Michelle McConnell

Slaughterhouse Blood: An Emerging Source of Bioactive Compounds

Slaughterhouse blood is an inevitable part of the meat production food chain and represents a rich source of protein. The physicochemical characteristics and utilization of animal blood in various food and industrial applications has been well explored. However, in recent years much attention has been paid to the generation of peptides with biological activities from food by-products including blood. This review examines the angiotensin-converting enzyme inhibitory, antioxidant, antimicrobial, and other bioactive peptides derived from various slaughterhouse animal blood sources. Furthermore, the effect of enzyme choice, degree of hydrolysis, and peptide sequence or size on the potency of these bioactivity is discussed.

Towards generation of bioactive peptides from meat industry waste proteins: Generation of peptides using commercial microbial proteases.

Five commercially available food-grade microbial protease preparations were evaluated for their ability to hydrolyse meat myofibrillar and connective tissue protein extracts to produce bioactive peptides. A bacterial-derived protease (HT) extensively hydrolysed both meat protein extracts, producing peptide hydrolysates with significant in vitro antioxidant and ACE inhibitor activities. The hydrolysates retained bioactivity after simulated gastrointestinal hydrolysis challenge. Gel permeation chromatography sub-fractionation of the crude protein hydrolysates showed that the smaller peptide fractions exhibited the highest antioxidant and ACE inhibitor activities. OFFGEL electrophoresis of the small peptides of both hydrolysates showed that low isoelectric point peptides had antioxidant activity; however, no consistent relationship was observed between isoelectric point and ACE inhibition. Cell-based assays indicated that the hydrolysates present no significant cytotoxicity towards Vero cells. The results indicate that HT protease hydrolysis of meat myofibrillar and connective tissue protein extracts produces bioactive peptides that are non-cytotoxic, should be stable in the gastrointestinal tract and may contain novel bioactive peptide sequences.

In vitro biocompatibility and cellular interactions of a chitosan/dextran‐based hydrogel for postsurgical adhesion prevention

In this paper, we report the in vitro biocompatibility and cellular interactions of a chitosan/dextran-based (CD) hydrogel and its components as determined by mutagenicity, cytotoxicity, cytokine/chemokine response, and wound healing assays. The CD hydrogel, developed for postsurgical adhesion prevention in ear, nose, and throat surgeries, was shown by previously published experiments in animal and human trials to be effective. The hydrogel was synthesized from the reaction between succinyl chitosan (SC) and oxidized dextran (DA). Cytotoxicity was assessed in an xCELLigence system and cytokine/chemokine responses were measured by ELISA in human macrophage, nasopharyngeal epithelial, and dermal fibroblast cells. A wound healing model utilized nasopharyngeal epithelial cells. CD hydrogel and DA were nonmutagenic in the Ames test. CD hydrogel showed moderate cytotoxicity for the cell lines, DA being the cytotoxic component. Some inhibition of wound healing occurred due to the cytotoxic nature of DA. Cells cultured with CD hydrogel showed no increase in TNF-α, IL-10, and IL-8 levels. It is hypothesized that the cytotoxicity of DA is moderated when reacted with SC and that CD hydrogel inhibits unwanted fibroblastic invasion preventing scarring and adhesions. Together with the previously published human and animal trial data, the results indicate CD hydrogel is biocompatible in the setting of endoscopic sinus surgery. This work represents the first study of CD hydrogel with human cell lines and provides essential information for its future application in biomedicine.

Antioxidant activities, sensory and anti-influenza activity of grape skin tea infusion

Grape skin extracts from pinot noir and pinot gris exhibited significant in vitro antiviral (influenza virus) activity. Five tea infusions from grape skins (Vitis vinifera var. pinot noir and pinot gris) without any additives (control pinot noir and control pinot gris) or by adding variable amounts of green tea and hibiscus were investigated as a means to utilise wine wastes. The antioxidant activities (DPPH scavenging capacity and superoxide anion radical scavenging capacity), total phenolics, the polyphenolics profile and objective colour measurements (CIELab) were determined on freeze-dried water extracts of all five tea infusions, hibiscus and green tea. The colour parameters, L(∗) and a(∗) values, varied widely (P<0.05) for all the infusions as a result of having different levels and variety of pigments. The tea infusions exhibited weak antioxidant activity and the antiviral activity in grape skin appears not related to phenolics contents.

Purification and characterization of a rhamnose-binding chinook salmon roe lectin with antiproliferative activity toward tumor cells and nitric oxide-inducing activity toward murine macrophages.

In this study, a rhamnose-binding lectin from the roe of chinook salmon (Oncorhynchus tshawytscha) was purified and characterized, and its biological activities were examined in several model systems. Chinook salmon roe lectin had a molecular mass of 30 kDa and agglutinated rabbit and bovine erythrocytes. The hemagglutination activity of the lectin was not affected by metal ions. The lectin was stable up to 70 °C and between pH 4 and pH 11. Chinook salmon roe lectin did not exert antifungal activity toward the fungal species tested and did not exhibit mitogenic response toward mouse splenocytes up to a concentration of 5 mg/mL. The lectin had selective antiproliferative activity toward human breast cancer MCF-7 cells and hepatoma Hep G2 cells. It also induced the production of nitric oxide from mouse peritoneal macrophages. This is the first report that demonstrates these biological activities from chinook salmon roe lectin.

A comparison of IgG and IgG1 activity in an early milk concentrate from non-immunised cows and a milk from hyperimmunised animals

Abstract This paper reports on the activity of bovine immunoglobulins IgG and IgG1 directed towards 19 microbial pathogens in an early milk concentrate from non-immunised cows and in a hyperimmune milk using an ELISA technique. Both products contained IgG and IgG1 which bound to all the microbial antigens tested. The early milk concentrate demonstrated a significantly higher IgG and IgG1 antibody titre (P

Production of bioactive peptide hydrolysates from deer, sheep, pig and cattle red blood cell fractions using plant and fungal protease preparations.

Protease preparations from plant (papain and bromelain) and fungal (FP400 and FPII) sources were used to hydrolyze the red blood cell fractions (RBCFs) separated from deer, sheep, pig, and cattle abattoir-sourced blood. After 1, 2, 4 and 24h of hydrolysis, the antioxidant and antibacterial activities of the peptide hydrolysates obtained were investigated. The increase in trichloroacetic acid-soluble peptides over the hydrolysis period was examined using the o-phthaldialdehyde (OPA) assay and the hydrolysis profiles were illustrated using SDS-PAGE. Papain generated RBCF hydrolysates exhibited higher ferric reducing antioxidant power (FRAP) and oxygen radical absorbance capacity (ORAC) compared to those generated with bromelain, FP400 and FPII. At certain concentrations, 24h hydrolysates of RBCF using FP400 and FPII were able to inhibit the growth of Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. The results indicated that the use of proteases from plant or fungal sources can produce animal blood hydrolysates with antioxidant and antimicrobial activities.

In-Depth Characterization of Sheep (Ovis aries) Milk Whey Proteome and Comparison with Cow (Bos taurus)

An in-depth proteomic study of sheep milk whey is reported and compared to the data available in the literature for the cow whey proteome. A combinatorial peptide ligand library kit (ProteoMiner) was used to normalize protein abundance in the sheep whey proteome followed by an in-gel digest of a 1D-PAGE display and an in-solution digestion followed by OFFGEL isoelectric focusing fractionation. The peptide fractions obtained were then analyzed by LC-MS/MS. This enabled identification of 669 proteins in sheep whey that, to our knowledge, is the largest inventory of sheep whey proteins identified to date. A comprehensive list of cow whey proteins currently available in the literature (783 proteins from unique genes) was assembled and compared to the sheep whey proteome data obtained in this study (606 proteins from unique genes). This comparison revealed that while the 233 proteins shared by the two species were significantly enriched for immune and inflammatory responses in gene ontology analysis, proteins only found in sheep whey in this study were identified that take part in both cellular development and immune responses, whereas proteins only found in cow whey in this study were identified to be associated with metabolism and cellular growth.

Production of bioactive peptide hydrolysates from deer, sheep and pig plasma using plant and fungal protease preparations

Plasma separated from deer, sheep and pig blood, obtained from abattoirs, was hydrolysed using protease preparations from plant (papain and bromelain) and fungal (FP400 and FPII) sources. Antioxidant and antimicrobial activities of the peptide hydrolysates obtained after 1, 2, 4 and 24h of hydrolysis, were investigated. The release of trichloroacetic acid-soluble peptides over the hydrolysis period was monitored using the o-phthaldialdehyde (OPA) assay, while the hydrolysis profiles were visualised using SDS-PAGE. The major plasma proteins in the animal plasmas were identified using MALDI-TOF-TOF MS. Hydrolysates of plasma generated with fungal proteases exhibited higher DPPH radical-scavenging, oxygen radical-scavenging capacity (ORAC) and ferric reducing antioxidant power (FRAP) than those generated with plant proteases for all three animal plasmas. No antimicrobial activity was detected in the hydrolysates. The results indicated that proteolytic hydrolysis of animal blood plasmas, using fungal protease preparations in particular, produces hydrolysates with high antioxidant properties.

Bovine Complex Milk Lipid Containing Gangliosides for Prevention of Rotavirus Infection and Diarrhoea in Northern Indian Infants

Rotavirus (RV) is a leading cause of morbidity and mortality in children under 5-yrs, presenting commonly with diarrhoeal symptoms. In a prospective 12-wk double-blind randomised controlled trial we assessed acceptability and efficacy of a high-ganglioside complex milk lipid (CML) for prevention of RV infection in 450 infants, aged 8-24 months, at 3 sites in Northern India. Prevalence of diarrhoea and RV was unseasonably low at baseline (all-cause diarrhoea, ACD, n = 16; RV-diarrhoea, RVD, n = 2; RV infection, RV, n = 20) and throughout the trial, with only 110 total episodes of ACD over 12-wks (CML, n = 62; Control, n = 48) of which 10 were RVD (CML, n = 4; Control, n = 6). Mean duration that RVD persisted was lower in the CML (2.3 ± 0.5 days) group than Control (3.8 ± 1.3 days, P = 0.03), but only 3 of 450 end of trial stool samples were identified as RV (<1%; CML, n = 2; Control, n = 1). This hampered the assessment of efficacy of CML, despite the large a priori determined sample size. During the trial similar numbers of infants reported adverse events (AEs: CML = 41%, Control = 46%), with the majority of events classified as mild and not related to the intervention. In conclusion, further clinical trials aginst a higher background of seasonal prevalence are necessary to assess efficacy of this nutritional intervention to prevent RVD. Importantly, however, high-ganglioside CML was acceptable for long-term consumption in infants aged 8-24 months.ABSTRACTRotavirus (RV) is a leading cause of morbidity and mortality in children younger than 5 years of age, presenting commonly with diarrhoeal symptoms. In a prospective 12-week double-blind randomised controlled trial we assessed acceptability and efficacy of a high-ganglioside complex milk lipid (CML) for prevention of RV infection in 450 infants, ages 8 to 24 months, at 3 sites in northern India. Prevalence of diarrhoea and RV was unseasonably low at baseline (all-cause diarrhoea [ACD], n = 16; RV diarrhoea [RVD], n = 2; RV infection, RV positive [RV+], n = 20) and throughout the trial, with only 110 total episodes of ACD for 12 weeks (CML, n = 62; control, n = 48) of which 10 were RVD (CML, n = 4; control, n = 6). Mean duration that RVD persisted was lower in the CML group (2.3 ± 0.5 days) than that in the control group (3.8 ± 1.3 days, P = 0.03), but only 3 of 450 end of trial stool samples were identified as RV+ (<1%; CML, n = 2; control, n = 1). This hampered the assessment of efficacy of CML, despite the large a priori determined sample size. During the trial similar numbers of infants reported adverse events (AEs: CML 41%, control 46%), with the majority of events classified as mild and not related to the intervention. In conclusion, further clinical trials against a higher background of seasonal prevalence are necessary to assess efficacy of this nutritional intervention to prevent RVD. More important, however, high-ganglioside CML was acceptable for long-term consumption in infants ages 8 to 24 months.