Effects of a ß-Glucan-Rich Blend of Medicinal Mushrooms and Botanicals on Innate Immune Cell Activation and Function Are Enhanced by a Very Low Dose of Bovine Colostrum Peptides.

Nutraceutical immune support offers potential for designing blends with complementary mechanisms of action for robust support of innate immune alertness. We documented enhanced immune activation when bovine colostrum peptides (BC-Pep) were added to an immune blend (IB) containing ß-glucans from yeast, shiitake, maitake, and botanical non-ß-glucan polysaccharides. Human peripheral blood mononuclear cells (PBMCs) were cultured with IB, BC-Pep, and IB + BC-Pep for 20 h, whereafter expression of the activation marker CD69 was evaluated on NK cells, NKT cells, and T cells. Cytokine levels were tested in culture supernatants. PBMCs were co-cultured with K562 target cells to evaluate T cell-mediated cytotoxicity. IB + BC-Pep triggered highly significant increases in IL-1ß, IL-6, and TNF-α, above that of cultures treated with matching doses of either IB or BC-Pep. NK cell and T cell activation was increased by IB + BC-Pep, reaching levels of CD69 expression several fold higher than either BC-Pep or IB alone. IB + BC-Pep significantly increased T cell-mediated cytotoxic killing of K562 target cells. This synergistic effect suggests unique amplification of signal transduction of NK cells and T cells due to modulation of IB-induced signaling pathways by BC-Pep and is of interest for further pre-clinical and clinical testing of immune defense activity against virally infected and transformed cells.

Graduate Student Literature Review: Role of antioxidants in calf immunity, growth, and health.

The neonatal period for dairy calves is crucial for immune, metabolic, and physical development, which opens a window of disease susceptibility. Although the industry has relied on tools such as colostrum and vaccination to support early life immunity, there are several challenges when vaccinating neonatal calves: (1) the inability to mount an effective immune response, (2) interference with maternal antibodies, and (3) oxidative stress (OS). Oxidative stress is characterized as the imbalance of pro-oxidants to antioxidants that results in cellular oxidative damage or dysfunction. Oxidative stress has become a topic of interest in the neonatal period because it negatively affects lymphocyte function, which might affect vaccine response. Widely studied in mature cattle, antioxidant supplementation has the potential to improve reduction-oxidation balance and immune response. Evidence supporting the use of antioxidants such as vitamins and minerals in neonatal calves is far scarcer but necessary to optimize immunity and disease resistance. This review summarizes research on the effect of antioxidant supplementation on calf immunity, health, and productivity and highlights remaining gaps in knowledge. Overall, micronutrient supplementation, including vitamins and minerals, in preweaning and postweaning calves improved immune responses but there is conflicting evidence supporting the subsequent positive effect on calf health and growth performance.

Risk factors related to the appearance of umbilical disorders in dairy calves / [Fatores de risco relacionados ao surgimento de afecções umbilicais em bezerros leiteiros]

The objective of this study was to determine the types of calve housing used in dairy farms, the prevalence of umbilical disorders and related risk factors. The 16 farms studied were visited to characterize the types of installation and possible risk factors, as well as information obtained from a questionnaire applied to the farmers. 806 Holstein calves were physically examined, in addition to collecting blood samples for the evaluation of Failures in Passive Immunity Transfer (FPIT), in animals that manifested inflammatory omphalopathies, and were also submitted to ultrasound examination. The prevalence of omphalopathies was assessed by Fisher's test, and multivariate logistic regression to assess risk factors. Eight types of installation were found: tropical house, suspended cage, collective stall, collective picket, Argentinean type, single-story cage, individual stall, and collective picket with chain. Omphalopathies accounted for 6.45% of the calves. Small size farms (up to 99 lactation cows) had high risk for umbilical disorders, ground floor collective calves, without side protection, with sand floor, in closed sheds and without heatstroke were considered risk factors for omphalopathies. Adequate colostrum and umbilical antisepsis are not associated with disease, its appearance being related to the housing conditions of the animals.(AU)

O objetivo deste estudo foi determinar os tipos de alojamento para bezerros leiteiros, a prevalência de onfalopatias e os fatores de risco relacionados. As 16 fazendas estudadas foram visitadas buscando-se caracterizar os tipos de instalação e os possíveis fatores de risco, além de informações obtidas de um questionário aplicado aos fazendeiros. Foram examinados fisicamente 806 bezerros da raça Holandesa, além da coleta de amostras de sangue, para avaliação da falha de transferência de imunidade passiva (FTIP), nos animais que manifestaram onfalopatias inflamatórias, sendo submetidos também ao exame ultrassonográfico. A prevalência das onfalopatias foi avaliada por teste de Fisher, e foi feita regressão logística multivariada a fim de se avaliarem os fatores de risco. Verificou-se oito tipos de instalação: casinha tropical, gaiola suspensa, baia coletiva, piquete coletivo, bezerreiro tipo argentino, gaiola térrea, baia individual e piquete coletivo com corrente. As onfalopatias corresponderam a 6,45% dos bezerros. Os bezerreiros coletivos térreos, sem proteções laterais, com piso de areia, borracha, concreto ou madeira, em galpões fechados, sem insolação, com alta densidade animal, antissepsia umbilical realizada por três dias e FTIP acima de 50% foram considerados fatores de risco para onfalopatias e possuem relação com o bezerreiro, sendo decisivas para evitar essas condições a colostragem e a antissepsia umbilical adequadas.(AU)

Diverse Immune Effects of Bovine Colostrum and Benefits in Human Health and Disease.

The health benefits of bovine colostrum have extensively been studied, including immune effects mediated by immunoglobulins, lactoferrin, and casein, as well as by certain growth factors. Some of these effects are not directly related to the absorption of proteins from the intestinal tract. The ingestion of BC can modulate the function of subsets of lymphocytes, macrophages, and dendritic cells and increase regulatory cytokines such as interleukin 10. In this review, we predominantly focused on evidence from human studies on benefits in health and disease. This review highlights that clear evidence of the prevention of infectious diseases in pre-term infants such as necrotizing enterocolitis, neonatal sepsis or prevention of cancer metastasis is lacking. This is clearly an area where translational science has to be strengthened, taking the considerable evidence from numerous ex vivo studies on cells and tissues and from animal interventions. The review focuses predominantly on human data.

Milk Exosomes Transfer Oligosaccharides into Macrophages to Modulate Immunity and Attenuate Adherent-Invasive E. coli (AIEC) Infection.

Exosomes are abundance in human body fluids like urine, milk and blood. They act a critical role in extracellular and intracellular communication, intracellular trafficking and physiological regulation. Multiple immune-modulatory components, such as proteins, RNAs and carbohydrates (glycoproteins), have been found in human milk exosomes, which play immune-regulatory functions. However, little is known about oligosaccharides in milk exosomes, the "free sugars", which act critical roles in the development of infant's immature mucosal immune system. In this study, the profile of milk exosomes encapsulated human milk oligosaccharides (HMOs) was calibrated with characteristic oligosaccharides in colostrum and mature milk, respectively. The exosomes containing human milk oligosaccharides were uptaken by macrophages, which were responsible for the establishment of intestinal immunity. Furthermore, mice pretreated with exosome encapsulated HMOs were protected from AIEC infection and had significantly less LPS-induced inflammation and intestinal damage. Exosome encapsulated milk oligosaccharides are regarded to provide a natural manner for milk oligosaccharides to accomplish their critical functions in modifying newborn innate immunity. The understanding of the interaction between a mother's breastfeeding and the development of an infant's mucosal immune system would be advantageous. The transport of milk oligosaccharides to its target via exosome-like particles appears to be promising.

Colostrum IgA1 antibodies recognize antigens from Helicobacter pylori and prevent cytoskeletal changesin human epithelial cells.

Helicobacter pylori is a Gram-negative bacterium found on the luminal surface of the gastric mucosa in at least 50% of the world's human population. The protective effect of breastfeeding against H. pylori infection has been extensively reported; however, the mechanisms behind this protection remain poorly understood. Human IgA from colostrum has reactivity against H. pylori antigens. Despite that IgA1 and IgA2 display structural and functional differences, their reactivity against H. pylori had not been previously determined. We attested titers and reactivity of human colostrum-IgA subclasses by ELISA, immunoblot, and flow cytometry. Colostrum samples from healthy mothers had higher titers of IgA; and IgA1 mostly recognized H. pylori antigens. Moreover, we found a correlation between IgA1 reactivity and their neutralizing effect determined by inhibition of cytoskeletal changes in AGS cells infected with H. pylori. In conclusion, colostrum-IgA reduces H. pylori infection of epithelial gastric cells, suggesting an important role in preventing the bacteria establishment during the first months of life. As a whole, these results suggest that IgA1 from human colostrum provides protection that may help in the development of the mucosal immune system of newborn children.

Humoral and Cell-Mediated Immune Response in Colostrum from Women Diagnosed Positive for SARS-CoV-2.

Objective: To evaluate the immune response to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in colostrum from women who tested positive for the virus. Methods: Between March and September 2020 we obtained bilateral colostrum samples collected on spot cards within 48 hours of delivery from 15 new mothers who had previously tested positive for SARS-CoV-2. Four of 15 women provided liquid colostrum, which was used for validating results obtained from spot cards. Archived bilateral colostrum samples collected from 8 women during 2011-2013 were used as pre-coronavirus disease 2019 (COVID-19) controls. All samples were tested for reactivity to the receptor binding domain (RBD) of the SARS-CoV-2 spike protein using an enzyme-linked immunosorbent assay that measures SARS-CoV-2 RBD-specific IgA, IgG, and IgM and for levels of 10 inflammatory cytokines (interferon-gamma [IFN-γ], tumor necrosis factor-alpha, interleukin [IL]-1ß, IL-2, IL-4, IL-6, IL-8, IL-10, IL-12, IL-13) using a multiplex electrochemiluminescent sandwich assay. Results: Our validation studies indicate that the levels of SARS-CoV-2-specific antibodies and the associated cytokines measured in liquid colostrum are comparable to levels eluted from spot cards. Bilateral colostrum samples from 73%, 73%, and 33% of the 15 COVID-19 mothers exhibited IgA, IgG, and IgM reactivity to RBD, respectively. In addition, symptomatic COVID-19 mothers had statistically significant elevated levels of 4 of the 10 inflammatory markers (IFN-γ, IL-4, IL-6, and IL-12) compared to asymptomatic COVID-19 mothers. Conclusions: A strong humoral immune response is present in the colostrum of women who were infected with SARS-CoV-2 before delivering. The evolution and duration of the antibody response, as well as dynamics of the cytokine response, remain to be determined. Our results also indicate that future large-scale studies can be conducted with milk easily collected on paper spot cards.

Potential Benefits of Bovine Colostrum in Pediatric Nutrition and Health.

Bovine colostrum (BC), the first milk produced from cows after parturition, is increasingly used as a nutritional supplement to promote gut function and health in other species, including humans. The high levels of whey and casein proteins, immunoglobulins (Igs), and other milk bioactives in BC are adapted to meet the needs of newborn calves. However, BC supplementation may improve health outcomes across other species, especially when immune and gut functions are immature in early life. We provide a review of BC composition and its effects in infants and children in health and selected diseases (diarrhea, infection, growth-failure, preterm birth, necrotizing enterocolitis (NEC), short-bowel syndrome, and mucositis). Human trials and animal studies (mainly in piglets) are reviewed to assess the scientific evidence of whether BC is a safe and effective antimicrobial and immunomodulatory nutritional supplement that reduces clinical complications related to preterm birth, infections, and gut disorders. Studies in infants and animals suggest that BC should be supplemented at an optimal age, time, and level to be both safe and effective. Exclusive BC feeding is not recommended for infants because of nutritional imbalances relative to human milk. On the other hand, adverse effects, including allergies and intolerance, appear unlikely when BC is provided as a supplement within normal nutrition guidelines for infants and children. Larger clinical trials in infant populations are needed to provide more evidence of health benefits when patients are supplemented with BC in addition to human milk or formula. Igs and other bioactive factors in BC may work in synergy, making it critical to preserve bioactivity with gentle processing and pasteurization methods. BC has the potential to become a safe and effective nutritional supplement for several pediatric subpopulations.

Innate and humoral immune parameters at delivery in colostrum and calves from heifers experimentally infected with Neospora caninum.

Neospora caninum is a leading cause of abortion in cattle worldwide. The study of the immune response against N. caninum is critical to understand its epidemiology, pathogenesis, diagnosis and, ultimately, in preventing and controlling bovine neosporosis. Herein, we determined the gene expression of innate immune components endosomal RNA-sensing TLRs, BMAP28 cathelicidin, TNF-α and IL-10 and characterized the variation in both IgG ratio and avidity at delivery in N. caninum-infected heifers challenged at day 210 of gestation, colostrum and their calves. Increased BMAP28 expression was observed not only in colostrum but also in peripheral blood mononuclear cells (PBMC) and umbilical cord of calves from N. caninum-infected heifers in comparison with mock-infected control group. In addition, statistically significant decrease of TLR7 and IL-10 expression levels were observed in umbilical cord, suggesting an attempt to avoid an exacerbated immune response against the parasite. At delivery, serum and colostrum samples from infected group evidenced specific IgG anti-N. caninum. Infected heifers showed IgG1/IgG2 ratios <1 and high avidity specific IgG. As expected, colostrum samples of these animals exhibited a high IgG1 concentration and elevated avidity values. Three out of four calves from N. caninum-infected heifers had specific IgG with IgG1/IgG2 ratios>1 and lower avidity values before colostrum intake. Interestingly, both IgG1/IgG2 ratios and avidity values increased in seropositive calves after colostrum intake. Overall, this study provides novel information on neonatal immunity in congenitally infected calves, which is essential to understand how the immune pathways could be manipulated or immune components could be employed in order to improve protection against neosporosis.

Active free secretory component and secretory IgA in human milk: do maternal vaccination, allergy, infection, mode of delivery, nutrition and active lifestyle change their concentrations?

BACKGROUND: Free secretory component (free SC) in human milk is a critical constituent of secretory IgA (SIgA) for immune exclusion, but its concentration in human milk is unknown. To evaluate the relationship between free SC and SIgA, the influence of maternal factors (vaccination during pregnancy, allergy, previous infections, nutrition, mode of delivery and active lifestyle) on the concentrations of those secretory immune components in human milk was investigated. METHODS: Concentration of active free SC and SIgA in 124 milk samples from 91 mothers were measured via ELISA. RESULTS: Free SC in milk from Tdap-vaccinated mothers was lower than the Tdap-flu-vaccinated, flu-vaccinated or Rhogam-vaccinated mothers. Free SC in mothers who had a cesarean delivery was higher than mothers who had a vaginal delivery. Free SC in the nonallergic group was higher than the allergic group. Free SC was higher in mothers who rarely/never eat junk food, than in mothers who always/frequently eat junk food. Free SC also was higher in the moderate exercise group (active lifestyle) compared with the group who rarely/never exercise (sedentary lifestyle). Free SC in human milk was not affected by previous maternal infection or probiotic supplementation whereas SIgA was not changed by all investigated maternal factors. CONCLUSION: This study suggests that active free SC is more impacted by maternal factors than active SIgA in human milk. IMPACT: Active free secretory component (free SC) is more impacted by maternal factors than active secretory IgA (SIgA) in human milk. Vaccination during pregnancy, allergy, nutrition, type of delivery and active lifestyle affect the secretion of free SC in human milk, but not SIgA secretion. Free SC in human milk is a critical constituent of secretory IgA (SIgA) for immune exclusion against pathogens and its active concentration in milk strongly varies between mothers, partially due to their specific maternal background.

Colostrum supplementation with n-3 fatty acids does not alter calf outcome on a healthy commercial farm.

Our objective was to supplement colostrum with n-3 fatty acids (FA) to provide anti-inflammatory mediators that may improve the immune response of neonatal calves. Elevated markers of inflammation have been associated with increased occurrence of calf disease in early life, thus decreasing animal productivity. We hypothesized that a colostrum supplement containing 60-mL of a 1:1 ratio fish:flaxseed oil blend with or without 200 mg of α-tocopherol might provide an advantageous start to early life by decreasing oxidative stress and regulating the inflammatory response. Calves were blocked by birth order and randomly assigned to 1 of 3 treatments: no supplement added to colostrum (control), 60 mL of 1:1 fish:flaxseed oil blend, and 60 mL of 1:1 fish:flaxseed oil blend with 200 mg of α-tocopherol. In total, 180 heifer calves (n = 60 per treatment) were enrolled on a commercial farm. After colostrum feeding, all calves were housed in individual hutches and fed milk replacer 3 times per day. Health was scored 3 times per week until weaning at 8 wk of age. Weight, wither height, and heart girth were measured after birth, 3 wk, and 8 wk of age to assess preweaning growth. A subgroup of 54 calves (18 blocks or 18 calves per treatment) were sampled 2 d (± 8 h) after birth to evaluate oxidant status, serum total protein, and inflammatory gene and cytokine protein expression in blood after an in vitro lipopolysaccharide (LPS) challenge as indicators of health and immunity. At 9 wk, calves were transported 18 h to another farm, and medical records were kept as an indicator of disease incidence up to 13 wk of age. Calf mortality was 1.8%, which is below industry average, and exceptional health scores were observed throughout the study. Health scores and growth were similar throughout the preweaning period regardless of treatment. Serum total protein indicated successful passive transfer in all calves, and oxidant status index was not affected by treatments on d 2 of age. Concentrations of tumor necrosis factor α increased with LPS stimulation, but the increase was not altered by treatment. Likewise, leukocyte gene expression of tumor necrosis factor α, IL-8 and IL-10, and cyclooxygenase-2 increased upon LPS stimulation, but the fold change did not differ with treatment. In conclusion, 60 mL of 1:1 ratio fish:flaxseed oil colostrum supplement did not enhance preweaning calf performance. Supplementing n-3 FA in a one-time meal may not provide the anti-inflammatory benefits observed with continuous feeding.

Heat treatment of bovine colostrum: II. Effects on calf serum immunoglobulin, insulin, and IGF-I concentrations, and the serum proteome.

In-depth analysis of colostrum components has identified hundreds of proteins, but data are sparse regarding their systemic uptake in the newborn calf. Moreover, heat treatment may influence these colostral components and their absorption. Our objectives were to describe the serum proteome of newborn calves before and after colostrum feeding and the possible effects of colostral heat treatment. Newborn Holstein heifer calves (n = 22) were randomized within pair and fed heat-treated (n = 11; 60°C, 60 min) or raw (n = 11) colostrum at 8.5% of birth body weight by esophageal feeder within 1 h of birth. After the single colostrum feeding, calves were not fed until after the 8-h time point, when milk was offered free-choice. Blood samples were taken immediately before feeding (0 h), as well as 4, 8, and 24 h after feeding. Whole blood packed cell volume (%), serum Brix percentage, and plasma glucose concentrations were determined for all time points. Plasma insulin and insulin-like growth factor-I concentrations were determined by radioimmunoassay for selected time points. Serum IgA and IgG were measured by radial immunodiffusion at 24 h. The serum proteome was analyzed using nano-scale reverse-phase chromatography and tandem mass spectrometry (nano LC-MS/MS) in 0- and 8-h samples. For proteomics analysis, ratios of results for 8-h to 0-h samples were analyzed with false discovery rate adjustment. For all other outcomes, repeated-measures ANOVA was performed with the fixed effects of group, time, and their interaction, and random effect of pair. Serum Brix percentage and glucose concentrations increased over time and were independent of colostrum treatment. Serum IgG and IgA concentrations at 24 h did not differ between groups. Nano LC-MS/MS identified a total of 663 unique proteins in serum, of which 261 increased in abundance, whereas 67 decreased in abundance after feeding in both groups. Among serum proteins that increased in abundance and that were previously identified in colostrum, many belonged to those involved in immune response, coagulation, the classical complement pathway, or the antimicrobial peptide class of cathelicidins. Serum proteins that decreased in abundance and that were identified in colostrum belonged to the alternative complement pathway and the membrane attack complex. Thirty-eight proteins differed in calves that were fed heat-treated colostrum compared with those fed raw colostrum. Decreased abundances in calves fed heat-treated colostrum included several enzymes involved in glycolysis or glycogenolysis, whereas the incretin gastric inhibitory polypeptide and serum insulin were increased in this group. Our findings point to important innate immune defense pathways associated with colostrum ingestion in newborn calves. Furthermore, calves fed heat-treated colostrum showed differences in serum proteins and enzymes associated with carbohydrate metabolism.

Effects of prepartum zinc-methionine supplementation on feed digestibility, rumen fermentation patterns, immunity status, and passive transfer of immunity in dairy cows.

The aim of this study was to determine the effects of prepartum supplementation of zinc-methionine (Zn-Met) on feed digestibility, rumen fermentation patterns, and immunity status in dams and passive immunity transfer in their calves. A randomized complete design was used in this study. Forty multiparous Holstein dairy cows in late pregnancy (60 d before the expected calving date) were blocked by parity (2.1 ± 0.3), body weight (651 ± 52 kg), and expected calving date, and randomly assigned to 1 of 4 treatments. Cows were supplemented with Zn as Zn-Met at 0, 20, 40, or 60 mg/kg of dry matter (DM) from 60 d before expected calving date to the calving day. Though the nutrient digestibility was not affected by Zn supplementation, DM intake, Zn digestibility, and Zn deposition increased linearly with increasing Zn-Met supplementation. Ruminal pH and molar proportion of individual volatile fatty acids were similar, whereas a linear decrease and increase were observed in ruminal ammonia and microbial crude protein concentration, respectively, with increasing Zn-Met supplementation. Maternal serum concentration of alkaline phosphatase, carboxypeptidase, Cu and Zn superoxide dismutase, and total antioxidant capacity were greater in cows supplemented with >40 mg of Zn/kg of DM compared with the control group. With increasing Zn-Met supplementation, maternal blood concentration of IL-1 decreased linearly, whereas IL-2 and IL-6 increased linearly, and no differences were observed in IL-4. Concentration of nonesterified fatty acids and ß-hydroxybutyric acids in maternal blood was similar between treatments. No difference was observed in colostrum composition with increasing Zn-Met supplementation. Concentration of Zn and immunoglobulins (including IgA, IgG, and IgM) in maternal blood did not differ among treatments. However, Zn concentration in colostrum and blood of calves increased linearly. The concentration of IgA and IgM in colostrum increased linearly with increasing Zn-Met supplementation, whereas no differences in immunoglobulins were observed in calf blood. In conclusion, Zn supplementation as Zn-Met at 40 of mg/kg of DM may improve antioxidant activity of dam and potentially increase passive immunity transfer in calves.

Impact of birth weight and neonatal nutritional interventions with micronutrients and bovine colostrum on the development of piglet immune response during the peri-weaning period.

Immune system development of piglets is influenced by birth weight and colostrum and milk intake. Moreover, the dam transfer to piglets of vitamins A and D and copper, which play important role in immunity, is limited during lactation. In this study, we evaluated the potential of maternal and neonatal supplementations with vitamins A and D and copper, with or without neonatal supplementation of bovine colostrum (BC), to modulate the immune system development of low birth weight (LBW) and high birth weight (HBW) piglets during the peri-weaning period. Litters from 23 control sows (CONT) were assigned to one of the following treatments: 1) control (C); 2) oral administration at 2 and 8 days (d) of age of retinol-acetate, 25-hydroxyvitamin D and CuSO4 and exposure to UVB light for 15 min every second day from d 5 to d 21 (ADCu); 3) oral administration of dehydrated BC (4 g/d) from d 5 to d 10 (BC); 4) ADCu + BC. This experimental design was repeated with 24 sows fed extra daily supplements of 25-hydroxyvitamin D (4,000 IU), ß-carotene (30,000 IU) and Cu-yeast (equivalent 45 mg of Cu) from 90 d of gestation until weaning at d 21 (SUPPL). Within each litter, 2 LBW and 2 HBW piglets were euthanized at d 16 and d 23 in order to characterize leukocyte subsets in mesenteric lymph nodes (MLN) and blood by flow cytometry, and to measure gene expression in the MLN and jejunal mucosa by qPCR. At d 16, results revealed that the percentages of γδ and cytotoxic T lymphocytes were significantly reduced in LBW compared to HBW piglets. The jejunal expression of interleukin (IL) 22 was also up-regulated, along with MLN expression of C-C Motif Chemokine Ligand 23, bone morphogenetic protein 2 and secreted phosphoprotein 1 (SPP1), whereas jejunal expression of tumor necrosis factor α was decreased in LBW piglets. At d 23, LBW piglets showed lower amounts of γδ T lymphocytes, higher percentages of CD3- and CD3-CD8α+CD16+ leukocytes (which include Natural killer cells) and lower jejunal expression of IL18. Furthermore, supplementation with BC increased the blood percentage of CD3-CD16+ leukocytes and reduced jejunal IL5 and MLN IL15 expression whereas supplementation with ADCu + BC increased jejunal TNF superfamily 13B and MLN SPP1 expression. Our results suggest that immune system development after birth differed between LBW and HBW piglets and that early dietary supplementation with BC and ADCu has the potential to modulate development of immune functions.

Does Caesarean Section or Preterm Delivery Influence TGF-ß2 Concentrations in Human Colostrum?

Human colostrum (HC) is a rich source of immune mediators that play a role in immune defences of a newly born infant. The mediators include transforming growth factor ß (TGF-ß) which exists in three isoforms that regulate cellular homeostasis and inflammation, can induce or suppress immune responses, limit T helper 1 cells (Th1) reactions and stimulate secretory immunoglobulin A (IgA) production. Human milk TGF-ß also decreases apoptosis of intestinal cells and suppresses macrophage cytokine expression. The aim of the study was to determine the concentration of TGF-ß2 in HC obtained from the mothers who delivered vaginally (VD) or by caesarean section (CS), and to compare the concentrations in HC from mothers who delivered at term (TB) or preterm (PB). In this study, 56% of preterm pregnancies were delivered via CS. The concentrations of TGF-ß2 were measured in HC from 299 women who delivered in the 1st Department of Obstetrics and Gynaecology, Medical University of Warsaw: 192 (VD), 107 (CS), 251 (TB), and 48 (PB). The colostrum samples were collected within 5 days post-partum. TGF-ß2 levels in HC were measured by the enzyme-linked immunosorbent assay (ELISA) test with the Quantikine ELISA Kit-Human TGF-ß2 (cat.no. SB250). Statistical significance between groups was calculated by the Student t-test using StatSoft Statistica 13 software. The mean TGF-ß2 concentration in patients who delivered at term or preterm were comparable. The levels of TGF-ß2 in HC were higher after preterm than term being 4648 vs. 3899 ng/mL (p = 0.1244). The delivery via CS was associated with higher HC concentrations of TGF-ß2. The levels of TGF-ß2 were significantly higher in HC after CS than VD (7429 vs. 5240 ng/mL; p = 0.0017). The data from this study suggest: caesarean section was associated with increased levels of TGF-ß2 in HC. The increased levels of TGF-ß2 in HC of women who delivered prematurely require further research. Early and exclusive breast-feeding by mothers after caesarean section and premature births with colostrum containing high TGF-ß2 levels may prevent the negative impact of pathogens which often colonize the gastrointestinal tract and may reduce the risk of chronic diseases in this group of patients.

Antigenic stimulation during pregnancy modifies specific iga1 and iga2 subclasses in human colostrum according to the chemical composition of the antigen

ABSTRACT Background: Several studies have evaluated the effect of infectious diseases and vaccine protocols during pregnancy on maternal milk immunoglobulin (Ig) levels, to understand the protection conferred by lactation on newborns. Colostrum is the primary source of maternal IgA for the newborn. IgA participates in protection mechanisms in the neonate's mucosa. In humans, IgA has two subclasses with differential anatomical distribution among mucosal compartments. Total IgA levels in maternal milk vary after antigen stimulation and have differential affinities in function of the chemical composition of the antigens. We studied the effect of antigenic stimulation during pregnancy on the concentrations of specific IgA1 and IgA2 subclasses in human colostrum. Methods: We analyzed data from 113 women in Mexico City and compared the amount of IgA subclasses in colostrum against three antigens: two from vaccine protocols (tetanus toxoid and pneumococcal polysaccharides) and lipopolysaccharide, a ubiquitous antigen in the gastrointestinal tract. Results: In agreement with the previous reports, we showed that IgA1 from colostrum mainly recognized protein antigens; in sharp contrast, IgA2 was mostly directed against polysaccharide antigens. These levels increased in women who had previous contacts through vaccination or infections during pregnancy. Conclusions: Antigen interaction during pregnancy increased the amount of specific IgA subclasses, depending on the chemical composition of the antigen.

Efficacy of orally administered porcine epidemic diarrhea vaccine-loaded hydroxypropyl methylcellulose phthalate microspheres and RANKL-secreting L. lactis.

Here, we examined the efficacy of are combinant subunit antigen-based oral vaccine for preventing porcine epidemic diarrhea virus (PEDV). First, we generated a soluble recombinant partial spike S1 protein (aP2) from PEDV in E. coli and then evaluated the utility of aP2 subunit vaccine-loaded hydroxypropyl methylcellulose phthalate microspheres (HPMCP) and RANKL-secreting L. lactis (LLRANKL) as a candidate oral vaccine in pregnant sows. Pregnant sows were vaccinated twice (with a 2 week interval between doses) at 4 weeks before farrowing. Titers of virus-specific IgA antibodies in colostrum, and neutralizing antibodies in serum, of sows vaccinated with HPMCP (aP2) plus LL RANKL increased significantly at 4 weeks post-first vaccination. Furthermore, the survival rate of newborn suckling piglets delivered by sows vaccinated with HPMCP (aP2) plus LL RANKL was similar to that of piglets delivered by sows vaccinated with a commercial killed porcine epidemic diarrhea virus (PED) vaccine. The South Korean government promotes a PED vaccine program (live-killed-killed) to increase the titers of IgA and IgG antibodies in pregnant sows and prevent PEDV. The oral vaccine strategy described herein, which is based on a safe and efficient recombinant subunit antigen, is an alternative PED vaccination strategy that could replace the traditional strategy, which relies on attenuated live oral vaccines or artificial infection with virulent PEDV.

Colostrum fails to prevent bovine/camelid neonatal neutrophil damage from AFB1.

Exposure to environmental toxicants that affect the immune system and overall health of many mammals is mostly unavoidable. One of the more common substances is the mycotoxins, especially carcinogenic aflatoxin (AF)B1 which also causes immune suppression/dysregulation in exposed hosts. The present study analyzed the effects of naturally occurring levels of AFB1 on apoptosis of healthy bovine and camelid neonatal neutrophils (PMN) that were isolated both before and after host consumption of colostrum. Cells from bovine and camel neonates (n = 12 sets of PMN/mammal/timepoint) were exposed for 24 h to a low level of AFB1 (i.e. 10 ng AFB1/ml) and then intracellular ATP content and caspase-3, -7, and -9 activities (determined by bioluminescence) were assessed. The results indicated a significant lessening of intracellular ATP content and equivalents of luminescence intensity in AFB1-treated PMN in all studied samples, i.e. isolated pre-and post-colostrum consumption. In contrast, caspase-3, -7, and -9 activities in both pre- and post-colostrum consumption bovine and camelid PMN were noticeably increased (∼>2-fold). The damaging effects of AFB1 were more pronounced in bovine neonate PMN than in camelid ones. These results showed that camelid or bovine neonatal PMN collected pre- and post-colostrum are sensitive (moreso after consumption) to naturally occurring levels of AFB1. While merits of colostrum are well known, its failure to mitigate toxic effects of AFB1 in what would translate into a critical period in the development of immune competence (i.e. during the first few days of life in bovine and camelid calves) is surprising. The observed in vitro toxicities can help clarify underlying mechanisms of immune disorders caused by AFs in animals/humans.

Oropharyngeal Colostrum Positively Modulates the Inflammatory Response in Preterm Neonates.

During the first days of life, premature infants have physiological difficulties swallowing, thereby missing out on the benefits of breastfeeding. The aim of this study is to assess the effects of oropharyngeal mother's milk administration in the inflammatory signaling of extremely premature infants. Neonates (n = 100) (<32 week's gestation and/or <1500 g) were divided into two groups: mother's milk group (n = 48), receiving 0.2 mL of oropharyngeal mother's milk every 4 h for the first 15 days of life, and a control group (n = 52), not receiving oropharyngeal mother's milk. Serum concentrations of interleukin (IL) IL-6, IL-8, IL-10, IL-1ra, tumor necrosis factor alpha (TNF-α), and interferón gamma (IFN-γ) were assessed at 1, 3, 15, and 30 days of postnatal life. Maternal and neonatal outcomes were collected. The rate of common neonatal morbidities in both groups was similar. The mother's milk group achieved full enteral feeding earlier, and showed a decrease in Il-6 on days 15 and 30, in IL-8 on day 30, and in TNF-α and INF-γ on day 15, as well as an increase in IL-1ra on days 3 and 15 and in IL-10 on day 30. Oropharyngeal mother's milk administration for 15 days decreases the pro-inflammatory state of preterm neonates and provides full enteral nutrition earlier, which could have a positive influence on the development of the immune system and inflammatory response, thereby positively influencing other developmental outcomes.

Zika Virus Alters the Viscosity and Cytokines Profile in Human Colostrum.

The resurgence of cases of Zika virus (ZIKV) infection, accompanied by epidemic of microcephaly in Brazil, has aroused worldwide interest in understanding the biological mechanisms of the virus that allow patient management and the viral dissemination control. Colostrum and human milk are possible sources of virus spread. Therefore, the objective of this study was to analyze the repercussions of ZIKV infection on rheological parameters and inflammatory cytokines of colostrum. The prospective cohort study included 40 puerperal donors of colostrum, divided into 2 groups: control (without ZIKV infection, n = 20) and a group infected with ZIKV during the gestational period (n = 20). Analyses were performed for the detection of ZIKV by polymerase chain reaction (PCR). In addition to obtaining the rheological parameters and quantification of IL-10 and IL-6 cytokines by flow cytometry, ZIKV and other flaviviruses were not detected in colostrum. However, maternal infection reflected increased viscosity, decreased levels of IL-10, and elevated levels of IL-6. The higher viscosity may represent a mechanical barrier that hinders the spread of the virus. The lower levels of anti-inflammatory mediators and higher inflammatory cytokines may possibly alter the viscosity, and it seems the higher viscosity represents a possible mechanism of adaptation of breastfeeding against a response to ZIKV.