Immunoglobulin G Concentrations Research Articles

An optimized method for plasma extracellular vesicles isolation to exclude the copresence of biological drugs and plasma proteins which impairs their biological characterization.

Extracellular vesicles (EVs) are cell membrane-derived phospholipid bilayer nanostructures that contain bioactive proteins, enzymes, lipids and polymers of nucleotides. They play a role in intercellular communication and are present in body fluids. EVs can be isolated by methods like ultracentrifugation (UC), polyethylene-glycol-precipitation (PEG) or size exclusion chromatography (SEC). The co-presence of immunoglobulins (Ig) in EV samples isolated from plasma (pEVs) is often reported and this may influence the assessment of the biological function and phenotype of EVs in bio- and immunoassay. Here, we studied the presence of an Ig-based therapeutic (etanercept) in pEV samples isolated from rheumatoid arthritis (RA) patients and improved the isolation method to obtain purer pEVs. From plasma of etanercept (Tumor-necrosis-factor (TNF)-α antibodies)-treated RA patients pEVs were isolated by either UC, PEG or SEC. SEC isolated pEVs showed the highest particle-to-protein ratio. Strong TNF-α inhibition determined in a TNF-α sensitive reporter assay was observed by pEVs isolated by UC and PEG, and to a lesser extent by SEC, suggesting the presence of functional etanercept. SEC isolation of etanercept or labelled immunoglobulin G (IgG) showed co-isolation of these antibodies in the pEV fraction in the presence of plasma or a high protein (albumin) concentration. To minimize the presence of etanercept or immunoglobulins, we extended SEC (eSEC) column length from 56mm to 222mm (total stacking volume unchanged). No effect on the amount of isolated pEVs was observed while protein and IgG content were markedly reduced (90%). Next, from six etanercept- treated RA patients, pEVs were isolated on a eSEC or standard SEC column, in parallel. TNF-α inhibition was again observed in pEVs isolated by conventional SEC but not by eSEC. To confirm the purer pEVs isolated by eSEC the basal IL-8 promoter activation in human monocytes was determined and in 4 out of 5 SEC isolated pEVs activation was observed while eSEC isolated pEVs did not. This study shows that extended SEC columns yielded pEVs without detectable biologicals and with low protein and IgG levels. This isolation method will improve the characterization of pEVs as potential biomarkers and mediators of disease.

Health status and infections in patients with symptomatic primary and secondary immunoglobulin G (IgG) deficiencies receiving intravenous IgG replacement

BackgroundThe effects of intravenous immunoglobulin G replacement on perceived health and infection susceptibility of patients suffering from immunoglobulin G (IgG) deficiencies should be evaluated in a prospective analysis.MethodsPatients with symptomatic primary or secondary IgG deficiencies were interviewed prior to the first IgG infusion (t0) and over the course of their treatment (t1 - t6). The respondents rated their current health using a 100 point scale (EQ-5D-5L), ranging from 0 (‘worst imaginable health’) to 100 (‘best imaginable health’). The patients also provided information on the frequency of infections and of infections requiring antibiotics in the past 8 weeks. A healthy control group (CG) without oncologic diseases answered the questions once.ResultsOne hundred six patients with a median age of 65 years (21–85 years) were investigated. The median serum IgG concentration changed from 500 mg/dl (t0) to 772 mg/dl (t6). The mean number of infections and of infections requiring antibiotics decreased during IgG replacement significantly. Current health according to EQ-5D-5L improved from 57 (t0) to 68 (t6), compared to 73 in the CG.ConclusionDuring the course of IgG replacement patients reported fewer and less severe infections. Their health assessment improved but still was inferior to the healthy CG.

Evaluation of Brix Refractometry for the Estimation of Colostrum Quality in Jennies

Donkey placenta does not allow the passage of immunoglobulins; thus, foals are born hypogammaglobulinemic and an adequate intake of high-quality colostrum in the first 24 hours of life is crucial for the surviving. The study aims to assess the relation between colostrum immunoglobulin G (IgG) concentration evaluated by the single radial immunodiffusion (SRID) test and the Brix refractometer in donkeys to establish a cutoff value for high quality of colostrum based on Brix refractometry. Colostrum was collected at foaling, and at 6, 12, and 24 hours after foaling from the left and the right half of nine Amiata jennies. A total of 72 colostrum samples were analyzed. A Friedman test with a Dunn’s test for multiple comparisons was used for assessing the differences between the left and right half at each sampling time. No differences were found between the left and right halves; the average value was used to analyze the effect of sampling time on the IgG concentrations and Brix values. The relationship between colostrum IgG concentrations (SRID test) versus Brix value and Brix value versus time was analyzed using two different mixed linear models. A strong statistically significant relation has been found between IgG concentrations and Brix values (R2 = 0.84). The relation between IgG concentrations and Brix refractometer showed a cutoff point of 17% Brix score for the identification of high-quality colostrum. The Brix value (%) decreased continuously from 16.29 by 0.29 × hour. Jennies’ and donkey foals’ management may be greatly improved using this simple and cheap device.

Emerging concept of vitreous concentrations of proteins and lipids as discriminant of fresh water drowning death

BackgroundDrowning is the leading cause of unintentional death in the world, with about 372,000 drowning deaths reported annually. Forensic opinion of drowning is considered one of the most difficult tasks in confirming the exact cause of death. Drowning is now possibly patronized to cover up intentional crimes. This study was aimed at utilizing concentrations of vitreous proteins and lipids in making inroad into the inquest of true drowning. Twelve albino rabbits constituted the sample size. The rabbits were divided into three groups; control death (CD), post-mortem submersion death (PSD) and truly drowned death (TDD). Vitreous humor was extracted from the eyes of all the rabbits and centrifuged for biochemical analysis. The biochemical parameters analyzed included vitreous total protein (TP), albumin (ALB), globulin (GLO), albumin/globulin (A/G) ratio, immunoglobulin G (IgG), immunoglobin M (IgM) total cholesterol (TC), triacylglycerol (TG), high density lipoprotein (HDL), low density lipoprotein (LDL), very low density lipoprotein (VLDL) and glucose (GLU) using standard methods. ResultThe mean concentrations of vitreous parameters amongst the groups were compared using one-way anova (pos hoc-LSD). The findings revealed that the concentrations of vitreous TP, GLO, IgG, IgM, TG, TC, TG, HDL, VLDL and GLU were significantly elevated (p < 0.05), whereas ALB and A/G ratio decreased significantly amongst the drowned death group compared to that either CD or PSD. ConclusionThe findings have indicated that some vitreous protein and lipid parameters could aid in discriminating deaths that truly emanated from true drowning from that of post-mortem drowning and suffocation.

Assessment of Brix refractometry to estimate immunoglobulin G concentration in beef cow colostrum.

BackgroundBrix refractometry can be used to assess colostral immunoglobulin G (IgG) concentration, but studies identifying Brix percentages to detect high‐ and low‐IgG colostrum are lacking for beef cows and interlaboratory agreement is unknown.ObjectivesEvaluate Brix refractometer performance and interlaboratory agreement for assessing beef cow colostrum IgG concentration, including determination of thresholds to identify colostrum containing IgG concentrations <100 g/L and ≥150 g/L.AnimalsBeef cows (n = 416) from 11 cow‐calf operations in Alberta, Canada.MethodsColostral IgG concentrations were measured using radial immunodiffusion (RID) and estimated by Brix refractometry for this retrospective study. Spearman correlation coefficients were assessed between RID and Brix refractometry. Likelihood ratios and misclassification cost‐term analysis were used to determine optimal Brix percentages for detecting colostrum containing IgG concentrations <100 g/L and ≥150 g/L. Concordance correlation coefficient (CCC) and Bland‐Altman analyses were performed for Brix percentages obtained at 3 different laboratories.ResultsBrix percentages obtained at 3 laboratories were positively correlated with IgG results (r = 0.72, 0.68, and 0.76, respectively). Colostrum Brix percentages of <24% and ≥30% were optimal for indicating IgG concentrations of <100 g/L and ≥150 g/L, respectively. Interlaboratory agreement was substantial, with CCC ranging from 0.89 to 0.96 and Bland‐Altman analysis showing small mean differences (−1.2% to 0.09% Brix) and narrow limits of agreements (−4.8% to 2.4% Brix) among laboratories.Conclusions and Clinical ImportanceBrix refractometry shows good potential for reliably estimating IgG concentrations in beef cow colostrum across multiple laboratories and can be recommended to aid colostrum management decisions on farms.

Kappa Free Light Chains and IgG Combined in a Novel Algorithm for the Detection of Multiple Sclerosis.

Background: It is well known that the cerebrospinal fluid (CSF) concentrations of free light chains (FLC) and immunoglobulin G (IgG) are elevated in multiple sclerosis patients (MS). Therefore, in this study we aimed to develop a model based on the concentrations of free light chains and IgG to predict multiple sclerosis. We tried to evaluate the diagnostic usefulness of the novel κIgG index and λIgG index, here presented for the first time, and compare them with the κFLC index and the λFLC index in multiple sclerosis patients. Methods: CSF and serum samples were obtained from 76 subjects who underwent lumbar puncture for diagnostic purposes and, as a result, were divided into two groups: patients with multiple sclerosis (n = 34) and patients with other neurological disorders (control group; n = 42). The samples were analyzed using turbidimetry and isoelectric focusing. The κIgG index, λIgG index, κFLC index, and λFLC index were calculated using specific formulas. Results: The concentrations of CSF κFLC, CSF λFLC, and serum κFLC and the values of κFLC index, λFLC index, and κIgG index were significantly higher in patients with multiple sclerosis compared to controls. CSF κFLC concentration and the values of κFLC index, λFLC index, and κIgG index differed in patients depending on their pattern type of oligoclonal bands. κFLC concentration was significantly higher in patients with pattern type 2 and type 3 in comparison to those with pattern type 1 and type 4. The κFLC index, λFLC index, and κIgG index were significantly higher in patients with pattern type 2 in comparison to those with pattern type 4. The κFLC index and κIgG index were significantly higher in patients with pattern type 2 in comparison to those with pattern type 1, and in patients with pattern type 3 compared to those with pattern type 4. The κIgG index was markedly elevated in patients with pattern type 3 compared to those with pattern type 1. In the total study group, κFLC, λFLC, κFLC index, λFLC index, κIgG index, and λIgG index correlated with each other. The κIgG index showed the highest diagnostic power (area under the curve, AUC) in the detection of multiple sclerosis. The κFLC index and κIgG index showed the highest diagnostic sensitivity, and the κIgG index presented the highest ability to exclude multiple sclerosis. Conclusion: This study provides novel information about the diagnostic significance of four markers combined in the κIgG index. More investigations in larger study groups are needed to confirm that the κIgG index can reflect the intrathecal synthesis of immunoglobulins and may improve the diagnosis of multiple sclerosis.

Intravenous immunoglobulin significantly reduces exposure of concomitantly administered anti-C5 monoclonal antibody tesidolumab.

Awareness of drug-drug interactions is critical in organ transplant recipient management. However, biologic agents interfering with monoclonal antibodies is not widely considered. We report the effect of high-dose intravenous immunoglobulin (IVIg) on safety, tolerability, pharmacokinetics (PK), and pharmacodynamics (PD) of the human anti-C5 monoclonal antibody tesidolumab (LFG316) in end-stage renal disease patients awaiting kidney transplant. In this single-center, phase 1, open-label, parallel-group study, 8 patients were assigned to receive either single-dose tesidolumab+IVIg or tesidolumab alone, with 56-day follow-up. Within-group PK parameters were consistent. Mean tesidolumab exposure decreased 34%, clearance increased 63%, and half-life decreased 41% comparing tesidolumab+IVIg to tesidolumab alone. IVIg influence on tesidolumab elimination was most evident in the first 3weeks. Complete suppression of both total and alternative complement activities was maintained for 4weeks in the tesidolumab alone group and for 2weeks in the tesidolumab+IVIg group. Tesidolumab was well tolerated. IVIg infused before tesidolumab affected tesidolumab PK and PD, resulting in a shortened period of full complement activity inhibition. These findings suggest a clinically relevant impact of IVIg on monoclonal antibody clearance and indirectly hint at an IVIg mechanism of action in treating autoimmune diseases and allosensitization by accelerating pathogenic IgG antibody degradation. Trial registration number: NCT02878616.

Effect of nano-ZnO, compared to ZnO and Zn-methionine, on performance, nutrient status, rumen fermentation, blood enzymes, ferric reducing antioxidant power and immunoglobulin G in sheep

This study was conducted to assess the effect of nano-ZnO, compared to ZnO and Zn-methionine, on dry matter (DM) intake (DMI), performance, nutrient status, digestibility, rumen fermentation, microbial nitrogen (N) synthesis (MNS), blood enzymes, ferric reducing antioxidant power (FRAP) and immunoglobulin G (IgG) in sheep. Thirty-six healthy male Shal sheep were divided into four experimental groups (n = 9) in a completely randomized design, and assessed for 40 days. The control animals received the basal diet without Zn supplementation, containing a native Zn content of 22.6 mg/kg DM. Other sheep were fed with the diets containing 28 mg supplementary Zn/kg DM in the form of ZnO, Zn-methionine or nano-ZnO. The DMI of sheep receiving Zn-methionine was higher (P = 0.035) than the control group, but was similar among the Zn-supplemented groups. Although supplementation of the basal diet with Zn increased (P < 0.05) the digestibility, rumen total volatile fatty acids (VFA), average daily gain and feed efficiency, the chemical forms of Zn did not affect these variables. The intake, daily absorption, retention and blood concentration of Zn were not affected by using nano-ZnO instead of ZnO and Zn-methionine. The animals receiving Zn-methionine had higher (P = 0.019) rumen fluid Zn than the ZnO fed sheep. Compared to the control diet, the Zn supplementations increased retained N (P = 0.037) and decreased rumen ammonia-N (P = 0.046). Using nano-ZnO instead of ZnO and Zn-methionine had no effect on rumen ammonia-N, retained N and MNS. The blood urea-N (BUN) concentration was lower (P = 0.027) for the Zn-methionine and nano-ZnO fed animals versus the control and ZnO groups. The nano-ZnO and Zn-methionine fed sheep had higher (P < 0.05) rumen and blood FRAP and Zn absorbability than the ZnO group. The blood IgG concentration was higher (P = 0.028) for the Zn-methionine and nano-ZnO fed animals versus the control sheep, and nano-ZnO group versus the ZnO fed animals. There were no differences among sheep in terms of the ruminal pH, individual VFA proportions and protozoa counts, Cu status and blood alkaline phosphatase, aspartate aminotransferase and alanine aminotransferase. Overall, using nano-ZnO in the diet of sheep had advantages over ZnO in terms of increasing the Zn absorbability, rumen and blood FRAP and blood IgG, and decreasing BUN. However, there were no differences between the animals consuming nano-ZnO or Zn-methionine.

The Association between Colostral density and immunoglubulin concentrations of Dairy Cattle

Abstract The colostrum contains diverse and numerous immunoglobulins. The neonatal calf obtains majority of passive immunity from colostrum, which especially has huge effects on calf health. Historically, the amount of immunoglobulins in colostrum has been predicted through the density of the colostrum, but this correlation has not yet been clearly determined. In this study, we determined the correlation between the density and the immunoglobulin concentration, the difference between Holstein and Jersey cattle, and the impact by parity number. Densities were measured with Cruise densimeter. The concentration of immunoglobulins was assayed by ELISA. The concentration of total immunoglobulin G (IgG) and antibody against foot-and-mouth disease virus (FMDV) tend to increase following the density of the colostrum. However, the level of antibody against rota-corona virus was not positively correlated with the density. Regarding species and parity effects, interestingly, Holstein cows had significantly higher values in total IgG and FMDV antibody than Jersey cows. Multiparous dairy cows produced colostrums with higher concentrations of total IgG, FMDV antibody than primiparous dairy cows.

Effect of Soy Lecithin Supplementation in Beef Cows before Calving on Colostrum Composition and Serum Total Protein and Immunoglobulin G Concentrations in Calves.

Simple SummaryUntil recently, fat supplements were considered merely as a source of energy for cows during transition. However, individual fatty acids included in fat supplements may clearly induce different production and metabolic responses, which in consequence change the nutritional value of bovine colostrum. Therefore, it is necessary to consider the type of fat additive to use in feed ration. This study aimed to determine an effect of soy lecithin supplementation on beef cow’s colostrum composition and selected blood parameters in their calves. Obtained results suggest that soy lecithin addition in cows before calving has a beneficial impact on colostrum composition. This concerns mainly an increase of linoleic acid in colostrum, which may influence IgG activity in calf serum. In turn, higher content of these components in colostrum may contribute to improve calves’ survival rate during the first weeks of their life.The aim of this study was to investigate the impact of soy lecithin supplementation in beef cow’s nutrition on colostrum composition and serum concentrations of immunoglobulin G (IgG) and serum total protein (STP) in calves. Twenty pregnant Charolaise cows were assigned to two groups. In the supplementation group (n = 10) during the last four weeks of pregnancy, soy lecithin was administrated in an amount of 20 g/cow/day. In both groups, basic composition (protein, fat, lactose, dry matter), somatic cell count (SCC), total bacteria count (TBC), IgG concentration, and fatty acids profile were determined in colostrum samples. Moreover, STP and IgG concentration were measured in calves’ blood samples on the 3rd, 7th, 14th, and 21st days of life, mothered by supplementation and control cows. Animals fed with soy lecithin before calving produced colostrum with a higher (p = 0.049) level of linoleic acid (C18:2 n-6). In addition, these results showed that soy lecithin supplementation has contributed to an increase (p = 0.029) of serum IgG in calves on the 14th day of life. The impact of such change in colostrum on IgG levels on calves serum and their half-life need further analysis.

Seroprevalences of antibodies against pertussis, diphtheria, tetanus, measles, mumps and rubella: A cross-sectional study in children following vaccination procedure in Guangzhou, China

This study investigated the concentrations and seroprevalence of immunoglobulin G (IgG) antibodies against pertussis, diphtheria, tetanus, measles, mumps and rubella among children in Guangzhou, China. We conducted a cross-sectional study focusing on the post-vaccination immune statuses of children on scheduled immunisation. Human IgG antibody against six diseases were measured using commercial enzyme-linked immunosorbent assay kits. Of 620 subjects, the male-to-female ratio was 2.04 (416/204). Seroprevalence (81.97% vs 90.20%) and IgG concentrations (686.55 IU/mL vs 884.26 IU/mL, P < 0.05) for measles, tetanus (0.94 IU/mL vs 1.21 IU/mL) and rubella (34.33 IU/mL vs 47.37 IU/mL) were all higher in females. No differences based on sex were observed in the seroprevalence and IgG concentrations for anti-pertussis antibodies, anti-diphtheria antibodies and anti-mumps. Slight increase in seroprevalence and IgG concentration occurred with anti-pertussis antibodies after primary and booster vaccinations (from 0.00% [1 m], 5.45% [6 m], to 17.14% [1.5 yr]; and from 8.57% [5 yr] to 15.79% [6 yr]). Although no booster vaccination was given after age 6 yr, the seroprevalence and IgG concentration for anti-pertussis antibodies remained relatively stable. For diphtheria, tetanus, measles and rubella, seroprevalence reached their peaks after the primary and first booster vaccination. A plateau occurred after age 1.5 yr with a declining trend in subjects >8–10 yr. The IgG concentrations of these 4 pathogens showed a dramatic increase after primary vaccination, with steadily declining trends thereafter. For mumps, subjects showed increased seroprevalence and IgG concentration after the primary mumps-containing vaccination in 1.5-yr-olds (from 7.14% to 57.14%; 52.13 IU/mL to 214.18 IU/mL); however, following that low seroprevalence levels (from 42.86% to 80.00%) were observed. The post-vaccination immune statuses against diphtheria, tetanus, measles and rubella were relatively satisfactory, compared to those against pertussis and mumps. Booster vaccination against pertussis and mumps at appropriate time should be considered.

Short communication: Red deer (Cervus elaphus) colostrum during its transition to milk

We studied changes in chemical composition, somatic cell count, and immunoglobulin G (IgG) and M (IgM) content in red deer (Cervus elaphus) colostrum during the transition to milk at different times after parturition (<5 h, 24 h, 48 h, 2 wk, and 4 wk). The production level was higher at 2 and 4 wk of lactation than during the first day after parturition, with intermediate values at 48 h postpartum. Fat content did not vary during the study period. However, total protein and casein contents were particularly high in the initial 5 h after parturition, decreasing to approximately 50% after 24 h postpartum. Conversely, lactose concentration was low in the beginning (<5 h), increasing gradually throughout the study. Similarly, dry matter dropped during the first 24 h and then remained constant throughout the study. Urea content decreased during the study, showing a slight recovery at 4 wk. Somatic cell count was higher during the first hours after parturition and gradually decreased throughout the study period. The IgG content was higher before 5 h postpartum than at 24 h postpartum. After 5 h, the level of IgG decreased progressively until it reached 0.18 mg/mL at 4 wk of lactation. We observed a similar pattern for IgM content, but it decreased more quickly than IgG and was not detected after 2 wk. In the case of deer, milk should be considered transitional from 24 to 48 h after parturition, and samples collected after 2 wk can be considered mature milk.

An Early Fecal Microbiota Transfer Improves the Intestinal Conditions on Microflora and Immunoglobulin and Antimicrobial Peptides in Piglets.

The goal of this study was to investigate the effects of early fecal microbial transfer (FMT) on the microflora of recipient piglets, where Yorkshire newborn piglets and Min sows (an indigenous pig breed in China) were used as the fecal recipients and donors, respectively, to reveal the changes in immunity and development-related functions of the intestinal mucosa driven by FMT. The recipient group was inoculated with fecal microbial fluids from days 1 to 10. On day 21, the relative abundance of the Proteobacteria was reduced; the concentrations of immunoglobulin M (IgM) and immunoglobulin G (IgG) in the jejunal mucosa, and that of IgG in the ileal mucosa of the recipient group, were increased (P < 0.05). On day 40, the relative abundance of the Firmicutes in the recipient group was increased, while that of Bacteroides was decreased. The concentrations of IgG and IgM in the ileal mucosa of the recipient group were increased. FMT protected the intestine by modulating the antimicrobial peptides of the intestinal mucosa (P < 0.05). The results of this study revealed that early FMT can improve the gut microbiota, intestinal mucosal immunity, and intestinal development-related functions of Yorkshire piglets.

Immunoglobulin G and immunoglobulin G subclass concentrations differ according to sex and race

BackgroundSerum immunoglobulin G (IgG) concentrations are integral to the workup of immune deficiencies and IgG4-related disease (IgG4-RD). Demographic differences in IgG concentrations are poorly described but can influence test interpretation, contribute to racial disparities in primary immunodeficiency diagnosis, and explain demographic differences in IgG concentrations in IgG4-RD. ObjectiveTo assess differences in IgG and IgG subclass concentrations according to sex and race. MethodsWe identified patients with IgG and IgG subclass concentrations measured in a large health care system. Multivariate-adjusted differences in IgG and IgG subclass concentrations and the proportion of subjects with results outside of reference ranges according to sex and race were estimated. ResultsOf the 12,851 patients, the mean age was 54.7 years and 7917 (62%) were female. Of these, 11,673 (91%) were white, 611 (5%) were black, and 302 (2%) were Asian. Compared with the mean concentrations of white patients, Asian and black patients had higher mean concentrations of IgG (1340.0 and 1504.4 vs 988.1 mg/dL, P < .001), IgG1 (782.0 and 938.4 vs 592.4 mg/dL, P < .001), IgG2 (493.5 and 384.2 vs 305 mg/dL, P < .001), IgG3 (76.6 and 91.9 vs 55.9 mg/dL, P < .001), and IgG4 (140.4 and 53.6 vs 41.6 mg/dL, P < .001). Immunoglobin G subclass 4 concentrations were higher in males than those in females (56.3 vs 37.4 mg/dL, P < .001). Similar observations were made when comparing the proportions of patients with results outside of reference ranges and after stratifying by diagnosis. ConclusionImmunoglobin G and IgG subclass concentrations differ according to sex and race. These findings may have implications for the interpretation of these test results but require confirmation in diverse, healthy populations.

The Influence of Moisture Content and Temperature on the Long-Term Storage Stability of Freeze-Dried High Concentration Immunoglobulin G (IgG).

High protein concentration products for targeted therapeutic use are often freeze-dried to enhance stability. The long-term storage stability of freeze-dried (FD) plasma-derived Immunoglobulin G (IgG) from moderate to high concentrations (10–200 mg/mL) was assessed. Monomer content, binding activity and reconstitution times were evaluated over a 12-month period under accelerated and real-term storage conditions. In the first case study it was shown that FD IgG from 10 to 200 mg/mL had minimal monomer/activity losses at up to ambient temperature after 12 months of storage. However, at 45 °C the sucrose-to-protein ratio played a significant impact on IgG stability above 50 mg/mL. All IgG concentrations witnessed moisture ingress over a 12-month period. The impact of moisture ingress from environmental exposure (between 0.1% and 5% w/w moisture) for IgG 50 mg/mL was assessed, being generated by exposing low moisture batches to an atmospheric environment for fixed time periods. Results showed that at −20 °C and 20 °C there was no significant difference in terms of monomer or antigen-binding activity losses over 6 months. However, at 45 °C, there were losses in monomer content, seemingly worse for higher moisture content samples although model binding activity indicated no losses. Finally, the difference between a low moisture product (0.1–1% w/w) and a moderately high moisture (3% w/w) product generated by alternative freeze-drying cycles, both stoppered under low oxygen headspace conditions, was evaluated. Results showed that at −20 °C and 20 °C there was no difference in terms of binding activity or monomer content. However, at 45 °C, the low moisture samples had greater monomer and binding activity losses than samples from the highest moisture cycle batch, indicating that over-drying can be an issue.

Influence of the medications containing phospholipids on the serum immunoglobulin g level in calves during formation of colostral immunity

Abstract. The results of the application of the medication “Membranostabil” and native phospholipid bilayer liposomes based on soybean lecithin developed by this research group for correction of serum immunoglobulin G (IgG) content in newborn calves during colostral immunity formation are presented. The indices of IgG content in blood serum of newborn calves have been investigated in dynamics - from the birth till the age of 11 days. Studies were performed on newborn calves of three groups (control, and two experimental ones) of the Ukrainian black-and-white dairy breed. The level of IgG was investigated by electrophoresis in polyacrylamide gel Quantitative estimation of protein fractions was performed by scanning the electrophoregram, with their subsequent graphical reconstruction and calculation by relative units or area using a computer program. It is established that “Membranostabil” medication and native liposomes from phospholipid bilayer based on soybean lecithin activate transport of immunoglobulins in small intestine and promote a significant increase of content of serum IgG compared to calves of the control group. The content of IgG in the serum of newborns calves of both experimental groups at 6 hours of age significantly increased and remained higher throughout the duration of the experiment, except for the calves of the first experimental group at the age of 7 days, compared with calves in the control group. The dynamics with comparative analysis of serum IgG content between calves of individual groups is shown. The increase of content of serum IgG in newborn calves is one of factors preventing early immunodeficiency, sepsis, development of digestive disorders, and other diseases of young animals. Keywords: colostral immunity, colostrum, immunoglobulin G (IgG), newborn calves, Membranostabil

Dietary β-sitosterol regulates serum lipid level and improves immune function, antioxidant status, and intestinal morphology in broilers

This research investigated effects of dietary β-sitosterol addition at different levels on serum lipid levels, immune function, oxidative status, and intestinal morphology in broilers. One-day-old broiler chicks were allocated to 5 groups of 6 replicates. Chickens in the 5 groups were fed a basal diet supplemented with 0 (control group), 40, 60, 80, and 100 mg/kg of β-sitosterol for 42 D, respectively. β-Sitosterol linearly decreased (P < 0.05) concentrations of serum total cholesterol, jejunal tumor necrosis factor α (TNF-α), and ileal interleukin 1β (IL-1β) and mRNA relative expressions levels of jejunal TLR4 and ileal MyD88, whereas it linearly increased (P < 0.05) contents of jejunal immunoglobulin G (IgG), ileal secreted IgA and glutathione, jejunal catalase activity and Nrf2 mRNA relative expression level, villus height (VH), and VH-to-crypt depth (CD) ratio (VH:CD) in the jejunum and ileum. Linear and quadratic increases (P < 0.05) in absolute and relative spleen weight were observed by dietary β-sitosterol, whereas malondialdehyde (MDA) concentration in the jejunum and ileum followed the opposite trend (P < 0.05). Compared with the control group, dietary β-sitosterol at higher than or equal to 60 mg/kg level decreased (P < 0.05) contents of serum total cholesterol, ileal MDA, and jejunal TLR4 mRNA relative expression level, whereas it increased (P < 0.05) absolute spleen weight and ileal glutathione content. Higher than or equal to 80 mg/kg level of β-sitosterol enhanced (P < 0.05) jejunal IgG concentration, VH, catalase activity, and Nrf2 relative expression level and ileal secreted IgA content, but reduced (P < 0.05) ileal IL-1β content and MyD88 mRNA relative expression level. β-Sitosterol addition at 60 and 80 mg/kg levels increased (P < 0.05) relative spleen weight, whereas it decreased (P < 0.05) jejunal MDA accumulation. Moreover, 100 mg/kg level of β-sitosterol reduced (P < 0.05) jejunal TNF-α level, but it increased (P < 0.05) VH in the jejunum and VH:CD in the jejunum and ileum. Accordingly, dietary β-sitosterol supplementation could regulate serum cholesterol level, promote immune function, and improve intestinal oxidative status and morphology in broilers.

Investigation of the Defatted Colostrum 1H-NMR Metabolomics Profile of Gilts and Multiparous Sows and Its Relationship with Litter Performance

Simple SummarySwine colostrum quality and quantity can influence the growth and survival of piglets and contribute to the differences in productive traits of gilts and multiparous sows. The aim of the study was to characterize the soluble metabolomics profile of defatted colostrum of sows at different parity number (PA) and to correlate the metabolomics profile with the colostrum Brix percentage as an estimate measure of immunoglobulin G (IgG) concentration and sow productive traits. The metabolomics profile of colostrum reveals a slight influence of parity, which can influence the quantity of specific metabolites including glycine and lactose. Specific metabolites including lactose creatine, myo-inositol, and O-phosphocholine partially explain the colostrum IgG Brix percentage. Sows’ productive traits performance, including the litter weight at birth and piglets’ mortality, can be influenced by the metabolites related to a sow’s metabolic condition. Increasing knowledge on the interplay between colostrum composition and litter performance can pave the way to define management strategies to provide piglets with good-quality colostrum, improving welfare and economic sustainability of pig rearing by reducing piglet mortality.The aim of the study was to characterize the soluble metabolomics profile of defatted colostrum of sows at different parity number (PA) and to correlate the metabolomics profile with the Brix percentage estimate of colostrum immunoglobulin G (IgG) and sow productive traits. A total of 96 Meidam (crossbreed Large White × Meishan) sows of PA from 1–4 (PA1: 28; PA2:26; PA3:12; PA4:26) were included, and their productive traits were recorded at 10 days post-farrowing. Colostrum IgG was quantified using a Brix refractometer, and metabolomics profile was assessed using 1H-NMR spectroscopy. Sows’ PA slightly influenced the metabolomics profile of colostrum. lactose and glycine were higher in PA1 compared with PA4 (p 0.05) and N-acetylglucosamine (GlcNAc) tended to be higher in PA2 than PA3 and PA4 (p < 0.10). The Brix percentage of IgG was negatively associated with lactose and positively with creatine, myo-inositol, and O-phosphocholine (p < 0.05). Taurine was positively related to litter weight at birth. GlcNAc and myo-inositol were linked to piglet mortality at day 10 with a negative and positive trend, respectively. In conclusion, colostrum of gilts and multiparous sows had a similar metabolomics profile. Specific metabolites contributed to explanation of the variability in colostrum Brix percentage estimate of IgG concentration and the sows’ productive performance.

Comparative humoral immune response against cytopathic or non-cytopathic bovine viral diarrhea virus infection

Bovine viral diarrhea virus (BVDV) infection causes immune dysfunction. The current study investigated the effect of cytopathic (cp) or noncytopathic (ncp) strains of BVDV on immunomodulation by the levels of total serum immunoglobulin G (IgG), the IgG1, IgG2, BVDV neutralizing antibodies and total white blood cell (WBC) count. Twenty (20) BVDV seronegative dairy calves (5–6 months old) were divided in two groups of ten. The animals were infected with either a cp or ncp virus isolated from the same animal (ncp BVDV1b-TGAN or cp BVDV1b-TGAC). One group of 10 was infected with ncp TGAN while the other group of 10 was infected with cp TGAC. Calves infected with cp BVDV had a significant decrease in total IgG as well as IgG1 concentration at 7 days post infection (DPI) that recovered by 21 DPI (total IgG) and 35 DPI (IgG1), respectively. There was no effect of ncp BVDV infection on total IgG concentration in the first 7 days of infection (DOI); however, IgG1 concentration was significantly reduced and IgG2 concentration was significantly increased at 7 DOI. At 35 DPI, ncp TGAN-infected calves had significantly higher total IgG, IgG1 as well as IgG2 compared to cp TGAC-infected calves. Ncp BVDV induced higher BVDV homologous and heterologous neutralizing antibodies compared to the cp BVDV strain. Calves infected with ncp BVDV had significantly reduced WBC counts at 7 DPI that recovered by 14 DPI. Overall, these findings indicate that humoral immunosuppression occurs early following BVDV infection with the largest effect on IgG1 levels.

Effect of immunoglobulin G concentration in dairy cow colostrum and calf blood serum on Cryptosporidium spp. invasion in calves.

Aim:The research aimed to test the association between the level of immunoglobulin G (IgG) in bovine colostrum and calf blood serum and to evaluate its relation to Cryptosporidium spp. invasion in calves.Materials and Methods:Fresh colostrum and fecal specimens from cows (n=114) as well as blood and fecal specimens from newborn calves (n=114) were collected in the dairy cattle farm. Investigated calves were separated from their mothers directly after birth and received 2 L of colostrum in two separate feedings within the first 24 h. Blood samples were taken from calves at the age of 2 days. Coprological samples were taken from calves at the age of 1, 10, and 15 days. Both colostrum and fecal samples from cows were taken on the 1st day after calf birth. Rectal fecal samples were collected separately from each calf and cow into plastic bags. The collected calf serum samples and bovine colostrum samples were tested for bovine IgG by competitive enzyme-linked immunosorbent assay kit bovine Ig. To record oocysts of Cryptosporidium spp. in feces, the flotation method was used. Binomial logistic regression was performed to ascertain the effects of IgG in bovine colostrum and calf blood serum on the likelihood of Cryptosporidium spp. infection in calves.Results:The concentration of IgG in bovine colostrum was higher (70.7±26.6 g/L, mean±standard deviation) than that in calf blood serum (13.2±6.1 g/L); the statistically significant difference was 57.4 g/L (95% confidence interval, 52.4-62.4), t (124.872)=22.536, p<0.001. Mann–Whitney’s U-test showed a significant difference between samples collected on days 10 and 15 of the experiment (U=1944, z=2.330, p=0.020). The higher number of oocysts in calf feces was recorded on day 15 (median=6.5) compared to day 10 (median=4). The prevalence of calf infection from days 10 to 15 increased from 26.3 to 45.6% and was at least 3 times higher than in cows. A statistically significant positive correlation was recorded between IgG concentration of cow colostrum and calf blood serum (r (114)=0.414, p=0.001), whereas a correlation between the concentration of IgG and the intensity of Cryptosporidium spp. infection was not recorded (p>0.05). The logistic regression model was not statistically significant (χ2(2)=0.013, p=0.99 (10 days) and χ2(2)=0.100, p=0.95 (15 days)).Conclusion:Mother passive transfer of immunity to the offspring through colostrum does not influence the susceptibility of calves to Cryptosporidium infestation.