Bovine colostrum whey: Postpartum changes of particle size distribution and immunoglobulin G concentration at different filtration pore sizes.

Bovine colostrum, as vital as it is for calves, is also a valuable source of functional components with rich health benefits for humans. Bovine colostrum whey consists of a large number of bioactive proteins and peptides. The most abundant of these is IgG. Particle size distribution (PSD) is an important feature of many of the processes in the dairy food industries. Despite this, scientific literature on PSD of colostrum whey is scarce. The goal of this research was to describe bovine colostrum whey PSD with an emphasis on postpartum milking time, filtration (pore size 450, 100, and 20 nm), IgG concentration, and lactation number. For this purpose, 4 postpartum milking colostrum samples were sequentially milked from 46 Holstein cows at 12 ± 1 h intervals. Colostrum whey was prepared by renneting and diluted (1:200) for PSD analyses by a Malvern Zetasizer Nano ZS (Malvern Instruments Ltd., Malvern, UK). Immunoglobulin G concentration of these diluted colostrum whey samples were analyzed by an Octet K2 (Molecular Devices LLC, San Jose, CA) system. Linear mixed model analysis revealed significant effects of filter pore size, postpartum milking, and lactation on colostrum whey IgG concentrations. The percentage of particles in the size interval 5 to 15 nm (the hydrodynamic diameter of IgG is around 10 nm) had an intermediate positive correlation (r = 0.50) with IgG concentration. Furthermore, we showed that PSD was associated with IgG concentration, postpartum milking time, and lactation number. The PSD measurement results showed the mean hydrodynamic diameter of 100 nm pore size filtered colostrum whey to be around 10 nm. This, with the IgG concentration results, suggests that even though the size of IgG is around 10 nm, a 100 nm pore size is adequate for membrane-involved IgG separations. In terms of energy efficiency of the filtration process, the use of a larger filter pore size can make a remarkable difference, for example, in pressurizing and cooling costs. Our work contributes to the development of sustainable and widely available colostrum-derived food and feed supplements.

Influence of a cell salvage washing system and leukocyte reduction filtration on bacterial contamination of canine whole blood ex vivo.

OBJECTIVE: To determine the ability of cell salvage washing and leukoreduction filtration to remove bacterial contamination from canine whole blood. STUDY DESIGN: Ex vivo nested cohort study. SAMPLE POPULATION: Commercially purchased fresh canine whole blood (n = 33 units). METHODS: Commercially obtained canine whole blood was inoculated with known concentrations of one of three species of bacteria, Escherichia coli (ATCC 25922), Staphylococcus pseudintermedius (quality control strain; Texas A&M University), or Pseudomonas aeruginosa (ATCC 27853). Negative controls were inoculated with sterile saline. The inoculated blood was processed through a cell salvage system and filtered through a series of two leukocyte reduction filters. Samples were aseptically collected at five points during processing (inoculum, prewash, postwash, post-first filtration, and post-second filtration) for bacterial enumeration. RESULTS: Bacterial concentrations were reduced by 85.2%, 91.5%, and 93.9% for E coli, S pseudintermedius, and P aeruginosa, respectively, after washing (P < .0001), and bacterial concentrations were reduced by 99.9%, 100%, and 100%, respectively, after the first filtration (P < .0001). After the second filtration, none of the three species of bacteria could be isolated (100% reduction). No bacterial growth was obtained from negative controls throughout the study. The type of bacteria (P = .29) did not allow prediction of bacterial reduction. CONCLUSION: Cell salvage washing combined with leukoreduction filtration eliminated bacterial contamination of whole dog blood (P < .0001). CLINICAL SIGNIFICANCE: Cell salvage washing and leukoreduction filtration could be applied to intraoperative autotransfusion in clinical animals, especially those treated for trauma or hemorrhage with concurrent bacterial contamination.

Evaluation of a filter system to harvest plasma for identification of failure of passive transfer in newborn calves.

The objective of this study was to evaluate a filter system to harvest plasma to assess failure of passive transfer (FPT) in newborn calves. Blood samples (n = 227) for serum and plasma harvesting were collected via jugular vein puncture from Holstein calves aged 1 to 7 d from 4 commercial dairy herds in Northeast Germany. Serum IgG concentrations were determined using a sandwich ELISA. Failure of passive transfer was defined as IgG concentrations <10 mg/mL and used as a gold standard. One handheld optical refractometer (Euromex Holland, Arnhem, the Netherlands) and 2 digital Brix refractometers (device 1: HI 96801 digital refractometer, Hanna Instruments, Woonsocket, RI; device 2: Misco PA201, Misco, Solon, OH) were used to analyze total proteins in serum or plasma. The colostrum uptake of the calf can thus be monitored and calves with FPT can be identified. Serum was obtained through centrifugation. Plasma was obtained through either a filter system or centrifugation. For plasma filtration, approximately 2 mL of lithium heparin blood was injected into the inlet reservoir of a plasma filter (2-Drop-Filter, Pharmadoc, Lübeck, Germany) using a disposable syringe. Receiver operating characteristic curve analyses were used to determine optimum thresholds for each of the 3 devices using different media. Sixty-seven (30%) calves had FPT. For the handheld optical refractometer, the optimum threshold was 5.6 g/dL [sensitivity 70.1%; specificity 80.0%; positive predictive value (PPV) 60.1%; negative predictive value (NPV) 86.2%; area under the curve (AUC) 0.85] using serum. For centrifuged plasma, the optimum threshold was 6.3 g/dL (sensitivity 82.1%; specificity 68.1%; PPV 52.5%; NPV 89.9%; AUC 0.84), and for filtered plasma, the threshold was 6.0 g/dL (sensitivity 56.7%; specificity 90.0%; PPV 70.9%; NPV 82.9%; AUC 0.80). For device 1, the optimum threshold was 8.9% Brix (sensitivity 82.1%; specificity 63.8%; PPV 48.7%; NPV 89.5%; AUC 0.81), 9.4% Brix (sensitivity 76.1%; specificity 73.7%; PPV 55.4%; NPV 87.8%; AUC 0.80), using serum and centrifuged plasma, respectively. For device 2, the optimum threshold was 8.7% Brix (sensitivity 74.6%; specificity 76.2%; PPV 57.4%; NPV 87.5%; AUC 0.83), 9.5% Brix (sensitivity 80.6%; specificity 70.6%; PPV 54.0%; NPV 89.5%; AUC 0.83), and 9.2% Brix (sensitivity 58.2%; specificity 87.5%; PPV 66.6%; NPV 83.0%; AUC 0.80) using serum, centrifuged plasma, and filtered plasma, respectively. Based on the AUC, the 3 devices yielded comparable test characteristics to identify calves with FPT. In conclusion, a filter system can be used to facilitate the evaluation of FPT as a point of care technique in calves without the need for serum centrifugation.

An improved method for a rapid determination of phytase activity in animal feed.

The current direct colorimetric assay for phytase activity in feeds has interference from high P background and other factors. Our objective was to develop a rapid and reliable spin column method to accurately determine phytase activity in feed ingredients or complete diets. After the feed sample was extracted by stirring in 0.2 M citrate buffer, pH 5.5, for 30 min at room temperature, the oily layer of the supernatant fraction was removed by passing through an acrodisc syringe filter (0.45-microm HT Tuffryn membrane, Gelman Laboratory, Ann Arbor, MI). The filtrate was then loaded onto a spin column (MW cutoff 30,000, Millipore, Bedford, MA) to remove free phosphate before the phytase activity assay. Compared with the direct assay, this new procedure improved both accuracy and reproducibility. When diets contained phytase at 0 to 1,500 U/kg (as fed), the CV for multiple assays of the same samples (n = 6) by the new method ranged from 1 to 6% compared with 28 to 39% by the direct method. A linear relationship was found between the added phytase activity in practical diets and the analyzed activity by the new method (r2 = 0.99; P < 0.01). In conclusion, the spin column method is an improved assay for phytase activity in animal feed, and may be used for quality control of phytase supplementation.