Effect of induction of parturition on immunoglobulin content of colostrum and calf serum

Thirty-four pregnant crossbred beef cows were injected with prostaglandin F(2) alpha (PGF group, n = 11), dexamethasone (DEX group, n = 11), or saline (control group, n = 12) on Day 270 of gestation. Immediately after calving, all colostrum was milked from each cow. A sample was taken, and the remainder was fed to that cow's calf within one hour of birth. Serum was collected from each calf at 0 and 24 h of age. Immunoglobulin G (IgG) content of colostrum and serum was determined with commercial radial immunodiffusion plates. The data from four PGF cows that did not calve until after 140 h post injection were excluded from the results. Mean (+/- SD) volumes (ml) of colostrum were 2086 (+/-1148.4) for the PGF group, 1336 (+/-583.7) for the DEX group, and 2404 (+/-1140.7) for the control group. Mean (+/- SD) concentrations (mg/dl) of IgG in colostrum were 6017 (+/-3351.2) for PGF, 10285 (+/-5370.7) for DEX and 10766 (+/-5098.3) for the control group. Mean (+/- SD) total quantities of IgG (g) in colostrum were 133.9 (+/-120.03) for PGF, 134.1 (+/-96.67) for DEX and 235.6 (+/-147.22) for the control. IgG concentrations were very low or were not detectable in serum of all calves prior to administration of colostrum. Mean (+/- SD) concentrations (mg/dl) of IgG in serum of calves at 24 h of age were 1469 (+/-905.8) for calves from PGF cows, 1819 (+/-1289.8) for calves from DEX cows, and 3317 (+/-1888.2) for calves from control cows. Calves from control cows had significantly more IgG at 24 h than calves from PGF cows or DEX cows (p<0.05). Calves born to cows induced to calve early may be at an increased risk of failure of passive transfer and so should be monitored for IgG concentrations.     

Prevention and treatment of influenza with hyperimmune bovine colostrum antibody

Background

Despite the availability of specific vaccines and antiviral drugs, influenza continues to impose a heavy toll on human health worldwide. Passive transfer of specific antibody (Ab) may provide a useful means of preventing or treating disease in unvaccinated individuals or those failing to adequately seroconvert, especially now that resistance to antiviral drugs is on the rise. However, preparation of appropriate Ab in large scale, quickly and on a yearly basis is viewed as a significant logistical hurdle for this approach to control seasonal influenza.

Methodology/Principal Findings

In this study, bovine colostrum, which contains approximately 500 g of IgG per milking per animal, has been investigated as a source of polyclonal antibody for delivery to the respiratory tract. IgG and F(ab'')2 were purified from the hyperimmune colostrum of cows vaccinated with influenza A/Puerto Rico/8/34 (PR8) vaccine and were shown to have high hemagglutination-inhibitory and virus-neutralizing titers. In BALB/c mice, a single administration of either IgG or F(ab'')2 could prevent the establishment of infection with a sublethal dose of PR8 virus when given as early as 7 days prior to exposure to virus. Pre-treated mice also survived an otherwise lethal dose of virus, the IgG- but not the F(ab'')2-treated mice showing no weight loss. Successful reduction of established infection with this highly virulent virus was also observed with a single treatment 24 hr after virus exposure.

Conclusions/Significance

These data suggest that a novel and commercially-scalable technique for preparing Ab from hyperimmune bovine colostrum could allow production of a valuable substitute for antiviral drugs to control influenza with the advantage of eliminating the need for daily administration.     

Heat treatment of bovine colostrum: effects on colostrum metabolome and serum metabolome of calves

Bovine colostrum is important for neonates' health due to its nutritive and non-nutritive components. Heat treatment of colostrum is a well-established management tool, but it may influence colostrum components and affect the health status of calves. In our previous studies, we had shown that colostrum proteome and serum proteome of calves were altered by heat treatment to different degrees. Our objectives in this study were to investigate the effects of heat treatment on colostrum metabolome and the effect of feeding heat-treated colostrum on the serum metabolome of newborn calves. Further, the changes in serum metabolome from before to after colostrum feeding were characterized. Newborn Holstein female calves (n = 10) were randomized within pairs and fed heat-treated (n = 5; 60 °C, 60 min) or raw (n = 5) colostrum at 8.5% of birth BW by esophageal feeder within 1 h of birth. After a single colostrum feeding, calves were not fed until after the 8 h time point. Blood samples were taken immediately prior to feeding (0 h) and 8 h after feeding. The colostrum and serum metabolome were first analyzed using reverse-phase chromatography and tandem MS, and serum metabolome was then further analyzed using hydrophilic interaction chromatography and tandem MS. In colostrum metabolome, 458 features were identified and 328 were annotated and a trend of separation between raw and heat-treated colostrum could be observed through multivariate analysis. In serum metabolome, 3 360 features were identified and 1 439 were annotated, but no trend of separation was observed between the two groups of calves fed raw colostrum vs. heat-treated colostrum. The serum metabolome presented substantial differences comparing before (0 h) and after colostrum feeding (8 h); in particular, a tripeptide, β-homovaline-β-homoalanine-β-homoleucine, and 1-(2-acetamido-2-deoxy-α-d-glucopyranosyl)-1D-myo-inositol had higher concentrations after colostrum feeding than before, along with other metabolites that were not fully annotated. Based on a relatively small sample size, our findings point to the effect of heat treatment on the change of colostrum metabolome, but not on the change of serum metabolome of calves fed raw colostrum vs. heat-treated colostrum. Further studies using larger sample size and complementary analytical techniques are warranted to further explore potential heat treatment-induced alterations in colostrum metabolome.     

Oxidation of methanol,formaldehyde and formic acid by methanol-utilizing yeast

Methanol-utilizing yeast,Candida boidini 11 Bh, characterized by high tolerance to methanol during growth, displays even higher tolerance when the oxidation rate by intact cells is tested. Low respiration activity is found even at 22% v/v of methanol. The half-saturation constant was 17–18mM. The half-saturation constants for the two oxidation intermediates, formaldehyde and formic acid were 3.6–4.0 and 30–33mM, respectively. When applied together with standard concentration of methanol, very low concentrations of both intermediates stimulated the oxidation rate. These results are discussed in connection with the relationship between growth and oxidation, the tolerance to high concentrations of inhibitory products and the mechanism of inhibition.     

Isolation and purification of lactoferrin and immunoglobulin G from bovine colostrum with serial cation-anion exchange chromatography

High-value dairy proteins such as lactoferrin (LF) and immunoglobulin (IgG) were separated from bovine colostrum. The whey was initially adjusted to pH 6.8 with 1 mol/L NaOH and then went through centrifugation, precipitation, and filtration to eliminate the fat and caseins in bovine colostrum. The treated whey was further ultra-filtrated to partially remove both other proteins and carbohydrates under 50 kD molecular weight. Then the ultra-filtrated whey was passed through cation and anion exchange columns in series. The LF and IgG were adsorbed on cation and anion exchanger, respectively, due to their different pI. Both the cation and anion exchange columns were washed with de-ionized water followed by successive elution with sodium chloride solutions of increasing molarities (0.27 and 0.85 mol/L; 17 and 51 mmol/L) in a stepwise manner, respectively. After desalted, the elution was freeze-dried. Finally, the LF and IgG with respective purities of 95.0% and 96.6% were obtained.     

Validation of Brix refractometer to estimate colostrum immunoglobulin G content and composition in the sow

Colostrum is an essential source of immunoglobulin G (IgG) for neonate piglets. However, colostrum IgG content and nutritional composition can vary considerably among sows due to age, parity, feeding regime and immunological background. Currently, there is no practical way to obtain information about colostrum IgG concentration at herd level. We evaluated sows’ colostrum IgG content on-farm using a Brix refractometer and its performance was compared with that of an IgG ELISA. In addition, nutritional compositions of the colostrum samples were analyzed using Fourier transform IR spectroscopy. Colostrum samples (5 to 6 ml) (n=153) were obtained within 0 to 3 h of farrowing. However, to obtain a 24 h IgG profile for 11 sows, colostrum samples were collected at 0, 2, 4, 6, 8, 10, 16 and 24 h after farrowing. A 0.3 ml of freshly drawn colostrum sample was used for the on-farm measurement of Brix percentages using a digital refractometer shortly after collection. The remaining fractions of the samples were frozen and submitted to laboratory analysis for total IgG, using a commercially available pig IgG ELISA kit. For nutritional composition analysis, a 35 ml colostrum sample (n=34) was obtained immediately after birth of first piglet from the first three pairs of frontal teats. Colostrum concentrations of IgG averaged 52.03±30.70 mg/ml (mean±SEM) at 0 to 3 h after farrowing. Concentration of IgG decreased on average by 50% during the 1st day of lactation (P<0.01). Sow parity did not influence colostrum concentrations of IgG. Differences in colostrum composition were recorded between two herds and among the parity groups (P<0.05). The Brix refractometer measurement of colostrum and the corresponding log transformed IgG measurements from the ELISA were moderately correlated (r=0.63, P<0.001, n=153). Based on the classification we suggest here, low levels of IgG (14.5±1.8 mg/ml) were recorded for colostrum samples with Brix readings below 20%. Borderline colostrum IgG content (43.8±2.3 mg/ml) had Brix readings of 20% to 24%, adequate colostrum IgG content (50.7±2.1 mg/ml) had Brix % readings of 25% to 29% and very good IgG colostrum content (78.6±8.4 mg/ml) had Brix readings >30%. Colostrum IgG concentration is highly variable among sows, Brix measurement of a sows’ fresh colostrum is an inexpensive, rapid and satisfactorily accurate method of estimating IgG concentration, providing indication of differentiation between good and poor IgG content of colostrum.     

Effects of bovine colostrum, foal serum immunoglobulin concentration and intravenous plasma transfusion on chemiluminescence response of foal neutrophils

Summary. The effects of bovine colostrum, absorption of equine colostral immunoglobulins and age on phagocytic and serum opsonizing activity of nine clinically healthy foals were examined. Cells and serum were collected prior to suckling and at 7, 14 and 28 days of age. Seven foals had serum IgG concentrations >600mg/dl whereas two foals had <350mg of IgG/dl. Phagocytic and serum opsonic activity of eight clinically ill foals with <400mg of IgG/dl of serum were also examined before and after plasma transfusion. Phagocytic and serum opsonizing activities were evaluated by an assay for chemiluminescence (CL) after addition of opsonized streptococci. Results showed that bovine colostrum stimulated CL of foal neutrophils. Preliminary characterization of opsonins in bovine colostrum by ammonium sulphate fractionating and heat inactivation indicated that opsonins generating CL were mainly associated with immunoglobulin G. Chemiluminescence generated by foal neutrophils varied with age with foal neutrophils collected at day 14 producing more CL than adult neutrophils ( P <0.05). Foal serum opsonizing activity was similar to adult opsonizing activity if serum IgG concentrations were >600mg/dl but it was less if IgG concentration was <350mg/dl ( P <0.05). Chemiluminescence generated by foal and adult neutrophils was higher when post-transfusion foal serum was used as the source of opsonin than when pre-transfusion foal serum was used ( P <0.05). When adult serum was the opsonin, chemiluminescence of foal neutrophils collected before and after plasma transfusion did not differ. The increase in CL following plasma transfusion was probably due to an increase in serum opsonizing activity.     

Studies on the immunoglobulin spectrum of porcine serum and colostrum

An analysis of the immunoglobulin region of porcine blood serum proved the presence of a number of protein fractions which is higher than in other animal species. The presence of antibody activity to tetanus anatoxin examined by radioimmunoelectrophoresis was found in three γ G-globulin fractions, which could be distinguished from each other, and also in two additional fractions. One of the latter was the γ-macroglobulin fraction and the other one might correspond to γ A-globulin even though its mobility was higher than the mobility of γ A-globulin of human serum. A comparison of the immunoglobulin spectrum of porcine serum, colostrum, and of the suckling pig serum after ingestion of the colostrum showed that the main component of porcine colostrum constitutes the γ G-immunoglobulin, even though the colostrum seems to contain other components of the latter which have a higher electrophoretic mobility. The antibody activity of colostrum was found only in fractions γ G and γ M. The fraction of colostrum which might correspond to γ A did not display any activity at all. The activity in suckling pig serum was also concentrated in fractions γ G and γ M.     

Late gestation diet supplementation of resin acid-enriched composition increases sow colostrum immunoglobulin G content,piglet colostrum intake and improve sow gut microbiota

Resin acid-enriched composition (RAC) mainly containing tall oil fatty acid with an active component of resin acid (RA) can improve the microbial population in the digestive system, change the microbial fermentation, and improve the feed conversion ratio. We investigated the effects of dietary supplementation of RAC on sow colostrum yield (CY), colostrum composition and gut microbiota. Tall oil fatty acid and RA are commonly termed RAC and CLA, pinolenic, abietic, dehydrobiotic acids are characteristic components of RAC. The experiment was conducted in three trials in three respective herds. Sows were fed with a control diet and the same diet supplemented with 5 g RAC/day per sow during the last week of gestation. The 16S ribosomal RNA gene sequencing technique was used to assess sows’ faecal microbiota populations at farrowing. Colostrum nutritional composition, acute phase proteins (APPs) and immunoglobulin (Ig) content were also assessed. Individual piglets were weighed at birth and 24 h after the birth of first piglets in order to calculate CY and later at 3 to 4 weeks to calculate average daily gain. The RAC-fed sows had significantly higher IgG levels (P<0.05) in all three herds but treatment did not influence colostrum IgA and IgM concentration. There were no significant differences in colostrum protein, lactose and fat content in sows of the two diet groups (P>0.05), but those fed RAC had higher levels of colostrum serum amyloid A. Colostrum yield was significantly higher in RAC-fed sows in herds 2 and 3 with heavier piglets between 3 and 4 weeks of age (P<0.05), but not in herd 1 (P>0.05). Resin acid-enriched composition supplementation significantly increased some beneficial and fermentative bacteria (Romboutsia and Clostridium sensu stricto) than the control diet (P<0.01) while some opportunistic pathogens (Barnesiella, Sporobacter, Intestinimonas and Campylobacter), including Proteobacteria, were suppressed. Therefore, RAC added to the sow diet at late pregnancy increases colostrum IgG, colostrum availability for neonate piglets, and seems to promote better maternal intestinal microbial sources.     

The effect of methanol, formaldehyde, and formic acid on growth of Candida boidinii 11 Bh.

The dependence of the specific growth rate of Candida boidinii 11 Bh on methanol concentration follows Monod-type kinetics only in a narrow concentration range (0--0.8 v/v), with a saturation constant of about 19 mmol. With a further increase in methanol concentration of up to 3% v/v, the specific growth rate remains constant. Beyond this level, the growth rate gradually drops to zero, reaching the threshold value at 7.8% v/v. The specific growth rate is also strongly dependent on the size of the inoculum. The biomass yield decreases with an increase in the methanol concentration. Formaldehyde and formic acid, oxidative products of methanol, approximately 100 and ten times, respectively, more toxic to growth than the original substrate.     

The gamma-carboxy glutamic acid content of human and bovine prothrombin following warfarin treatment.

A form of prothrombin induced by Warfarin therapy, has been isolated which is adsorbed onto insoluble barium salts, but has a reduced biological activity. This protein contains, on average, seven out of a possible ten gamma-carboxy glutamic acid residues. A second form of prothrombin is also described, which is not adsorbed into barium slats, and has less than 1% the activity of the normal protein, contains only four gamma-carboxy glutamic acid residues. The significance of these results is discussed.     

The relationship between the insulin content and inhibitory effects of bovine colostrum on protein breakdown in cultured cells

Protein degradation in ten mammalian cell lines is markedly inhibited by small amounts of bovine colostrum. This response is consistent with the growth-promoting activity of colostrum that has been reported previously. Fractionation of colostrum on DEAE cellulose showed that most of the inhibitory activity against protein breakdown in H35 cells coeluted with insulin. Insulin concentrations in different batches of bovine colostrum ranged from 0.67 nM to 5.7 nM, approximately 100-fold higher than in blood. The sensitivity of protein breakdown in H35 or MH₁C₁ hepatoma lines to these colostrum samples was proportional to their insulin concentrations and could largely be accounted for by the amount of insulin present. Removal of insulin from colostrum by means of a protein A-anti-insulin antibody affinity column was accompanied by a loss of the ability of colostrum to inhibit protein breakdown in H35 or MH₁C₁ cells. However, in IMR90 fibroblasts, a cell line with a similar sensitivity to colostrum as the two hepatomas but very insensitive to insulin, protein breakdown was still inhibited by the insulin-free colostrum. These results suggest that, whereas the effect of bovine colostrum in H35 or MH₁C₁ cells is actually a response to insulin, different growth factors in colostrum account for the inhibition of protein breakdown in other cell lines.     

Use of immunoglobulin enriched bovine colostrum against oral challenge with enterohaemorrhagic Escherichia coli O157:H7 in mice

An immunoglobulin enriched bovine colostrum preparation, IMMULAC (New Zealand Dairy Group, Cambridge, New Zealand), contains antibodies against various bacterial antigens. In the present study, we investigated the protective effects of a commercial bovine colostrum preparation against infections with enterohaemorrhagic Escherichia coli (EHEC) O157:H7 in a murine model. Balb/c mice were given drinking water containing streptomycin for 3 days before and following oral challenge with streptomycin-resistant EHEC O157:H7 strain (O157-SM(R)). In mice pretreated with streptomycin, EHEC O157:H7 maintained stable levels of bacterial colonization in the intestines for the 3-week experimental time period. Oral administration of colostrum resulted in rapid decrease in the bacteria numbers compared with administration of skim-milk. Colostrum showed no direct in vitro bactericidal properties against either EHEC O157:H7. When sections prepared from cecum walls of streptomycin-pretreated mice were incubated in vitro with EHEC O157:H7, the colostrum significantly prevented the attachment of the organisms to the sections when compared with skim-milk. These results indicate that oral administration of bovine colostrum effectively protects mice against food-borne infections by inhibiting bacterial attachment to the intestinal mucous membrane, colonization and growth in the intestinal tract.     

Successful hyperimmune bovine colostrum treatment of Savanna monitors (Varanus exanthematicus) infected with Cryptosporidium sp

Therapy based on the protective passive immunity of hyperimmune bovine colostrum (HBC) (raised against Cryptosporidium parvum in cows) was applied to 4 Savanna monitors (Varanus exanthematicus) with gastric Cryptosporidium sp. infections. All lizards were moderately emaciated, and their fecal and gastric lavage samples contained moderate numbers of Cryptosporidium sp. oocysts. The first 3 of 7 gastric HBC treatments at 1-wk interval each decreased the numbers of oocysts in the fecal and gastric samples to undetectable levels. Neither feces nor lavages of the HBC-treated lizards contained Cryptosporidium sp. oocysts after the HBC therapy, whereas such samples of a single control lizard remained positive for oocysts. Two of the HBC-treated lizards died spontaneously due to metastasized carcinoma and septicemia of unknown etiology, respectively, and 2 lizards treated and killed during the experiment were histologically negative for developmental stages of Cryptosporidium sp. The control lizard died spontaneously of septicemia of unknown etiology and contained developmental stages of Cryptosporidium sp. in the gastric region. The HBC therapy was efficacious in V. exanthematicus and is recommended for lizards with gastric cryptosporidiosis.     

Effect of oak acorn level on colostrum composition and plasma immunoglobulin G of late-pregnant goats and their kids

Oak acorn contains high levels of tannins which have capacity to form complexes with proteins and consequently reduce their availability and as a result it could reduce colostrum quality and immunoglobulin (Ig) synthesis. Thus, the aim of this experiment was to investigate the effects of dietary oak (Quercus persica) acorn (OA) level during the last 60 days of pregnancy on colostrum composition and plasma metabolites and IgG level of goats and their kids. In all, 24 multiparous pregnant goats (41.7±2.3 kg BW) were assigned into one of the three experimental diets consisted of control diet (without OA) and diets containing 20% (OA₂₀) or 40% (OA₄₀) OA on dry matter basis. Goats fed OA₄₀ had lower BW change compared with other groups (P⩽0.05). Kids birth weight was not affected by experimental diets (P>0.05). Goats fed OA containing diets had lower plasma glucose, triglyceride and Fe concentrations compared with those fed the control diet (P<0.01). Plasma IgG (P<0.01) and urea (P<0.05) concentrations were lower in goats fed OA₄₀ compared with others. Animals fed OA containing diets had higher plasma alanine aminotransferase than those fed the control diet (P<0.05). Goats fed OA₄₀ produced colostrum with lower fat (P⩽0.05) and lactose (P<0.01) contents compared with other animals. Goats fed OA containing diets had lower colostrum IgG level than those fed the control diet (P⩽0.05). Kids plasma total protein concentration was not affected by experimental diets (P>0.05), whereas kids born from goats fed OA containing diets had lower plasma IgG level compared with the control diet (P<0.01). It is concluded that feeding OA during the last 60 days of pregnancy decreased colostrum quality which may have adverse effect on kid’s survival.     

Enzymic characteristics of fat globule membranes from bovine colostrum and bovine milk

Fat globule membranes have been isolated from bovine colostrum and bovine milk by the dispersion of the fat in sucrose solutions at 4 degrees C and fractionation by centrifugation through discontinuous sucrose gradients. The morphology and enzymic characteristics of the separated fractions were examined. Fractions comprising a large proportion of the total extracted membrane were thus obtained having high levels of the Golgi marker enzymes UDP-galactose N-acetylglucosamine beta-4-galactosyltransferase and thiamine pyrophosphatase. A membrane-derived form of the galactosyltransferase has been solubilized from fat and purified to homogeneity. This enzyme is larger in molecular weight than previously studied soluble galactosyltransferases, but resembles in size the galactosyltransferase of lactating mammary Golgi membranes. In contrast, when fat globule membranes were prepared by traditional procedures, which involved washing the fat at higher temperatures, before extraction, galactosyltransferase was not present in the membranes, having been released into supernatant fractions, When the enzyme released by this procedure was partially purified and examined by gel filtration, it was found to be of a degraded form resembling in size the soluble galactosyltransferase of milk. The release is therefore attributed to the action of proteolytic enzymes. Our observations contrast with previous biochemical studies which suggested that Golgi membranes do not contribute to the milk fat globule membrane. They are, however, consistent with electron microscope studies of the fat secretion process, which indicate that secretory vesicle membranes, derived from the Golgi apparatus, may provide a large proportion of the fat globule membrane.     

Effect of nitrogen fertilization and cropping on the composition and amount of the soil-organic-matter extracted with formic acid

Summary A calcareous meadow soil was treated with urea at three levels in pots with and without a crop of maize. Results showed no significant change in the amounts or composition of the first organic-matter fraction (extracted with 98% formic acid), but urea addition caused significant decrease (at 5% level) in the yield and nitrogen of the second fraction (extracted with hydrofluoric acid). As expected the yields and nitrogen contents of the maize crop grown for seven weeks generally increased with increasing rates of nitrogen fertilization.     

Interaction of trypsin with a trypsin inhibitor from bovine colostrum]

Kinetics of trypsin association with trypsin inhibitor from colostrum (IC) was studied. The association rate constant is 3-10-5 M- minus 1 sec- minus 1 at pH 7,8, 25 degrees C. The rate constant for the complex dissociation was determined from the kinetics of the IC displacement from the complex with trypsin by a specific substrate and was found to be 5-10- minus 6 sec- minus 1 (pH 7,8; 25 degrees C). The equilibrium constant (Ki) was measured in a special experiment and was equal to 4-10- minus 12 M (p H 7,8; 25 degrees C). The similarity of this reaction and the association of trypsin with other protein inhibitors was discussed.