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PubMed1986

Naturally occurring IgA antibodies to ocular and oral microorganisms in tears saliva and colostrum: evidence for a common mucosal immune system and local immune response.

In this study, levels of naturally occurring secretory IgA (sIgA) antibodies to bacterial isolates representative of the microflora of the human eye (Staphylococcus epidermidis and a Corynebacterium species) and the oral cavity (Streptococcus mutans serotypes c and d) were assessed in three different human external secretions. Tears and parotid saliva samples collected at the same time from 22 healthy subjects and colostrum from 11 healthy women (1-14 days post partum) were assessed for sIgA anti-bacterial antibody levels by an enzyme-linked immunosorbent assay. Significantly higher levels (P less than 0.001) of IgA antibodies to Staphylococcus epidermidis and the Corynebacterium sp. were found in tears than in parotid saliva. Furthermore, higher levels of sIgA antibodies to Streptococcus mutans serotype d occurred in parotid saliva than in tears of these subjects. Although levels of salivary sIgA antibodies to S. mutans serotype c were lower than those seen to serotype d, they were not significantly different from those in tears. However, absolute sIgA anti-serotype c antibodies per mg IgA were higher in saliva than in tears. When sIgA antibody levels to the four bacterial strains were assessed in colostrum, the proportion of sIgA antibodies per mg IgA were much lower than seen in tears or saliva. These results suggest that natural sIgA antibodies which occur in human external secretions are induced by antigen ingestion and stimulation of the common mucosal immune system. However, the local presence of antigen at a mucosal site induces greater clonal expression and results in higher levels of sIgA antibodies than at mucosal sites not exposed to local antigenic stimulation.

Adolescent↗

PubMed2001

Elevated extrahepatic expression and secretion of mammary-associated serum amyloid A 3 (M-SAA3) into colostrum.

Mammary-associated serum amyloid A 3 (M-SAA3) was secreted at highly elevated levels in bovine, equine and ovine colostrum and found at lower levels in milk 4 days postparturition. N-terminal sequencing of the mature M-SAA3 protein from all the three species revealed a conserved four amino acid motif (TFLK) within the first eight residues. This motif has not been reported to be present in any of the hepatically-produced acute phase SAA (A-SAA) isoforms. Cloning of the bovine M-Saa3 cDNA from mammary gland epithelial cells revealed an open reading frame that encoded a precursor protein of 131 amino acids which included an 18 amino acid signal peptide. The predicted 113 residue mature M-SAA3 protein had a theoretical molecular mass of 12,826Da that corresponded with the observed 12.8kDa molecular mass obtained for M-SAA3 in immunoblot analysis. The high abundance of this extrahepatically produced SAA3 isoform in the colostrum of healthy animals suggests that M-SAA3 may play an important functional role associated with newborn adaptation to extrauterine life and possibly mammary tissue remodeling.

Amino Acid Motifs↗

PubMed1999

Apelin, the natural ligand of the orphan receptor APJ, is abundantly secreted in the colostrum.

By using a strategy that we have developed to search for the ligands of orphan seven-transmembrane-domain receptors [S. Hinuma et al., Nature 393 (1998) 272-276], we have recently identified a natural ligand, apelin, for the orphan 7TMR, APJ [K. Tatemoto et al., Biochem. Biophys. Res. Commun. 251 (1998) 471-476]. In this paper, we isolated rat and mouse apelin cDNAs, and analyzed the tissue distribution of apelin mRNA in rats. Although apelin mRNA was widely detected in a variety of tissues, the highest expression of apelin mRNA was detected in the mammary gland of pregnant rats. In the mammary gland, biologically active apelin and its mRNA considerably increased during pregnancy and lactation, and reached a maximal level around parturition. Moreover, a large amount of apelin (14-93 pmol/ml) was found to be secreted in the bovine colostrum, and it was still detectable even in commercial bovine milk. Since apelin partially suppressed cytokine production by mouse spleen cells in response to T cell receptor/CD3 cross-linking, the oral intake of apelin in the colostrum and milk might modulate immune responses in neonates.

Adipokines↗

PubMed1997

Generation of bovine immune colostrum against Streptococcus mutans and Streptococcus sobrinus and its effect on glucose uptake and extracellular polysaccharide formation by mutans streptococci.

Due to potential side-effects of active immunization by cariogenic mutans streptococci, oral administration of passively-derived antibodies could be a more acceptable way to reduce colonization and virulence of these microorganisms in human dentition. The aim of this study was to produce antistreptococcal immunoglobulins into bovine colostrum and explore the possible antibacterial mechanisms of these immunoglobulins against mutans streptococci. Specific serum IgG antibodies to whole cell antigens of both Streptococcus mutans and Streptococcus sobrinus increased rapidly in cows during immunization and were high also in the final whey-product. Low concentration (0.5% w/v) of bovine immune preparation inhibited significantly the incorporation of [14C]glucose by both S. mutans and S. sobrinus. Higher concentration (> 1%) was needed to inhibit the glucosyltransferase or fructosyltransferase activities of these bacteria. No such inhibitory effects were observed with the control preparation from the non-immunized cows. Our results indicate that bovine immune colostrum has a significant inhibitory potential against mutans streptococci, apparently dependent on the presence of specific IgG antibodies against S. mutans and S. sobrinus.

Animals↗

PubMed1998

Therapeutic efficacy of hyperimmune bovine colostrum treatment against clinical and subclinical Cryptosporidium serpentis infections in captive snakes.

Therapy based on the protective passive immunity of Hyperimmune Bovine Colostrum (HBC) (raised against Cryptosporidium parvum in dairy cows immunized during gestation) was tested for heterologous efficacy in subclinical and clinical infections of 12 captive snakes with C. serpentis. Six gastric HBC treatments of 1% snake weight at 1-week intervals each, have histologically cleared C. serpentis in three subclinically infected snakes, and regressed gastric histopathological changes in one of these snakes. In all snakes, each subsequent HBC treatment significantly decreased the number of oocysts recovered in gastric lavage eluants (P < 0.03). The treatments induced oocyst-negative gastric eluants and stools in all snakes, and improved clinical signs of infection. Clinically infected snakes displayed severe histopathological changes in the gastric region; however, the numbers of developmental stages of C. serpentis were moderate. Considering the severity of pathology, much lower than expected pathogen numbers were observed, and it is believed that clinically infected snakes did not have enough time to repair tissue damage that had occurred over the years of infection. As the HBC treatment was safe and highly efficacious, it is recommended to gastrically administer the HBC therapeutically to snakes that are clinically or subclinically infected with C. serpentis. Hyperimmune bovine colostrum can also be used in snake supportive therapy or prophylaxis.

Animals↗

PubMed2001

Occurrence of an unusual phosphorylated N-acetyllactosamine in horse colostrum.

The colostrum of horses (thoroughbreds) was extracted and fractionated to yield Gal(beta1-4)GlcNAcalpha1-phosphate, which has not previously been detected in any mammalian milk or colostrum, as well as Neu5Ac(alpha2-3)Gal(beta1-4)Glc. The structures of these saccharides were established by NMR spectroscopy and MALDI-TOF mass spectrometry.

Amino Sugars↗

PubMed1998

Sialyllactose occurs as free lactones in ovine colostrum.

Three sialyl oligosaccharide fractions were separated from ovine colostrum by gel filtration, anion exchange chromatography and normal-phase HPLC. They were characterized by 1H-NMR spectrometry as follows: Neu5Acalpha2-->3Galbeta1-->4Glc, Neu5Gcalpha2-->6Galbeta1-->4Glc and three forms of Neu5Gcalpha2-->3Galbeta1-->4Glc, namely Neu5Gcalpha2-->3Galbeta1-->4Glc itself, its lactone derivative between the carboxyl group of Neu5Gc and Gal OH-2 and another lactone derivative between the carboxyl group and Gal OH-4. In this study, Neu5Gc-lactose lactones, in their free form, have been isolated for the first time from any natural sources including milk or colostrum.

Animals↗

PubMed2001

Necrotoxigenic Escherichia coli type-2 invade and cause diarrhoea during experimental infection in colostrum-restricted newborn calves.

There exists experimental evidence that necrotoxigenic Escherichia coli (NTEC) strains producing the cytotoxic necrotising factor 1 cause intestinal and extra-intestinal disease in piglets. On the other hand, no experimental model has been developed with NTEC strains producing the cytotoxic necrotising factor 2. In all, 14 colostrum-restricted calves were orally challenged with two strains isolated from the faeces of a diarrheic calf (B20a) or from the heart blood of a septicaemic calf (1404). All calves had diarrhoea which lasted until euthanasia in eight of them. In those calves, diarrhoea was correlated with the faecal excretion of the challenge strains. At necropsy, vascular congestion of the intestinal mucosa, hypertrophy of the mesenteric lymph nodes (MLN) and some congestion of the lungs were observed. Bacteriology confirmed the colonisation of the intestine by the challenge strains which were also recovered from the heart blood, the lungs and/or the liver. Histological sections confirmed enterocolitis, lymphadenitis and limited bronchopneumonia. In the intestinal tissue sections, bacteria testing positive in an in situ DNA hybridisation assay with a CNF2 probe were observed. Those results were confirmed by immunohistochemistry with a polyclonal anti-O78 and a monoclonal anti-F17b antisera. Three of the five control calves receiving either saline or a CNF(-), F17a strain (25KH09) had no clinical signs or lesions. The other two presented a profuse liquid diarrhoea but those calves were positive for the presence of K99(+) E. coli. In this model, both NTEC2 strains were thus, able to colonise the intestine, to cause long-lasting diarrhoea and to invade the blood stream with localisation in various internal organs in colostrum-restricted conventional newborn calves.

Animals↗

PubMed1998

Simplified method for purification of colostrum to obtain secretory component of immunoglobulin A, using secretory component as a reference protein in tracheal aspirate fluid.

Many studies employ bronchoalveolar lavage fluid for assessment of biologically active substances secreted from the lung. However, investigators continue to search for a useful reference standard to correct for the inevitable but variable degree of dilution of this fluid. The glycoprotein, soluble secretory component of IgA, may serve as a valid reference protein. We report a simplified method for the purification of secretory component from colostrum. Soluble secretory component was isolated from human colostrum using serial centrifugation, size-exclusion fractionation and ion-exchange chromatography. Secretory component rich fractions were assayed by enzyme immunoassay. They were also evaluated for total amino acid content and distribution and sequence determination with satisfactory agreement with published results. We then demonstrated that soluble secretory component concentration in tracheal aspirate fluid did not correlate with either albumin or with total protein measured in the same samples. Therefore, we conclude that the secretory component of IgA serves as a useful reference marker because its use may avoid errors resulting from leakage of plasma proteins into epithelial lining fluid. Advantages of this method for establishing a standard for secretory component include ready availability of soluble secretory component, simplicity of the method and relative rapidity of the techniques.

Chemical Fractionation↗

PubMed1999

Chemical characterisation of six oligosaccharides in a sample of colostrum of the brown capuchin, Cebus apella (Cebidae: primates).

Carbohydrates were extracted from a sample of brown capuchin colostrum and six of the component oligosaccharides were separated and purified by gel filtration and preparative thin layer chromatography. Their structures were determined by 1H-NMR to be as follows: Gal beta 1-->4[Fuc alpha 1-->3]Glc (3-fucosyllactose) Gal beta 1-->3Gal beta 1-->4Glc (beta-3'-galactosyllactose) Gal beta 1-->6Gal beta1-->4Glc (beta-6'-galactosyllactose) Gal beta-->3[Gal beta 1-->4GlcNAc beta 1-->6]Gal beta 1-->4Glc (lacto-N-novopentaose I) Gal beta 1-->4GlcNAc beta 1-->3[Gal beta 1-->4GlcNAc beta 1-->6]Gal beta 1-->4Glc (lacto-N-neohexaose) Neu5Ac alpha 2-->3Gal beta 1-->4Glc (3'-N-acetylneuraminyllactose) Of these, all except lacto-N-novopentaose I have been previously found in human milk or colostrum.

Animals↗

PubMed1981

Leucocytes of sheep colostrum, milk and involution secretion, with particular reference to ultrastructure and lymphocyte sub-populations.

The leucocytes in sheep colostrum, milk and involution secretion were studied using the electron microscope. The predominant cell type in colostrum was the polymorphonuclear leucocyte (PMNL) (41--84%), followed by the macrophage (8--49%), and the lymphocyte (6--11%). Plasma cells were present in low numbers (1--2%) while no secretory epithelial cells were observed. In mid-lactation the cell components changed so that the macrophage was the predominant cell (83--86%) followed by the lymphocyte (10--17%). In early involution secretion, PMNL reappeared but declined in secretion obtained 21 d after weaning, when the macrophage was again the predominant cell. Associated with the whole cells were membranous extracellular materials and "sunburst' fragments of the secretory epithelial cells. These were phagocytosed by both PMNL and macrophages, which also engulfed fat droplets from the milk. The lymphocytes were examined for plasma membrane markers and the T-cell percentage (E+ rosettes) fluctuated within wide limits, (0--80%). However, there was no discernible trend associated with stage of lactation. The B-cell markers (C1 and Fc gamma) also fluctuated widely and it was evident that contaminating monocytes reduced the accuracy of the count. However, it appeared that more B-cells were present in milk than in blood.

Animals↗

PubMed1991

Ovine lactoferrin: isolation from colostrum and characterization.

Highly purified lactoferrin was isolated from ovine colostrum by sequential purification on CM-Sephadex C-50 and Blue-Sepharose, with overall yield of 55%. The ovine lactoferrin was characterized by SDS-PAGE, its amino acid composition and N-terminal sequence to residue 30. Homology with bovine and human lactoferrins was greater than 80 and 50% respectively. Antibodies to ovine lactoferrin were raised in rabbits and used to develop an enzyme-linked immunosorbent assay (ELISA). The antiserum was not cross reactive with other colostrum proteins.

Amino Acid Sequence↗

PubMed1992

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.

Animals↗

PubMed1995

Assessment of mammary gland metabolism in the sow. I. Development of methods for the measurement of cellular metabolites in milk and colostrum.

We have modified deproteinization methods and a number of spectrophotometric and bioluminescent methods in order to measure the concentrations of cellular metabolites in small volumes (< 0.3 ml) of sows' colostrum and milk. For the majority of the assays, recoveries ranged from 92 to 105%. The binding of ATP and UTP to a calcium phosphate-citrate-caseinate complex in milk, and the decrease in ATP (92%/h) and UTP (18.1%/h) concentrations during in vitro incubation of the whey fraction suggested that it was unlikely ATP and UTP (both < 1 microM) could exist free in sows' milk. The mean concentrations (range) of cellular metabolites in milk (6-11 d post partum) were: glucose, 669 microM (220-1367); glucose 6-phosphate, 63.0 microM (27.6-101.4); glucose 1-phosphate, 18.3 microM (13.1-24.8); UDPglucose, 296 microM (170-494); UDPgalactose, 635 microM (230-945); lactose, 162 mM (124-187); UDP, 105 microM (85-130); UMP, 1760 microM (1326-2587); inorganic phosphate, 13.5 mM (1.4-29.3); ATP, < 0.5 microM; ADP, 53.6 microM (10.5-171.25); AMP, 215 microM (61.6-491.6); cAMP, 22.3 microM (3.5-61.6); galactose, 198 microM (118-474) and fructose, 226 microM (172-283). Differences in the concentrations of glucose, glucose 6-phosphate, glucose 1-phosphate, UDPgalactose and cAMP between fore and hind milk samples indicated postsecretory changes in the concentrations of certain metabolites. Changes in the concentrations of metabolites during in vitro incubation of milk and of colostrum suggested that these postsecretory changes were probably due to the actions of enzymes present in mammary secretion.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine Triphosphate↗

PubMed2004

Screening and identification of acidic carbohydrates in bovine colostrum by using ion/molecule reactions and Fourier transform ion cyclotron resonance mass spectrometry: specificity toward phosphorylated complexes.

A screening method was developed for the identification of acidic saccharides from biological mixtures utilizing gas-phase derivatization and mass spectrometry. Phosphorylated compounds were differentiated from other acidic species by exploiting the selective reactivity of chlorotrimethylsilane with the phosphate ions (phosphorylated compounds shift by 72 Da, allowing rapid compound detection). A 13-component mock mixture was used to demonstrate the viability of the method, and a detection limit of 600 nM (30 fmol) was determined. This method was applied to the identification of acidic compounds from bovine colostrum. To further verify the selectivity of the ion/molecule reaction, exact mass measurements were used to determine the elemental composition of 14 compounds. Eight novel acidic carbohydrate species were observed in bovine colostrum, six of which have never been reported previously in milks. Tandem mass spectrometric experiments allowed compound characterization for two of these components.

Animals↗

PubMed1999

C3/C4 concentration ratio reverses between colostrum and mature milk in human lactation.

The levels of complement fractions C3 and C4 were assayed in human milk in a classic nephelometric assay adapted to this secretion. Concentrations of these molecules were measured in 667 milk samples obtained sequentially from 76 volunteer lactating mothers during the first 12 weeks of lactation. Immunonephelometry was performed using skimmed milk samples diluted 10 times and yielded reproducible (coefficients of variation in within- and between-run precision lower than 9% for C3 and than 14% for C4) and accurate (linear recovery in dilution-overloading assay) data. High concentrations (mean +/- SE) were found for C3 (199.32+/-16.35 mg/L) and C4 (113.42+/-11.16 mg/L) in colostrum samples (n = 159; days 1-5). A significant (P<0.001) and rapid decrease was observed in transitional milk samples (n = 198; days 6-14), containing 57.71+/-5.18 and 72.39+/-4.98 mg/L of C3 and C4, respectively. Stable lower levels were noted in mature milk samples (n = 310; days 15-84) at 30.36+/-1.57 mg/L for C3 (P<0.001) and 53.38+/-3.61 mg/L for C4 (P<0.05). The decrease rate was different for C3 and C4, yielding a reversal of the C3/C4 ratio between colostrum and more mature milk.

Calibration↗

PubMed1999

Trace element transfer from the mother to the newborn--investigations on triplets of colostrum, maternal and umbilical cord sera.

OBJECTIVE: To investigate the trace element transfer from the mother to the newborn. DESIGN: The concentrations of the eight essential elements calcium (Ca), cobalt (Co), copper (Cu), magnesium (Mg), manganese (Mn), molybdenum (Mo), tin (Sn), and zinc (Zn), and of the non-essential and toxic elements barium (Ba), beryllium (Be), bismuth (Bi), cadmium (Cd), cesium (Cs), lanthanum (La), lithium (Li), lead (Pb), rubidium (Rb), antimony (Sb), strontium (Sr), and thallium (Tl) were determined in umbilical cord (n = 29) and corresponding maternal sera (n = 29) as well as in colostrum (n = 27). RESULTS: Umbilical cord serum concentrations of Ca, Mn, and Zn were 120%, 150%, and 148% of the maternal value, respectively. Maternal sera had twice the Cu concentrations found in healthy adults and five-times higher Cu than umbilical cord sera. Concentration ratios colostrum/maternal serum and colostrum/umbilical cord serum were approximately one for Co, 1.4 for Mg, two for Ca, Mn, and Sn, five for Cu (maternal serum), eight for Mo, and ten for Zn. Concentrations of the toxic elements Cd and Pb decreased in the order colostrum (Pb 2.6 microg/L; Cd 0.6 microg/L), maternal sera (0.8 microg/L; 0.3 microg/L), umbilical cord sera (0.4 microg/L; 0.2 microg/L). Maternal serum Ba and Rb was 182% and 66% of the umbilical cord value. For Sr and Li, an almost perfect correlation between umbilical cord and maternal sera was found. For Ba, Co, Cu, Mn, Zn none, and for Ca, Cs, Mn, Mo, Rb only weak positive correlations between these two compartments could be established. CONCLUSIONS: The results of this study indicate that an active transport mechanism for the transport of Ca, Mn, Rb, and Zn from the mother to the newborn exists, whereas Cs, Li, and Sr follow concentration gradients. As regards Cu, the placenta showed to have a blocking effect on the transfer from the mother to the baby.

Adult↗

PubMed1984

Partial purification and characterization of a growth factor present in goat's colostrum. Similarities with platelet-derived growth factor.

A factor in goat's colostrum which stimulates DNA synthesis and cell proliferation in Swiss 3T3 fibroblasts has been purified approx. 350-fold by a sequence of acid precipitation, cation-exchange chromatography and gel filtration. The growth factor is a highly basic, heat stable (100 degrees C for 5 min) polypeptide with Mr approx. 35000. The polypeptide resists denaturation by guanidinium chloride or urea but is totally inactivated by treatment with reducing agents. The factor, which we have termed colostric basic growth factor ( CBGF ), inhibits the binding of 125I-labelled epidermal growth factor (125I-EGF) to Swiss 3T3 fibroblasts but does not inhibit 125I-EGF binding to epidermoid A431 cells. CBGF interacts synergistically with plasma in stimulating DNA synthesis in quiescent Swiss 3T3 cells. The chemical and biological properties of CBGF are thus very similar to the properties reported for the human platelet-derived growth factor. Although high concentrations of CBGF are present in the colostrum of goats, cows, and sheep, the milk of these species contains little or no factor. The origin and possible functions of CBGF are unknown.

Animals↗