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Levels of immunoreactive insulin, neurotensin, and bombesin in porcine colostrum and milk.

High concentrations of insulin (411 +/- 214 microU/ml), neurotensin-like (265 +/- 72 pg/ml), and bombesin-like immunoreactivities (1995 +/- 288 pg/ml) were detected in porcine colostrum using radioimmunoassay, as compared to the levels found in sow blood serum at farrowing (5 microU/ml, less than 12 pg/ml, and 17 pg/ml, respectively). After 72 h of lactation, the levels of insulin and neurotensin-like immunoreactivities had decreased to 28 +/- 17 microU/ml and 89 +/- 23 pg/ml, respectively, while the bombesin-like activity remained constant. Characterization with reversed-phase high performance liquid chromatography showed that the insulin immunoreactivity eluted at the same position as the insulin standard, while the elution patterns of the neurotensin-like and bombesin-like immunoreactivities (eluted in three separate peaks) did not correspond to that of their respective standards. The biological function of the peptide hormones in colostrum/milk may be as triggers of the developmental changes taking place in the nursing neonate, especially in the gastrointestinal tract.

Animals↗

Casein and casein subunits in preterm milk, colostrum, and mature human milk.

Human milk proteins have both nutritional and physiological roles for the breast-fed infant. While the biochemistry and developmental patterns for many whey proteins are well known, our knowledge of human casein and its subunits is still limited. We have recently developed a method to isolate casein from whey proteins in human milk and to separate the casein subunits by fast protein liquid chromatography. In this study we have applied this methodology to study the casein subunit pattern in preterm milk, colostrum, and mature milk. Casein concentration increased with lactation time, largely due to an increase in glycosylated forms of casein (kappa-caseins). Thus, the relative proportion of beta-casein to kappa-casein decreased during the lactation period. The patterns of phosphorylated and glycosylated casein subunits were found to vary during lactation, showing that both synthesis and posttranslational modification of beta- and kappa-casein are regulated by different mechanisms.

Adult↗

Reduction in virus-neutralizing activity of a bovine colostrum immunoglobulin concentrate by gastric acid and digestive enzymes.

Bovine milk immunoglobulin concentrates have been proposed for inducing passive immunity against various enteric pathogens. In vitro digestion studies were conducted to evaluate the effect of gastrointestinal secretions on the virus-neutralizing activity of a concentrate prepared from the colostrum of cows that were immunized with rotavirus. The proteolytic activity of human gastric and duodenal fluid specimens was used to design a two-stage in vitro digestion model with commercial enzymes for estimating the individual impact of pepsin, gastric acid, and select pancreatic enzymes on antirotavirus activity in bovine milk immunoglobulin concentrates. The rotavirus-neutralizing titer of concentrate was decreased by incubation with pepsin at pH 2, a pool of pancreatic enzymes at pH 7.5, or sequential digestion with pepsin (pH 2) and pancreatic enzymes (from initial titer of 55,210 to 2,030, 19,500, and 320, respectively). Reduction in rotavirus-neutralizing titer after gastric-phase digestion was primarily due to acidic conditions and not to proteolytic cleavage by pepsin. Although both trypsin and carboxypeptidase caused significant proteolysis of concentrate during duodenal-phase digestion, only trypsin caused a significant reduction in rotavirus-neutralizing titer. The extent of digestion was the same for concentrate suspended in water or skim milk. The results demonstrate that the biological activity of bovine milk antibodies is reduced by exposure to acid and trypsin in vitro and suggest that neutralization of both gastric acid and pancreatic trypsin may enhance the effectiveness and economic feasibility of passive oral immunoprophylaxis with bovine milk immunoglobulins.

Animals↗

Colostrum protein concentrate enhances intestinal adaptation after massive small bowel resection in juvenile pigs.

OBJECTIVES: Short bowel syndrome (SBS) usually results from the surgical removal of a large segment of small intestine. Patient outcome depends on the extent of intestinal resection and adaptation of the remaining intestine. We evaluated the impact of colostrum protein concentrate (CPC) on intestinal adaptation after massive small bowel resection in a porcine model of infant SBS. METHODS: Four-week-old piglets underwent an approximate 75% small bowel resection (R, n = 23) or a control transection operation (C, n = 14). Postoperatively, animals from both groups received either pig chow (R = 6, C = 5), polymeric infant formula (R = 6, C = 3) or polymeric infant formula supplemented with CPC (R = 11, C = 6) for 8 weeks until sacrifice. Clinical outcome measures included weight gain and stool consistency. Morphologic measures were intestinal villus height and crypt depth. Functional outcome measure was mucosal disaccharidase activity. RESULTS: Resected animals fed polymeric infant formula alone had reduced weight gain compared with controls fed the same diet (P < 0.005). Despite massive small bowel resection, animals fed pig chow or polymeric infant formula supplemented with CPC grew at an equivalent rate to controls fed polymeric infant formula alone. Resected animals supplemented with CPC had increased villus length and crypt depth in the jejunum (P < 0.001) and ileum (P < 0.001) compared with resected animals fed either pig chow or polymeric infant formula alone. CONCLUSION: In an animal model of SBS, CPC supplementation of polymeric infant formula resulted in normal weight gain and features of enhanced morphologic adaptation.

Adaptation, Physiological↗

Passive protection of lambs against enteropathogenic Escherichia coli: role of antibodies in serum and colostrum of dams vaccinated with K99 antigen.

Lambs from suckling ewes vaccinated with the K99 antigen were resistant to challenge with K99-positive enteropathogenic Escherichia coli. Serum and colostrum from these ewes were compared with samples from control ewes to establish methods for monitoring vaccination and to determine the mechanism of protection. Vaccination stimulated production of K99 antibodies. These could be detected by an indirect haemagglutination test and a haemagglutination-inhibition test. Antiglobulin and gel-diffusion tests were less reliable. Experiments with brush-border cells from calf intestine showed that the antibodies were associated with anti-adhesive activity. The antibodies were predominantly IgG and did not neutralise the activity of heat-stable enterotoxin. It was concluded that neutralisation of the adhesive properties of the K99-positive E. coli by colostral antibodies significantly contributed to the resistance of the lambs from vaccinated ewes.

Animals↗

Structure of the asialyl oligosaccharide chains of kappa-casein isolated from ovine colostrum.

Three oligosaccharide alditols, a di (44-63%), tetra (32-46%), and a pentasacchariditol (5-10%) have been isolated from desialylated caseinomacropeptide obtained from kappa-casein of ewe colostrum. The disacchariditol is 2-acetamido-2-deoxy-3-O-(beta-D-galactopyranosyl)galactitol; in the tetrasaccharide this unit is substituted in position 6 of the galactosaminitol moiety by 2-acetamido-2-deoxy-4-O-(beta-D-galactopyranosyl)-beta-D-glucopyranose, while the pentasacchariditol is derived from the tetrasaccharide by addition of a D-galactopyranosyl unit to position 3 of the galactopyranose unit substituting the glucosamine unit.

Animals↗

Anti-acrosin antibodies in sera, colostrum and genital organs of vaccinated ewes: estimation and correlation with fertility.

Three ewes were vaccinated with boar sperm acrosin. Despite the presence of antiacrosin antibodies which crossreact with ram acrosin in the sera of the immunized sheep, no significant reduction in fertility was observed. Incubation of ejaculated or capacitated boar spermatozoa with anti-acrosin serum or IgG did not reduce the halos of digestion in the gelatinolysis test. However, an unexplained decrease of the forward progression and number of lytically active spermatozoa was observed after this treatment. We found antiacrosin antibodies and determined their titers in the sera of the vaccinated ewes, in colostrum and in aqueous extracts of cervix, uterus, oviduct and ovary. No antiacrosin antibodies could be detected in the sera of new born lambs. The immunoglobulins IgA, IgG and IgM were isolated from the sera of the ewes. The acrosin-specific antibodies belonged exclusively to the IgG class. In genital extracts the amount of IgG, IgA and IgM was determined. IgG was found in higher amount in every extract. IgA is present in similar amount only in the cervix, in the other parts only in traces. In all extracts IgM is found in a very low level. The specificity of the antibodies for acrosin was proven by active enzyme staining of the precipitin lines obtained by immunodiffusion.

Acrosin↗

Actinobacillus pleuropneumoniae serotype-2 antibodies in sow colostrum in Finnish pig-health-scheme herds.

A direct ELISA with phenol-extracted antigen for Actinobacillus pleuropneumoniae serotype 2 was developed. The test was specific when tested with rabbit antisera prepared against different A. pleuropneumoniae serotypes. It had better-than-moderate repeatability and it made a clear distinction between positive and negative samples. A total of 5477 colostrum samples from breeding sows from herds participating in the Finnish Pig-health Scheme were tested using the ELISA test. A total of 1307 positive samples were found in 129 out of 154 herds, thus indicating that most of the disease-control herds in Finland are infected with A. pleuropneumoniae serotype 2. These infections were almost entirely subclinical.

Actinobacillus Infections↗

Shedding of porcine circovirus into colostrum of sows.

The purpose of the present study was to investigate porcine circovirus (PCV) shedding into the milk of sows. Colostrum was collected from 33 sows. PCV1 was isolated from four of 33 milk whey samples. PCV1 DNA was detected by polymerase chain reaction in three of these samples and in three of 10 milk cell samples. PCV2 was also isolated and detected from every single milk whey sample. These results showed that PCV1 and PCV2 were shed into the milk of sows and suggest that PCV can be transmitted to offspring by an oral route through milk.

Animals↗

Effect of postpartum maternal infection on proteins and trace elements in colostrum and early milk.

In developing countries, maternal infections during lactation are common. In this study, we evaluated the effect of acute maternal postpartum infection on the composition of colostrum and early milk with special emphasis on milk proteins and trace elements. The study was carried out in two maternity hospitals in Lima, Peru. Subjects were normally nourished women (body mass index (BMI) > 20.0) who intended to exclusively breastfeed their child and who had fever and clinical symptoms of infection within the first 48 h postpartum (n = 34). Non-ill women of similar characteristics were selected as controls (n = 23). Blood and milk samples were taken on days 1 and 14 postpartum. An acute phase response was confirmed by significantly increased serum levels of C-reactive protein in infected women. Serum zinc levels increased significantly from day 1 to day 14, but were not affected by infection. Serum copper levels were significantly higher in ill women than in non-ill women on day 1. All participating women were breastfeeding on day 14. Whey protein levels, the whey/casein ratio and total protein levels decreased significantly with time, but were not affected by infection. There were no differences in milk iron or copper levels with time or infection. Milk zinc levels decreased significantly with time, but were not affected by infection. Maternal infection during the early postpartum period does not appear to adversely affect the initiation of lactation or milk protein and trace element contents.

Adolescent↗

Changes in colostrum composition and in the permeability of the mammary epithelium at about the time of parturition in the goat.

1. Changes in milk composition and in mammary permeability to labelled sucrose and monovalent ions have been studied in late pregnancy and at the time of parturition. These data have been compared with those obtained previously in lactating goats.2. Colostrum contained more sodium, chloride, protein, immunoglobulins and less potassium and lactose than milk. The composition of the aqueous phase changed near term and normal milk composition was reached either several days before or several days after parturition. The main decrease in the immunoglobulin concentration occurred when the animal was first milked or suckled.3. In contrast to lactating animals, [(14)C]sucrose passed from blood to milk and [(14)C]lactose from milk to blood. There was a significant correlation between the entry of labelled sucrose into milk and the sodium concentration.4. The rate of passage of (24)Na and (36)Cl into milk from blood was higher in late pregnancy than in lactation.5. In late pregnancy, the concentrations of both sodium and chloride were inversely correlated with the concentration of lactose while there was a positive correlation between potassium and lactose.6. Hydrostatic pressure in the lumen of the mammary gland increased during late pregnancy but there was no significant correlation with the composition of the aqueous phase of milk.7. When one gland of 4 animals was milked twice-daily starting 9-19 days before parturition, the composition of the secretion changed to that of normal milk only in the gland that was being milked. Similarly, [(14)C]sucrose, (24)Na and (36)Cl entry from blood decreased. Milk yield also increased but a further increase occurred post-partum.8. Comparison of the passage of (3)HOH, (24)Na and (36)Cl into different fractions of milk during I.A. infusions indicates that the mammary ducts are less permeable to ions than the alveoli, but it was not possible to decide whether the ducts are as impermeable as during lactation.9. It is suggested that ion and lactose movements occur via a para-cellular route and these results are discussed in relation to transport mechanisms and in terms of local and systemic controlling mechanisms.

Animals↗

[Bacteriostatic activity of bovine colostrum].

Succeeding the previous report on antivirus activity of non immunized bovine colostrum immunoglobulin, we studied the bacteriostatic activity of the immunoglobulin, lactoferrin and lactoferrin Fe by using the automated antimicrobial susceptibility testing system. We have presented the representative positive and negative reactions. Then, we have reported that the immunoglobulin was effective in 10 of a total of 20 species which we examined. In some species, the lactoferrin was slightly more effective than the immunoglobulin was. These results indicated that the immunoglobulin, lactoferrin and lactoferrin Fe were useful for the bacteriostatic reaction.

Animals↗

Oral bovine colostrum supplementation enhances buffer capacity but not rowing performance in elite female rowers.

A randomized, double-blind, placebo controlled design was used in which 13 elite female rowers, all of whom had competed at World Championships, were supplemented with 60 g day-1 of either bovine colostrum (BC; n = 6) or concentrated whey protein powder (WP; n = 7) during 9 weeks of pre-competition training. All subjects undertook the study as a group and completed the same training program. Prior to, and after 9 weeks of supplementation and training, subjects completed an incremental rowing test (ROW1) on a rowing ergometer consisting of 3 3 4-min submaximal workloads and a 4-min maximal effort (4 max), each separated by a 1-min recovery period. The rowing test was repeated after a 15-min period of passive recovery (ROW2). The 4 max for ROW1 provided a measure of performance, and the difference between the 4 max efforts of ROW1 and ROW2 provided an index of recovery. Blood lactate concentrations and pH measured prior to exercise and at the end of each workload were used to estimate blood buffer capacity (beta). Food intake was recorded daily for dietary analysis. There were no differences in macronutrient intakes (p >.56) or training volumes (p >.99) between BC and WP during the study period. Rowing performance (distance rowed and work done) during 4 max of ROW2 was less than ROW1 at baseline (p <.05) but not different between groups (p >.05). Performance increased in both rows by Week 9 (p <.001), with no difference between groups (p >.75). However, the increase was greatest in ROW2 (p <.05), such that by Week 9 there was no longer a difference in performance between the two rows in either group (p >.05). b was not different between groups for ROW1 at baseline (BC 38.3 5.0, WP 38.2 7.2 slykes; p >.05) but was higher in BC by Week 9 (BC 40.8 5.9, WP 33.4 5.3 slykes; p <.05). b for ROW2 followed the same pattern of change as for ROW1. We conclude that supplementation with BC improves b, but not performance, in elite female rowers. It was not possible to determine whether b had any effect on recovery.

Administration, Oral↗

Variability of the immunological state of germfree colostrum-deprived Minnesota miniature piglets.

Minnesota miniature piglets obtained by hysterectomy and deprived of colostrum were examined for the presence of immunoglobulin by immunoelectrophoresis, double-gel diffusion, and radial immunodiffusion techniques with specific anti-immunoglobulin chain sera. A large amount of variability existed between different litters of piglets and between different piglets within the same litter, ranging from no detectable immunoglobulin in the serum to very high immunoglobulin levels approaching that of the adult pig. All known classes of porcine immunoglobulin including immunoglobulin G, immunoglobulin M, and immunoglobulin A could be found in the sera from litters where there was extensive placental damage. This contaminating immunoglobulin was shown to have antibody activity to actinophage MSP-2 even when present in very low concentrations. The low level contamination with immunoglobulin G, which was the most frequently encountered type of contaminant, was demonstrated to be similar to sow immunoglobulin G both antigenically and in its molecular size. The data demonstrates that individual piglets must be tested for immunoglobulin content rather than being assumed to be immunologically "virgin" and emphasizes the need for an intact placenta barrier to obtain piglets free from maternal immunoglobulin and devoid of antigenic stimulation.

Animals↗

Antibacterial activity in colostrum and milk associated with protection of piglets against enteric disease caused by K88-positive Escherichia coli.

Piglets suckled by dams that had been vaccinated with K88 antigen were significantly more resistant to deaths caused by neonatal diarrhea after challenge with a large dose of a K88-positive enteropathogenic strain of Escherichia coli than piglets suckled by control dams. The factors most likely to be involved in protection of the piglets were investigated by comparing the antibacterial activities of serum and mammary secretions from the two groups of dams. Vaccination stimulated the production of K88 antibodies, which were associated with anti-adhesive activity directed against the adhesive properties of the K88 antigen, and of O8 antibodies; the latter antibodies were attributed to traces of O8 antigen in the vaccine. Neutralizing activity against heat-labile enterotoxin was present in several dams before vaccination but was not stimulated by bacteriostatic activities were similar in serum and mammary secretions from both groups of dams and appeared to play no significant role in the protective after parturition were atrributed to exposure of the dams to the challenge strain excreted by the piglets. It was concluded that neutralization of the adhesive properties of K88 antigen by K88 antibodies in colostrum and in milk contributed significantly to the protection of piglets from vaccinated dams. However, the contribution of antibacterial activities associated with the greater levels of O8 antibodies in colstrum from the vaccinated group cannot be entirely excluded.

Agglutinins↗

Use of specific antibody to demonstrate glycocalyx, K99 pili, and the spatial relationships of K99+ enterotoxigenic Escherichia coli in the ileum of colostrum-fed calves.

The attachment of enterotoxigenic Escherichia coli (ETEC) strain B44 (O9:K30:K99:F41:H-) to the ileal epithelium of newborn colostrum-fed calves was studied by electron microscopy. Stabilization of the bacterial glycocalyx (K30) and pili (K99) by fixation of tissue sections in specific antibody and staining with ruthenium red were used so that the bacterial surface structures could be clearly visualized and their spatial relationship to the intestinal brush border defined. When sections of ileum from infected calves were neither fixed in antibody nor stained with ruthenium red, the ETEC cells colonizing the small intestine were separated from each other and from the brush border by an electron-translucent halo; neither the glycocalyx nor the pili could be clearly resolved. When ruthenium red staining was used, the halo was partially filled by a net of electron-dense fibers composed of pili and condensed glycocalyx which extended to the brush border. Tissue sections reacted with anti-K30 antibody before staining with ruthenium red revealed microcolonies of ETEC surrounded by a discrete electron-dense glycocalyx 0.3 to 1.0 micrometers thick and in tight contact with the epithelial cell surface. When ileal tissue was treated with K99 antibody, the K99 pili were visible as discrete fibers extending from the bacterial cell surface through the glycocalyx. We discuss the role of these cell surface components in pathogenic adhesion and in the formation of protected microcolonies at the surface of the infected ileal epithelium.

Adhesiveness↗

Infection of cesarean-derived colostrum-deprived 1-day-old piglets with Arcobacter butzleri, Arcobacter cryaerophilus, and Arcobacter skirrowii.

Neonatal piglets have been used as models to study human campylobacteriosis and helicobacteriosis. The purpose of this study was to determine the relative pathogenicities, on the basis of the duration of fecal shedding and colonization of tissues, of three Arcobacter species in 1-day-old cesarean-derived colostrum-deprived piglets. Two experiments were conducted. In experiment 1, two piglets each were infected per os with either Arcobacter butzleri ATCC 49616, Arcobacter cryaerophilus 1B ATCC 43159, Arcobacter skirrowii CCUG 10374, or the three field strains of A. butzleri (approximately 5 X 10(9) CFU per piglet). Rectal swab samples were taken prior to infection and daily thereafter for up to 7 days. Arcobacter spp. were detected at least once in rectal swab samples of all but one of the experimentally infected piglets but not in the control. At necropsy, A. butzleri was recovered from the lung, kidney, ileum, or brain tissues of the four infected piglets which had received either the field strain or the ATCC type strain of A. butzleri. A. cryaerophilus 1B was detected in rectal swab samples for up to 7 days postinfection but was not cultured from tissues at necropsy. Arcobacters were detected in the rectal swab sample of the A. skirrowii-infected piglet only on day 3 postinfection; no isolates were obtained from tissues at necropsy. No gross pathological lesions were consistently noted in the experimentally infected piglets. In experiment 2, two piglets each were infected per os with A. butzleri ATCC 49616, A. cryaerophilus 1A ATCC 43158, A. skirrowii CCUG 10374, or the single A. butzleri field strain Yard J/c (approximately 5 X 10(9) CFU per piglet). Arcobacter spp. were cultured from rectal swab samples of all but one of the experimentally infected piglets at least once. At necropsy Arcobacter spp. were cultured from the liver, kidney, ileum, or brain tissues of two of the four A. butzleri-infected piglets. However, no severe gross pathology was noted. These data suggest that Arcobacter spp., especially A. butzleri, can colonize neonatal pigs.

Animals↗

Recognition of enteropathogenic Escherichia coli virulence determinants by human colostrum and serum antibodies.

Human colostra and sera collected from Mexican mothers and their children at birth and 6 months thereafter were studied for the presence of antibodies against the bundle-forming pilus and several chromosomal virulence gene products (intimin and secreted proteins EspA and EspB) of enteropathogenic Escherichia coli (EPEC). Among 21 colostrum samples studied, 76, 71.5, 57, and 47% of them contained immunoglobulin A (IgA) antibodies against EspA, intimin, EspB, and BfpA, respectively. Interestingly, there was a difference in IgG response to EPEC antigens between the sera from neonates and sera from the same children 6 months later. While the number of neonates reacting to Esps and intimin diminished when they reached 6 months of age, those reacting with BfpA increased from 9 to 71%. Intimin from an enterohemorrhagic E. coli strain was also recognized by most of the samples reacting with EPEC intimin. These data suggest that Bfp and Esps elicit an antibody response during the early days of life of neonates and support the value of breast-feeding in areas of the world where bacterial diarrheal infections are endemic.

Adhesins, Bacterial↗