Porcine Blood Plasma Research Articles

In-house validation and factorial effect analysis of a LC-MS/MS method for the determination of steroids in bovine and porcine blood plasma

Anabolic steroids are banned from use in food-producing animals in the European Union (Council Directive 96/22/EC). To control the zero-tolerance concept, an LC-MS/MS method for the screening and confirmation of most of the relevant natural and synthetic estrogenic and androgenic steroids in bovine and porcine blood plasma was developed and validated. The method permits confirmation and quantification of all analytes above a concentration of 0.65 µg L–1. The validation was carried out according to Commission Decision 2002/657/EC, Chapter 3.1.3 “Alternative Validation”, by applying a matrix-comprehensive in-house validation concept. Decision limit CCα, detection capability CCβ, recovery, repeatability, within-laboratory reproducibility and the uncertainty of measurement were calculated. Furthermore, a factorial effect analysis was carried out to identify factors that have a significant influence on the method. Factors considered to be relevant for the method in routine analysis (e.g., operator, storage duration of the extracts before measurement and different cartridge lots) were systematically varied on two levels.

Changes in plasma orexin A and orexin B concentrations during the estrous cycle of the pig

Orexin A (OXA) and orexin B (OXB) are neuropeptides synthesized mainly in the lateral hypothalamus, which are involved in the control of various physiological functions such as energy homeostasis, sleep, wakefulness and feeding behavior. The present study analyzes orexins A and B levels in the porcine plasma during the estrous cycle. The highest plasma concentrations of orexin A were observed on days 2–3 of the estrous cycle (p<0.05 relative to days 10–12 and 14–16) and the lowest (p<0.05) on days 14–16. The highest orexin B levels in the blood plasma were noted on days 17–19 (p<0.05 vs. days 14–16). We demonstrated the presence of OXA and OXB in porcine blood plasma and the impact of the phase of the estrous cycle on the observed changes in plasma orexin levels.

Porcine plasma as polyphosphate and caseinate replacer in frankfurters

The aim was to replace polyphosphate and caseinate by porcine blood plasma as functional ingredients in frankfurters. Three trials, each consisting of one control, formulated with caseinate and tripolyphosphate, and one test, formulated with plasma, were carried out in a pilot plant. The frankfurters with plasma were compared to their respective controls by determining the composition, water holding capacity, cooking losses, internal colour, texture, microstructure, sensorial characteristics, and overall acceptance. No significant differences were found in proximate analysis, WHC, and cooking losses. Texture was not affected by the replacement, according to both sensorial and instrumental measurements. Nevertheless, the panellists detected the presence of animal taste and odour in plasma-containing sausages. Despite this, their overall acceptance was scored as 6.3 in a 10 maximum scale, so plasma could be considered as an interesting alternative to produce healthier and cheaper frankfurters.

Chemical composition and biological value of spray dried porcine blood by-products and bone protein hydrolysate for young chickens

1. The chemical composition of spray dried porcine blood by-products is characterised by wide variation in crude protein contents. In spray dried porcine blood plasma (SDBP) it varied between 670–780 g/kg, in spray dried blood cells (SDBC) between 830–930 g/kg, and in bone protein hydrolysate (BPH) in a range of 740–780 g/kg. Compared with fish meal, these feeds are poor in Met and Lys. Moreover, in BPH deep deficits of Met, Cys, Thr and other amino acids were found. 2. The experiment comprised 7 dietary treatments: SDBP, SDBC, and BPH, each at an inclusion rate of 20 or 40 g/kg diet, plus a control. The addition of 20 or 40 g/kg of the analysed meals into feeds for very young chickens (1–28 d post hatch) significantly decreased the body weight (BW) of birds. Only the treatments with 40 g/kg of SDBP and SDBC showed no significant difference in BW as compared with the control. There were no significant differences between treatments and type of meal for feed intake, haematocrit and haemoglobin concentrations in blood. 3. Addition of bone protein and blood cell meals to feed decreased the IgG concentration in blood and caused shortening of the femur and tibia bones. However, changes in the mineral composition of bones were not significantly affected by the type of meal used. 4. The blood by-products, which are rich in microelements, improved retention of Ca and Cu only. In comparison to control chickens, significantly better accretion of these minerals was found in treatments containing 20 g/kg of SDBP or 40 g/kg of SDBC. 5. Great variability in apparent ileal amino acid digestibility in chickens was determined. In this respect, some significant differences related to the type of meal fed were confirmed for Asp, Pro, Val, Tyr and His. In general, the apparent ileal digestibility of amino acids was about 2–3 percentage units better in chickens fed on diets containing the animal by products than in control birds.

Use of spray‐dried porcine blood by‐products in diets for young chickens

Spray-dried porcine blood plasma (SDBP) or blood cells (SDBC) at amounts of 20 or 40 g/kg were included to the feed mixtures that were given to young chickens within 1-28 (Exp. 1) or 1-30 (Exp. 2) days post-hatch. In comparison with the group fed mixtures containing plant components, chickens fed mixtures supplemented with 40 g/kg of SDBP significantly (p < 0.01) increased the body weight estimated on 14 day of life (Exp. 1). At the age of 28 or 30 days post-hatch, the body weight was improved significantly (p < 0.01 or 0.05) in both experiments. Significant differences (one-factorial anova) in feed conversion among particular feeding groups were stated in Exp. 1 only; however, calculations using two-factorial anova show insignificant differences depending on the used animal meal. In selected blood parameters (IgG, Ht, Hb), insignificant differences between feeding groups were stated. The use of SDBP in feed mixture significantly increased the Na retention in both experiments, and K accretion in Exp. 1 only. Application of SDBC and 40 g/kg of SDBP significantly or insignificantly improved Fe retention. Insignificant diversification of apparent ileal digestibility of nutrients was stated; the crude fat was significantly better digested in treatments fed mixtures with animal meals but kind of animal meal was without any significant effect. Significant differences in digestibility of amino acids were recorded for Pro, Cys, Val, His, Lys and Arg. In chickens fed mixture with SDBC, higher coefficients of apparent digestibility of Cys, Val and His (Exp. 1) and Cys and His (Exp. 2) than in other feeding groups were obtained. The kind of used blood by-products has not affected the histological structure of intestine wall.

Isolation of a Calcium-binding Peptide from Enzymatic Hydrolysates of Porcine Blood Plasma Protein

A calcium-binding peptide was purified from hydrolysates of porcine plasma protein. Porcine plasma protein was hydrolyzed using Flavourzyme, and the hydrolysates were filtered using YM-3 membrane. The membrane filtered solution was fractionated using Sephadex G-15, normal phase high-performance liquid, ion exchange chromatography, and fast protein liquid chromatography for a calcium-binding peptide. The sequence of the purified calcium-binding peptide was analyzed using liquid chromatography/electron spray ionization tandem mass spectrum and identified to be Val-Ser-Gly-Val-Glu-Asp-Val-Asn. The calcium- binding peptide can be used as a food ingredient or feed for calcium deficiency.

Antioxidant activity and functional properties of porcine plasma protein hydrolysate as influenced by the degree of hydrolysis

Antioxidant activity and functional properties of porcine blood plasma protein hydrolysates (PPH) prepared with Alcalase at 6.2%, 12.7% and 17.6% of degree of hydrolysis (DH) were investigated. The PPH showed stronger radical-scavenging ability and possessed stronger Cu2+-chelation ability and a reducing power compared to non-hydrolysed plasma protein (P < 0.05). The antioxidant activity of PPH, indicated by thiobarbituric acid-reactive substance (TBARS) values in a liposome-oxidising system, increased with increasing DH (P < 0.05). The Alcalase hydrolysis increased protein solubility from its original 68.46–81.79% (non-hydrolysed) to 82.95–94.94% (hydrolysed) over a broad pH range (3.0–8.0). However, hydrolysis decreased surface hydrophobicity and suppressed emulsifying and foaming capacity of the plasma protein. To identify antioxidant peptide, PPH was subjected to ultrafiltration, ion-exchange chromatography and reverse-phase high performance liquid chromatography (RP-HPLC), and the amino acid sequences of isolated peptides were determined by liquid chromatography/tendem mass spectrometry (LC–MS/MS). The peptide with the strongest antioxidant activity had the amino acid sequence of His-Asn-Gly-Asn. The results indicated that PPH could be used as a novel antioxidant but may be of limited utility as an emulsifying or foaming agent.

Purification of an iron-binding nona-peptide from hydrolysates of porcine blood plasma protein

To isolate a novel iron-binding peptide, porcine plasma protein (PPP) was hydrolyzed using a commercial protease. The degree of hydrolysis and iron-binding capacity was determined using the trinitrobenzenesulfonic acid and orthophenanthroline method, respectively. The hydrolysates of blood plasma protein were then filtered using YM-3 membrane, and an iron-binding peptide was isolated using gel permeation, ion exchange, and normal phase high-performance liquid chromatography. The purified iron-binding peptide was identified to be a nona-peptide, Asp–Leu–Gly–Glu–Gln–Tyr–Phe–Lys–Gly (1055 Da) based on liquid chromatography/electrospray ionization (LC/ESI) tandem mass spectrum and a sequence of porcine plasma protein.

In vitro study of drug-eluting stent coatings based on poly(l-lactide) incorporating cyclosporine A – drug release, polymer degradation and mechanical integrity

In this study, absorbable polymer stent coatings for localized drug delivery based on poly(L-lactide) (PLLA) and cyclosporine A (CsA) were developed and tested in vitro. Metallic stents were coated with different compositions of PLLA/CsA (70/30, 60/40, 50/50% w/w) and beta-sterilized. The specimens were used to assess the drug release kinetics with HPLC. Sterilization influenced polymer degradation was measured with GPC. Mechanical integrity of the stent coatings was studied with SEM. The interconnection of the coated stents with a balloon-catheter was characterized by the measurement of stent dislodgment force. A migration assay was used to determine the inhibitory effect of the model drug CsA on smooth muscle cell (SMC) migration. The release of CsA was established over time periods up to 24 days in sodium chloride solution and in porcine blood plasma. An inhibition of SMC migration (max. 26-33%) was found for CsA concentrations of 4 x 10(-5) to 4 x 10(-7) mol/l. Marked molecular weight reduction (70-80%) of the PLLA matrix occurred after beta-sterilization. We also observed a substantial decrease of in vitro degradation time. The maintenance of the mechanical integrity of the polymer coating during crimping and dilation of the specimens could be verified, and a sufficient stent dislodgment force of 0.8-0.9 N was measured.

Heat-induced gelation of porcine blood plasma proteins as affected by pH

Porcine plasma is a by-product of the meat industry that can be used as a food ingredient. It is a protein mixture, hence its composition can be modified to meet specific functionality requirements. In the present paper, the gelation properties of plasma and its two major fractions (serum and albumin) have been studied at pH 4.5, 6.0 and 7.5. Polyacrylamide gel electrophoresis (SDS–PAGE) revealed that albumin was the constituent that remained soluble to a larger extent during heat-treatments, and that acidic coagulation occurred at pH 4.5, making weak interactions the predominating ones between protein aggregates. Differential scanning calorimetry (DSC) and rheological tests showed that both the thermal stability and the gelation point of protein solutions were lower as pH decreased. The textural properties and water-holding capacities of plasma and albumin gels were more pH-dependent than serum. Albumin gels were the weakest and those of plasma at pH 7.5, the strongest. It has been determined that interactions between protein fractions play a key role in the gelling properties due to synergistic effects. This knowledge should be useful in the engineering of a plasma derivative product designed for specific food requirements, by reformulating its natural composition and enhanced by controlling pH.

Surface functional properties of blood plasma protein fractions

The solubility, foaming and emulsifying properties of porcine blood plasma and its main protein fractions (serum, globulins and albumin) were investigated at pH 4.5, 6.0 and 7.5 in order to clarify the contribution of each fraction and encourage the optimisation of plasma-derived products. Soluble protein contents above 85% were obtained in all samples. Plasma, serum and albumin showed good foaming capacities, reasonably similar at different pH conditions, although the highest foam stability corresponded to both albumin and plasma at pH 4.5 and 6.0. All protein fractions showed good emulsifying activities, but the stability of the formed emulsions decreased with acidification, being emulsions of albumin and globulins at pH 7.5 the most stable ones. In addition, the interaction indexes calculated to investigate protein–protein interactions revealed synergistic interactions between albumin and globulins when in co-occurrence in their foaming capacity at pH 6.0 and 7.5, and in the stability of emulsions at pH 4.5 and 6.0, but slightly negative effects in the solubility of the mixture, and a great decrease in the stability of emulsions at pH 7.5. On the other hand, the elimination of fibrinogen improved the stability of emulsions and foams at acidic conditions.

Porcine Blood Plasma Transgluataminase Combined with Thrombin and Fibrinogen as a Binder in Restructured Meat

The purpose of this study was to use pig blood plasma transglutaminase (TGase) combined with thrombin and fibrinogen as a binder, which was applied to restructured meat, and to investigate its effect on the restructured meat quality. Pig meat was obtained 10 h post mortem from a traditional market was ground using a 10 mm aperture plate. A binder admixture was added (TGase:thrombin:fibrinogen mixed as 0.5:1:20 (v/v/v) to which was added 12% of its volume of 0.25 M calcium chloride) atO, 5, 10, 15 and 20% of meat weight. Measurements included cooking loss, shrinkage rate, shear value, total plate count, pH value, TBA value, color difference, tension strength and sensory evaluation. The results showed that ground meat containing 20% w/w of binder admixture had higher cooking loss, shrinkage rate and shear value (p 0.05). Tension strength was increased with increased level of binder admixture. Addition up to 15% binder admixture to restructured meat showed better scores of sensory texture, flavor and total acceptability (p<0.05).

Improvement of gelling properties of porcine blood plasma using microbial transglutaminase

The effect of microbial transglutaminase ( MTGase) on the texture and water-holding capacity (WHC) of heat-induced gels prepared from porcine blood plasma at pH 5.5 was investigated. Different concentrations of commercial MTGase were added to plasma and incubated for several times under specific conditions of temperature and pH. From the results obtained, it can be postulated that enzymatic treatment enhances textural properties and WHC of plasma gels at pH 5.5, especially when incubated with 3% of the commercial product for 3 h at 30 °C and pH 7. This treatment increased by 0.4 N the hardness of gels and reduced by 3% the water released after gel centrifugation with respect to the control samples. SDS–PAGE confirmed that cross-linking took place when MTGase was added to plasma solutions. However, the results suggest that the sole addition of MTGase was not effective enough to improve the gelling properties of plasma proteins under acidic conditions.

Study on a Binder by Using Porcine Blood Plasma Transglutaminase, Thrombin and Fibrinogen

The purpose of this study was to prepare a binder containing porcine blood transglutaminase (TGase), thrombin and fibrinogen. Extracted TGase, thrombin and fibrinogen were used alone or mixed with different proportions of their volume (v/v/v) by nine combinations as follows were 0.5:1:15, 0.5:1:20, 0.5:1:25, 1:1:15, 1:1:20, 1:1:25, 1.5:1:15, 1.5:1:20 and 1.5:1:25, respectively. Five ml of each combination were mixed with 0.6 ml of 0.25 M calcium chloride before experiment. After storage at 4C for 0, 1, 2, 3, 4 and 5 weeks, enzyme activity, total plate count, pH value, and SDS-PAGE of TGase, thrombin and fibrinogen were tested and pH value, clotting time and gel strength of the nine combination binders were determined. The results showed that total plate count of thrombin and pH value of TGase were significantly higher (p<0.05) than in other treatments. SDS-PAGE results showed that purified TGase, thrombin and fibrinogen from porcine blood plasma compared with commercial products (Sigma) had the same band patterns and nine different combination binders had no significant effect. Enzymatic activity of TGase and thrombin decreased as storage time increased. Total plate count of TGase, thrombin and fibrinogen and clotting time of the binder increased as storage time increased. The higher amount of fibrinogen in combinations, the stronger the gel strength. (Asian-Aust. J. Anim. Sci. 2006. Vol 19, No. 1 : 137-143)

Effect of gamma-irradiation on the physicochemical properties of porcine and bovine blood plasma proteins

To elucidate the effect of γ-irradiation on the physicochemical properties of blood plasma proteins, bovine and porcine blood, released from a slaughterhouse, was collected and plasma proteins were prepared. Physicochemical properties of blood plasma protein powders and solutions, such as molecular weight distribution, secondary structure, solubility, and viscosity, were examined after γ-irradiation at 1, 5, 7, and 10 kGy. Oxygen radicals, such as hydroxyl and superoxide anion radicals, generated by γ-irradiation, affected the physicochemical properties of the blood plasma protein solutions, but not the plasma protein powders. Circular dichroism and sodium dodecyl sulphate–polyacrylamide gel electrophoresis studies showed that an increase of γ-irradiation decreased the ordered structure of plasma protein solutions and caused initial fragmentation of the polypeptide chains and subsequent aggregation. However, solubility and viscosity of the irradiated plasma protein powders, as well as secondary structure and molecular weight profile, were not changed significantly with radiation dose.

Emulsifying and gelling properties of porcine blood plasma as influenced by high-pressure processing

Abstract The emulsifying properties and the characteristics of heat-induced gels (texture and water-holding capacity) prepared from pressurised blood plasma solutions of different pH (5.5–7.5) were investigated. Changes in the plasma, that affected its behaviour as an emulsifier, occurred after pressurisation. The highest emulsifying activity was found for samples treated at 400 MPa. At pressures above 400 MPa, the emulsifying activity and stability, at all pHs, decreased with increasing pressure. The hardness of heat-induced gels decreased significantly as the pressure increased above 400 MPa, this effect being more noticeable with decreasing pH. However, although a 600 MPa pressurisation induced a further decrease in the firmness of gels from solutions at pH 6.5 and 7.5, it increased the hardness of gels at pH 5.5. For treatments up to 500 MPa, the highest elasticity was found in gels from solutions at pH 7.5. Pressure treatments above 400 MPa improve the water-holding capacity of heat-induced gels prepared from plasma solutions at pH ⩾6.5.

High Hydrostatic Pressure as a Method to Reduce Microbial Contamination of Porcine Blood Plasma

The suitability of high hydrostatic pressure as an alternative method to produce porcine blood plasma with a sufficient microbial stability without affecting its functional properties was evaluated. The effects of high pressure on plasma microorganisms were highly dependent on processing temperature. Treatments of 15 min at 450 MPa carried out at 5 'C led to reductions of about 90% in microbial counts and to 20-50% decreases in the growth ability of the survivors. At 25 'C and 40 'C, the efficiency was increased to reduction values of 99.82 and 99.97%, respectively. The lowering of the growth capacity was about 50% at 25 'C and up to 80% at 40 'C. The most efficient treatment (450 MPa, for 15 min at 40 'C) did not lead to appreciable negative effects on the functional properties of heat-induced gels from porcine blood plasma.

High Hydrostatic Pressure as a Method to Reduce Microbial Contamination of Porcine Blood Plasma

High Hydrostatic Pressure as a Method to Reduce Microbial Contamination of Porcine Blood Plasma

High Hydrostatic Pressure as a Method to Reduce Microbial Contamination of Porcine Blood Plasma

High Hydrostatic Pressure as a Method to Reduce Microbial Contamination of Porcine Blood Plasma

Purification and Characterization ofBeauveria bassiana Proteinases

Two chymotrypsin-like serine proteinases are produced by B. bassiana 278 when grown on different carbon and nitrogen sources. By employing acetone precipitation, gel filtration and ion-exchange chromatographies, the enzymes were separated from the culture filtrate after propagation of the fungus on medium enriched either with ground larvae of Apis mellifera (Proteinase I) or porcine blood plasma (Proteinase II). The purified enzymes with a molecular mass of approximately 32 kDa hydrolyzed natural protein substrates: casein, hide powder azure (HPA), azocoll and much less elastin Congo Red and collagen. They differ from each other in the optimum pH value, amino acid composition, MICHAELIS constant and susceptibility to natural chymotrypsin inhibitors. Both proteinases hydrolyze suc-Ala-Ala-Pro-Phe-p-NA with an apparent K m of 2.03 × 10 -3 M and 1.04 × 10 -4 M. respectively. The turkey ovomukoid (OMTKY) and cathepsin G/chymotrypsin inhibitor inhibit only Proteinase II from the larvae hemolymph of Apis mellifera (AMCI). The association constant of the interaction of this enzyme with AMCI was estimated to be very high (4.1 1 × 10 9 M -1 ).