로 반응시키고 NO생성량을 측정한 결과 초유 유청 단백질이 농도 의존적으로 NO의 생성을 억제하였다. 또한 LPS 자극에 의한 염증성 사이토카인(TNF-<TEX>${\alpha}$</TEX>, IL-<TEX>$1{\beta}$</TEX>, IL-6)이 생성되는 과정에서 초유 유청 단백질의 효과를 확인하였다. 그 결과, LPS를 단독으로 첨가했을 때 TNF-<TEX>${\alpha}$</TEX>, IL-<TEX>$1{\beta}$</TEX>, IL-6의 농도가 모두 증가하였으나 초유 유청 단백질을 첨가한 경우에는 염증성 사이토카인의 생성이 농도 의존적으로 감소하는 경향을 보였다. 초유 유청 단백질에 의해 NO를 비롯한 염증성 사이토카인의 생성이 억제되는 것이 heme oxygenase-1의 발현과 관련하는지 알아보고자 초유 유청 단백질을 농도별(0-10 mg/mL)로 처리하여 heme oxygenase-1의 발현여부를 측정한 결과 대조군과 비교하여 3-4배 이상의 발현 증가를 보였다. 이상의 결과들로 미루어보아 LPS로 자극된 RAW 264.7 세포에서 TGF-<TEX>${\beta}$</TEX> 등을 함유한 초유 유청 단백질은 10 mg/mL 이하의 농도에서 농도 의존적으로heme oxygenase-1의 발현을 유도하여 염증성 사이토카인인 TNF-<TEX>${\alpha}$</TEX>, IL-<TEX>$1{\beta}$</TEX>, IL-6 및 NO의 생성을 억제하는 것으로 판단되었다. This experiment was carried out in order to separate bovine colostral whey protein from Imsil province and to test the effect of immunological activity on RAW 264.7 cells. The colostral whey protein contained TGF-<TEX>${\beta}$</TEX> 7, 475 pg/g in total. We first tested the effect of the colostral whey protein on the proliferation of RAW 264.7 cells and it demonstrated cytotoxicity at concentrations greater than 20 mg/mL. Therefore, the immunological activities of colostral whey protein were investigated in maximum concentration of 10 mg/mL on LPS-induced RAW 264.7 cells. Results indicated that colostral whey protein inhibited the LPS-induced nitric oxide (NO) production in a dose-dependent manner. The colostral whey protein also suppressed the productions of proinflammatory cytokines (TNF-<TEX>${\alpha}$</TEX>, IL-<TEX>$1{\beta}$</TEX>, IL-6) in a dose-dependent manner. In addition to the immunological activity, colostral whey protein led to the expression of heme oxygenase-1 (HO-1) in RAW 264.7 cells. In conclusion, colostral whey protein containing TGF-<TEX>${\beta}$</TEX> inhibited the production of NO, TNF-<TEX>${\alpha}$</TEX>, IL-<TEX>$1{\beta}$</TEX>, and IL-6 via expression of HO-1.

Oral administration of insulin-like growth factor-I from colostral whey reduces blood glucose in streptozotocin-induced diabetic mice

The aim of the present study was to investigate the effects of oral administration of the insulin-like growth factor-I-rich fraction (IGF-I-RF) from bovine colostral whey on the regulation of blood glucose levels in streptozotocin (STZ)-induced diabetic mice. We obtained a peptide fraction containing IGF-I (10 ng/mg protein) from Holstein colostrum within 24 h after parturition by using ultrafiltration. The blood glucose levels of STZ-induced diabetic mice fed with IGF-I-RF (50 μg/kg per d) were significantly reduced by 11 and 33 % at weeks 2 and 4, respectively (P < 0·05). The body weights of STZ-induced diabetic mice increased following the oral administration of the IGF-I-RF. The kidney weights of STZ-induced diabetic mice decreased significantly (P < 0·05) following the administration of the IGF-I-RF, and the liver weights of STZ-induced diabetic mice decreased significantly (P < 0·05) following the administration of 50 μg/kg per d of the IGF-I-RF. The present results indicate that the IGF-I-RF obtained from Holstein colostrum could be a useful component for an alternative therapeutic modality for the treatment of diabetes in insulin-resistant patients.

Effect of IGF-I Rich Fraction from Bovine Colostral Whey on Murine Immunity

Insulin-like growth factor-I (IGF-I) rich fraction, collected components between 1 kDa and 30 kDa, was fractionated from bovine colostral whey using an ultrafiltration membrane. IGF-I was confirmed in the collected IGF-I rich fraction by both SDS- PAGE and Western blotting. The concentration of IGF-I in the IGF-I rich fraction was 10 ng/mg protein. One hundred microliters of the reconstituted IGF-I rich fraction was intraperitoneally injected into ICR male mice for 2 weeks at 24 h intervals. The functions of peritoneal macrophages, including phagocytosis, interleukin (IL)-6 and tumor necrosis factor (TNF)-α production, and nitric oxide and hydrogen peroxide production, were enhanced significantly by the administration of the IGF-I rich fraction in a dose-dependent manner (p<0.01). The proliferation of Concanavalin (Con) A-stimulated and Lipopolysaccharide (LPS)-stimulated splenocytes was also determined to have been enhanced significantly by the administration of the IGF-I rich fraction in a dose-dependent manner (p<0.01). Our results indicate that the administration of IGF-I rich fraction obtained from bovine colostral whey enhances both innate and acquired immunity for ICR male mice. (Asian-Aust. J. Anim. Sci. 2006. Vol 19, No. 2 : 297-304)

Colostral whey concentrate supplement increases complement activity in the sera of neonatal calves.

We evaluated the effect of a commercial bovine colostral whey on the complement-mediated immune responses of calves. Two groups of neonatal calves were fed, in addition to whole milk (WM) and pooled colostrum (PC), different amounts of a commercial immunoglobulin concentrate made from pooled colostral whey (Ig-C) for the first two feedings post natum. The control group was fed WM and PC only. Serum samples were obtained at the ages of 2, 7, 14 and 30 d. Bacteriolytic activity against complement-sensitive Escherichia coli JM103 and opsonic activity against complement-lysis-resistant E. coli IH3080 strains were studied, as well as the levels of C3 complement component and E. coli JM103 specific antibodies in the sera. Groups fed Ig-C had 2-3 times higher bacteriolytic activity than the control group of both the classic (P < 0.005) and alternative pathways (P < 0.0001) at days 2 and 7 post natum. This effect is obviously not caused solely by the antibodies ingested but also involves other unknown colostral factors, possibly lectins. The opsonisation capacities of the sera correlated well with the amounts of immunoglobulins ingested (P < 0.05) at days 2-14. The levels of C3 component in sera did not differ between the groups. In the group fed the largest amount of immunoglobulins levels of E. coli JM103-specific antibodies were highest (P < 0.0001). It can thus be concluded that the antibody independent complement activities of serum can be increased substantially by feeding colostral whey concentrate to calves during their first days of life.

Colostral proteins from cows immunised with Streptococcus mutans/S. sobrinus support the phagocytosis and killing of mutans streptococci by human leucocytes.

Passive immunisation, based on bovine colostral preparations, is an area of active research. Specific bovine antibodies inhibit the virulence factors of target pathogens but the interactions between whey preparations and human immune defence cells are not well known. Bovine colostrum inhibits the phagocytic activity of bovine leucocytes and this may reflect the biological activity of immunoglobulins in it. Therefore, this study aimed to examine the effects of bovine whey protein preparations from the colostrum of Streptococcus mutans/S. sobrinus-immunised and sham-immunised cows on binding, ingestion and killing of these bacteria by human leucocytes. Binding and ingestion of FITC-labelled bacteria were estimated by flow cytometry and leukocyte activation was measured as chemiluminescence. Killing rate was estimated by plate counting and by measuring bioluminescence from S. mutans- containing the insect luciferase gene. Colostral whey protein preparation from hyperimmunised cows activated human leucocytes by opsonising specific bacteria. Neutrophils, eosinophils and monocytes weakly phagocytosed non-opsonised bacteria and bacteria opsonised with control product. On the contrary, binding and ingestion were efficient in the presence of the preparation from immunised cows. Thus, these results show that bovine colostral whey proteins are able to support the activation of human phagocytes against pathogenic microbes and that this property is related to specific antibodies in whey preparations. These whey proteins may also be clinically useful, especially in preventing the colonisation of newly erupted teeth by mutans streptococci.

Concentrated bovine colostral whey proteins from Streptococcus mutans/Strep. sobrinus immunized cows inhibit the adherence of Strep. mutans and promote the aggregation of mutans streptococci.

The aim of this study was to examine the effect of bovine colostral whey proteins from cows immunized with Streptococcus mutans/Strep. sobrinus on the adherence and aggregation of caries-inducing bacteria, i.e., mutants streptococci. Both adherence and aggregation are important phenomena in the bacterial colonization of the human oral cavity. In all adherence experiments there was a significant difference between treatments by immune product (IP; from immunized cows) and a control product (CP; a similar product from non-immunized cows). The adherence of 35S-labelled Strep. mutans cells (serotype c) to parotid saliva-coated hydroxyapatite (SHA) was dose-dependently inhibited by both IP and CP if SHA was coated with either product before exposure to bacteria, but markedly lower concentrations of IP than CP were effective. When instead of SHA the bacterial cells were pretreated with IP or CP, only IP strongly and dose-dependently inhibited streptococcal adherence. When bacteria, IP or CP, and SHA were incubated simultaneously, a significant difference between IP and CP treatments was again found. Further, IP effectively aggregated both Strep. mutans and Strep. sobrinus cells, whereas hardly any effect was seen with CP. Both IP and CP aggregated the control bacterium Strep. sanguis, which affected the adherence of the pretreated bacteria.

Evidence Supporting the Mechanisms of Enteric Protection Provided by Colostral Whey Fed Supplements

With the increasing public concern over the use of antibiotics in the production of meat and dairy animals, we decided to examine bovine colostral whey as a source of safe, effective biological agents to control enteric pathogens and promote the growth of neonatal and weaning animals. Colostral whey, its antibody rich and/or antibody poor fractions were found to have bactericidal activity against 0157 E.coli and bacteriostatic activity against a wide range of enteropathogenic bacteria. These same colostral whey preparations did not inhibit the growth of non-pathogenic strains tested. The colostral whey preparations inhibited cytotoxic activity of Shiga-like toxin, the infection of cultured cells with enteric viruses and exhibited some anabolic activity on cultured cells. In an in vivo experiment with gnotobiotic animals, the colostral whey reduced the severity of an experimental infection and partly blocked the induction of systemic immunosuppression. We feel these effects were mediated by inhibition of the growth of the challenge E. coli and inhibition of the action of the Shiga-like toxin, which appears to be involved in the development of the immunosuppression. These studies suggest that colostral components may be good candidates for biological replacements of antibiotics as supplements to young animals.

Factors in ruminant colostrum that influence cell growth and murine IgE antibody responses.

Bovine colostrum was investigated as a source of biologically active molecules capable of stimulating the growth of mammalian cells in culture and modifying the immune response in a murine model. An extract prepared from bovine colostral whey by cation exchange and reversed-phase chromatography stimulated the growth of L6 rat myoblasts, Balb/c-3T3 mouse fibroblasts and BHK-21 baby hamster kidney cells with equal or greater potency than fetal bovine serum. Fractionation of the bovine colostral extract by gel-permeation chromatography in M-acetic acid identified a number of cell-growth factors for each cell type. Bovine colostral extract was compared with an ovine colostral whey preparation for its ability to modulate IgE antibody responses in mice. Doses of 8 and 4 mg/d of ovine colostral whey or bovine colostral extract specifically suppressed IgE antibody responses, whereas at lower doses suppression did not occur. We conclude that bovine colostrum contains cell-growth factors as well as immunomodulatory factors that are able to regulate the IgE response in a heterologous species.

Kinetics of Cryptosporidium parvum sporozoite neutralization by monoclonal antibodies, immune bovine serum, and immune bovine colostrum

Monoclonal antibodies, immune bovine serum, and immune bovine colostral whey neutralized infectivity of Cryptosporidium parvum sporozoites for mice in a time-dependent manner. Immune colostral whey neutralized sporozoites more rapidly and completely than immune serum, monoclonal antibody (MAb) 18.44, or a combination of MAb 18.44 and MAb 17.41. Mice were partially protected against oral challenge with C. parvum oocytes when treated with immune colostral whey, MAb 17.41, or a combination of MAb 17.41 and MAb 18.44.

A new approach to preparation and purification of colostrokinin from bovine colostrum

A simple and rapid method is reported for preparation and partial purification of colostrokinin from bovine colostral whey. The active substance obtained at the described stage of purification appears to be a basic polypeptide of low molecular weight. This peptide proved to be undestroyed by pepsin, only partially by trypsin and completely by papain.

Vacuolated Villous Epithelium of the Small Intestine of Young Pigs (with color plate I)

The villous epithelium of the small intestine from healthy pigs, killed during their first month of life, was examined. There were occasional vacuolated epithelial cells throughout the small intestine in most pigs; however, the occurrence of vacuolated epithelium (vacuolated epithelial cells in a continous layer along at least the distal 10% of all villi on the mesenteric side of a histological section) was related to area in the intestine and age of the pig. Ileal epithelium was vacuolated in nearly all pigs examined during their first 2 weeks of life; however, the frequency of vacuolated ileal epithelium was less in pigs examined during their third and still less in those examined during their fourth week of life. Jejunal epithelium was vacuolated less frequently than ileal epithelium and was not seen in pigs more than 11 days old. Vacuolated duodenal epithelium was not seen. Vacuolated ileal epithelium, in suckling pigs during their second week of life, was morphologically similar to that associated with absorption of colostrum in the neonate and took up bovine colostral whey pinocytotically. In contrast to unvacuolated epithelium, vacuolated epithelium stained supravitally with Evans blue dye given orally. Vacuolated ileal epithelial cells examined by autoradiography after exposure to tritiated thymidine had synthesized DNA more than 3 days previously. These observations were compared with reports that considered vacuolated epithelium, in 2 to 3-week-old pigs, to be pathologic. A hypothesis relating vacuolation to age of ileal cells was proposed.

Isolation and characterization of IgM from bovine colostral whey

Bovine IgM was isolated from colostral whey, by molecular sieve and DEAE-cellulose chromatography. It was characterized using physico-chemical and immuno-chemical techniques. The molecular weight of the intact molecule was found to be 1,030,000, while that of the computer μ and light chains was 76,000 and 22,500, respectively.

Isolation of the gamma-C-globulin of bovine cerebrospinal fluid.

The γc-globulin of bovine cerebrospinal fluid (CSF) was isolated in immunologically pure form from the globulin fraction of CSF or milk whey by gel filtration on Sephadex G-75. The globulin fraction was obtained by precipitation with sodium or ammonium sulfate rather than by DEAE chromatography because salt precipitation resulted in greater yields of the γc-globulin. When the protein was isolated from bovine colostral whey, about 90% of the immunoglobulin G (IgG) globulin was removed by a preliminary filtration of the whey on Sephadex G-75. The fraction containing the γc-globulin admixed with IgG globulin was then re-chromatographed on Sephadex G-75 to remove the IgG globulin. On gel filtration through a column of Sephadex G-75 that had been calibrated with proteins of known molecular weight, the elution volume of the γc-globulin corresponded to a molecular weight of 30 000. Different lots of γc-globulin that contained no detectable impurities by gel-diffusion tests were compared on a weight basis by a quantitative immunochemical method, and were found to have equal immunological activity.

Electrophoretic and immunological studies on sera from calves from birth to weaning. II. Electrophoretic and serological studies with special reference to the normal and induced agglutinins to Trichomonas foetus.

Summary1. The lacto-globulin in bovine colostral whey, and the γ globulin in the serum contain the normal and induced agglutinins to the protozoonTrichomonas foetus.2. A decline in γ globulin in sera from cattle approaching calving may be associated with the concentration of the lacto-globulin in the udder.3. The elimination of the passive normal and induced agglutinins acquired by the neonatal calf was logarithmic. The autogenous production of γ globulin by the calf occurred soon after birth, resulting in an apparent decline in the rate of elimination of the lacto-globulins.4. The autogenous production of γ globulin was not necessarily accompanied by that of detectable normal agglutinin.5. Calves injected withT. foetusantigen within the first 3 weeks of life, and in which there was evidence of autogenous γ globulin, developed normal autogenous but not induced agglutinins.6. Calves which received colostrum containing maternal induced and normal agglutinins, calves partially deprived of colostrum, and calves immunized in early life, produced the normal autogenous agglutinin between the 30th and 60th days, reaching the normal adult titre of 1/48–1/96.7. The properties of the normal agglutinins were investigated. They possess some characteristics significantly different from those of the induced agglutinins. These suggested that the normal agglutinins were γ globulin molecules with an ‘accidental’ genetic configuration complementary toT. foetus.The author wishes to thank Dr M. Robertson, F.R.S., and Sir Alan Drury, F.R.S. for their interest and encouragement during the course of this work, and Dr R. C. Campbell for the statistical analysis.