Hepcidin and Erythroferrone Correlate with Hepatic Iron Transporters in Rats Supplemented with Multispecies Probiotics.

The influence of probiotic supplementation on iron metabolism remains poorly investigated. However, a range of studies, especially on Lactobacillus plantarum 299v (Lp229v), have indicated a possible positive impact of probiotics on iron absorption. The aim of the study was to determine the effect of multistrain probiotic supply on iron balance. Thirty Wistar rats were randomized into three groups: placebo (KK group), and multistrain probiotic per os in a daily dose of 2.5 × 109 colony forming units (CFU) (PA group) or 1 × 1010 CFU (PB group). Multistrain probiotic consisted of nine bacterial strains: Bifidobacterium bifidum W23, B. lactis W51, B. lactis W52, Lactobacillus acidophilus W37, L. brevis W63, L. casei W56, L. salivarius W24, Lactococcus lactis W19, and Lc. lactis W58, in equal proportions. After six weeks, blood and organ samples were collected. No differences were found between the three groups in terms of serum concentrations of hepcidin (HEPC), lactoferrin (LTF), homocysteine (HCY), ferritin (Ft), or erythroferrone (ErFe), or in liver content of divalent metal transporter 1 (DMT1), transferrin receptors 1 and 2 (TfR), or ZRT/IRT-like protein 14 (ZIP14) proteins. In the overall sample, positive correlations were noted between the serum concentrations of hepcidin and lactoferrin, and hepcidin and ferritin; serum concentration of hepcidin and DMT1 and TfR1 in the liver; and serum concentration of erythroferrone and TfR2 in the liver. The correlations of serum hepcidin and erythroferrone with liver DMT1 and TfR represent significant mechanisms of Fe homeostasis. Our study has shown that multistrain probiotic supplementation used in the experiment did not disrupt the biochemical and hepatic regulatory processes of Fe balance and did not demonstrate significant influence on selected parameters of Fe metabolism.

Enhancement of immune response mediated by oropharyngeal colostrum administration in preterm neonates.

BACKGROUND: The immune system of preterm infants is immature, being a significant cause of morbidity and mortality, particularly in the preterm infant. Oropharyngeal colostrum administration could be an immunomodulatory aid. Our aim was to evaluate the effect of oropharyngeal colostrum on the serum levels of immunoglobulins, lactoferrin, and resistin during the first month of life and to track the clinical outcome of the neonates. METHODS: One hundred preterm neonates born at <32 weeks of gestation and/or weighing < 1500 g and assisted in the Neonatal Intensive Care Unit were enrolled and divided into two groups: colostrum (n = 48) and control (n = 52). The subjects assigned to the colostrum group received 0.2 mL of colostrum (oropharyngeal route) every 4 hours for the first 15 days of life, and if mothers have inability to breastfeed, they were included in the control group (no oropharyngeal colostrum). Serum concentrations of IgA, IgM, and IgG1, lactoferrin, and resistin were assessed in both groups at 1, 3, 15, and 30 days of life. Clinical data during hospitalization were collected. RESULTS: IgA and IgM increased in preterm neonates who were administered colostrum for 15 and 30 days. Lactoferrin increased after 30 days, and resistin increased after 15 days of supplying oropharyngeal colostrum. The colostrum group underwent full enteral nutrition before, and no differences were observed in the common neonatal morbidities. CONCLUSION: Oropharyngeal colostrum administration is safe in preterm neonates and improves their immunologic profile, showing a potential role as an immunomodulatory agent.

In vivo study of the effect of lactoferrin on iron metabolism and bioavailability from different iron chemical species for formula milk fortification.

Iron fortification in infant formulas is a common practice for providing iron to newborns in order to avoid its deficiency (anemia). Depending on the physicochemical species used, its bioavailability might be insufficient to meet iron requirements. In this vein, the influence of Lactoferrin (Lf) presence on iron bioavailability in 2-week-old wistar rats fed with formula milk fortified with 57 Fe(III)2 -Lf or 57 Fe(II)SO4 (in presence of Lf) using quantitative speciation (by HPLC-ICP-MS) and Isotope Pattern Deconvolution (IPD) is studied here. Results obtained were compared among fortifiers and also with the maternal group. In RBCs, iron was mainly bound to hemoglobin in all the assayed groups in the same extent. Regarding serum samples, several iron-proteins were observed (such as transferrin and albumin). In both samples, iron content in the fractions studied was similar in all groups compared and exogenous 57 Fe incorporation of intaked iron was always above 50%, showing no significative differences between physicochemical forms but related to the dose administered. Regarding iron stores (liver) the group fed with formula milk fortified with the higher dose of 57 FeSO4 in presence of Lf presented the highest values of total iron even superior than those found in the maternal group, and also the highest exogenous (57 Fe) incorporation. In conclusion, it was proved that iron fortification is required to ensure proper iron levels in all body compartments. No significative differences were observed between different physicochemical species when iron is administered at low doses. However, higher iron doses lead to a greater incorporation in all the iron-proteins studied.

Impact of human milk pasteurization on gastric digestion in preterm infants: a randomized controlled trial.

BACKGROUND: Holder pasteurization has been reported to modify human milk composition and structure by inactivating bile salt-stimulated lipase (BSSL) and partially denaturing some of its proteins, potentially affecting its subsequent digestion. OBJECTIVE: We sought to determine the impact of human milk pasteurization on gastric digestion (particularly for proteins and lipids) in preterm infants who were fed their mothers' own milk either raw or pasteurized. DESIGN: In a randomized controlled trial, 12 hospitalized tube-fed preterm infants were their own control group in comparing the gastric digestion of raw human milk (RHM) with pasteurized human milk (PHM). Over a 6-d sequence, gastric aspirates were collected 2 times/d before and after RHM or PHM ingestion. The impact of milk pasteurization digestive kinetics and disintegration was tested with the use of a general linear mixed model. RESULTS: Despite inactivating BSSL, instantaneous lipolysis was not affected by pasteurization (mean ± SD at 90 min: 12.6% ± 4.7%; P > 0.05). Lipolysis occurred in milk before digestion and was higher for PHM than for RHM (mean ± SD: 3.2% ± 0.6% and 2.2% ± 0.8%, respectively; P < 0.001). Pasteurization enhanced the proteolysis of lactoferrin (P < 0.01) and reduced that of α-lactalbumin (only at 90 min) (P < 0.05). Strong emulsion destabilization was observed, with smaller aggregates and a higher specific surface for PHM (P < 0.05). Pasteurization did not affect gastric emptying (∼30-min half time) or pH (mean ± SD: 4.4 ± 0.8) at 90 min. CONCLUSIONS: Overall, pasteurization had no impact on the gastric digestion of lipids and some proteins from human milk but did affect lactoferrin and α-lactalbumin proteolysis and emulsion disintegration. Freeze-thawing and pasteurization increased the milk lipolysis before digestion but did not affect gastric lipolysis. Possible consequences on intestinal digestion and associated nutritional outcomes were not considered in this study. This trial was registered at clinicaltrials.gov as NCT02112331.

[The influence of low-intensity laser irradiation of blood on the lactoferrin level in the patients presenting with community-acquired pneumonia].

UNLABELLED: The objective of the present study was to estimate the influence of low-intensity laser irradiation of blood on the levels of lactoferrin in the patients presenting with community-acquired pneumonia. MATERIALS AND METHODS: All the patients were divided into two groups. Those comprising the control group received only medicamental therapy. The standard treatment of the patients in the second group was supplemented by intravenous laser irradiation of blood (IVLIB-405 technique). Each irradiation session performed in a continuous mode lasted 5-7 minutes, with the total treatment course consisting of 7 daily procedures. Lactoferrin was determined before and after the treatment by the enzyme immunoassay with the use of "Lactoferrin Strip" kits ("Vektor-Best- Yug" JSC, Russia). RESULTS: Analysis of the results of the study revealed a significant decrease in the blood lactoferrin level down to the normal range in the patients treated with the use of laser therapy. CONCLUSION: The application of intravenous laser irradiation of blood for the combined treatment of the patients with community-acquired pneumonia was accompanied by normalization of the blood lactoferrin level and the improvement of the clinical course of the disease.

Oropharyngeal colostrum administration in extremely premature infants: an RCT.

OBJECTIVE: To determine the immunologic effects of oropharyngeal colostrum administration in extremely premature infants. METHODS: We conducted a double-blind, randomized, placebo-controlled trial involving 48 preterm infants born before 28 weeks' gestation. Subjects received 0.2 mL of their mother's colostrum or sterile water via oropharyngeal route every 3 hours for 3 days beginning at 48 to 96 hours of life. To measure concentrations of secretory immunoglobulin A, lactoferrin, and several immune substances, urine and saliva were obtained during the first 24 hours of life and at 8 and 15 days. Clinical data during hospitalization were collected. RESULTS: Urinary levels of secretory immunoglobulin A at 1 week (71.4 vs 26.5 ng/g creatinine, P = .04) and 2 weeks (233.8 vs 48.3 ng/g creatinine, P = .006), and lactoferrin at 1 week (3.5 vs 0.9 µg/g creatinine, P = .01) were significantly higher in colostrum group. Urine interleukin-1ß level was significantly lower in colostrum group at 2 weeks (55.3 vs 91.8 µg/g creatinine, P = .01). Salivary transforming growth factor-ß1 (39.2 vs 69.7 µg/mL, P = .03) and interleukin-8 (1.2 vs 4.9 ng/mL, P = .04) were significantly lower at 2 weeks in colostrum group. A significant reduction in the incidence of clinical sepsis was noted in colostrum group (50% vs 92%, P = .003). CONCLUSIONS: This study suggests that oropharyngeal administration of colostrum may decrease clinical sepsis, inhibit secretion of pro-inflammatory cytokines, and increase levels of circulating immune-protective factors in extremely premature infants. Larger studies to confirm these findings are warranted.

Iron metabolism in infants: influence of bovine lactoferrin from iron-fortified formula.

OBJECTIVE: The aim of this study was to evaluate whether an iron-fortified formula with a concentration of lactoferrin would significantly improve the hematologic indexes and iron status in term infants compared with those same values in infants fed an iron-fortified formula without lactoferrin. METHOD: In this prospective, multicenter, controlled intervention study, 260 infants ages 4 to 6 mo were selected from six maternal and children's health care hospitals in the area. All infants were divided into two groups with the sequence of outpatient: lactoferrin-fortified formula milk group (fortified group, FG, containing lactoferrin 38 mg/100 g milk and iron element 4 mg/100 g milk) and no lactoferrin fortified milk (control group, CG, containing lactoferrin 0 mg/100 g milk and iron element 4 mg/100 g milk) for 3 mo. The levels of weight, height, and head circumference and the concentration of hemoglobin (Hb), serum ferritin (SF), and serum transferring receptor (sTfR) were measured and sTfR-SF index (TFR-F index), total body iron content (TBIC) and low height for age (HAZ), low weight for age (WAZ), and low weight for height (WHZ) were computed before and after the intervention, respectively. RESULTS: In all, 213 (115 in FG and 98 in CG) infants completed the intervention trial and all measurements of biochemical indicators. There were no significant differences in the average amount of daily intake of formula milk (94.3 ± 9.8 g versus 88.2 ± 8.7 g for FG and CG; P > 0.05) and iron element (3.8 ± 0.4 mg versus 3.7 ± 0.6 mg for FG and CG; P > 0.05). The average amount of daily intake of lactoferrin for infants in FG group was 35.8 ± 3.7 mg. The levels of weight, WAZ, WHZ, Hb, SF, TFR-F index, and TBIC after intervention of infants in FG were all significantly higher than those of infants in CG weight, 8723 ± 245 g versus 8558 ± 214g; WAZ, 1.02 ± 0.31 versus 0.44 ± 0.18; WHZ, 0.98 ± 0.31 versus 0.41 ± 0.12; Hb, 125.5 ± 15.4 g/L versus 116.9 ± 13.1 g/L; SF, 44.7 ± 17.2 µg/L versus 31.6 ± 18.4 µg/L; TFR-F index, 1.88 ± 0.41 versus 1.26 ± 0.39; TBIC, 6.12 ± 0.78 mg/kg versus 5.26 ± 0.55 mg/kg for FG and CG; P < 0.05), but significantly lower (P < 0.05) for the prevalence of anemia (4.1% versus 7.5%), iron deficiency (13.9% versus 24.4%), and iron-deficient anemia (1.7% versus 6.1%). CONCLUSION: When infants who were exclusively breastfed were supplemented with lactoferrin-fortified milk, significant increases in TBIC and iron absorption in the intestine were seen.

Lactoferrin, myeloperoxidase, and ceruloplasmin: complementary gearwheels cranking physiological and pathological processes.

Copper-containing plasma protein ceruloplasmin (Cp) forms a complex with lactoferrin (Lf), an iron-binding protein, and with the heme-containing myeloperoxidase (Mpo). In case of inflammation, Lf and Mpo are secreted from neutrophil granules. Among the plasma proteins, Cp seems to be the preferential partner of Lf and Mpo. After an intraperitoneal injection of Lf to rodents, the "Cp-Lf" complex has been shown to appear in their bloodstream. Cp prevents the interaction of Lf with protoplasts of Micrococcus luteus. Upon immunoprecipitation of Cp, the blood plasma becomes depleted of Lf and in a dose-dependent manner loses the capacity to inhibit the peroxidase activity of Mpo, but not the Mpo-catalyzed oxidation of thiocyanate in the (pseudo)halogenating cycle. Antimicrobial effect against E. coli displayed by a synergistic system that includes Lf and Mpo-H2O2-chloride, but not thiocyanate, as the substrate for Mpo is abrogated when Cp is added. Hence, Cp can be regarded as an anti-inflammatory factor that restrains the halogenating cycle and redirects the synergistic system Mpo-H2O2-chloride/thiocyanate to production of hypothiocyanate, which is relatively harmless for the human organism. Structure and functions of the "2Cp-2Lf-Mpo" complex and binary complexes Cp-Lf and 2Cp-Mpo in inflammation are discussed.

[Effect of transcranial electrostimulation on clinical and laboratory characteristics in patients with gastric ulcer].

The study included 100 patients with gastric ulcer aged from 16 to 60 years. In 58 patients, traditional treatment was supplemented by transcranial electrotherapy using a TRANSAIR-02 apparatus. Laboratory studies included measurements of lactoferrin (LF) and tumour necrosis factor-alpha (TNF-alpha), besides standard analyses. Combined treatment (traditional therapy plus electrostimulation) resulted in a significant reduction in duration of the main symptoms of the gastric ulcer depending on the severity of ulceration process, patients' age and sex. Moreover, it facilitated ulcer scarring and had beneficial effect on dynamics of serum LF and TNF-alpha levels.

Oral bovine lactoferrin improves bone status of ovariectomized mice.

The aim of the present study was to evaluate the effect of dietary lactoferrin on bone metabolism in vivo using a postmenopausal animal model. We investigated whether bovine lactoferrin (bLF) ingestion could prevent bone loss in ovariectomized mice. Twelve-week-old female C3H mice either ovariectomized or sham operated were fed for 27 wk with the control diet (AIN-93M with 140 g of total milk protein as a protein source per kg of diet). Four groups of ovariectomized mice received diets including different concentrations of bLF (1, 5, 10, or 20 g of total milk protein were replaced by bLF). Ovariectomy induced a decreased uterine weight and a smaller gain of bone mineral density. Immunoreactive bLF was detected in the peripheral blood, and its concentration was related to the amount of bLF ingestion. bLF supplementation to the diet improved bone mineral density (BMD) and femoral failure load in a dose-dependent manner. We confirmed the direct effects of bLF in vitro using established and primary cultures of murine bone cells. Addition of bLF to the culture medium at a concentration of between 1 and 1,000 microg/ml stimulated both cell growth and differentiation of osteoblastic MC3T3 cells while inhibiting the growth of preosteoclastic RAW 267.4 cells. In primary culture of mixed bone cells, an enhanced osteoblast differentiation was associated with an inhibition of osteoclast differentiation at lower bLF concentrations (1-10 microg/ml). In conclusion, these findings suggest that dietary lactoferrin supplementation can have a beneficial effect on postmenopausal bone loss by modulating bone formation and resorption.

Granulocyte activation in humans is modulated by psychological stress and relaxation.

Our aim was to study the possible relationship between psychological stress and granulocyte activation primarily in healthy students during an examination period (n = 11) and also in chronically anxious patients (n = 15). We employed cell surface markers: lactoferrin, L-selectin, alphaMbeta2-integrin and CD15s and flow cytometry to detect changes in the activation state of granulocytes, with the start of the stressed state in students at the beginning of an examination period, which was associated with elevated blood plasma cortisol level, and following relaxation hypnosis in both students, during their examination term, and patients. The ratios of all four types of marker-carrier granulocytes increased at the start of the examination period in students; an especially dramatic (ca. 5-fold) enhancement was observed in the proportion of lactoferrin-bearing cells relatively to the pre-examination term value. After hypnosis, the percentage of lactoferrin-exposing granulocytes decreased considerably both in students and in patients, by about half; a similar decrease was observed in the ratio of CD15s-carrier cells in patients. No significant alteration was observed during the study in state or trait anxiety levels, and in total or differential leukocyte counts. Thus, granulocyte activation could be associated with stress, while relaxation may facilitate reducing activation of these cells. In both groups of subjects, granulocyte surface lactoferrin appeared to be a sensitive "stress indicator". This needs further evaluation.

Serum lactoferrin and immunoglobulin G concentrations in healthy or ill neonatal foals and healthy adult horses.

BACKGROUND: Lactoferrin is a colostral glycoprotein with antimicrobial properties. HYPOTHESES: (1) Serum lactoferrin and immunoglobulin G (IgG) concentrations are correlated and increase in healthy foals after ingestion of colostrum; (2) compared to healthy foals, ill foals will have lower lactoferrin concentrations that correlate with their IgG concentration, neutrophil count, the diagnosis of sepsis, and survival; and (3) plasma concentrations of lactoferrin will be less than serum concentrations. ANIMALS: Healthy foals (n = 16), mature horses (n = 10), and ill foals 1-4 days old (n = 111) that were examined for suspected sepsis were used for blood collection. Colostrum was obtained from 10 healthy mares unrelated to the foals. METHODS: Blood was obtained from the healthy foals at birth and 1-3 days of age and from the ill foals at admission. Serum IgG was quantified by single radial immunodiffusion (SRID). Lactoferrin concentrations in colostrum and blood were determined by an enzyme-linked immunosorbant assay. The sepsis score, blood culture results, neutrophil counts, and survival were obtained on ill foals. RESULTS: The mean colostral lactoferrin concentration was 21.7 microg/mL. Compared to values at birth, serum IgG (18+/-2 versus 2,921+/-245 mg/dL, SEM) and lactoferrin (249+/-39 versus 445+/-63 ng/mL, SEM) concentrations were significantly greater in healthy foals 1-3 days old. Serum lactoferrin concentration in 1-3-day-old healthy foals was not different from mature horses or ill foals. IgG and lactoferrin concentrations were significantly correlated only in healthy foals. Serum lactoferrin concentrations were significantly lower in ill neutropenic foals. The serum IgG concentration was significantly lower in ill foals as compared to healthy foals. Only serum IgG was significantly less in ill foals with a positive sepsis score and in nonsurvivors, Plasma lactoferrin concentrations were lower than serum concentrations, although values were significantly correlated. CLINICAL IMPORTANCE: Although both serum IgG and lactoferrin concentrations increase in healthy foals after ingestion of colostrum, only serum IgG is significantly correlated with the sepsis score and outcome.

Hematological, metabolic, and endocrine effects of feeding vitamin A and lactoferrin in neonatal calves.

There are reports of important interactions among vitamin A, lactoferrin (Lf), and components of the insulin-like growth factor (IGF) system. Newborn calves are deficient in vitamin A and have a low Lf status. Colostrum contains vitamin A, Lf, and members of the IGF system, including IGF binding proteins (IGFBP), and these compounds may exert mutual interactions in neonatal calves. Effects of feeding different amounts of vitamin A and Lf on hematological, metabolic, and endocrine traits during the first 5 d of life were studied in neonatal calves. Calves were fed a milk-based formula without (F) or with added vitamin A [F(A); 351, 402, 490, and 490 micromol/kg dry matter (DM) on d 1, 2, 3, and 4, respectively], Lf (F(L); 3850, 1990, 660, and 660 mg/kg DM on d 1, 2, 3, and 4, respectively), or vitamin A + Lf (F(AL)) and colostrum (C). Blood samples were taken preprandially on d 1 to 5 and postprandially on d 1, 2, and 4 for the study of hematological, metabolic, and endocrine traits. As intestinal absorption of Lf in formula-fed calves was obviously lower than in colostrum-fed calves, Lf supplementation did not influence the metabolic and endocrine status. Plasma vitamin A concentrations increased from d 1 to 5 in F(A), F(AL), and C, but remained low in F and F(L). Addition of vitamin A to F affected concentrations of vitamin A from d 2 to 5; hemoglobin and triglycerides on d 3; and tended to influence IGFBP-3 levels on d 5. On d 1, preprandial insulin concentrations in F and F(L) were higher than those in F(A) and F(AL) and growth hormone concentrations on d 3 in F(A) were higher than in F(AL). In C, plasma Lf concentrations increased on d 1 and then decreased until d 5, whereas plasma urea and IGF-I concentrations remained stable. Compared with F-fed calves, C-fed calves had higher plasma concentrations of Lf from d 1 to 4, higher concentrations of vitamin A on d 5, higher concentrations of total protein from d 2 to 5, higher concentrations of triglycerides on d 3 and 4, and higher concentrations of IGF-I on d 4 and 5, but lower urea concentrations on d 4 and 5 and lower basal growth hormone concentrations than in F, F(L), and F(A). In conclusion, metabolite and hormone concentrations were influenced by differences in feeding. Addition of vitamin A influenced concentrations of vitamin A, hemoglobin, and triglycerides and tended to affect IGFBP-3 levels. Lactoferrin and vitamin A did not interact with respect to the IGF-I and IGFBP-3 status.

Zinc, iron, copper, selenium, lactoferrin, and ferritin in human pus.

BACKGROUND: Restriction of zinc and iron available for microbial growth in tissues are well-recognized host defense mechanisms. The present studies were performed to characterize some constituents of human pus that may affect these important host defenses. METHODS: Zinc, iron, copper, calcium, and magnesium in pus were measured using an atomic absorption spectrophotometer; selenium was measured fluorometrically. Ferritin was measured with a fluorometric enzyme immunoassay, and lactoferrin was measured with a radial diffusion assay. The growth of Escherichia coli at 37 degrees C was measured in pus supernate adjusted to pH 5.5 or 7.4, in boiled supernate, or in supernate adjusted with 1.3 mM iron or 0.9 mM zinc singly or together. RESULTS: Zinc and iron concentrations in pus exceeded normal serum. Calcium and magnesium levels were 2- to 3-fold lower and higher, respectively, than normal serum values. Lactoferrin concentrations of were 880 +/- 48 microg/mL and ferritin levels were 20,726 +/- 2,667 ng/mL. Growth of an E coli strain was inhibited in pus at pH 5.5 but not at pH 7.4, and growth was enhanced by addition of iron or zinc to E coli suspended in pus at pH 6.7. CONCLUSIONS: To our knowledge, this is the first report of the zinc, iron, copper, selenium, lactoferrin, and ferritin levels of human pus. These studies provide additional insight into host defense mechanisms mediated by the restriction of the bioavailability of zinc and iron in suppurative infection.

Effects of supplemental lactoferrin on serum lactoferrin and IgG concentrations and neutrophil oxidative metabolism in Holstein calves.

Lactoferrin (LF) is an iron-binding protein present in both colostrum and secondary granules of polymorphonuclear neutrophils (PMNs). We hypothesized that supplemental LF enhances neutrophil function in neonatal calves. Newborn calves were assigned to receive colostrum (C), colostrum + LF (CLF, 1 g/kg), or milk replacer + LF (MRLF, 1 g/kg). Serum (LF and IgG) and whole blood (neutrophil isolation) samples were obtained prior to treatment (day 0) and at 24 hours and 9 days of age. Serum IgG concentrations (mean +/- SD) in C, CLF, and MRLF calves at 24 hours were 1,911 +/- 994 mg/dL, 2,181 +/- 625 mg/dL, and 0 mg/ dL, respectively. Serum LF concentrations in C, CLF, and MRLF calves on day 0 were 324 +/- 334 ng/mL (range 0-863 ng/mL), 135 +/- 158 ng/mL (range 0-429 ng/mL), and 318 +/- 337 ng/mL (range 0-964 ng/mL), respectively. LF concentrations in C, CLF, and MRLF calves at 24 hours were significantly higher (P < .05), at 1,564 +/- 1,114 ng/mL (range 335-3,628 ng/mL, 2,237 +/- 936 ng/mL (range 31-3,287 ng/mL), and 3,189 +/- 926 ng/mL (range 1,736-4,120 ng/mL), respectively. Cytochrome c reduction in opsonized zymosan-treated or phorbol ester-treated cells was not significantly affected by supplemental LF provided at birth. Oral LF is absorbed in calves but does not alter PMN superoxide production and does not alter IgG absorption.

C-reactive protein and antibacterial activity in blood plasma of colostrum-fed calves and the effect of lactulose.

Several milk proteins are very important for immunological defense and can be absorbed in the intestine of calves in the first hours after birth. The influence of colostrum intake and the effect of additional lactulose application on the concentration of C-reactive protein (CRP) in blood were investigated. The CRP is known as a mediator of innate immunity. Results were compared to the bovine acute phase protein haptoglobin, and to lactalbumin, lactoferrin, and immunoglobulins in plasma from calves. After colostrum intake, the concentration of most proteins were strongly increased. The data show, for the first time, a significant increase of CRP in the blood of calves 1 d after colostrum intake (nonlactulose group, n = 10), and an even more significant increase in CRP concentration (1 d postpartum) was measured in the group of animals with additional application of lactulose (lactulose group, n = 10) when compared to the nonlactulose group. In an in vitro assay with the plasma of these animals, an increased bactericidal activity was detected against Morganella morganii (1 d postpartum) in both groups, but again a higher activity occurred in the lactulose group. The results of these investigations emphasize the importance of colostrum intake during the first hours after birth for the defense potential of newborn calves. In addition, lactulose may have a positive effect in the period of passive transfer of colostrum proteins and in the immune defense.

Characteristic transfer of colostrum-derived biologically active substances into cerebrospinal fluid via blood in natural suckling neonatal pigs.

The characteristic transfer of colostral components into cerebrospinal fluid (CSF) via serum after natural suckling has been studied by sodium dodecyl sulphate (SDS) electrophoresis, two-dimensional electrophoresis, immunoblot and enzyme linked immunosorbent assay methods in non-suckling pigs. Total protein concentrations in the serum increased immediately after first suckling, reached a peak value at 12 h, corresponding to a 2.3-fold compared with pre-suckling level. The protein concentration in CSF also increased and reached peak value at 6 h corresponding to 1.6-fold compared with presuckling level. IgG in serum not detected before suckling, increased steeply after suckling, IgG, IgM and IgA transported into the serum were observed in completely intact form by immunoblot method. The IgG transported into serum was quickly transferred into CSF after natural suckling in contrast to the case of bovine IgG. Serum concentration of transferrin was maintained at high level before suckling and was not changed by suckling. Transferrin also detected in CSF was not changed by suckling. Bovine lactoferrin (Lf) administered into the intestinal lumen was transported into serum (0.01%) and also detected in CSF after 6 h as undegraded form (3.1%). Thus, homologous IgG and bovine Lf are transported into CSF, suggesting that the transport of macromolecules into CSF is selective in neonatal pigs.

Serum lactoferrin concentrations in colostrum-fed calves.

OBJECTIVE: To determine serum lactoferrin concentrations (SLFC) in neonatal calves before and after ingestion of colostrum and to develop models that predict SLFC as a function of colostral lactoferrin concentrations (CLFC) in calves. ANIMALS: 13 Holstein calves. PROCEDURE: Calves were fed 4 L of colostrum via oroesophageal feeder within 3 hours after birth. Serum samples were collected before ingestion of colostrum (day 0) and 2, 4, 6, and 7 days after birth. Colostrum and serum IgG concentrations were measured by use of radial immunodiffusion. The CLFC and SLFC were determined by use of an ELISA. RESULTS: Mean +/- SD SLFC on days 0, 2, 4, 6, and 7 were 2.5+/-1.6 (range 0.47 to 71), 6.0+/-3.0 (range 2.0 to 16.6), 12.0+/-12.4 (range 0.0 to 43.5), 171+/-13.6 (range 2.2 to 39.4), and 13.6+/-16.4 (range 0.0 to 43.8) mg/ml, respectively. The SLFC on days 6 and 7 differed significantly from SLFC on day 0. The model that best estimated SLFC on day 6 predicted that (SLFC)2 was a function of the logarithm of relative efficiency of passive transfer (REPT) and ([CLFC]2 x [REPT]2), where R2 = 0.4. The model for SLFC on day 7 predicted that (SLFC)2 was a function of log(REPT), where R2 = 0.44. CONCLUSIONS AND CLINICAL RELEVANCE: Definitive evidence for passive transfer of lactoferrin via colostrum is lacking, because SLFC on day 2 or 4 were not significantly different than day 0. Relative efficiency of lactoferrin absorption was directly related to SLFC on day 6 but inversely related to SLFC on day 7.

Effects of pasteurization of colostrum on subsequent serum lactoferrin concentration and neutrophil superoxide production in calves.

OBJECTIVE: To determine the effects of pasteurization of colostrum on serum lactoferrin concentration and neutrophil oxidative function by comparing values from calves given pasteurized (76 C, 15 minutes) colostrum versus calves given fresh frozen colostrum. ANIMALS: 8 Holstein bull calves were used to study the effects of pasteurization of colostrum on the absorption of lactoferrin and neutrophil oxidative burst. Three additional calves were used to study the effect of exogenous lactoferrin on neutrophil oxidative burst. METHODS: Calves were fed fresh frozen or heat pasteurized colostrum (76 C for 15 minutes) via esophageal feeder within 4 hours of birth. Neutrophils were isolated from whole blood samples. Neutrophil oxidative burst was induced by phorbol ester (300 ng/ml) stimulation of cells (1 X 10(6) cells) at 37 C. Serum lactoferrin concentrations were compared, using immunoblot analysis. Serum IgG concentrations were determined by radial immunoassay. Comparisons were made between the use of the 2 types of colostrum in calves by measuring subsequent serum IgG and lactoferrin concentrations and neutrophil superoxide production. RESULTS: Serum IgG and lactoferrin concentrations increased more in calves receiving fresh frozen colostrum. Neutrophil superoxide production was higher in neutrophils prepared from calves receiving fresh frozen colostrum. Colostral lactoferrin addition to neutrophil incubations resulted in increased oxidative burst. CONCLUSIONS AND CLINICAL RELEVANCE: Compared with calves given fresh frozen colostrum, calves given pasteurized colostrum had decreased serum IgG and lactoferrin concentrations and neutrophil superoxide production 24 hours after administration. These results suggest that pasteurizing bovine colostrum at 76 C for 15 minutes has substantial effects on passive transfer of proteins and neutrophil function.

[Immunoenzyme method of lactoferrin analysis]. / Immunofermentnyi metod opredeleniia laktoferrina.

The authors propose a highly effective method for isolation of lactoferrin from female colostrum. Lactoferrin is measured by the sandwich enzyme immunoassay. Its concentrations in the serum, urine, amniotic and cerebrospinal fluid in health are measured. The advantages of the method for lactoferrin purification and the practical significance of this protein are discussed.