Characterization of lyophilized equine colostrum.

Foals require maternal colostrum in the first hours of life to prevent failure of transfer of passive immunity (FTIP). Innovative storage methods such as lyophilization may enable conservation of colostrum immunoglobulins by a differentiated process of dehydration. The current study aimed to compare the quality of equine colostrum after freezing and after the lyophilization process. Thirty-one pregnant Quarter Horse mares were used. The IgG concentration of frozen and lyophilized colostrum was determined by simple radial immunodiffusion (SRID) and Brix refractometry. The physical-chemical composition (pH, total protein (TP), fat, lactose, salts, total solids (TS), and density) of the samples was evaluated and the lyophilized colostrum reconstitution test was performed. There were no significant differences (P > 0.05) in the variables IgG, fat, lactose, salts, TS, density, and pH between samples measured before and after lyophilization. There was a significant difference (P < 0.05) between the Brix average and the TP of the frozen and lyophilized colostrum samples. Lyophilization resulted in a small reduction (6.55%) in the IgG concentration measured by SRID. A strong positive correlation was observed between colostrum density and IgG concentration by SRID (r = 0.76) and between Brix and IgG concentration by SRID (r = 0.77). In the reconstitution test, the lyophilized colostrum was easily rehydrated in water, with full dilution, and remained stable. Lyophilization could be an alternative for the conservation of mare colostrum, since it is a very efficient process for retaining the physicochemical characteristics of the product, with minimal loss, particularly of IgG.

Effect of seaweed feeding before calving on colostral IgA concentration in Japanese Black cows.

In this study, we investigated the effect of feeding seaweed to Japanese Black cows before calving on IgA concentrations in colostrum. Seven Japanese Black breeding cows were used as test animals, with three cows in the seaweed-fed group (seaweed group) and four in the seaweed-non-fed group (control group). Each cow was fed 6 kg of sudangrass hay and 2.5 kg of compound feed twice daily (09:00 a.m. and 04:00 p.m.) as basal diets. Both groups had free access to water. In the seaweed group, commercially available seaweed feed was fed from 2 months before calving until the day of calving. The seaweed of 150 g/head/day was added to the basal diet at the morning feeding. Colostrum collected immediately after calving was used to measure IgA concentrations by ELISA. The IgA concentration in colostrum was significantly higher in the seaweed group than in the control group (P < 0.05). This suggested that feeding seaweed to Japanese Black cows before calving may increase IgA concentration in colostrum.

An Investigation of the Suitability of Viscosity Detection in Estimating IgG Content in Mare Colostrum.

An adequate supply of colostrum is important for the prevention of hypogammaglobulinaemia in foals. In addition to the quantity of colostrum consumed and the time of consumption, the quality of the colostrum, the immunoglobulin (Ig) G concentration, is crucial. The aim of this study was to determine whether the viscosity of equine colostrum was a suitable estimate of IgG concentration. IgG content of colostrum was measured by ELISA and viscosity directly measured with a cone plate viscometer and indirectly assessed with a funnel. Analysis of 56 colostrum samples obtained from 40 mares at different postpartum time points was conducted to assess colostrum samples with varying levels of quality. The range of IgG concentrations determined by ELISA was 0.83 to 245.5 mg/mL (30.69 ± 41.92 mg/mL). The range of viscosity values determined by the cone plate method was 1.84 to 110.00 cP (7.86 ± 17.48 cP) at a shear rate of 3 rpm. Colostrum drainage from the funnel (drainage time), varied between 7.9 and 30.0 s, with an average of 9.96 ± 4.48 s. As the data were not normally distributed, Spearman's rank correlation analyses were calculated and significant correlation found between viscosity and IgG content (ρ = 0.71, P < .001), as well as between drainage time and IgG content (ρ = 0.75, P < .001). These correlations indicate that determining the viscosity of equine colostrum by cone plate or drainage time, may be an effective proxy measurement of IgG content.

Comparison of turbidometric immunoassay and brix refractometry to radial immunodiffusion for assessment of colostral immunoglobulin concentration in beef cattle.

BACKGROUND: Colostral immunoglobulin G (IgG) concentration is critical to the attainment of adequate transfer of passive immunity in cattle, however, studies comparing available tools for measurement of colostral IgG concentration in beef cattle are limited. OBJECTIVES: To report the agreement between 3 commercially available tests for evaluating IgG concentration in beef colostrum. ANIMALS: Two hundred six beef-breed cows hospitalized for calving management or dystocia. METHODS: Retrospective study to assess IgG of whole colostrum measured stall-side via turbidimetric immunoassay (TI) and brix refractometry (BRIX), compared to fat separated (FS) analysis via single radial-immunodiffusion (RID; reference standard), TI-FS and BRIX-FS. Test performance was assessed using Passing Bablock regression, Bland-Altman analysis, and area under the curve to determine optimal thresholds. RESULTS: Correlation between RID and TI-FS, BRIX-FS, or BRIX was similar (Spearman's ρ = 0.717, 0.715, 0.716, respectively) but correlation for TI was poor (ρ = 0.586). Regression analysis identified a substantial constant (-214.75 [CI: -272.03 to -178.07]) and proportional (13.24 [CI: 11.81-15.37]) bias between the RID and TI-FS which was similar for TI. TI-FS concentrations of 28.47, 38.75, and 50.62 g/L, BRIX-FS of ≤21.9%, ≤24.0%, and ≤27.4%, and BRIX of ≤21.3%, ≤23.8%, and ≤26.4% indicated IgG concentrations <50, <100, and <150 g/L, respectively; appropriate cutoffs for TI could not be generated. CONCLUSIONS AND CLINICAL IMPORTANCE: Both TI and TI-FS demonstrated a large constant and proportional bias compared to RID; BRIX and BRIX-FS were well correlated with RID and remain a reliable method for estimation of colostral IgG concentration in beef cattle.

Minerals and essential amino acids of bovine colostrum: Phenotypic variability and predictive ability of mid- and near-infrared spectroscopy.

Colostrum quality and volume are fundamental for calves because it is the primary supplier of antibodies and the first source of energy, carbohydrates, lipids, proteins, minerals, and vitamins for the newborn. Assessing the detailed composition (i.e., AA and mineral content) of bovine colostrum (BC) on-line and at a reasonable cost would help dairy stakeholders such as farmers or veterinarians for precision feeding purposes and industries producing preparations containing BC such as foodstuff, supplements, and medicaments. In the present study we evaluated mid- (MIRS) and near-infrared spectroscopy (NIRS) prediction ability for AA and mineral composition of individual BC. Second, we the investigated the major factors affecting the phenotypic variability of such traits also evaluating the correlations with the Ig concentration. Results demonstrated that MIRS and NIRS were able to provide sufficiently accurate predictions for all the AA. The coefficient of determination in external validation (R2V) fell, in fact, within the range of 0.70 to 0.86, with the exception of Ile, His, and Met. Only some minerals reached a sufficient accuracy (i.e., Ca, P, S, and Mg; R2V ≥ 0.66) using MIRS, and also S (R2V = 0.87) using NIRS. Phenotypically, both parity and calving season affected the variability of these BC composition traits. Heifers' colostrum was the one with the greatest concentration of Ca and P, the 2 most abundant minerals. These minerals were however very low in cows calving in summer compared with the rest of the year. The pattern of AA across parities and calving season was not linear, likely because their variability was scarcely (or not) affected by these effects. Finally, samples characterized by high IgG concentration were those presenting on average greater concentration of AA. Findings suggest that infrared spectroscopy has the potential to be used to predict certain AA and minerals, outlining the possibility of implementing on-site analyses for the evaluation of the broad-sense BC quality.

Evaluation of laboratory and on-farm tests to estimate colostrum quality for dairy cows.

The objectives of this study were to evaluate different analytical methods to determine colostrum quality in dairy cattle, including one laboratory-based method (ELISA) and 4 on-farm tests. We hypothesized that the colostral IgG concentration using different analytical methods, such as ELISA (mg/mL), digital Brix refractometer (% Brix), colostrometer (specific gravity and mg/mL), an outflow funnel (seconds), and a lateral flow assay (mg/mL), were highly correlated with the reference method, radial immunodiffusion (RID; mg/mL) and would generate comparable results. Colostrum samples were collected from 209 Holstein Friesian cows on 2 commercial dairy farms in Germany. Colostrum weight and colostrum temperature were measured. Test characteristics, such as optimum thresholds, sensitivity, specificity, and area under the curve (AUC) were determined using a receiver operating characteristic curve analyses for each test. Out of 209 colostrum samples assessed by RID, 186 (89%) samples had high quality (≥50 mg IgG/mL), while 23 colostrum samples (11%) showed poor quality with IgG concentrations less than 50 mg/mL. The mean IgG concentration (±SD) was 101.3 ± 45.9 mg/mL and the range was 6.0 to 244.3 mg/mL. The Pearson correlation coefficient (r) between RID and ELISA was r = 0.78. In comparison to RID, Pearson correlation coefficients for the on-farm tests were: r = 0.79 (digital Brix refractometry), r = 0.58 (colostrometer: specific gravity), r = 0.61 (colostrometer: temperature corrected), r = 0.26 (outflow funnel) and r = 0.43 (lateral flow assay), respectively. The optimal threshold to identify high-quality colostrum using ELISA was 50.8 mg/mL with sensitivity 91.3%, specificity 92.3%, and AUC of 0.94. For the on-farm tests sensitivity ranged from 95.7% (Brix refractometry) to 60.9% (lateral flow assay). Specificity ranged from 88.6% (lateral flow assay) to 75.9% (colostrometer: temperature corrected). The AUC ranged from 0.93 (Brix refractometry) to 0.73 (outflow funnel). Based on the AUC, ELISA (0.94) and Brix refractometry (0.93) can be considered highly accurate. In conclusion, the ELISA is accurate to assess colostrum quality. Regarding the on-farm tests only the digital Brix refractometer and the colostrometer were adequate to determine colostrum quality.

Dietary embelin supplementation during mid-to-late gestation improves performance and maternal-fetal glucose metabolism of pigs.

This study aimed to evaluate the effects of dietary embelin supplementation during late gestation (from days 60 to 110) on performance and maternal-fetal glucose metabolism of pigs. Sixty sows (Duroc × Yorkshire × Landrace; parity = 1.68 ±â€…0.03; N = 20) were randomly divided into three gestation (day 60 of pregnancy) treatments, Control pigs (CON) were fed a basal diet, and the other animals were fed a basal diet supplemented with 200 or 600 mg/kg embelin per kg of feed. The body weight, backfat thickness and litter size of the sows, and birth weight and mortality of piglets were recorded. Sows' blood and piglets' umbilical cord blood were collected for the measurements of hematological parameters and anti-oxidative and immune indexes, and maternal-fetal glucose metabolism parameters, respectively. The colostrum and milk and fecal samples of the sows were also collected for analysis of milk composition and apparent total tract nutrient digestibility. Dietary embelin had no effect on the BW and backfat thickness of the sows but significantly increased the birth weight of piglets (P < 0.05) and decreased the mortality (P < 0.05). Moreover, the white blood cell counts (day 90), neutrophil count and mean cell hemoglobin (day 110), total anti-oxidant capacity (T-AOC), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT) content of the sows were increased significantly (P < 0.05) in the embelin groups than that in the CON group, whereas the malondialdehyde (MDA) content was decreased (P < 0.05). Embelin significantly increased immunoglobulin A (IgA) and immunoglobulin G (IgG) content in plasma of piglets as well as those in colostrum and milk of sows than the CON treatment (P < 0.05). In addition, dry matter, ash, and ether extract in the colostrum were similar between groups (P > 0.05), whereas the embelin significantly increased the crude protein in the milk. The apparent total tract nutrient digestibility was similar between treatments (P > 0.05). The embelin treatment significantly increased the glucose levels and lactate dehydrogenase B (LDHB) activity in sows plasma, and decreased the lactate levels in both sows and fetuses plasma (P < 0.05). Collectively, this study indicates that sows fed with embelin in mid-to-late gestation showed improved maternal health and anti-oxidative status, milk protein content, and maternal-fetal glucose metabolism, showing promise in natural plant extract nutrition for sows.

Abnormal glucose metabolism in sows in late gestation can lead to incapacity of sow production, and even reproductive disorders. It has been confirmed that inefficient glucose utilization and oxidative damage are intimately related. Thus, studies about alleviating oxidative stress and facilitating glucose metabolism in pregnant sows can be relevant. As an excellent anti-oxidative plant extract, embelin has been widely used in dietary supplementation of rodents, however, the effect of dietary supplementation with embelin on the performance of sows and newborn piglets, as well as on the glucose metabolism has rarely been reported. The present study provides the first evidence that dietary supplementation with embelin during mid-to-late gestation improved maternal immune and oxidative status, the milk quality as well as the glucose metabolism of both sows and piglets, suggesting that embelin may be a promising natural plant extractive of nutrition for sows especially during mid-to-late pregnancy and lactation.

Effect of Supplementation with Saccharomyces cerevisiae and ß-glucans to Mares During Late Gestation on Colostrum Quality and Passive Transfer of Immunity in Foals.

The objective of this study was to determine whether supplementation with Saccharomyces cerevisiae or ß-glucan, in the maternal diet during late pregnancy affects the concentration of total IgG in the colostrum of mares and influences the concentration of IgG in its foals. A total of 21 pregnant mares were used, aged 6±2 years, 3±1 pregnancies, 450±50kg in weight, and they were distributed into three groups: the control group (n=7); the S. cerevisiae group (n=7), which received 1010CFU of S. cerevisiae orally; and the ß-glucan group (n=7), which received 0.35g of ß-glucan orally. All groups started from the 300th day of their pregnancies until delivery. Samples of colostrum and serum from the mares were collected immediately after delivery. Blood samples from their foals were collected 12h after birth. The IgG measurement was performed using radial immunodiffusion. The results underwent a variance analysis. Higher concentrations of IgG were observed in the colostrum of mares that were supplemented with ß-glucans (74.14±15.25 g/L) when compared to the control group (53.80g±10.95g/L). Serum IgG concentrations of foals born to mares supplemented with Saccharomyces cerevisiae (11.57±5.05 g/L) showed a significant difference, with a higher concentration of IgG in the serum compared to the control group. Therefore, this study provides evidence that manipulation of the mares' diets in late gestation to add ß-glucan increased the IgG concentration in their colostrum. The addition of S. cerevisiae appears to improve the concentration of IgG in their foals within 12h after birth.

Formula supplementation with human and bovine milk oligosaccharides modulates blood IgG and T-helper cell populations, and ex vivo LPS-stimulated cytokine production in a neonatal preclinical model.

Introduction: Human milk contains structurally diverse oligosaccharides (HMO), which are multifunctional modulators of neonatal immune development. Our objective was to investigate formula supplemented with fucosylated (2'FL) + neutral (lacto-N-neotetraose, LNnt) oligosaccharides and/or sialylated bovine milk oligosaccharides (BMOS) on immunological outcomes. Methods: Pigs (n=46) were randomized at 48h of age to four diets: sow milk replacer formula (CON), BMOS (CON + 6.5 g/L BMOS), HMO (CON + 1.0 g/L 2'FL + 0.5 g/L LNnT), or BMOS+HMO (CON + 6.5 g/L BMOS + 1.0 g/L 2'FL + 0.5 g/L LNnT). Blood and tissues were collected on postnatal day 33 for measurement of cytokines and IgG, phenotypic identification of immune cells, and ex vivo lipopolysaccharide (LPS)-stimulation of immune cells. Results: Serum IgG was significantly lower in the HMO group than BMOS+HMO but did not differ from CON or BMOS. The percentage of PBMC T-helper cells was lower in BMOS+HMO than the other groups. Splenocytes from the BMOS group secreted more IL-1ß when stimulated ex vivo with LPS compared to CON or HMO groups. For PBMCs, a statistical interaction of BMOS*HMO was observed for IL-10 secretion (p=0.037), with BMOS+HMO and HMO groups differing at p=0.1. Discussion: The addition of a mix of fucosylated and sialylated oligosaccharides to infant formula provides specific activities in the immune system that differ from formulations supplemented with one oligosaccharide structure.

Neutrophil function and antibody production during the transition period: Effect of form of supplementary trace minerals and associations with postpartum clinical disease and blood metabolites.

Our objectives were to evaluate the effect of the form of supplementary trace minerals-inorganic salts (STM: Co, Cu, Mn, and Zn sulfates and Na selenite) or organic (OTM: Co, Cu, Mn, Zn proteinates, and selenized yeast)-fed at 100% of recommended levels in both pre- and postpartum diets on in vitro phagocytic activity of neutrophils, and in vivo IgG responses to an ovalbumin challenge during the transition period. In addition, we investigated the associations of these immunological responses with incidence of postpartum clinical diseases and the dynamic changes of metabolic markers during the transition period. Pregnant heifers and cows (n = 273) were enrolled at 45 ± 3 d before expected calving, blocked by parity and body condition score, and allocated randomly to STM or OTM supplementation. Cows in both treatments were fed the same diet, except for the form of supplementary trace minerals. Automatic feeding gates were used to assign treatments to individual cows. Blood was collected on d -7 ± 3 and 7 ± 3 relative to calving in a subgroup of cows (n = 131 and 133, respectively) to measure phagocytic activity of neutrophils in vitro using flow cytometry. Subcutaneous immunization with 0.5 mg of chicken egg ovalbumin was performed in a subgroup of cows (n = 181) on d -45, -21, and 3 relative to calving. Concentration of anti-ovalbumin IgG in serum was measured by ELISA on d -45, -21, 3, 7, and 21. Trace mineral concentrations in blood were measured by inductively coupled plasma mass spectrometry on d -45, -21, -7, 0, 7, and 21 relative to calving. Selected metabolites were measured on d -21, -10, -3, 0, 3, 7, 10, 14, and 21 relative to calving. Treatment did not affect the percentage of neutrophils performing phagocytosis on d -7 or 7 but the median fluorescence intensity of phagocytosis on d 7 was greater for OTM than STM. We found no differences between treatments in the level of anti-ovalbumin IgG in serum on any of the sampling days. Changes in neutrophil function from prepartum to postpartum were associated with incidence of postpartum clinical disease, postpartum feed intake and milk production, concentrations of Ca, K, Se, Mn, Co, and total protein in serum. Immunoglobulin G responses to ovalbumin injections were not associated with incidence of postpartum clinical disease but were associated with body weight, feed intake, energy balance, and concentrations of nonesterified fatty acids, aspartate aminotransferase, gamma-glutamyl transpeptidase, albumin, Na, P, and Cu in serum. In conclusion, replacement of STM by OTM improved one measure of phagocytic capacity of neutrophils in vitro, which was also greater in cows that did not develop postpartum clinical disease. The associations of innate and acquired immune responses with feed intake, energy balance, and circulating concentrations of key macro and micronutrients reinforce the importance of nutritional management for the health of dairy cows during the transition period.

Implications of placentation type on species-specific colostrum properties in mammals.

Maternal care is essential to optimally support survival of the offspring. During evolution of mammalian species, different phenotypes have evolved in relation to gestation length, number, size, and maturation stage of the offspring at parturition, as well as colostrum and milk composition. The aim of the present review is to describe relationships between placental function and colostrum and milk composition in different mammalian species. Species covered in this article include humans, rabbits, rodents (rat and mouse), carnivores (cats and dogs), and a variety of ungulate species (cattle, sheep, goats, pigs, and horses). Species-specific aspects are elucidated with a special focus on the transfer of passive immunity. In this regard, the structure and thus the capability of the placenta to transport immunoglobulins from maternal to fetal circulation in utero dictates the necessity of the passive transfer of immunity via colostrum. Consequently, species with exclusive postpartal transfer of immunity such as in all ungulate species have greater immunoglobulin G concentrations in colostrum than species with a prepartal transfer in utero, where especially immunoglobulin A with its local immune function in the gastrointestinal tract is present in colostrum (e.g., rabbit and human). In terms of the nutritional purpose, suckling frequency is an important factor determining the gross composition of colostrum as well as in the mature milk of these species. Milk of nidicolous animals with long intervals in-between suckling events contains more fat than milk of nidifugous animals with constant access to their mother. However, the importance of colostrum and milk consumption for newborn animals and human babies goes beyond nutrition and the transfer of immunity. Numerous bioactive components such as growth factors, hormones, and oligosaccharides are enriched in colostrum and transition milk, which support the development of the intestinal tract and local immune system.

During evolution of mammalians, intrinsic strategies and components of maternal care during the transition from pregnancy to lactation have evolved into a broad variety in gestation length, number, size, and maturation stage of the offspring at parturition, and colostrum and milk composition. The original purpose of immuno-protective glandular secretions is still conserved in many mammalian species, where colostrum, that is, the very first milk obtained after parturition, contains the greatest amounts of immunoglobulins, leukocytes, lysozyme, lactoferrin, oligosaccharides, etc. Apart from its nutritive function, bioactive components in colostrum and milk support the development of gastrointestinal structures and intestinal microflora. Depending on the placentation type and intrauterine transfer of immunoglobulins, the survival of the neonate depends more or less on the passive transfer of immunoglobulins via colostrum. The aim of the present review is to describe relationships between placenta function and colostrum (and milk, respectively) composition in different mammalian species. Special attention is paid to the transfer of passive immunity from the dam to the offspring.

Impact of prepartum administration of a vaccine against infectious calf diarrhea on nonspecific colostral immunoglobulin concentrations of dairy cows.

Passive transfer of colostral immunoglobulins from the cow to the calf is essential for calf health. The objective of this study was to determine if prepartum administration of a vaccine stimulates increased concentrations of colostral immunoglobulins of dairy cows beyond what is explained by vaccine-specific immunoglobulins. A prospective cohort study was conducted on a spring-calving commercial dairy farm that had a policy of only vaccinating cows with even ear tag numbers with a calf diarrhea vaccine, whereas cows with odd ear tag numbers were left unvaccinated. Cows in the vaccinated group (even ear tag numbers, n = 204) received a sensitizer and booster vaccination with a vaccine against bovine rotavirus (serotypes G6 and G10), bovine coronavirus, and E. coli having the K99 pili adherence factor. A sensitizer was given because the study vaccine was different from the vaccine previously used. Cows in the control group (odd ear tag numbers, n = 194) received a 2-mL subcutaneous sterile saline solution. Both groups received two treatments at a 3-wk interval, completing the treatments approximately 2 wk prior to the planned start of calving. During the calving period, technicians separated calves from cows immediately after parturition and prior to suckling, and cows were completely milked out within 6 h of parturition. Vaccine-specific, total, and nonvaccine-specific (total minus vaccine-specific) concentrations of immunoglobulin classes A, G1, G2a, and M (IgA, IgG1, IgG2a, and IgM, respectively) were quantified by mass spectrometry for 20 colostrum samples from each treatment group. Predicted mean non-vaccine-specific colostral IgM concentrations were 8.76 (95% CI = 7.18-10.67) and 5.78 (95% CI = 4.74-7.05) mg/mL for vaccinated and control cows, respectively (P = 0.005). Predicted mean non-vaccine-specific colostral IgG1 concentrations were 106.08 (95% CI = 92.07-120.08) and 95.30 (95% CI = 81.30-109.31) mg/mL among vaccinated and control cows, respectively; however, these means were not significantly different (P = 0.278). It is thus possible that the vaccine, in addition to specifically managing infectious calf diarrhea, may also have non-specific benefits by improving colostrum quality through increased non-vaccine-specific colostrum IgM concentrations. Further research is necessary to determine the mechanism for these preliminary findings, whether the effect may occur in other immunoglobulin classes, and what impacts it may have on calf health outcomes.

Unlike human babies, calves do not receive protective immune proteins (immunoglobulins) from the mother before birth, so a sufficient volume of immunoglobulin-rich colostrum of adequate quality must be consumed within hours of birth. It can be a challenge to meet this requirement for all dairy calves. Prior to calving, cows can be vaccinated with a vaccine against specific infectious causes of calf diarrhea to stimulate elevated concentrations of specific immunoglobulins in their colostrum, which is consumed by their calves to protect them until their own immune systems develop. We enrolled cows that were either vaccinated or not with a calf diarrhea vaccine and, using novel laboratory techniques, measured concentrations of immunoglobulin classes A, G, and M in their colostrum. As expected, vaccinated cows had elevated concentrations of vaccine-specific immunoglobulins in their colostrum. However, they also had elevated non-vaccine-specific concentrations of immunoglobulin M. The vaccine may therefore have stimulated a nonspecific increase in colostral immunoglobulin M concentrations. Further research is necessary to confirm the preliminary findings of the present study and determine the mechanism for this apparent nonspecific increase in colostral immunoglobulin M concentrations, whether it may occur in other immunoglobulin classes, and whether it may benefit calf health and growth.

Comparison of Different Methods to Determine the Absorption of Colostral IgG in Newborn Foals.

The timely diagnosis of abnormalities in the passive transfer of colostral immunoglobulins is important for the health and development of newborn foals. This study investigated three different methods for measuring immunoglobulin G concentration in neonatal foals. Comparison of a commercial SNAP assay, total protein concentration determination, and total globulin calculation by subtracting the albumin fraction from total protein as an indirect parameter was performed on a quantitative ELISA, which served as a reference method. The study included 119 samples from 148 foals between the age of 1 and 6 days. A blood concentration of 800 mg/dL was considered to indicate adequate absorption of immunoglobulins, and a concentration of less than 400 mg/dL was considered to be hypogammaglobulinemia. The sensitivity of the SNAP test was 64.5% and specificity was 94.7% for diagnosing sufficient absorption of immunoglobulin G at a value of 800 mg/dL. A value of 54 g/L was found to be most appropriate for the use of total protein and provided a sensitivity of 67.3% and specificity of 84.2%. For total globulins, the most appropriate value was 27 g/L, which yielded a sensitivity of 74.5% and specificity of 81.6%. At values under 400 mg/dL, the sensitivity of the SNAP test was 89.4% and the specificity was 83.0%. Here, the most suitable value for the total protein was 51 g/L. This provides a sensitivity of 65.2% and a specificity of 76.8%. The most suitable concentration for the use of total globulin was determined to be 24 g/L, which provided a sensitivity of 75.8% and a specificity of 78.1%. The study and its results show that the SNAP test, the TP, and the TP-A method perform similarly well compared to the ELISA in determining IgG concentration of ≥800 mg/dL. Based on the 95% confidence intervals, however, the Snap test and the TP-A method appear to perform similarly well but better than the TP approach for IgG concentrations <400 mg/dL.

Effect of vitamin K3 supplementation on immunoglobulin G concentration in colostrum of periparturient Holstein dairy cows.

This study was to examine the effects of dietary vitamin K (VK) 3 supplementation on immune-related substances in milk, oxidative stress indices in plasma and VK1, and menaquinone 4 (MK-4) in plasma and milk in periparturient dairy cows. Forty healthy perinatal Holstein-Friesian dairy cows were used in this study. Twenty-one animals were randomly selected and categorized into the VK3 supplemented (50 mg/day/head as VK3) group; the remaining 19 were categorized into the control group. On day 3 after calving, blood and milk were sampled, and their chemical components were determined. The VK3 supplemented group had significantly higher menaquinone 4 levels in plasma and milk on day 3 postpartum than the control group. In addition, there was a significant increase in the immunoglobulin G (IgG) level in milk. VK3 may be absorbed from the gastrointestinal tract and converted to MK-4, the biologically active form of VK, in the mammary gland and other tissues. It was thought that the increase in MK-4 level in plasma and milk induced an increase in the concentration of IgG in milk. VK3 supplementation to periparturient dairy cows may contribute to the production of colostrum with high concentrations of IgG and MK-4.

Detection of nucleotide variants in FCGRT (Fc fragment of IgG, receptor, transporter, alpha) gene and their influence on colostral IgG concentration in Indian water buffalo (Bubalus bubalis).

BACKGROUND: The neonatal Fc receptors (FcRn) mediate the transfer of immunoglobulin G (IgG) molecules from a dam's circulation to the colostrum produced by it immediately after parturition. In ruminants, the calves born are agammaglobulinemic therefore, ingestion of colostrum with high concentration of IgG imparts passive immunity to the newborn. The FcRn molecule is a heterodimer, coded by FCGRT (Fc fragment of IgG Receptor Transporter neonatal) and B2M (Beta 2 microglobulin) genes. Present study attempted to identify single nucleotide polymorphisms (SNPs) in the FCGRT gene in 40 buffaloes of Murrah breed and evaluated the association of these nucleotide variations and haplotypes with IgG concentration in their colostrum. METHODS AND RESULTS: Animals producing colostrum with high IgG and low IgG levels were identified by indirect ELISA and selected. SNPs were detected in the FCGRT gene sequence of selected animals by amplifying it in nine fragments covering all exons (with flanking introns) followed by PCR-single strand conformational polymorphism (PCR-SSCP). A total of nine SNPs were observed of which seven were present in flanking introns and two in exon 4 of the gene. The SNP A75G was non-synonymous and produced an amino acid change from isoleucine to valine. The exonic SNPs and corresponding haplotypes were found to be significantly (P < 0.01 and 0.05 respectively) associated with colostral IgG concentration based on Odds ratios at 95% confidence interval. CONCLUSION: Polymorphism in FCGRT gene is found to be associated with IgG concentration in colostrum and identification of females with desirable variations may prevent failure of passive transfer in neonatal ruminants.

Use of blood matrices and alternative biological fluids for antibody detection in animal tuberculosis.

Bovine tuberculosis (bTB) control programs can be improved by implementation of advanced ante-mortem testing algorithms. Serodiagnostic methods using traditional blood or blood-derived specimens may benefit from the use of less invasive alternative biological fluids, provided those mirror systemic antibody responses. In the present study, we used Dual Path Platform (DPP) and Multiantigen Print Immunoassay (MAPIA) to compare antibody levels in ten sample types including whole blood (fresh and hemolyzed), plasma (fresh and leftover from Bovigam testing), serum, saliva, broncho-alveolar lavage, urine, diaphragm extract, and bile collected from cattle aerosol-infected with Mycobacterium bovis. High correlation (r = 0.97-0.99) in measurements of IgG antibodies to MPB70/MPB83 fusion antigen by DPP assay was found between all blood-derived specimens, supporting matrix equivalency. Broncho-alveolar lavage and diaphragm extract yielded positive results in all the infected animals tested, showing high correlation with matching serum data (r = 0.94 and r = 0.95, respectively) and suggesting their potential use in antibody assays. Characterized by MAPIA, the antigen reactivity patterns obtained with paired sera and alternative specimens were nearly identical, with slight differences in intensity. Antibodies were also found by DPP assay in saliva, urine, and bile from some of the infected animals, but the titers were relatively low, thus reducing the diagnostic value of such specimens. The proposed approach was evaluated in a pilot field study on warthogs diagnosed with M. bovis infection. Relative levels of antibody in tissue fluid obtained from lymph nodes or lungs were consistent with those detected in sera and detectable in all infected warthogs. The findings support the diagnostic utility of non-traditional biological fluids and tissue samples when used as alternative test specimens in serologic assays for bTB.

Evaluation of Brix refractometer as an on-farm tool for colostrum IgG evaluation in Italian beef and dairy cattle.

In this study it is hypothesized that there are differences between immunoglobulin G (IgG) content in colostrum from beef (Chianina, Podolica) and dairy (Holstein Friesian) cows and that variables such as breed, and parity can influence IgG content. The further objective was to determine if these factors may vary in terms of sensitivity, specificity and the cut point when data obtained with the digital Brix refractometer is compared with the gold standard radial immunodiffusion assay (RID). A total of 90 samples of first-milking colostrum were collected within 2 h after parturition. IgG concentration was determined indirectly by digital Brix refractometer and directly by RID. Results obtained by RID were compared among breed and parity. For the digital Brix refractometer, sensitivity and specificity to detect colostrum with an IgG concentration lower than 50 g/l were calculated and the optimal cut-point was selected for each breed. Samples containing less than <50 g/l IgG accounted for 15.9% of the total. Parity influenced colostral IgG concentration and beef cows had a higher mean concentration of IgG (101.1 g/l in Chianina and 90.6 g/l in Podolica) than dairy cows (71.1 g/l in Holstein Friesian) First parity Chianina cows had the highest IgG mean content (116.1 g/l). At the optimal cut-point for Brix refractometer (20%) sensitivity and specificity were 0.93 (0.84-0.97) and 0.81 (0.70-0.88), however, a breed-related cut-point could be used to reduce evaluation error. Linear regression modeling showed that refractometer data were related to RID (r = 0.78). Results obtained suggest that breed and parity can influence IgG content of colostrum and, despite the Brix refractometer being an excellent on-farm tool, a breed-based definition of optimal cut point is needed.

Distribution difference of colostrum-derived B and T cells subsets in gilts and sows.

Piglets are highly vulnerable to infections, but colostrum provides them with some protection. The function of colostrum components is unknown, as is if the amount and subsets of leukocytes in colostrum differ between gilts and sows. This study serially characterized leukocyte populations in colostrum for differential leukocyte counts. Differences in humoral and cellular composition of colostrum between 40 gilts and 40 sows (parities orders 3-4) from a commercial herd were examined. Flow cytometry is a useful tool to identify and quantify leukocyte subsets in sow colostrum. Overall, there were no (p ≥ 0.05) parity differences in total macrophages, granulocytes, and T and B cells. However, the sows' colostrum presented significantly higher (p ≤ 0.05) T lymphocyte subsets than gilts, such as central memory CD4+T cells, effector memory CD4+T cells, and central memory CD8+T cells. Among B-lymphocytes, percentages of SWC7+CD5+ cells were significantly higher in sow colostrum than in that of gilts. As expected, IgG concentrations were significantly higher in sows than in gilts. Colostrum from sows had significantly greater mitogenic activity than colostrum from gilts and this fact can be associated with the potential to accelerate the maturation of a newborn's gastrointestinal tract. Our findings suggest that parity order may be one among other factors influencing the cell population and, consequently, the immune adaptive response in piglets that induces neutralizing antibodies and cellular immune responses to antigens.

Predicting colostrum and calf blood components based on refractometry.

Provision of good quality colostrum is essential for the passive immunity and nutrition of newborn calves. In order to better predict the quality of colostrum and the transfer of passive immunity, the relationships between colostrum components and between calf serum components were examined in this study. Samples of bulk tank milk, colostrum pooled from several cows 0-4 d postpartum, and colostrum collected from individual cows twice daily for 3 d post-partum were compared. With the exception of fat percentage, there were strong correlations between the levels of the components in the pooled colostrum and in the individual cow colostrum collected 0-1 d postpartum. The correlations between total solids as measured by Brix refractometry and total protein, immunoglobulin G (IgG), lactose % and protein % in colostrum within 1 d postpartum and pooled colostrum were 0.92, 0.90, -0.88 and 0.98, respectively. These high correlations enabled these colostrum components to be accurately predicted from Brix % and therefore, the volume of colostrum required to feed neonate calves can be optimised based on Brix refractometry to avoid failure of passive immunity transfer. To assess whether the components obtained from colostrum were correlated in calf blood, newborn calves were separated from their dams before suckling and blood sampled before feeding (day 0), and on days 1 and 7, after receiving colostrum or milk twice a day. The correlations between glucose, total protein, IgG, and gamma-glutamyl transferase (GGT) levels in the calf blood were lower than the correlations observed between the colostrum components. The highest correlation was between serum protein measured by refractometer and serum IgG within one week postpartum. GGT activity was not a good indicator of serum IgG levels. However, serum protein refractometer measurements predicted serum IgG level with high accuracy, providing an on-farm test to determine that calves have received sufficient passive immunity and colostrum components.

Lactoferrin and Immunoglobulin Concentrations in Milk of Gestational Diabetic Mothers.

Gestational diabetes mellitus (GDM) is associated with an increased risk of having a high-care newborn and has an impact on maternal wellbeing. This study aimed to assess the effect of GDM on the lactoferrin (LF), secretory immunoglobulin A (SIgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) concentrations in early colostrum, colostrum, and transitional milk samples of hyperglycemic (n = 53) and normoglycemic (n = 49) mothers using enzyme-linked immunosorbent assay (ELISA). The concentrations of milk lactoferrin and SIgA, but not IgG and IgM, from hyperglycemic and normoglycemic mothers, showed a similar negative correlation with lactation from the first to the fifteenth day. Apart from early colostral IgG, there were no differences in concentrations of LF and immunoglobulins in milk from hyperglycemic and normoglycemic mothers. For hyperglycemia compensated by diet (GDM G1) or insulin treatment (GDM G2), slight differences were seen for LF and IgG, but not for SIgA and IgM, during an early stage of lactation only. Early colostral IgG and colostral LF of insulin-treated mothers were higher (10.01 ± 4.48 mg/L and 11.50 ± 0.58 g/L, respectively) than for diet-control diabetic mothers (7.65 ± 5.67 mg/L and 8.05 ± 1.38 g/L, respectively). GDM of mothers does not have a significant impact on immunological quality of early milk.