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Toxoplasmosis, caused by Toxoplasma gondii, poses a significant threat to sheep flocks, affecting reproductive performance and meat quality, and leading to economic losses. This study aimed to evaluate the correlation between anti-T. gondii IgG antibodies in the serum and colostrum of naturally infected ewes and to assess passive immunity in newborn lambs. Blood and colostrum samples were collected from 162 ewes and 182 lambs across 20 sheep farms in Paraíba, Brazil. Samples were tested for anti-T. gondii and anti-Neospora caninum IgG using indirect fluorescence antibody tests (IFATs), with titers ≥ 1:64 considered positive. Among the ewes, 45.1% tested positive for anti-T. gondii IgG in serum, with titers ranging from 1:64 to 1:16,384. The colostrum from 94.6% of the ewes also tested positive, although 74% had higher titers in their serum than in their colostrum. Concordance between serum and colostrum was high, with a kappa coefficient of 0.950. Lamb serum showed a perfect agreement with maternal colostrum (kappa = 0.962), demonstrating effective passive transfer of antibodies. This study confirms that colostrum is a reliable matrix for detecting anti-T. gondii antibodies and assessing passive immunity in lambs. The high concordance between serum, colostrum, and lamb titers suggests that IFATs on colostrum can be a practical tool for monitoring maternal antibody transfer, contributing to the better management of T. gondii infections in sheep flocks. [ABSTRACT FROM AUTHOR]

Toxoplasmosis, caused by Toxoplasma gondii, poses a significant threat to sheep flocks, affecting reproductive performance and meat quality, and leading to economic losses. This study aimed to evaluate the correlation between anti-T. gondii IgG antibodies in the serum and colostrum of naturally infected ewes and to assess passive immunity in newborn lambs. Blood and colostrum samples were collected from 162 ewes and 182 lambs across 20 sheep farms in Paraíba, Brazil. Samples were tested for anti-T. gondii and anti-Neospora caninum IgG using indirect fluorescence antibody tests (IFATs), with titers ≥ 1:64 considered positive. Among the ewes, 45.1% tested positive for anti-T. gondii IgG in serum, with titers ranging from 1:64 to 1:16,384. The colostrum from 94.6% of the ewes also tested positive, although 74% had higher titers in their serum than in their colostrum. Concordance between serum and colostrum was high, with a kappa coefficient of 0.950. Lamb serum showed a perfect agreement with maternal colostrum (kappa = 0.962), demonstrating effective passive transfer of antibodies. This study confirms that colostrum is a reliable matrix for detecting anti-T. gondii antibodies and assessing passive immunity in lambs. The high concordance between serum, colostrum, and lamb titers suggests that IFATs on colostrum can be a practical tool for monitoring maternal antibody transfer, contributing to the better management of T. gondii infections in sheep flocks.

Vitamin B-6 status, assessed by plasma pyridoxal phosphate (PLP) concentrations, and vitamin B-6 concentrations in breast milk were examined in 47 lactating mothers supplemented with different amounts of pyridoxine.HCl (PN.HCl) during pregnancy and the first 6 mo of lactation. PLP concentrations in cord blood and maternal plasma at 2 d postpartum and vitamin B-6 concentration in colostrum were positively correlated with the amount of PN.HCl supplementation prenatally (r = 0.71, p less than 0.001; r = 0.74, p less than 0.001; and r = 0.78, p less than 0.001, respectively). Correlations between the amounts of PN.HCl supplementation postnatally and plasma PLP concentrations increased with the length of supplementation. Plasma PLP concentrations were also correlated with vitamin B-6 concentrations of milk samples, which were obtained on the same day as plasma. PN.HCl supplements between 2.5 and 4.0 mg/d (2.1-3.4 mg PN equivalents) ensured vitamin B-6 adequacy of the mother and maintained relatively saturated concentrations of vitamin B-6 in breast milk.

The composition of colostrum and milk and of sows fed either 3.4 kg or 1.0 kg daily of a conventional sow diet during the last 14 days of gestation was investigated. The fat content of colostrum of the very restrictedly fed sows was higher compared with the standard fed sows. There was a significantly decrease in the crude protein and immunoglobulin contents of colostrum during the first 4 h post farrowing. The milk composition 14 days after farrowing was not affected by the late pregnancy feeding level. The composition of milk was significantly correlated with the 3 week litter weight. The colostrum IgG content was not correlated to the mortality of piglets during the first week post favouring. The content of crude protein and immunoglobulins in the milk was not correlated with the frequency of post weaning diarrhoea.

Lactoferrin content of colostrum obtained from cows within 24 h after parturition was measured using a single radial immunodiffusion test and was compared among cows of two dairy breeds (Holstein-Friesian, Jersey) and two beef breeds (Japanese Black and Japanese Brown). Average lactoferrin content in colostrum of dairy breeds was 2 mg/ml and in colostrum of beef breeds was .5 mg/ml. Lactoferrin content of colostrum due to lactation number was also different among breeds. In dairy breeds, multiparous cows had lactoferrin content two to three times higher than that of primiparous cows; beef breeds showed no obvious differences between lactation years. Lactoferrin content also varied considerably within breed. In beef breeds, half the cows had values of nearly zero. Transferrin content in colostrum was fairly constant (.9 mg/ml) and was not as variable among and within breeds. There was no correlation between lactoferrin and transferrin contents in colostrum. Examination of cows lacking lactoferrin suggested that transferrin plays an important role as an iron carrier from a cow to her newborn calf.

Immunoaffinity columns made with monoclonal antibodies to either human or bovine lactoferrins were prepared to isolate human lactoferrin or bovine lactoferrin from milks by a single chromatographic step. Recoveries of human lactoferrin and bovine lactoferrin were 98 and 97%, respectively. The human lactoferrin recovered from defatted human colostrum was 98% pure with 93% iron-binding capacity. Amount of recovered bovine lactoferrin, as well as purity and iron-binding capacity, varied widely depending on the source of bovine milks and pretreatments (particularly pasteurization temperature). The best source to isolate bovine lactoferrin was raw skim milk yielding a protein 97% pure and with a 99% iron-binding capacity. Thus, immunoaffinity chromatography provides an effective one-pass isolation of highly pure human or bovine lactoferrin with reasonable recovery and iron-binding capacity.

Growth-promoting activity in bovine colostrum has been detected as the capacity to stimulate protein synthesis in L6 myoblasts. By using this assay as a measure of bioactivity, a growth factor has been purified to near homogeneity from centrifuged colostrum by a series of steps including acid extraction, chromatography on sulphopropyl-Sephadex, followed by adsorption to, and elution from, C18 columns using acetonitrile and propan-1-ol gradients. The purified growth factor has a low solubility at neutral and alkaline pH and has an Mr of 7800 by gel-permeation chromatography. Sequence analysis of the first 30 amino acids from the N-terminus indicated complete identity in this region with human insulin-like growth factor-1. Accordingly we conclude that the purified growth factor is bovine insulin-like growth factor-1.

Colostrum removed from mammary glands of parturient cows before letdown contained no greater concentration of immunoglobulins than colostrum collected after letdown and complete milking. Greater absorption of colostral immunoglobulins in calves suckling their dams initially postpartum over those hand fed by buckets or bottles is not from higher concentration of immunoglobulins in the gland and teat cistern. Colostral concentrations of three immunoglobulin classes (G, M, and A) obtained from one partial and six consecutive complete milkings at 12-h intervals postpartum decreased at different rates over time or number of milkings. Potential prophylactic value is discussed of continued lacteal secretion of immunoglobulins A and M available to the calf after intestinal closure to systemic absorption of colostral immunoglobulins.

Protease inhibitors (inhibiting trypsin, chymotrypsin, and elastase) were demonstrated in human milk from birth to 4 months after delivery. No pepsin inhibitor was found. The protease inhibitors were localized in the alpha 1-region--inhibiting trypsin, chymotrypsin, and elastase--and in a more cathodal region--inhibiting chymotrypsin--in agarose gel electrophoresis of human milk. alpha 1-antitrypsin and antichymotrypsin were demonstrated by crossed immunoelectrophoresis. Electroimmunoassay showed the concentration of alpha 1-antitrypsin in 1st day milk to be 10.9% and the concentration of antichymotrypsin was 116% of that of adult serum. The concentrations decreased during the 1st wk; from 1 wk to 4 months they were 1.6% for alpha 1-antitrypsin and 3.8% for antichymotrypsin of those of adult serum. One ml of human milk from the first 3 days inhibits, by enzymatic methods, 0-150 microgram (mean value, 70 microgram) trypsin. Milk from 1 week after delivery and later inhibits 0-65 microgram (mean value, 38 microgram) trypsin per ml.

Colostrum was inoculated with Streptococcus lactis or yogurt culture or preserved with .1% (vol/vol) formalin in two separate experiments. All preparations then were stored at ambient temperature for 24 days. With increasing storage time, a larger proportion of the total colostrum nitrogen was not precipitated in 10% (wt/vol) trichloroacetic acid. By day 24, this represented 30 to 35% for the fermented samples and 10 to 15% for the formalin preserved samples. The majority of this nonprecipitable nitrogen was amino acids and small peptides. Most of the nitrogen in colostrum even after 24 days of storage should be nutritionally useful to the calf because even the nonprecipitable portion is amino acids and peptides.

Bovine beta 2-microglobulin (beta 2-m), the light chain of the histocompatibility antigen, was isolated in crystalline form from colostrum. Previous studies from this laboratory on the solution properties of this protein suggest the existence of a time-dependent multiple aggregation phenomenon. To clarify the molecular states of beta 2-m, its solution properties were studied by ultracentrifugation and spectropolarimetry. Sedimentation equilibrium experiments at pH 5.0 (0.08 M NaCl, 0.02 M sodium phosphate) at concentrations less than 0.3 mg/ml give Mr = 11,800. From sedimentation velocity results, we conclude that bovine beta 2-m is a much more symmetrical and compact molecule than either guinea pig or human beta 2-m. At concentrations above 0.4 mg/ml under the same conditions, sedimentation equilibrium experiments show that a monomer to tetramer reversible self-association occurs. Also, the tetramerization increases with decreasing temperature. beta 2-Microglobulin undergoes an irreversible temperature-dependent association to a much larger aggregate over a period of 7 days, as evidenced by sedimentation equilibrium and velocity results. The rate of this aggregation decreases as the pH approaches the isoelectric point (pH 7) from either side. Furthermore, circular dichroism measured at pH 5.0 under time-dependent aggregating conditions showed a marked increase in the percentage of disordered structure, leading to the conclusion that this effect is a denaturation phenomenon.

Use of a microwave oven to thaw frozen colostrum was evaluated. Colostrum was collected from nine cows, four of which were immunized to produce specific colostral antibodies. Colostrum from each cow was frozen, subsequently thawed, and pooled. One-liter aliquots of the pooled colostrum were frozen and assigned randomly to three thawing treatments. Colostrum was thawed using one of three regimens: 10 min in a microwave oven at full power (650 W), 17 min in a microwave oven at half power (325 W), and 25 min in 45 degrees C water. Colostrum thawed in the microwave oven was slightly coagulated and had lower volume and total protein content than colostrum thawed in water. Casein and pH were not different among treatments. Both concentration and total content of immunoglobulin A were higher in the control than in microwave treatments. Neither amount nor concentration of immunoglobulin G and immunoglobulin M were different among treatments. Immunological activity, measured by a hemolytic test, was lower for microwave treatments than the control but did not differ between microwave treatments. Frozen colostrum thawed in a microwave oven should provide a reasonable source of colostrum when fresh high quality colostrum is not available.

Milk urea nitrogen (UN) concentration was examined as a possible index to protein-energy intake in female collared peccaries (Tayassu tajacu). Captive adults were bred and assigned to one of four experimental diets through gestation and lactation. Females fed a high protein diet produced milk with UN concentrations exceeding those of low-protein-fed females. A low energy intake tended to elevate UN concentrations in milk.

New neutral oligosaccharides from cow colostrum kappa-casein were identified and characterized by 500-MHz 1H-NMR spectroscopy. Their structures are Gal beta(1----3)GalNAc-ol, Gal beta(1----3)[GlcNAc beta(1----6)]GalNAc-ol, Gal beta(1----3)[Gal beta(1----4)GlcNAc beta(1----6)]GalNAc-ol, Gal beta(1----3)[Fuc alpha(1----3)[Gal beta(1----4)]GlcNAc beta(1----6)]GalNAc-ol. The tetrasaccharide and the cow colostrum kappa-caseinoglycopeptide which contains this oligosaccharide inhibit the hemagglutination of blood group I human erythrocytes. In cow mature milk only the disaccharide is characterized. The variability of these neutral oligosaccharides in cow kappa-casein as a function of time after calving is studied.

A simplified and accurate method for determination of naturally occurring vitamin D3 in bovine milk was established by high-performance liquid chromatography (HPLC) using successively reversed-phase and straight-phase columns. Exactly 25.0 ml of a sample of bovine milk was taken and the lipid was extracted with a solvent mixture of petroleum ether and ethyl ether (1:1) with small amounts of ethanol and Triton X-100, present. The extracted lipid was subjected to the first preparative HPLC using a Nucleosil 5C18 column (reversed-phase type) with acetonitrile-methanol (1:1) as the mobile phase, and a fraction containing vitamin D3 was isolated. The fraction was subsequently subjected to the second analytical HPLC using a Zorbax SIL column (straight-phase type) with 0.4% isopropanol in n-hexane as the mobile phase. Vitamin D3 was assayed by estimating the peak height on the chromatogram. The overall recovery and CV values were 92.1 +/- 8.7% and 9.4%, respectively, which were satisfactory. The proposed method was applied to several kinds of colostrum, early and later bovine milk in pairs. The assayed values in the colostrum of the group with large amounts of vitamin D3 administered before delivery to prevent parturient paresis were higher than those in the group with no administration. However, the values of the former group generally decreased in the respective early and later milk in step and there were few differences among those in the later milk of the two groups. The assayed values in later milk were 30-80 IU/liter.

A 22-mo experiment was conducted to determine effects of mineral supplementation of cows on mineral elements in milk, colostrum, and blood serum. Purebred Brahman beef cows grazing pastures grown on sandy and well-drained soils were allotted to three treatments of 12 cows each as follows: control, a complete commercial mineral mixture, control plus dietary and injected Se, and control plus fortified mineral mixture. Milk and serum were analyzed for minerals 2 consecutive yr at parturition and 3 mo postpartum. Mean cow serum, Ca, Mg, P, and packed cell volume values were higher at parturition than 3 mo later, whereas serum Cu was highest at 3 mo. Mean serum concentrations were affected by year with higher Ca and hemoglobin and lower Cu concentrations in 1982 than in 1981. Increasing dietary Se resulted in higher serum Se. Colostrum was higher in Mg, P, Cu, Fe, Se, and Zn than milk, whereas milk was higher than colostrum in Mn. Means for treatment X time showed cows fed dietary and dietary plus injected Se and vitamin E had more Se in milk than milk from cows fed no supplemental Se.

Samples of milk were taken at intervals during lactation from 35 mothers of term and 26 mothers of preterm infants and assayed for 8 B complex vitamins. Both term and preterm milks varied widely in vitamin content between mothers. Mean concentrations of thiamin, vitamin B6, nicotinic acid, pantothenic acid, biotin, and folic acid increased progressively over several weeks after parturition but vitamin B12 concentrations declined generally and riboflavin values showed little change. Preterm milk was not richer in vitamins than term milk of the corresponding stage of lactation and it appeared that intake of B vitamins differed widely among preterm infants given their own mothers' milk. These infants may have meagre body reserves and an increased need for vitamins, and breast milk whether from their own mother or from the milk bank may not meet their needs. There is a strong case for supplementing breast milk given to preterm babies with the B complex vitamins.

Porcine colostrum casein contained at least one major polypeptide band in both acid gel and sodium dodecyl sulfate gel electrophoresis that did not appear in patterns of casein prepared from porcine milk. In sodium dodecyl sulfate gel patterns, this polypeptide possessed a molecular weight of 62,000 and stained positively with "Stains-all" but not with periodic acid-Schiff reagent. Several minor caseins that appeared in acid and alkaline gel patterns from colostrum could not be detected in casein prepared from milk. Seven minor polypeptides in sodium dodecyl sulfate gel patterns of whey proteins prepared from porcine colostrum could not be detected in milk. Identical acid and alkaline gel patterns were obtained for whey proteins prepared from colostrum and milk. Only the sodium dodecyl sulfate gel electrophoretic pattern of casein prepared from milk, and possibly the alkaline gel electrophoretic pattern of whey protein prepared from milk, contained polypeptides not in patterns from colostrum. Total phosphorus contents of colostrum and milk casein were 2.96% and 3.78%. Casein prepared from porcine colostrum contained almost twice as much hexosamine and slightly elevated N-acetylneuraminic acid as casein prepared from milk. Hexose content was nearly equivalent. Whey protein prepared from milk contained more total hexose than casein prepared from colostrum, whereas N-acetylneuraminic acid and hexosamine contents were nearly equivalent.

With a simple, sensitive, and rapid SDS-PAGE procedure, developed during the course of this study, calmodulin was not found in cows' milk. The absence of calmodulin at the level of detection provided by this procedure (.4 micrograms) was unexpected. Because milk is a repository for many protein and enzyme systems, and because the mammary gland is known to contain calmodulin, it was conjectured that at least some "leakage" of the protein into milk would occur, especially into colostrum.

A disaccharide was isolated from rat mammary tissue and determined to be 6-O-beta-galactopyranosyl myo-inositol (6-beta-galactinol) on the basis of combined gas-liquid chromatography-mass spectrometry of the permethylated and peracetylated derivatives as well as previously reported chemical and enzymatic evidence. 6-beta-Galactinol, also found in rat milk, increased during lactation, and on the 18th day represented approximately 17% of the total non-lipid neutral myo-inositol. The sugar was absent in all other rat tissues examined, suggesting a unique association with the process of lactation. This novel galactoside of one human subject on the basis of paper and gas-liquid chromatography.

Antigenic components of buffalo (Bos bubalis) serum, which were also components of buffalo colostrum, seminal plasma, milk whey, saliva, and tears, were investigated by the ager gel diffusion test and immunoelectrophoresis. Immunoglobulins of buffalo serum were identified by immunoelectrophoresis employing rabbit-anti-buffalo serum and rabbit-anti-buffalo gamma-globulin. Based on immunoelectrophoretic patterns immunoglobulin G (IgG), IgGA, and IgM were detected both in the serum and colostrum of buffaloes. Tears contained both IgG and IgM. Cross-reactions of buffalo serum with seminal plasma, saliva, and milk whey were observed only in the IgG region. By polyacrylamide gel electrophoresis, lipoprotein (5.2% +/- 0.41), IgM (11.4% +/- 3.1), IgG (9.4% +/- 0.98), haptoglobin 21.8% +/- 3.73), transferrin (10.4% +/- 2.15), ceruloplasmin (7.8% +/- 1.3), postalbumin (20.8% +/- 2.09), and albumin (13.7% +/- 0.75) were identified provisionally.

Colostral immunoglobulin concentrations were compared among five breeds of dairy cows. Colostrum samples (3 to 4 kg) were obtained immediately after parturition from 19 Ayrshire, 17 Brown Swiss, 12 Guernsey, 19 Holstein, and 5 Jersey cows. Average total colostral immunoglobulin was 8.1, 6.6, 6.3, 5.6, and 9.0% for them. Colostrum from Jerseys had the highest immunoglobulin G (6.65%), immunoglobulin A (1.86%), and immunoglobulin M (.53%) of all breeds. Colostrum from Holsteins was lowest in immunoglobulin G (4.12%) whereas that from Guernseys was lowest in immunoglobulin A (.90%) and immunoglobulin M (.39%). Breeds may differ, and the high calf mortality and health problems frequently reported for Guernseys may be related to the low immunoglobulins A and M in colostrum. Concentration of colostral immunoglobulin by calving number was examined over breeds for 23, 16, 17, and 14 cows by first, second, third, and fourth or later calvings. Colostral immunoglobulin was lower from first-calf heifers (5.68%) than from third (7.91%) and fourth (7.53%) parity cows. Colostral immunoglobulins A and G tended to be lower in first lactations.

Cows from five dairy herds were used to determine persistence of antibiotic residues in colostrum and milk following dry cow therapy. Cows were treated in all quarters at drying off with antibiotics approved for use for nonlactating cows. Antibiotics procaine penicillin G plus dihydrostreptomycin, novobiocin, cloxacillin, or cephapirin were compared with no treatment. Composite colostrum samples were collected from each cow at first milking after parturition. Samples were screened for residues by Delvotest P. Colostrum samples positive by Delvotest also were tested by Bacillus stearothermophilus disc assay. Four of 186 colostrum samples from cows treated with antibiotics at drying off were positive for residues by Delvotest. Only one was confirmed positive by disc assay following heat treatment. All colostrum samples from 48 cows not treated were negative. Samples of first marketable milk also were collected. Over 96% of milk samples from cows treated at drying off and 100% of milk samples from cows not treated were negative for residues by Delvotest. If manufacturer's recommendations are followed, antibiotic residues in colostrum and milk following dry cow therapy with products in our study should not be a significant problem.

Pre-colostrum and colostrum from goats cause a marked inhibition of the binding of 125I-labelled epidermal growth factor (125I-EGF) to Swiss 3T3 cells. The ability of these secretions to inhibit 125I-EGF binding is closely correlated with the ability to stimulate DNA synthesis in quiescent 3T3 cell cultures, suggesting that goat mammary secretions may contain an EGF-related mitogen. However, the material in colostrum which inhibits 125I-EGF binding to Swiss 3T3 cells is a basic protein with Mr greater than 20000 and is thus quite different from mouse and human EGF. Furthermore, the colostral-mediated inhibition of 125I-EGF binding, although rapid and apparently competitive, differs from the inhibition of binding induced by native, unlabelled EGF. Thus, the inhibitory effect of colostrum is markedly decreased when the assay temperature is shifted from 37 degrees C to 4 degrees C whereas unlabelled EGF is an effective competitive inhibitor at both 37 degrees C and 4 degrees C. Incubation of cells with EGF causes a reduction in cell surface EGF receptors whereas exposure to colostrum does not induce down-regulation of the EGF receptor. Our results suggest that the colostral factor does not bind directly to EGF receptors but inhibits 125I-EGF binding by an indirect mechanism which involves a temperature-sensitive step.

The half-life of IgG1 immunoglobulin was measured in six neonatal calves following a meal of iodine-125 labeled IgG1 in colostrum derived from their dams. The half-life as measured by the decrease in plasma concentration of IgG1 was 19.9 +/- 1.9 days. However, the half-life as measured by the disappearance of [iodine-125]IgG1 from the plasma was 11.5 +/- .6 days. The latter value is closer to the true half-life because it is not affected by endogenous production of IgG1 by the tissues of the young calf. A decrease in the specific activity of plasma [iodine-125]IgG1 with time representing the body "pool" of IgG1 (half-life 25.8 +/- 6.1 days) suggests that the calf from birth to about 20 days of age is capable of synthesizing a significant amount of IgG1 immunoglobulin.

The concentrations of iron, copper, zinc, calcium, magnesium, sodium, potassium, protein, carbohydrate, and fat were analyzed in milk from rhesus monkeys (Macaca mulatta) during the course of lactation. Concentrations of iron, copper, zinc, magnesium, sodium, potassium, and protein were higher in milk of early lactation (colostrum) than in mature milk, while concentrations of calcium increased with lactation time. Concentrations of zinc in monkey colostrum and mature milk were similar to that of human milk, while iron, copper, calcium, and protein concentrations were higher than in human milk.

Twenty Holstein calves received 2 L of colostrum twice within 12 h after birth; the first feeding occurred within 2 h of parturition. The increase in adsorption efficiency was related to the gamma globulin provided in the first colostrum feeding. Absorption efficiency ranged from 2.4 to 46.1%. The number of sheddings of rotavirus and Cryptosporidium by the calves during their first 4 wk of life was associated with serum gamma globulin concentration 24 h after birth and absorption efficiency. Absorption efficiency and body weight combined accounted for 60.4% of the variation in the number of sheddings; heavier calves shed more than lighter calves. During the first 4 wk, calves that shed more frequently gained less weight; weight gain was also associated with serum gamma globulin levels 24 h after birth. Colostrum composition varied between quarters of the same cow. Total protein and gamma globulin content of colostrum from the rear quarters was higher than from the front quarters. The association between number of sheddings and absorption efficiency suggest that calves should not be fed colostrum containing less than 9 g/100 ml of total protein.

Human lactoferrin has been purified to apparent homogeneity with high recovery (greater than 95%) in one chromatographic step using immobilized single-stranded DNA-agarose. Human colostral whey, at neutral pH, was loaded onto columns of DNA-agarose; a single purified protein, lactoferrin, was eluted using elevated ionic strength buffers. The lactoferrin purified in this manner was shown to be homogeneous by high-performance ion-exchange chromatography (Mono-S), immobilized metal ion (Cu2+) affinity chromatography, and reverse-phase (C18) chromatography. Electrophoresis on SDS-polyacrylamide gradient (10-20%) gels and silver staining showed the purified lactoferrin preparation to contain a single protein of 78 kD with intact immunologic determinants. Similar results were obtained before and after iron saturation of the colostral whey proteins. Apolactoferrin purified in this manner was shown to bind iron with high affinity. These results demonstrate the effectiveness of immobilized DNA as one of the most rapid and complete lactoferrin purification procedures described thus far.

A practical field method for measuring immunoglobulin concentration in bovine colostrum has been developed from the linear relationship between colostral specific gravity and immunoglobulin concentration. Fourteen colostrums were collected within 24 h postpartum from nursed and unnursed cows and were assayed for specific gravity and major colostral constituents. Additionally, 15 colostrums were collected immediately postpartum prior to suckling and assayed for specific gravity and immunoglobulin concentration. Regression analysis provided an equation to estimate colostral immunoglobulin concentration from the specific gravity of fresh whole colostrum. From this, a colostrometer was developed for practical field use.

beta 2-Microglobulin has been isolated and crystallized from bovine colostrum and represents the only crystalline form of this protein reported. The complete sequence of bovine beta 2-microglobulin was determined using only one proteolytic enzyme, Staphylococcus aureus V8 protease. Automated degradation of the intact molecule and two large peptides produced by enzymic digestion provided unequivocal placement of all residues. Bovine beta 2-microglobulin, molecular weight 11,630, contains 98 residues as compared with 99 for the human, rabbit, guinea pig, and murine proteins. The valine residue at position 49 in all the aforementioned species is deleted in the bovine variant. This crystalline protein is also uniquely characterized by three di-prolyl sequences in the first third of the molecule. beta 2-Microglobulin is a highly conserved protein, and in a comparison of absolute amino acid sequence differences among species, bovine beta 2- microglobulin ranges from a low of 24 substitutions as compared with the rabbit homologue and 26 for human to 32 for mouse and guinea pig.

Samples of colostrum from both Ethiopian and Norwegian women contained antirotavirus activities of immunoglobulin and non-immunoglobulin nature. No significant differences in rotavirus immunoglobulin A or in rotavirus-inhibiting activity were found between samples from the two countries. The non-immunoglobulin inhibitory activity was trypsin sensitive and heat stable (100 degrees C for 10 min). Escherichia coli heat-labile enterotoxin antibodies were measured in the colostrum samples by enzyme-linked immunosorbent assay. No E. coli enterotoxin-specific immunoglobulin A was detected, possibly due to the high background caused by the nonspecific adsorption of immunoglobulin A to the enzyme-linked immunosorbent assay plates in the absence of toxin. A total of 5 of 15 Ethiopian colostrum samples and 0 of 11 Norwegian colostrum samples neutralized the effect of E. coli heat-labile enterotoxin on YI adrenal cells. Both the Ethiopian and the Norwegian colostrum samples contained a non-immunoglobulin enterotoxin-inhibitory activity when the toxin was measured by enzyme-linked immunosorbent assay. This inhibitory activity was not trypsin sensitive, and extraction by chloroform-methanol indicated that the inhibitor was of a lipid nature.

Using a combined indirect immunoperoxidase (PAP)/direct immunofluorescence (IF) staining technique, pregnancy associated alpha 2-glycoprotein (alpha 2-PAG) has been shown to be present solely in the IgA producing plasma cells. alpha 2-PAG was undetectable in plasma cells synthesizing any of the other immunoglobulin isotypes (IgG, IgM, IgD and IgE). Furthermore, we have observed increased numbers of alpha 2-PAG positive cells in sites recognized for their local IgA production e.g. lactating breast, salivary gland, respiratory and gastrointestinal mucosae. In support of this cellular association with IgA, alpha 2-PAG was measured by enzyme immunoassay in colostrum, breast milk, saliva, jejunal secretions, bile and urine. The relevance of this novel and preferential association of immunosuppressive alpha 2-PAG with IgA is discussed with regard to the regulation of gut immunoresponsiveness.

Human milk protein was determined by three colorimetric methods and by Kjeldahl analysis. The distribution of nitrogen (N) and protein was determined within various milk compartments. Total N, whey, casein, nonprotein nitrogen (NPN), cell N and N in the fat fraction were analyzed by micro-Kjeldahl analysis after a series of centrifugation and ultracentrifugation separations. Fresh milk samples (colostrum, transitional milk and mature milk) were centrifuged at 500 X g to separate milk cells and at 5000 X g to skim the milk. Decelled milk and skimmed milk were ultracentrifuged at 189,000 X g to separate fat and casein micelles from whey. NPN was analyzed after trichloroacetic acid precipitation. Whole milk, decelled milk, skimmed milk and whey were analyzed for protein with the Lowry method, modified for fat-containing samples, the Bradford dye-binding assay (Bio-Rad) and the Pierce bicinchoninic acid (BCA) assay. Cell nitrogen had a tendency to be lower in mature milk than in colostrum. Colostrum contained only 6% casein protein, whereas mature milk contained 13%. Fat from skimming was lower in N than fat from ultracentrifugation. Average NPN levels were similar for milk from all three lactation periods, and constituted 10% of colostrum N and 25% of mature milk N. Protein determined by the Bio-Rad method on whole milk samples had the lowest variability (square root MSE) when correlated to Kjeldahl values. All three assays had lower variability when analyzing whey and skimmed milk than when analyzing whole milk. The Lowry method and the Bio-Rad method had low variability for whey and skimmed milk samples, but the Lowry method yielded analytical values closest to Kjeldahl protein values. The BCA method consistently overestimated Kjeldahl protein by 30%.

The concentration of urogastrone (URO) and lactose was measured in breast secretions from four women. URO levels were high in precolostrum and fell markedly around the time of parturition to relatively constant low levels in mature milk. When milk samples were fractionated on a Biogel P10 column, the major peak of immunoreactivity occurred at the point at which pure URO eluted. There was good correspondence between the fall in URO levels at birth and the increase in lactose levels, suggesting that the decrease in URO was a dilution effect as the osmotic influence of lactose increased.

The occurrence of phosphate-containing sialyl oligosaccharides in bovine colostrum was investigated. Two major sialyl oligosaccharide phosphates were identified, one of which was structurally similar to the previously characterized sialyl oligosaccharide 1-phosphates of human urine. The second sialyl oligosaccharide phosphate of bovine colostrum was found to be of a novel type. Structural studies including monosaccharide and phosphate analysis, glycosidase and phosphatase treatments, methylation analysis, and periodate treatment indicated the structure of this compound to be NeuAc alpha 2-6Gal beta 1-4GlcNAc-6-P. This provides the first evidence for the occurrence of N-acetylglucosamine 6-phosphate as an integral component in complex carbohydrates.

Proteins in colostrum and skimmed milk from humans and mice were separated by electrophoresis on polyacrylamide gels and stained with Coomassie blue (CB), Ethyl-Stains-all (ESA), and periodic acid-Schiff (PAS) to investigate changes that may occur in milks throughout lactation. In mouse colostrum but not in mature mouse milk, a PAS-positive protein of apparent molecular weight of 60,000 stained prominently blue with ESA. A protein in human milk with a molecular weight of 68,000 stained similarly but was present throughout lactation. The intensity of blue staining of these minor proteins in milk approached that obtained with casein phosphoproteins. The metachromatic dye ESA stains phosphoproteins and sialic acid-rich glycoproteins blue to blue-green. Removal of phosphorus from the former and sialic acid from the latter results in those proteins staining red with ESA. The intensity of blue staining of the 60,000 and 68,000 Mr proteins was diminished but not lost following treatment with phosphatase. It was eliminated following neuraminidase digestion of the mouse protein and mild acid hydrolysis of the human protein. Coomassie blue staining of the proteins was not affected by these procedures. Following electrophoresis of milk and milk fractions in a non-sodium dodecyl sulfate-containing system, the proteins were identified by their characteristic staining properties with ESA and isolated.

The authors measured polychlorinated biphenyls (PCBs) and dichlorodiphenyl dichloroethene (DDE) in maternal serum, cord blood, placenta, and serial samples of breast milk from 868 women. Almost all samples of breast milk showed detectable levels of both chemicals. Overall, values for DDE in this study are within the range of those found previously, whereas those for PCBs are somewhat higher. Possible causes of variation in levels were investigated. For DDE, older women, Black women, cigarette smokers, and women who consumed sport fish during pregnancy had higher levels; only age and race showed large effects. For PCBs, older women, women who regularly drink alcohol, and primiparae had higher levels. In addition, both chemicals showed modest variation across occupational groupings. Casual exposure to a PCB spill did not result in chemical levels different from background. In general, women have higher levels in their first lactation and in the earlier samples of a given lactation, and levels decline both with time spent breast-feeding and with number of children nursed. These striking declines are presumably a measure of exposure to the child.

Human milk contained a factor or factors that protected suckling mice from lethal fluid loss due to Escherichia coli heat-stable toxin. This protective effect was present throughout lactation. The factor was heat stable, acid and alkali stable (pH 2 and 12), and of low molecular weight. Ion-exchange chromatography indicated that it was uncharged at neutral pH. The factor was insensitive to proteinase K digestion and partitioned in the aqueous phase in 2:1 (vol/vol) chloroform-methanol. Acid hydrolysis (2 N HCl at 100 C for 3 hr) destroyed the protective effect. Neither an amino acid mix simulating human milk nor lactose in phosphate-buffered saline was protective. These data indicate that the protective factor in human milk is neither a lipid nor a protein but appears to be a carbohydrate.

There is a decrease in the specific activity of labeled IgG1 of serum over 3 wk following the feeding of iodine-125 labeled immunoglobulin IgG1 in colostrum to calves at birth. This decrease indicated the appearance of new IgG1 from some source. To determine if this new IgG1 came from endogenous production in the calf or from continued small amount of intestinal absorption from milk, labeled IgG1 was added to normal milk and fed to calves of various ages up to 3 wk after an initial feeding of colostrum at birth. Labeled IgG1 was also added to colostrum fed to calves at birth, and the calves were maintained on a normal milk diet or fed a synthetic milk diet. Determination of iodine-125 in the serum protein fractions of these calves indicated that there was no apparent intestinal absorption of labeled IgG1 from the milk in the period from 2 days to 3 wk. Furthermore, comparable decreases occurred in the specific activity of labeled IgG1 in serum in the calves fed the labeled IgG1 in colostrum at birth and subsequently maintained either on a diet including milk or on the synthetic milk diet devoid of IgG1. The results support the conclusion that the origin of new IgG1 in the calf after about 36 h and up to about 3 wk of age arises from endogenous production at a rate of about 1 g of IgG1 per day.

By a procedure based on ion-exchange chromatography, five chromium-containing fractions were separated from the ethanol extract (50-90%) of cows' colostrum which should provide infant with all or most of the chromium needed in available form. Three of the fractions are anionic (A-1, A-2 and A-3) and two are cationic (C-1 and C-2). Two major fractions, A-1 and A-3, comprise together about 77% of the total chromium recovered in the effluent after ion-exchange chromatography and C-1 and C-2 comprise together only about 21%. Approximate molecular weight was determined by the Sephadex gel chromatography to be 1,500 for A-1 and C-2 and 2,000 for C-1. A-3 was eluted from the Sephadex gel column at the elution position corresponding to that of inorganic trivalent chromium. Among five fractions, A-1 shows glucose tolerance factor (GTF) activity as measured by the stimulation of [U-14C] glucose oxidation to 14CO2 in rat epididymal adipocytes. A-1 stimulates glucose oxidation at chromium concentration as low as 100 pg/ml only when it is incubated with insulin. These findings suggest that A-1 fraction separated from cow milk contains a low-molecular-weight, chromium complex which plays a role in glucose metabolism in close relation with the action of insulin.

The complete amino acid sequence of bovine colostrum cysteine proteinase inhibitor was determined by sequencing native inhibitor and peptides obtained by cyanogen bromide degradation, Achromobacter lysylendopeptidase digestion and partial acid hydrolysis of reduced and S-carboxymethylated protein. Achromobacter peptidase digestion was successfully used to isolate two disulfide-containing peptides. The inhibitor consists of 112 amino acids with an Mr of 12787. Two disulfide bonds were established between Cys 66 and Cys 77 and between Cys 90 and Cys 110. A high degree of homology in the sequence was found between the colostrum inhibitor and human gamma-trace, human salivary acidic protein and chicken egg-white cystatin.

Total (free plus protein-bound) and free cortisol concentrations in paired samples of serum and skim milk or colostrum were determined by specific radioimmunoassay after extraction or ultrafiltration. For 11 cows in established lactation mean total cortisol concentrations +/- SE in serum and milk were 4.5 +/- .7 ng/ml and .35 +/- .06 ng/ml, respectively. Concentrations of free cortisol in serum and milk were about .3 ng/ml, which is at the lower limit of the present assay. Following 200 IU Synacthen injection to 5 cows, total cortisol in serum and milk rose to 67 +/- 6.0 ng/ml and 11.4 +/- 1.5 ng/ml by 8 h postinjection, while free concentrations increased to 4.3 +/- .7 ng/ml and 6.5 +/- .8 ng/ml in serum and milk, respectively. Eight additional cows were sampled at 1 to 2 d after parturition, and total cortisol in serum and colostrum was 16.6 +/- 1.6 ng/ml and 4.4 +/- 1.3 ng/ml, and free cortisol was 2.4 +/- .4 ng/ml and 1.8 +/- .5 ng/ml, respectively. Free cortisol concentrations in colostrum or in milk from cows in established lactation, before or after adrenal stimulation, are directly related to free cortisol concentrations in the blood, while the bound cortisol in milk is more related to these two stages of lactation rather than the bound hormone in the blood. Measurement of free cortisol in milk should allow the monitoring of changes in plasma free cortisol in studies of stress in dairy cows.

Relative accuracy of the single radial immunodiffusion technique to measure immunoglobulin concentrations of colostral preparations (whey, whole, or fat-free) has been assessed. Fresh colostrum samples were analyzed for major constituents. Gammaglobulin as a standard was compared to total immunoglobulin concentration derived from single radial immunodiffusion analysis of colostral preparations with no differences except between standard and whey. Differences were in part from either enhancement or interference of immunoglobulin diffusion by colostral constituents. Removal of casein and fat during whey preparations caused a concentrating effect upon immunoglobulin constituents resulting in exaggerated precipitin rings. Whey has produced unreliable results: therefore, whole colostrum is recommended for single radial immunodiffusion analysis.

Alpha-1-antitrypsin concentration was analyzed by immunoelectrophoresis in samples of human colostrum (n = 3) and of mature milk from mothers between 2 to 52 wk postpartum (n = 39), one of whom was known to be PiMZ with a PiZZ infant. All milk samples tested contained alpha-1-antitrypsin. The three colostrum samples contained 140, 520, and 250 mg/liter. The mature milk of women who had been lactating less than 6 months had a higher concentration (7.2 +/- 3.6 mg/liter) (mean +/- SD) than in the women who had been lactating 6-12 months (4.8 +/- 1.8 mg/liter) (p less than 0.03). The milk of the woman of Pi type MZ had an alpha-1-antitrypsin concentration of 7.0 mg/liter at 7 wk postpartum and 4.1 mg/liter at 52 wk. It has been previously demonstrated that enhanced absorption of intact proteins occurs in early infancy. The presence of antiproteases in human milk provided during early infancy may serve to inhibit the absorption of intact proteases, limiting their entry into the portal circulation.

A method for preparing purified IgA from bovine mammary secretions is described. Whey was initially fractionated by gel filtration and fractions containing IgA were pooled, concentrated and digested with pepsin. The digest was rechromatographed on the same type of gel twice and the resulting IgA preparation tested for purity by an enzyme immunoassay procedure. Five different preparations tested were found to contain no measureable IgM or IgG2 and 0.8% to 1.1% IgG1 on a weight basis. If colostral whey was digested with pepsin prior to chromatography, the IgA preparations contained 1.1% to 2% IgG1 and no measureable IgM and IgG2. The procedure provides a reasonably easy method of eliminating most of the contaminating IgG1 (dimeric) and allows preparation of quantities of IgA for immunochemical studies and standardization of serological techniques.