Effect Of Bovine Lactoferrin Research Articles

Evaluation of in vitro antirotaviral activity of lactoferrin from different species using a human intestinal model

Rotaviruses are the leading agents of severe diarrheal disease in infants and young children and complementary strategies to vaccination are needed. Therefore, we evaluated the activity of lactoferrin (LF) from different species against rotavirus in differentiated Caco-2/TC7 cells. The mechanism of rotavirus inhibition and the influence of iron saturation and sialic acid content were also investigated. Furthermore, the effect of bovine LF on Toll-like receptor expression (TLRs) during rotavirus infection was studied. LF presented high inhibitory activity against rotavirus above 8 mg mL−1, the bovine protein being the most potent. The antirotaviral mechanism of LF was mainly related to the virus neutralisation at an early step of infection. Interestingly, a direct relationship between rotavirus neutralisation activity and content of sialic acid in LFs was observed. Finally, the neutralisation of rotavirus by LF leaded to a decreased expression of TLRs in Caco-2/TC7 cells, with a more evident effect on TLR7.

Effect of bovine lactoferrin on recurrent urinary tract infections: in vitro and in vivo evidences.

Uropathogenic Escherichia coli (UPEC) strains are the primary cause of urinary tract infections (UTIs). UPEC strains are able to invade, multiply and persisting in host cells. Therefore, UPEC strains are associated to recurrent UTIs requiring long-term antibiotic therapy. However, this therapy is suboptimal due to the increase of multidrug-resistant UPEC. The use of non-antibiotic treatments for managing UTIs is required. Among these, bovine lactoferrin (bLf), a multifunctional cationic glycoprotein, could be a promising tool because inhibits the entry into the host cells of several intracellular bacteria. Here, we demonstrate that 100μg/ml bLf hinders the invasion of 2.0 ± 0.5 × 104CFU/ml E. coli CFT073, prototype of UPEC, infecting 2.0 ± 0.5 × 105 cells/ml urinary bladder T24 epithelial cells. The highest protection (100%) is due to the bLf binding with host surface components even if an additional binding to bacterial surface components cannot be excluded. Of note, in the absence of bLf, UPEC survives and multiplies, while bLf significantly decreases bacterial intracellular survival. After these encouraging results, an observational survey on thirty-three patients affected by recurrent cystitis was performed. The treatment consisted in the oral administration of bLf alone or in combination with antibiotics and/or probiotics. After the observation period, a marked reduction of cystitis episodes was observed (p < 0.001) in all patients compared to the episodes occurred during the 6months preceding the bLf-treatment. Twenty-nine patients did not report cystitis episodes (87.9%) whereas the remaining four (12.1%) experienced only one episode, indicating that bLf could be a worthwhile and safe treatment in counteracting recurrent cystitis.

Effect of bovine lactoferrin on seroconversion following polio vaccine administration in children: protocol for a double-blinded randomised controlled trial

IntroductionThe oral polio vaccine (OPV) has substantial results in eliminating wild poliovirus and the vaccine of choice in polio eradication. However, the mucosal immunity induced by the OPV is still...

Transcriptome analysis of Lactobacillus rhamnosus GG strain treated with prebiotic - bovine lactoferrin under a cold environment.

Lactoferrin (LF) is secreted by mammals and displays extensive biological effects. We previously reported that bovine LF (BLF) can boost the cold tolerance of a well-applied probiotic strain, Lactobacillus rhamnosus GG (LGG), to grow robustly under a cold environment, but the molecular mechanism is not clear. Here, RNA-seq analysis was conducted to ascertain molecular pathways underlying cold tolerance exerted by BLF. LGG was cultured in a cold environment (22 °C) in the presence or absence of BLF. Transcriptome analysis indicated that BLF significantly elicited 1.2–3.2 fold (log2 Fold change) higher expression levels of genes related to stress, defense, cell division, and transporter in LGG, including the genes CspA, LytR, XRE, MerR, and GpsB. The KEGG pathway and GO analyses confirmed that BLF can modulate a few central pathways to boost the growth of LGG. BLF also reduced metabolic pathways involved in purine, amino acid, pyrimidine, one-carbon metabolism, and secondary metabolites in LGG. We speculate that the reduction of the above pathways may play key roles to reduce energy requirement and maintain carbon metabolism balance in LGG for surviving and growing in a cold state, and BLF can be an excellent prebiotic to LGG cultured in this cold condition (22 °C). Overall, this study uncovers the molecular effects of BLF on LGG.

Effects of bovine lactoferrin and chitosan nanoparticles on serum biochemical indices, antioxidative enzymes, transcriptomic responses, and resistance of Nile tilapia against Aeromonas hydrophila

The present study was carried out to investigate the effects of dietary bovine lactoferrin (BLF) or chitosan nanoparticles (CHN) alone or in combinations on serum biochemical indices, antioxidative capacity, transcriptomic responses, non-specific immunity, and resistance of Nile tilapia (Oreochromis niloticus) against challenge with Aeromonas hydrophila. Fish were fed on the basal diet with no supplements and served as control (CTR), and six other experimental diets containing different levels of BLF (800 and 1200 mg per kg diet), CHN (500 and 1000 mg per kg diet), and their combinations (400 mg BLF plus 250 mg CHN per kg diet, and 600 mg BLF plus 500 mg CHN per kg diet) for 45 days. At the end of the experiment, serum, and tissue specimens (liver and kidney) were collected, fish in all groups were challenged with A. hydrophila and then observed for another ten days to calculate the RPS. Compared to the CTR group, no significant differences were recorded in TP, ALB, GLO, BUN, and CREAT values among all treatments. Serum LYZ, ALT, AST, and ALP enzyme activities were significantly increased in all experimental groups over the CTR (P < 0.05), and their highest values were recorded in the combined treatments. Moreover, dietary supplementation with CHN (1000 mg/kg) and combined treatments significantly increased the SOD, CAT, and GSH-Px enzyme activities compared to other groups (P < 0.05). The highest mRNA expression levels of IGF-1 gene in liver, and IL-1β, and IFN-γ genes in kidneys were found in CHN (1000 mg/kg) group and combined treatments more than other groups. Interestingly, no, or mild histopathological alterations were noticed in the hepatopancreas and posterior kidney of the treated groups. A significantly higher RPS was identified in the combined treatments challenged with A. hydrophila compared with the CTR group. This study exemplifies the positive impacts of dietary supplementation with BLF or CHN alone or combinations on the antioxidative status, immunity, and disease resistance of Nile tilapia.

Effect of bovine lactoferrin on prevention of late-onset sepsis in infants &lt;1500 g: a pooled analysis of individual patient data from two randomized controlled trials

We previously conducted two randomized controlled trials with bovine lactoferrin (bLF) for the prevention of late-onset sepsis (LOS) in infants with a birth weight <2500 g (Study 1) and <2000 g (Study 2). The aim of this study was to determine the preventative effects of bLF on culture-proven or probable LOS in infants with a birth weight <1500 g from both studies, and to determine the effect of bLF in relation to intake of human milk. Both trial designs had similar inclusion and exclusion criteria, the same dose of bLF [200 mg·(kg body mass)-1·day-1], and used the same control (maltodextrin). We fitted multivariate Cox regression models to estimate the effect of bLF on the risk of development of the composite outcome, adjusting for covariates. We included 335 neonates with a mean birth weight of 1162 ± 244 g; 27.5% were <1000 g. There were 33 first episodes of LOS in the bLF treatment group and 48 in the control group (19.5% vs. 28.9%). bLF had a protective effect on the risk of development of LOS [hazard ratio (HR) = 0.64; %95 CI = 0.41-0.99; p= 0.048]; particularly among infants weighing <1000 g [HR = 0.46; %95 CI = 0.22-0.96; p= 0.039] and infants with a low intake of human milk [HR = 0.40; %95 CI = 0.19-0.84; p= 0.015]. Therefore, bLF supplementation protects infants <1500 g from LOS, particularly those infants not receiving human milk.

Effect of bovine lactoferrin as a novel therapeutic agent in a rat model of sepsis-induced acute lung injury

Sepsis is a serious clinical condition resulting from severe infection. High rates of mortality and tissue damage have been reported in intensive care unit (ICU) patients with sepsis. Bovine lactoferrin (BLF) is a well-known 80-kDa glycoprotein in the transferrin family that inhibits sepsis in low-birth-weight neonates. The present study investigated the protective effects of BLF in a rat model of sepsis-induced acute lung injury (ALI). The wet/dry ratio, lipid peroxidation, antioxidant markers, total protein, total cell count, inflammatory markers and myeloperoxidase (MPO) levels were assessed. Histopathological analysis was also carried out. BLF treatment reduced the wet/dry ratio of lung tissue by 30.7% and 61.3%, and lipid peroxidation by 22.3% and 67%, at concentrations of 100 and 200 mg/kg, respectively. Superoxide dismutase (SOD), reduced glutathione (GSH), glutathione peroxidase (Gpx) and catalase were increased by more than 50% under treatment with 200 mg/kg BLF. Inflammatory markers, neutrophils, lymphocytes and total cell count were reduced by more than 50% under treatment with 200 mg/kg BLF. BLF treatment significantly reduced MPO activity, by 28.2% and 74.3%, at concentrations of 100 and 200 mg/kg, respectively. Neutrophilic infiltration and edema were observed in control rats. However, BLF treatment restored intestinal microvilli to the normal range and reduced inflammatory cell invasion. Collectively, these results suggest that BLF is an effective therapeutic agent against sepsis-induced ALI.

Is Lactoferrin More Effective in Reducing Late-Onset Sepsis in Preterm Neonates Fed Formula Than in Those Receiving Mother's Own Milk? Secondary Analyses of Two Multicenter Randomized Controlled Trials.

Lactoferrin is the major antimicrobial protein in human milk. In our randomized controlled trial (RCT) of bovine lactoferrin (BLF) supplementation in preterm neonates, BLF reduced late-onset sepsis (LOS). Mother's own milk (MM) contains higher concentrations of lactoferrin than donor milk or formula, but whether BLF is more effective in infants who receive formula or donor milk is uncertain. To evaluate the incidence of LOS in preterm infants fed MM and in those fed formula and/or donor milk. This is a (A) post hoc subgroup analysis, in our RCT of BLF, of its effects in preterm infants fed MM, with or without formula, versus those fed formula and/or donor milk (no-MM) and (B) post hoc meta-analysis, in our RCT of BLF and in the ELFIN (Enteral Lactoferrin in Neonates) RCT, of the effect of BLF in subgroups not exclusively fed MM. (A) Of 472 infants in our RCT, 168 were randomized to placebo and 304 were randomized to BLF. Among MM infants, LOS occurred in 22/133 (16.5%) infants randomized to placebo and in 14/250 (5.6%) randomized to BLF (relative risk or risk ratio (RR): 0.34; relative risk reduction (RRR): 0.66; 95% confidence interval (95% CI) for RR: 0.18-0.64; p < 0.0008). Among no-MM infants, LOS occurred in 7/35 (20.0%) randomized to placebo and in 2/54 (3.7%) randomized to BLF (RR: 0.19; RRR: 0.81; 95% CI for RR: 0.16-0.96; p = 0.026). In multivariable logistic regression analysis, there was no interaction between BLF treatment effect and type of feeding (p = 0.628). (B) In 1,891 infants not exclusively fed MM in our RCT of BLF and in the ELFIN RCT, BLF reduced the RR of LOS by 18% (RR: 0.82; 95% CI: 0.71-0.96; p = 0.01). Adequately powered studies should address the hypothesis that BLF is more effective in infants fed formula or donor milk than those fed MM. Such studies should evaluate whether a specific threshold of total lactoferrin intake can be identified to protect such patients from LOS.

Effect of bovine lactoferrin and human lactoferrin on the proliferative activity of the osteoblast cell line MC3T3-E1 in vitro

We conducted a comparative in vitro study on the proliferative effects of natural human lactoferrin (nhLF) and bovine lactoferrin (bLF) on osteoblasts. We investigated cell proliferation, cell survival, cell cycle, and mRNA and protein expression of proliferating cell nuclear antigen. Results indicated that treatment with 100 μg/mL of bLF or nhLF promoted the proliferation and sustenance of osteoblasts, and increased the length of the G2/M and S phases compared with the untreated osteoblasts. Results of real-time quantitative PCR and Western blot showed that mRNA and protein expression of proliferating cell nuclear antigen by osteoblasts treated with bLF or nhLF were greater than those of the untreated control. At the same concentration, bLF demonstrated a greater effect on osteoblast proliferation than did nhLF. This study provides insights of significance in the utlization of bLF in healthy food formulas.

Effects of bovine lactoferrin on cell viability, proliferation and inflammation in intestinal epithelial cells treated by lipopolysaccharide

Objective To evaluate the effects and mechanisms of bovine lactoferrin (bLF) on cell viability, proliferation, and the protective roles in intestinal epithelial cell-6 (IEC-6) treated by lipopolysaccharide(LPS). Methods The rat jejunum epithelial cell lines IEC-6 were cultured in vitro.The effects of bLF on cell viability and proliferation in IEC-6 cells were detected by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay and 5-Bromo-deoxyuridine (Brdu) assay, respectively.Inflammatory cytokines and their mRNA of interleukin-1β(IL-1β), interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and interleukin-8(IL-8)were analyzed by real-time PCR and enzyme-linked immunosorbent assay(ELISA). Western blot was used to measure the levels of mitogen-activated protein kinase(MAPK)activation and nuclear factor kappa β (NF-κB) nuclear translocation. Results Dose dependent effects of bLF on cell viability and proliferation were observed in IEC-6 cells in vitro(F=3.825, 5.861, all P<0.05), especially in a dose of 100 mg/L, and bLF significantly stimulated cell viability and proliferation compared with non-treatment group(q=5.240, 3.765, all P<0.05). The mRNA levels of IL-1β, IL-6 and TNF-α were decreased by co-stimulation of bLF and LPS compared with the LPS treatments alone in IEC-6 cells in vitro(q=14.28, 10.12, 16.45, all P<0.001). The secretion of IL-6 and TNF-α was also decreased by co-stimulation of bLF and LPS (q=15.06, 6.74, all P<0.01). In vitro, bLF treatment at dose of 100 mg/L could inhibit the activation of MAPK/NF-κB signal pathway induced by LPS (q=12.96, 18.54, all P<0.001). Conclusion In vitro, bLF can promote IEC-6 viability and proliferation, and have anti-inflammatory effects via inhibited activation of MAPK/NF-κB nuclear translocation. Key words: Bovine lactoferrin; Intestinal epithelial cell; Proliferation; Inflammation

Lactoferrin inhibits melanogenesis by down-regulating MITF in melanoma cells and normal melanocytes.

The aim of this study was to evaluate the effect of bovine lactoferrin (bLf) on melanin-producing cells and to elucidate its mechanism of action. We tested the anti-melanogenic effect of bLf on a 3-dimensional cultured pigmentation skin model and confirmed a 20% reduction in pigmentation, suggesting that bLf was transdermally absorbed and it suppressed melanin production. Treatment of human melanoma cells with bLf resulted in a significant, dose-dependent suppression of melanin production. Apo-bLf and holo-bLf suppressed melanogenesis to the same degree as bLf. The key feature behind this anti-melanogenic effect of bLf was the down-regulation of the microphthalmia-associated transcription factor (MITF), leading to the suppression of tyrosinase activity. Treatment with bLf resulted in both decreased expression of MITF mRNA and enhanced degradation of MITF protein. However, the primary effector was enhanced phosphorylation of extracellular signal-regulated kinase (ERK), leading to the phosphorylation and degradation of MITF. Our finding that bLf suppresses melanin production in melanocytes indicates that bLf is a possible candidate for application as a skin-whitening agent.

Potential use of bovine lactoferrin as adjuvant for the modulation of the specific immune response in rainbow trout (Oncorhynchus mykiss W.)

The present experiment was carried out in order to evaluate the potential adjuvant effect of bovine lactoferrin (bLF) in modulating the specific antibody (Ab) response to an exogenous protein, the human-γ-globulins (HγG), injected intraperitoneally in rainbow trout Oncorhynchus mykiss (Walbaum 1792). Moreover, serum natural antibody (NatAb) level, serum total immunoglobulin (IgM) level and specific Ab response to bLF were also analysed. Experimental data showed that HγG is able to stimulate a specific Ab response when not adjuvanted with Freund’s complete adjuvant (FCA), whereas bLF induces a specific Ab response only if adjuvanted with FCA. Statistical analyses revealed that neither bLF or FCA administered singly had significant effects on serum NatAb and IgM levels, whereas the specific immune response observed against HγG when adjuvanted with bLF was comparable to that achieved when FCA was used as the adjuvant. These results provide evidence of the potential for bLF to be used as an adjuvant for salmonids.

Effect of bovine lactoferrin from iron-fortified formulas on diarrhea and respiratory tract infections of weaned infants in a randomized controlled trial

ObjectiveThe aim of this study was to evaluate the effect of supplementation with bovine lactoferrin (bLf) from iron-fortified formulas on diarrhea and respiratory tract infections (RTIs) in weaned infants. MethodsIn this prospective, multicenter, controlled intervention study, 260 infants ages 4 to 6 mo who previously were exclusively breastfed but weaned were randomized into two groups: a lactoferrin-fortified formula milk group (fortified group, FG, containing lactoferrin 38 mg/100 g milk) and a no lactoferrin-fortified milk (control group, CG); breastfed infants were enrolled and served as a reference group (breastfed group, BG). The intervention duration was 3 mo. The morbidity of diarrhea and RTIs were collected during supplementation. ResultsThe results of the study demonstrated evidence of a lower incidence rate of respiratory-related illnesses and fewer symptoms of running nose, cough, and wheezing for infants in the FG and BG groups compared with those in the CG (P < 0.05). Despite the undistinguished incidence rate of vomiting, nausea, and colic, the occurrences of diarrhea-related illnesses were significantly lower for children in the FG and BG than for those in CG (P < 0.05). ConclusionThe beneficial effects on infectious morbidity over 3 mo highlighted the potential of bLF supplementation for previously weaned infants; these findings may be applicable to other infants living in similar socioeconomic districts.

Bovine Lactoferrin Decreases Cholera-Toxin-Induced Intestinal Fluid Accumulation in Mice by Ganglioside Interaction

Secretory diarrhea caused by cholera toxin (CT) is initiated by binding of CT’s B subunit (CTB) to GM1-ganglioside on the surface of intestinal cells. Lactoferrin, a breast milk glycoprotein, has shown protective effect against several enteropathogens. The aims of this study were to determine the effect of bovine-lactoferrin (bLF) on CT-induced intestinal fluid accumulation in mice, and the interaction between bLF and CT/CTB with the GM1-ganglioside receptor. Fluid accumulation induced by CT was evaluated in the mouse ileal loop model using 56 BALB/c mice, with and without bLF added before, after or at the same time of CT administration. The effect of bLF in the interaction of CT and CTB with GM1-ganglioside was evaluated by a GM1-enzyme-linked immunosorbent assay. bLF decreased CT-induced fluid accumulation in the ileal loop of mice. The greatest effect was when bLF was added before CT (median, 0.066 vs. 0.166 g/cm, with and without bLF respectively, p<0.01). We conclude that bLF decreases binding of CT and CTB to GM1-ganglioside, suggesting that bLF suppresses CT-induced fluid accumulation by blocking the binding of CTB to GM1-ganglioside. bLF may be effective as adjunctive therapy for treatment of cholera diarrhea.

Eff ects of Bovine Lactoferrin on the Healing of Tooth Sockets in Rats

Eff ects of Bovine Lactoferrin on the Healing of Tooth Sockets in Rats

Effect of bovine lactoferrin in a therapeutic hamster model of hepatic amoebiasis1This article is part of a Special Issue entitled Lactoferrin and has undergone the Journal's usual peer review process.

Entamoeba histolytica is the causative agent of amoebiasis, a disease that produces dysentery as a result of the perforation of the large intestine. This parasite often invades other organs, primarily the liver, leading to an amoebic liver abscess (ALA), which can cause death. Metronidazole is the drug of choice for the treatment of ALA; however, it produces toxic side effects in patients. Lactoferrin (Lf) is a glycoprotein of the innate immune response that sequesters iron in the mucosae. Lf possesses immune-regulatory properties, such as antiinflammatory and antioxidant activities. Moreover, the microbicidal activity of apoLf, which lacks bound iron, has been shown. In this study, we evaluated the therapeutic effect of bovine Lf (bLf) against ALA in a model of hepatic amoebiasis in hamsters. Interestingly, hamsters treated intragastrically with Lf (2.5 mg/100 g mass) over a period of 8 days showed no clinical signs of disease and ALA was effectively decreased, with only 0.63% detectable lesion, compared with 63% in untreated animals. Furthermore, liver function and blood cells approached normal levels among those receiving bLf treatment. These results suggest that bLf may aid in the therapy of amoebiasis, likely without producing undesirable effects in patients.

Effect of bovine lactoferrin on the minimum inhibitory concentrations of ampicillin and trimethoprim–sulfamethoxazole for clinical Shigella spp. strains1This article is part of a Special Issue entitled Lactoferrin and has undergone the Journal’s usual peer review process.

Here, we determined the effect of bovine lactoferrin (bLF) on the minimum inhibitory concentration (MIC) of ampicillin and trimethoprim–sulfamethoxazole in Shigella . Using a microdilution method, the MIC was determined in the presence or absence of bovine lactoferrin (10 mg/mL) on 88 Shigella strains (56 Shigella flexneri , 15 Shigella boydii , 13 Shigella sonnei , and 4 Shigella dysenteriae ) previously isolated from peruvian children &lt;2 years old. A fold change of 2 or more in MIC values was considered significant. For ampicillin, 67 (76%) strains were highly resistant; one-third of the strains (32%) showed a decrease in ampicillin MIC in the presence of LF. This was more typical of MIC values in less resistant strains. For 7 (8%) ampicillin-resistant strains, the decrease in the MIC resulted in the strains reaching the cutoff for susceptible in the presence of bLF. For trimethoprim–sulfamethoxazole, 93% of the isolates (n = 82) were highly resistant and only 4 isolates (5%) decreased their MIC in the presence of bLF. None of the trimethoprim–sulfamethoxazole resistant strains became susceptible in the presence of LF. The decrease in the MIC in the presence of bLF seems to depend on the mechanisms of action of each antibiotic. In vivo studies are needed to further evaluate bLF as a coadjuvant to antibiotic treatment of resistant Shigella.

Inhibitory Effect of Bovine Lactoferrin on Human Hepatitis B Virus Replication in HepG2.2.15 Cells

In this study, the effect of bovine lactoferrin on HBV replication in HBV-producing HepG2.2.15 cells was analyzed. MTT assay was used to determine the non-toxic concentrations of BLF to the culture cells. The results showed that cyto- toxicity increased as the concentration of BLF increased. However, no apparent cytotoxicity was observed with concentration of BLF less than 6.25 mg/ml. The HepG2.2.15 cells were subsequently incubated with BLF of two non-toxic concen- trations. mRNA expression level of HBV core protein and surface protein were examined by real-time RT-PCR. The results showed that the mRNA level of both HBV core protein and surface protein were reduced in cells incubated with both non-toxic concentrations of BLF. Our findings suggested that BLF was able to inhibit HBV repli- cation. BLF could be a new candidate of anti-HBV drugs.

Effect of Bovine Lactoferrin and Casein Peptide Powder on Microbial Growth and Glucose Utilization by Microorganisms in Pork Meat During Storage at 4oC

Effect of Bovine Lactoferrin and Casein Peptide Powder on Microbial Growth and Glucose Utilization by Microorganisms in Pork Meat During Storage at 4oC

Lactoferrin promotes hyaluronan synthesis in human dermal fibroblasts

Bovine lactoferrin (BLF) enhanced production of hyaluronan in normal human dermal fibroblasts. The elevation of hyaluronan was accompanied by elevation of HAS2 (hyaluronan synthase 2) mRNA transcription and HAS2 protein expression. The promoting effect of BLF was not observed for HAS1. In addition, COL1A1 transcription and collagen synthesis were enhanced by BLF. These observations suggest that BLF promotes wound healing by increasing hyaluronan and type-I collagen synthesis.