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Antibody dependent cytotoxicity of human colostrum phagocytes.

Monocytes and polymorphonuclear neutrophils of human colostrum were identified using cytochemical staining procedures. The ability of colostral phagocytes to function in antibody dependent cellular cytotoxicity was found to be satisfactory at high effector/target cells ratio (20:1) Colostral monocytes showed a diminished cytotoxicity when compared to monocytes of adult or neonatal peripheral blood at low effector/target cells ratio (1:1). This could be due to the extensive fat ingestion by these cells or other inhibitory factors found in colostrum.

Antibody-Dependent Cell Cytotoxicity↗

Intestinal macromolecule absorption in the fetal pig after infusion of colostrum in utero.

The ability of the fetal pig intestine to absorb large proteins was investigated in utero. Six pregnant sows were anesthetized (Na pentobarbital) at 99-102 d of gestation (term = 115 +/- 2 d), and a catheter was inserted into the esophagus of two to three fetuses per sow. Via these catheters, sterile solutions (10.0 mL) of colostrum whey (CW, n = 5), milk whey (MW, n = 5), or amniotic fluid (AF, n = 4) were infused into the fetal pig stomachs every 6 h for 6-8 d starting on the day after surgery (d 0). Levels of IgG in the three fluids were 120, 0.5, and 0 mg/mL, respectively. During the first 2-3 d of infusion, plasma IgG levels rose rapidly in the CW fetuses (to 7.5 +/- 0.8 mg/mL), whereas IgG remained absent in plasma from MW and AF fetuses. Absorption of a macromolecule marker, BSA, was also higher when the marker was given with CW rather than with MW or AF. However, when all three treatment groups were given CW + BSA on the last experimental day (d 6-8), the mean BSA increment in the CW group was only 5-8% of that in the AF group, with intermediate values for the MW group. Neither at the beginning nor at the end of the experiment was macromolecule uptake in individual CW fetuses correlated with their cortisol level in plasma. The prenatal pig intestine is similar to the neonatal pig intestine in that colostrum stimulates both the macromolecule absorption and the cessation of macromolecule uptake (intestinal closure). However, fetal pigs have a lower protein absorptive capacity and a longer preclosure period than newborn pigs; this may be related to an immature structure and function and a slow enterocyte proliferation rate in the prenatal pig intestine.

Amniotic Fluid↗

Purification and characterization of a bovine colostrum-derived growth factor.

A growth factor in bovine colostrum was purified to homogeneity by a combination of acid extraction, boiling, cation exchange chromatography, isoelectric focusing, and reverse phase HPLC. The bovine colostrum growth factor (BCGF) had an isoelectric point of about 10, a native mol wt of about 30,000, was resistant to inactivation by boiling and exposure to pH 1, but was inactivated by dithiothreitol. BCGF appeared to be structurally related to human platelet-derived growth factor (PDGF) and competed with human PDGF in a radioreceptor assay. However, while human PDGF appeared to be a heterodimer of 17,000 and 14,000 mol wt subunits, BCGF appeared to be a homodimer of 20,000 mol wt subunits. Purified BCGF had a specific activity in stimulating 3T3 cell proliferation of about 3-6 U/ng and was active at about 1-2 ng/ml.

Animals↗

Identification of a soluble UDP-Gal: Gal (beta 1-4)Glc (or GlcNAc) (alpha 1-3) galactosyltransferase of bovine colostrum.

A soluble UDP-Gal: Gal (alpha 1-3) galactosyltransferase was first detected in bovine colostrum and this enzyme activity was simply assayed by using rho-nitrophenyl-beta-lactoside (Gal(beta 1-4)Glc-C6H5NO2, rho NP-lactoside) as an acceptor. Treating the radioactive product with alpha- or beta-galactosidase, the radioactivity (greater than 95%) was released by only alpha-galactosidase and was identified as [3H]galactose. This shows that galactosyl residue was alpha-linked to rho-nitrophenyl-beta-lactoside. Methylation, hydrolysis, thin layer chromatography and fluorography of the reaction product (Gal(alpha 1-)-[3H]Gal(beta 1-4)Glc-rho NP) yielded 2,4,6-tri-O-methyl[3H]galactose, indicating that galactosyl residue had been transferred to the carbon-3 position of the terminal nonreducing beta-galactosyl residue in rho-nitrophenyl-beta-lactoside. These results confirmed that the structure of the reaction product was Gal(alpha 1-3)Gal(beta 1-4)Glc-rho NP. The enzyme requires Mn2+ for its activity, and shows pH optimum from 6.5 to 7.5. rho-Nitrophenyl-beta-lactoside and asialo alpha 1-acid glycoprotein were more effective as an acceptor than N-acetyllactosamine. The bovine colostrum (alpha 1-3) galactosyltransferase could not convert human O red cells into B active cells, indicating that this enzyme preparation did not contain the activity to synthesize human blood group B erythrocytes.

Animals↗

The carbohydrate linkage site of cow colostrum trypsin inhibitor.

Cow colostrum trypsin inhibitor (chromatographic form AIV[7])was subjected to basic conditions that favour beta-elimination of carbohydrates from O-glycosidic linkages. The unchanged carbohydrate composition and the unchanged values for serine and threonine indicate the presence of an alkali-stable N-glycosidic bond to asparagine. From a tryptic digest of S-aminoethylated inhibitor the glyco-decapeptide (residues 24 through 33) was isolated. The carbohydrate composition was identical with that of the S-aminoethylated inhibitor. Further degradation of this peptide by carboxypeptidase C (and aminopeptidase) produced the glycopeptide Asn(CHO)-Ser-(Thr) with the same carbohydrate composition. Thus, a single carbohydrate moiety is bound by a N-glycosidic linkage to asparagine-27 of the colostrum inhibitor. It is located opposite to the reactive site at the base of a pear-shaped molecule.

Animals↗

On the carbohydrate composition of bovine colostrum trypsin inhibitor.

The trypsin inhibitor from bovine colostrum has been separated into several forms by CM-Sephadex and DEAE-cellulose chromatography. These forms differ in the amount and composition of the carbohydrate they contain, which has been quantitated for four components by gas-liquid chromatography and standrad colorimetric procedures. The monosaccharides fucose, mannose, galactose, galactosamine, glucosamine and sialic acids have been determined. A microheterogeneity was establish ed in the carbohydrate moiety, which amounts to about 40% of the total molecular weight (Mr 11 000 - 14 000) of bovine colostrum inhibitor.

Animals↗

The amino-acid sequence of beta-lactoglobulin II from horse colostrum (Equus caballus, Perissodactyla): beta-lactoglobulins are retinol-binding proteins.

beta-Lactoglobulin isolated from horse colostrum is heterogeneous and contains two components: beta-lactoglobulin I and beta-lactoglobulin II. These two proteins are monomeric and show differences in their electrophoretic mobilities, chain lengths and primary structures. The complete amino-acid sequence of beta-lactoglobulin II was determined by automated Edman degradation of the intact protein and of the peptides derived from these by digestion with trypsin or chymotrypsin and by chemical cleavage with cyanogen bromide. Unlike other beta-lactoglobulins which contain 162 amino acids, horse beta-lactoglobulin II is unique in that it contains 166 amino acids. The additional four amino acids represent an insertion between positions 116 and 117 of other beta-lactoglobulins so far sequenced, including horse beta-lactoglobulin I. Sequence comparison of beta-lactoglobulins I and II from horse colostrum reveals 48 amino acid substitutions (30%). Such a diversity between members of the beta-lactoglobulin gene family has not been encountered before. Sequence comparison with bovine beta-lactoglobulin A shows 85 amino acid replacements accounting for 53% of the residues. The structural homology with human retinol-binding protein may reveal similar biological functions and clues to the origin of milk proteins.

Amino Acid Sequence↗

Measurement of betacellulin levels in bovine serum, colostrum and milk.

Betacellulin, a member of the epidermal growth factor (EGF) family, was originally isolated and identified from the conditioned medium from a murine pancreatic beta-cell carcinoma cell line. Recently, we isolated bovine betacellulin from a growth factor enriched cheese whey extract, but there is no information on the presence of betacellulin in other biological fluids. We have cloned the cDNA for bovine betacellulin, produced recombinant betacellulin and shown that it has a similar potency to the purified native molecule in stimulating the proliferation of Balb/c3T3 fibroblasts. We have produced a polyclonal antiserum to bovine betacellulin which did not cross-react with EGF or transforming growth factor-alpha (TGF-alpha). The antibody was used in a homologous RIA that was able to detect betacellulin in pooled bovine colostrum sampled during the first 3 days after calving (2.30+/-0.11 ng/ml mean+/-s.e.m.; n=6), in bovine milk soluble fraction (1.93+/-0.64 ng/ml mean+/-s.e.m.; n=5) and in bovine cheese whey (2.59+/-0.16 ng/ml mean+/-s.e.m.; n=3). The betacellulin concentration in foetal bovine serum (FBS) (3.68+/-0.59 ng/ml mean+/-s.e.m.; n=6) greatly exceeded that of betacellulin in serum from male calves 1 and 5 weeks of age (0.53+/-0.15 ng/ml and 0.70+/- 0.09 ng/ml respectively; mean+/-s.e.m.; n=9). Betacellulin measured in the serum of these same animals when aged between 27 and 43 weeks was below the detection limits of the RIA. Sera from 10 out of 36 unmated heifers contained betacellulin levels within the detection limits of the assay (0.433+/-0.06 ng/ml mean+/-s.e.m.; n=10). The presence of betacellulin in bovine colostrum and milk suggests that it plays a role in the growth and development of the neonate and/or mammary gland function. The results also show that betacellulin is undetectable in the castrated adult male circulation. Additionally, although present in very low amounts, serum betacellulin could be under hormonal regulation in the female, since betacellulin was detected in sera from 27% of the unmated heifers examined in this study. The high levels of betacellulin detected in FBS relative to newborn and adult serum suggests a possible endocrine role for this growth factor in the bovine foetus.

3T3 Cells↗

Evaluation of treatment of colostrum-deprived kittens with equine IgG.

OBJECTIVE: To evaluate equine IgG as a treatment for kittens with failure of passive transfer of immunity (FPT). ANIMALS: 13 specific pathogen-free queens and their 77 kittens. PROCEDURE: Kittens were randomized at birth into 9 treatment groups. One group contained colostrum-fed (nursing) kittens; the other groups contained colostrum-deprived kittens that were administered supplemental feline or equine IgG PO or SC during the first 12 hours after birth. Blood samples were collected at serial time points from birth to 56 days of age for determination of serum IgG concentrations. The capacity of equine IgG to opsonize bacteria for phagocytosis by feline neutrophils was determined via flow cytometry. RESULTS: Kittens that received feline or equine IgG SC had significantly higher serum IgG concentrations than those of kittens that received the supplements PO. In kittens that were administered supplemental IgG SC, serum IgG concentrations were considered adequate for protection against infection. The half-life of IgG in kittens treated with equine IgG was shorter than that in kittens treated with feline IgG. Feline IgG significantly enhanced the phagocytosis of bacteria by feline neutrophils, but equine IgG did not. CONCLUSIONS AND CLINICAL RELEVANCE: Serum concentrations of equine IgG that are considered protective against infection are easily attained in kittens, but the failure of these antibodies to promote bacterial phagocytosis in vitro suggests that equine IgG may be an inappropriate treatment for FPT in kittens.

Administration, Oral↗

Effects of vitamin E and selenium on immune responses of peripheral blood, colostrum, and milk leukocytes of sows.

This study was designed to assess how dietary vitamin E (E) and (or) selenium (Se) concentrations affect immune responses of gestating and peripartum sows. Multiparous sows (24), assigned to one of four groups at breeding, were fed ensiled, shelled corn-soybean meal-based diets without supplemental E or Se (-E-Se), with .3 mg of Se/kg (-E+Se), with 60 IU of E/kg (+E-Se), or with both supplemental E and Se (+E+Se) during gestation and to d 4 of lactation. Blood was obtained on 0, 30, 60, and 90 d of gestation and at parturition for serum E and Se assays. Lymphocytes and polymorphonuclear cells (PMN) were isolated from the blood, colostrum, and 4-d milk samples for immune studies. Compared with the control (+E+Se) diet, the -E-Se diet reduced (P < .05) the serum tocopherol and Se concentrations, the mitogenic responses of lymphocytes of peripheral blood (PBL) and colostrum (CL), the phagocytic activity of blood and colostral PMN, and the microbicidal activity of blood, colostral, and milk PMN. The -E+Se diet reduced (P < .05) the serum tocopherol concentrations, the mitogenic responses of PBL and CL, and the phagocytic activity of PBL. The +E-Se diet reduced (P < .05) serum Se concentrations and the phagocytic activity of PMN. The data indicated that E restriction depressed PBL and PMN immune functions, whereas Se restriction depressed mainly PMN function.

Animals↗

Essential fatty acid status and characteristics associated with colostrum-deprived gnotobiotic and conventional lambs. Growth, organ development, cell membrane integrity and factors associated with lower bowel function.

A factorial experiment involving gnotobiotic (GN) and conventional (CV) colostrum-deprived lambs and diets formulated to be adequate or deficient in linoleic acid was conducted to determine the effect(s) of the intestinal microflora on the essential fatty acid (EFA) status of the host and subsequent physiological consequences, i.e., growth, organ development, cell membrane integrity and lower bowel function. Lambs were obtained by sterile surgical procedures and housed in sterile isolators or in conventional metabolism stalls for 60 d. Skimmed cow's milk with 6% hydrogenated coconut oil and vitamins A, D and E added with and without .32% of the total calories as linoleic acid was homogenized, bottled and autoclaved, then fed to appetite three to four times daily. The GN lambs supplemented with linoleic acid gained significantly faster between 13 and 41 d of age and more efficiently between 27 and 41 d than the other treatment groups. The absence of dietary linoleic acid decreased liver and spleen weights and, in general, suppressed development of organs except the brain. Red blood cell hemolysis was not affected by treatment. Although showing signs of chronic mild diarrhea, the GN neonatal ruminant differed in Cl- concentration and dry matter percentage of its lower bowel contents from the "classic rodent model." The results indicate that neonatal colostrum-deprived lambs have an EFA requirement, as evidenced by decreased growth and performance characteristics in the GN linoleic deficient vs GN supplemented group, and suggests that the required level is in excess of .32% of the total caloric intake as linoleic acid.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Influence of body condition and lasalocid during late gestation on blood metabolites, lamb birth weight and colostrum composition and production in Finn-cross ewes.

The objective of this experiment was to examine the effects of body condition (BC) and of lasalocid (L) the last 4 wk of gestation on blood metabolite profiles, lamb birth weight and colostrum composition and production. Twenty-eight 3-yr-old Finn-Targhee ewes (73 kg) were assigned randomly within BC grouping (2.5 or 3.5) and individually fed a diet of 90% alfalfa pellets and 10% of a supplement containing either no L (C = control) or L. Gestation and 24-h postlambing weights were higher (P less than .05) for 3.5 BC versus 2.5 BC ewes. Lasalocid had no effect (P greater than .05) on ewe weight. Average number of lambs born within treatment groups were similar (P greater than .05). Total kilograms of lamb born were greater (P less than .10) for 3.5 BC ewes. Body condition 3.5 ewes had greater concentrations of total protein (P less than .10) and albumin (P less than .05) the last 4 wk of gestation than those in the 2.5 BC group. Feeding L decreased (P less than .05) blood urea-N in comparison with C ewes. Colostrum composition and production were not influenced (P greater than .05) by BC, L or number of lambs born. Serum 3-hydroxybutyrate seemed to be a good indicator of energy metabolism; albumin and blood urea-N concentrations reflected dietary protein intake. Lasalocid had a minimal effect on nutrient metabolism and productivity of ewes fed in excess of the NRC (1985) protein and energy requirements.

Animals↗

Isolation of two new proteins from bovine colostrum which stimulate epidermal growth factor-dependent colony formation of NRK-49F cells.

Defatted bovine colostrum and decaseinated whey contain biological activity similar to that of TGF-beta in that they stimulate the anchorage-independent growth of NRK-49F cells in the presence of EGF. Two new proteins with such activity, termed BC-1 and BC-2, were isolated from decaseinated colostrum by a sequence of DEAE-Sephacel chromatography, Sephadex G-100 gel filtration in 1 M acetic acid, Sephadex G-50 re-gelfiltration in 8 M urea-1 M acetic acid and reverse-phase FPLCs. The two proteins showed distinctly different molecular weight from that of TGF-beta 1. BC-1 was a small Mr peptide of 8.5 kD. BC-2 had molecular mass of 46 kD, which yielded peptides of 46,40, and 6 kD on reduction. In contrast to TGF-beta, both proteins did not lose their biological activity on reduction. Both BC-1 and BC-2 suppress DNA synthesis in concanavalin A-stimulated thymocytes. These results suggest that BC-1 and BC-2 belong to a new class of mitogen/inhibitors, though their biological activities resemble those of TGF-beta.

Animals↗

Absorption of protein and IgG in calves fed a colostrum supplement or replacer.

Newborn Holstein bull calves (n = 32) were assigned to receive a colostrum supplement (CS) containing defibrinated bovine plasma or a colostrum replacer (CR) containing an immunoglobulin concentrate obtained by concentrating the immunoglobulin (Ig)G fraction of bovine plasma. The CS and CR contained 11.1 and 21.2% of dry matter as IgG, respectively. Each animal was fed two 454-g feedings at 1 and 8 h of age. The two feedings of CS and CR provided 95 and 187 g of IgG, respectively. Mean plasma IgG at 24 h of age was 8.0 and 13.6 g/L in calves fed CS and CR, respectively, indicating acceptable absorption of Ig from both sources. Mean apparent efficiency of IgG absorption in calves fed CS and CR were 33 and 30%, respectively, and did not differ between treatments. Mean plasma total protein at 24 h in calves fed CS and CR were 4.99 and 4.98 g/dl and did not differ between treatments. Increased plasma protein concentration from 0 to 24 h (4.5 g/L) was lower than the mean increase in plasma IgG concentration during the same period (10.3 g/L), indicating altered protein profile in the blood during the first 24 h of life. Correlation between plasma IgG and total protein at 24 h of age was significant within treatment, but the relationship between IgG and protein in plasma at 24 h varied between treatments. Predicted plasma total protein concentrations at 10 g of IgG/L of plasma at 24 h were 5.4 and 4.2 g/dl, in calves fed CS and CR, respectively. Prediction of plasma IgG concentration using total plasma protein may be inappropriate when calves are fed CS or CR.

Absorption↗

Insulin-like growth factor-I and its binding proteins in colostrum compared to measures in serum of Holstein neonates.

Colostral insulin-like growth factor-I (IGF-I) may be beneficial in the development of gastrointestinal tracts of bovine neonates. Thus, the purpose of this study was to examine relationships among concentrations of IGF-I and IGF-binding proteins (IGFBP) in colostrum used at two initial feedings and serum concentrations of IGF-I, IGFBP, total protein, gamma glutamyltransferase (GGT), and immunoglobulin G at 0 and 48 h after birth in Holstein neonates. Calves (n = 22) were separated from dams immediately after birth. Blood samples were taken before initial feeding and at 48 h after birth. Calves were fed 2 L of colostrum twice and milk replacer thereafter. Linear regression of serum IGF-I at 48 h and colostral IGF-I revealed a significant positive relationship (R2 = 0.204). Serum IGFBP-3 at 48 h and colostral IGFBP-3 also had a positive relationship (R2 = 0.143). However, linear regression of colostral IGF-I on the difference in serum IGF-I at 48 and 0 h was not significant. Calves were assigned to group 1 (0-h serum IGF-I < 10 ng/ml; n = 11) or group 2 (0-h serum IGF-I > or = 10 ng/ml; n = 11) for further analysis. There were no differences in serum IGF-I or IGFBP-2, -3, -4, and -5 concentrations at 48 h between groups 1 and 2. Correlation coefficients revealed negative relationships of serum IGF-I at 0 h to the difference between serum IGF-I at 48 and 0 h (r = -0.824), as well as birth weight of the calf to the amount of GGT at 48 h (r = -0.604). Females had lower birth weights than males, but sex of calf did not affect serum measures. At 0 h, but not 48 h, total serum protein was correlated to serum GGT concentrations (r = 0.573). From indirect evidence, absorption of colostral IGF-I and IGFBP-3 into systemic circulation may occur, but relative importance compared to endogenous sources is uncertain.

Animals↗

Effect of suckling followed by bottle feeding colostrum on immunoglobulin absorption and calf survival.

Dairy claves (983 head) were left with their dams to suckle for 1 day following birth. They then were fed 1 liter of pooled colostrum and placed in individual pens. Blood samples from each calf were taken prior to and 24 h following colostral feeding. Analysis of blood for immunoglobulins G and M indicated that 42% of the calves had failed either to suckle or to absorb colostral immunoglobulins from suckling. Concentrations of IgG and IgM in serum of calves prior to bottle feeding ranged from 0 to 63 mg/ml and 0 to 15 mg/ml with means of 11.3 and 2.9 mg/ml. From bottle feeding 1 liter of colostrum, 70% of the calves absorbed colostral immunoglobulins, an average increase of 5.0 +/- 4.5 mg/ml for IgG and 1.3 +/- 1.3 mg/ml for IgM. Some calves did not absorb IgG from either suckling or bottle feeding (11.9%) and 20.8% failed to absorb IgM. Mortality rate for all experimental calves was 3.9%; for 83 agammaglobulinemic calves, 13.3%. The high rate of survival (86.7%) of the 83 agammaglobulinemic calves could be due to the local prophylactic effect of colostral immunoglobulins in the intestine even though closure had occurred.

Animals↗

Early variations of blood plasma gamma-glutamyl transferase in newborn calves--a test of colostrum intake.

Fourteen newborn calves had low gamma-glutamyl transferase catalytic activity concentration in blood plasma (10 to 31 U/liter). Colostrum intake induced a dramatic increase of plasma gamma-glutamyl transferase activity, which was maximal on the 1st or 2nd day (370 to 5000 U/liter) after birth; then plasma enzyme activity slowly decreased and was stable about 100 U/liter on the 20th day. In two calves given either boiled or synthetic milk, plasma gamma-glutamyl transferase activity was not higher than 25 U/liter. Gamma-glutamyl transferase activity of blood plasma in newborn calves is an easy and inexpensive test of colostrum intake.

Animals↗

Distribution of supplemental selenium in the serum, hair, colostrum, and fetus of parturient dairy cows.

A selenium supplementation study was conducted with Holstein cows to evaluate amounts and methods of administration, namely, oral supplementation (1 vs. 2 mg/head/d, last 60 d of gestation) and intramuscular injection (50 mg at 40 and 20 d prepartum vs. 50 mg at 60, 40, and 20 d prepartum). Blood was collected every 10 d, starting 60 d prepartum and extending 20 d postpartum for cows, and at birth to 20 d for calves. Hair samples of cows were obtained at 60 to 40 and 20 d prepartum, and of the calves at birth. Colostrum samples were obtained for assay. Oral (2 mg/d) and both injection treatments resulted in increased selenium in serum of cows, but selenium in serum of calves at birth was greater only for those whose dams received selenium by injection. Selenium in hair of cows was increased by both injection treatments as well as by the oral treatment of 2 mg/d. Selenium in hair of calves was increased by all methods of administering selenium to the dams. Selenium in colostrum was not affected by any treatment. Thus, pregnant dairy cattle respond to selenium supplementations of a daily oral intake of 2 mg/head/d, the last 60 d of gestation, or by injection of 50 mg each on d 40 and 20 prepartum.

Animals↗