Protein Pellet Research Articles

Development of third generation protein rich snacks from lentil and egg powder through microwave assisted extrusion cooking

PurposeThe purpose of this study was to formulate a complete protein food from lentil flour (LF) and egg powder (EP) through microwave-assisted extrusion technology.Design/methodology/approachIn the first part of the hybrid technology, the feed proportion and extrusion conditions were optimized through design expert using central composite rotatable design. In the second part of hybrid technology, the optimized protein pellets (PP) obtained were subjected to microwave heating (MH) for 50,100, 150, 200 and 250 s.FindingsThe optimum predicted conditions for development of pellets using extrusion cooking were feed proportion (85% LF and 15% EP), barrel temperature (140°C), screw speed (340 rpm) and feed moisture content (12%). When these pellets were subjected to MH, 150 s of heating time was considered as prudential to induce desirable quality changes in PP. The increase in sectional expansion index, crispness and overall acceptability from 0.637 to 0.659, 4.51 to 6.1 and 3.27 to 3.59 with corresponding decrease in bulk density and breaking strength from 73.33 to 69.75 kg/cm3 and 6.24 to 5.13 N during 150 s of MH indicated that quality characteristics of extruded PPs were improved after MH.Practical implicationsNowadays, consumers have become more health conscious than ever, and the demand for nutritious snacks has increased many folds. However, the high protein content restricts expansion of snacks, which was overcome by subjecting extruded pellets to MH to produce third generation pellets. Furthermore, the PP has a protein content of 31.62%, which indicates that if an average person consumes 100 g of these snacks, it will suffice 60% of total recommended dietary intake (0.75 g/kg body weight/day). Lentil-based pellets expanded by use of such hybrid technology (microwave-assisted extrusion cooking) can help to provide a feasible, low cost and protein-rich diet for malnourished population besides being a value addition to lentils.Originality/valueLF in combination with EP was tested for the first time for development of nutrient dense pellets. Moreover, use of microwave-assisted extrusion cooking offers a workable and innovative technique of developing protein-rich pellets with improved physico-chemical and sensory attributes.

Malaria Box Compounds against Anopheles gambiae (Diptera: Culicidae) Carboxypeptidase B Activity to Block Malaria Transmission.

Carboxypeptidase B (CPB) plays an important role in blood digestion in mosquitos, aiding the release of free amino acids. Anopheles CPB is a target to block malaria transmission because it facilitates Plasmodium invasion of the mosquito midgut. Our study aimed to discover inhibitors of Anopheles CPB to prevent Plasmodium development in the mosquito. The Anopheles gambiae cpb (Agcpb) gene without a signal sequence was cloned into the pET28b expression vector. The recombinant AgCPB protein was expressed in E. coli BL21(DE3) within inclusion bodies after induction with 0.5 mM isopropyl β-D-1-thiogalactopyranoside at 37°C for 4 h. The protein pellet was dissolved in 6 M urea, purified by affinity chromatography, and dialyzed in reaction buffer. The refolded recombinant AgCPB could digest the hippuryl-arginine substrate similarly to that of the commercial porcine pancreas CPB. The 20 top-scoring malaria box compounds from the virtual-screening results were then chosen for an in vitro inhibition assay against AgCPB. Four of the 20 malaria box compounds could inhibit AgCPB activity. The compound MMV007591 was the most potent inhibitor with an IC50 at 0.066 µM. The results indicate that these candidate compounds may be utilized in drug development against mosquito CPB activity to curb malaria transmission.

Lignosulfonate properties and reaction conditions enhance precipitation and affect ensuing quality of proteins from green biomass juice for monogastric animal feed

This is the first study investigating the effects of lignosulfonate (LS) properties and reaction conditions on the precipitation of proteins from green biomass juice for producing monogastric animal feed. Both the improvement in the protein precipitation yield and the resulting impact on the quality of the protein pellets were investigated with new methods and approaches. High molecular weight (MW) LS gave the highest precipitation yield, 21–33% higher than traditional heat and isoelectric methods. Electrostatic interaction between LS and soluble proteins was observed to be higher in high MW LS than others, and likely improved the precipitation. A response surface design experiment showed that pH was more important than LS concentration for precipitation. The optimum conditions predicted by the model improved precipitation by up to 61% compared to traditional methods. Using high MW LS at near optimum conditions resulted in more LS being retained in the pellet, with a final LS content of up to 60 g/kg dry matter (DM). The improved precipitation yield came at the expense of lower crude protein (CP) content of the pellet. Despite reduced pellet protein purity, the total CP recovery from the juice was improved by up to 800 g/kg DM. The in vitro protein digestibility (IVPD) and amino acid composition were not significantly affected by the presence of LS. Using high MW LS at optimum reaction conditions can significantly improve protein precipitation yield from green biomass juice without compromising digestibility. However, finding the compromise between yield and protein purity is necessary to improve feasibility.

A Simplified Method to Assay Protein Carbonylation by Spectrophotometry.

Protein carbonylation is an irreversible oxidation process leading to a loss of function of carbonylated proteins. Carbonylation is largely considered as a hallmark of oxidative stress, the level of protein carbonylation being an indicator of the oxidative cellular status. The method described herein represents an adaptation to the commonly used 2,4-dinitrophenylhydrazine (DNPH)-based spectrophotometric method to monitor protein carbonylation level. The classical final sample precipitation was replaced by a gel filtration step avoiding the tedious and repetitive washings of the protein pellet to remove free DNPH while allowing optimal protein recovery.This improved protocol here implemented to assay protein carbonylation in plant leaves can potentially be used with any cellular extract.

A water-only process to fractionate yellow peas into its constituents

To meet the consumer demand for minimally processed foods and clean labels, the potential of processes where chemicals are omitted and only water is used needs to be explored. Mild wet fractionation of yellow pea, a water-only process, is investigated on maximum separation and efficient water use. By only using water, starch and protein from pea could be successfully separated, resulting in fractions high in yield and purity. Multiple washing steps of both the starch and the non-soluble protein fraction were performed to enhance separation. As a result of starch- and non-soluble protein pellet washing, the starch fraction was further depleted in protein and the protein solubility in the non-soluble protein fraction decreased. Ultrafiltration of the soluble protein fraction served to concentrate and the extracted water has potential to be reused in the process. Small solutes were concurrently extracted, which resulted in a higher protein purity in the soluble protein concentrate of 75%. The presented method has potential for upscale use in industry to produce protein fractions comparable to protein isolates obtained through conventional fractionation.

Body Condition Score, Rumination, Intake, Milk Production and Milk Composition of Grazing Dairy Cows Supplemented with Rumen-Protected Lysine and Methionine

The study utilised a pasture grazing based, voluntary traffic automatic milking system to investigate milk production of cows fed a pasture-based diet and supplemented with a pellet formulated with vs. without rumen-protected lysine and methionine (RPLM). The study adopted a switch-over design (over two periods of 5 and 10 weeks, respectively) and used 36 cows and equally allocated them into two experimental groups. The RPLM (Trial) pellet had 2% lower crude protein, but similar metabolizable energy content compared to the Control pellet. Pellet intake was 10.0 and 9.4 kg/day/cow. Milk yield was 36.2 and 34.4 kg/day/cow (p = 0.23), and energy corrected milk was 35.1 and 33.8 kg/day/cow (p = 0.076), and milk solids was 2.55 and 2.46 kg/cow/day (p = 0.073) in the Control and Trial groups, respectively. Milk fat%, milk protein%, milk fat: protein ratio, milking frequency and rumination time were not different between the two groups (p > 0.05). In period 1, plasma glucose was 3.1 mmol/L for both groups and milk urea were 150 and 127 mg/L in the Control and Trial groups, respectively. Both plasma glucose (as a proxy for energy supply) and milk urea (as a proxy for nitrogen use efficiency; NUE) were not different between groups (p > 0.05). This study showed that under a grazing pasture system, feeding lactating dairy cows a low protein pellet with RPLM supplementation, maintained milk production performance and NUE, compared with cows fed a high protein Control pellet diet with no RPLM. Further research should assess the long-term (seasonal) effects of feeding a diet formulated with RPLM on cow intake, health and reproductive performance.

Proteomic and Metabolomic Analyses of Maggots in Porcine Corpses for Post-Mortem Interval Estimation

Post-mortem interval (PMI) estimation is a critical task in forensic science. In this study, we used maggots collected from pig carcasses and applied an integrated proteomics and metabolomics approach to determine potential candidate substances for the estimation of PMI. After methanol precipitation, the supernatant containing metabolites and the protein pellet were separated and subjected to metabolomic and proteomic analyses using liquid chromatography-tandem mass spectrometry (LC-MS/MS). MS/MS data were analyzed for identification and quantification using Proteome Discoverer and Compound Discoverer software. A total of 573 metabolites and more than 800 porcine proteins were identified in maggots. This is the first dataset of proteins and metabolites in maggots collected from porcine carcasses. In this study, guanosine monophosphate, xanthine, inosine, adenosine, and guanine were detected with a similar tendency to increase during early days of maggot development and then decreased gradually. We broadly profiled various biomolecules through analysis in the spot of incident. Especially, we confirmed that proteome and metabolome profiling could be performed directly and indirectly.

51 Observing the Effect of Creep Feeding on Calf Behavior Before and After Weaning

Abstract The objective was to determine the effect of creep feeding on calf behavior before and after weaning. Cow-calf pairs (n = 54) were stratified by cow age, cow and calf BW, and allotted to 9 pastures. Pastures were randomly assigned to 3 treatments: unsupplemented control (CON), lower protein pellet (14.3% CP; LP), and a higher protein pellet (18.6% CP; HP) for 82 d. Pre-weaning calf behaviors were observed for 9 h on d 60, 64, and 74: position (standing or lying), location (pen, pasture, shade, creep), and consumption (creep, grass, milk, water, none). Post-weaning calf behaviors were observed for 12 h on d 83 and 84: position, walking, drinking, eating, and vocalizations. Prior to weaning, an interaction (P £ 0.01) occurred for consumption behaviors; CON calves spent the greatest percentage of time grazing at h 7 and 8 and not consuming at h 11. Also, LP calves had the greatest percentage of time nursing at h 8 and 11, with CON calves being intermediate, and HP calves being the least. On the day after weaning (d 83), an interaction (P = 0.02) was observed for time spent eating; CON calves spent the greatest time eating at h 10 and LP calves were the greatest at h 11. An interaction (P < 0.01) was observed for time spent walking on d 83; HP calves spent the greatest percentage of time walking at h 9 and 13. On d 84, an interaction (P < 0.01) was observed for time eating on d 84; CON calves spent the greatest percentage of time eating at h 7, 9, 11, 14, and 16 as well as overall (P = 0.05). An interaction (P < 0.01) was observed for number of vocalizations on d 84; CON calves had the greatest number of vocalizations at h 16–18 as well as overall (P < 0.01). Overall, creep feeding altered calf behaviors before and after weaning.

98 Growth and mineral content of tall fescue grown in manure extracts from horses fed varying trace mineral sources and levels

Horse manure spread on pasture can provide nutrients and minerals to plants, but the effect of horse diet on fecal mineral availability to plants is unknown. This study's objective was to determine if the amount and source of trace minerals (TM) fed to horses would affect the growth and mineral content of plants grown hydroponically in manure extracts. Nine horses were used in a replicated 3 × 3 Latin square design for the study's feeding portion. Horses had ad libitum access to forage and were fed a protein pellet containing one of 3 dietary treatments: a low TM treatment with no added TM (CON) or industry relevant TM level treatments with TM added as either organic TM (ORG) or inorganic TM (ING). Added TM were Co, Cu, Mn, Se, and Zn at equivalent amounts for ORG and ING. There were 3 21-d feeding periods, including a 5-d washout; each horse received each treatment. Feces were collected on d 21 of each period. Manure extracts were made from each horse by mixing wet feces with distilled water (30 g DM/L), incubating for 20 h, and straining. Extracts were combined by treatment and TM concentrations reflected dietary levels, with ING and ORG being similar and greater than CON. Manure extracts, distilled water/negative control (WAT) and Steinman's nutrient solution/positive control (NUT) were applied as liquid treatments. Tall fescue seeds were germinated in a growth chamber, and seedlings transferred to rockwool in pans (40 seedlings/pan) on d 9. Pans were randomly assigned to treatment with 4 replicate pans/treatment. Plants were grown in an environmentally controlled room, and watered with the liquid treatment to keep fluid amount consistent in the pans. Plant height was recorded every 10 d. At the experiment's end (d 41), plants were harvested to determine plant DM biomass and TM content using ICP-MS. Biomass, final height and TM data were analyzed using ANOVA and growth rate data were analyzed using regression and orthogonal contrasts. The NUT and ORG plants had greater final DM biomass and height compared with WAT plants, while ING and CON were intermediate ( P < 0.05). Growth was quadratic for WAT, with a plateau in growth at d 25. Growth in the remaining treatments was linear and rate was greatest for NUT, followed by ORG, lowest in CON, and ING did not differ from ORG and CON ( P < 0.05). Cu and Mn was greatest in NUT ( P < 0.05). Zn was lower in WAT compared with all treatments ( P < 0.055) and Co and Se were not different among treatments ( P > 0.05). The level and source of TM in horse diets affected plant availability of nutrients in the manure, which can influence pasture growth and composition.

Validation of a high-throughput method for analysis of pesticide residues in hemp and hemp products

Hemp has been an agricultural commodity for millennia, and it has been undergoing a resurgence in interest and production due to its high content of cannabinoids, protein, fiber and other ingredients. For legal possession and use throughout the USA, hemp and hemp products must have delta-9-tetrahydrocannabinol (THC) concentration < 0.3%. As with most crops, pesticides may be applied when farming hemp, which need to be monitored in food, feed, and medicinal products. The aim of this work was to evaluate and validate the recently developed “quick, easy, cheap, effective, rugged, safe, efficient, and robust” (QuEChERSER) sample preparation mega-method to determine pesticide residues in hemp plants, flowers, powders, oils, and pellets. High-throughput analysis of final extracts for 106 targeted pesticides and metabolites from North American monitoring lists entailed: 1) ultrahigh-performance liquid chromatography - tandem mass spectrometry (UHPLC-MS/MS) with column back-flushing, and 2) instrument-top sample preparation + low-pressure gas chromatography (ITSP+LPGC-MS/MS). In QuEChERSER, 2 g sample is extracted with 10 mL 4/1 (v/v) acetonitrile/water by mechanical shaking for 10 min, followed by 3 min centrifugation. For LC, 0.2 mL of extract is taken and solvent exchanged into initial mobile phase followed by 5 min ultra-centrifugation prior to the 10 min analysis. For GC-amenable pesticides, the remaining initial extract is partitioned with 4/1 (w/w) anh. MgSO4/NaCl, and 1 mL is taken for automated ITSP cleanup in parallel with 10 min LPGC analysis. In the former case, the UHPLC column is back-flushed with 1/1 (v/v) methanol/acetonitrile for 3 min between each injection to keep the system clean and avoid ghost peaks. Multi-level, multi-day validation results achieved 70–120% recoveries with RSDs < 20% for more than 80% of the analytes in hemp protein powder, oil, pellets, and fresh plant (dried hemp plant and flower were too complex). Limits of quantification (LOQs) were < 10 ng/g were achieved for nearly all pesticides, yielding 2.8% false negatives among >13,000 analyte results in the spiked samples. The QuEChERSER method was demonstrated to meet the challenge for several complex hemp matrices.

In-Vitro and In-Silico Assessment of Per- and Polyfluoroalkyl Substances (PFAS) in Aqueous Film-Forming Foam (AFFF) Binding to Human Serum Albumin.

Drinking water contaminated by fluorosurfactant-based aqueous film-forming foams (AFFF) is a source of human exposure to poly- and perfluoroalkyl substances (PFAS). However, assessment of bioaccumulation potentials of diverse PFAS in commercial products such as AFFF have been insufficient and challenging, especially due to a lack of analytical standards. Here we explore the value of suspect screening, equilibrium dialysis, and molecular-docking simulations to identify potentially bioaccumulative PFAS. We exposed human serum albumin (HSA) protein to dilutions of a legacy AFFF produced by 3M in 1999 using equilibrium dialysis and screened in-vitro protein-binding affinities using high-resolution mass spectrometry (HRMS). Through suspect screening, we identified 32 PFAS and 18 hydrocarbon surfactants in the AFFF that bound to HSA. Quantification of noncovalent association constants for 26 PFAS standards confirmed that many PFAS, including the short-chain perfluoropropane sulfonic acid (log Ka= 4.1 ± 0.2 M−1), exhibit strong binding affinities with HSA. At least five PFAS in AFFF (including three PFAS with less than five perfluorocarbons) remained bound to the precipitated HSA pellet after extensive solvent washing—an indication of high PFAS binding potential. Three PFAS (PFBS, PFOS, and PFOA) were confirmed in the protein pellet with analytical standards and quantified after acid digestion—this sample fraction accounted for 5 to 20% of each compound mass in the sample. We calculated pseudo-bioconcentration factors (BCFpseudo) for PFAS that suspect screening flagged as noncovalently bound or potentially covalently bound. Most PFAS exhibiting high BCFpseudo, especially those with seven perfluorocarbons, contained a carboxylic acid or a sulfonic acid. Finally, we used molecular docking to simulate HSA binding affinities for 62 ligands (26 PFAS targets, 18 PFAS qualified in AFFF, and 18 hydrocarbon surfactants qualified in AFFF). We found that molecular docking can effectively separate HSA-binding and -nonbinding compounds in AFFF. In-vitro and in-silico approaches described in this study provide replicable, high-throughput workflows for assessing bioaccumulation potentials of diverse PFAS in commercial products.

Biofloc formation without occurrence of cultured organisms (P. vannamei post-larvae) to sustain high density of biofloc concentration - A preliminary study

Carbon-nitrogen ratio, micro-fauna abundance, water flow rate, water quality parameters were several factors studied by researchers. Unfortunately, main nutrient source and occurrence of organisms were as critically important to be investigated for biofloc formation. In this study, biofloc was formed without the occurrence of organisms (P. vannamei post-larvae) with the additional of an economical nutrient source and inoculum to speed-up and increase the concentration of biofloc formation. The triplicate experimental set-up was prepared under two condition - Treatment A (30% protein pellet feed and molasses) and Treatment B (30% protein pellet feed, molasses, wheat flour, urea, superphosphate and clay) where P. vannamei post-larvae was used as control of each treatment. Water quality parameters, biofloc volume and P. vannamei post-larvae survival rate were observed within 30 days. There was no significant difference (P < 0.05) between the water quality parameters, however, biofloc volume showed a significant difference with and without the occurrence of P. vannamei. Treatment B shows a progressive result where biofloc volume was indicated up to 90.0±4.58 mL/L at day 30. Formation of biofloc without the addition of P. vannamei post-larvae was successful within 5 days with the addition of inoculum and can be used as a starter stock in forming biofloc.

Dietary choices in a generalist herbivore, the eastern cottontail Sylvilagus floridanus, in urban landscapes

Abstract We tested for dietary choices of foods varying in nutrient composition by cottontail rabbits on two college campuses in midwestern USA. We quantified choices among pellets of varying nutritional quality at artificial food patches. Dietary choices differed between seasons and locations. Spring giving-up densities (GUDs: food left behind) did not show differences in food choices and were lower than summer GUDs. In Appleton, the cottontails favoured both high protein and fibre pellets, whereas the medium protein and fibre pellets were favoured in Chicago. The cottontails maintained their choice of high protein, high fibre pellets at three spatial scales. The cottontails varied food intake to balance their protein, salt and fibre needs at different times and locations. Studying dietary choices and the effect of resource quality on foraging responses by urban wildlife provides a useful tool to study ecological interactions and can help minimize damage in urban environments such as parks.

Effect of increasing the amount of hay fed on Holstein calf performance and digestibility from 2 to 4 months of age

A good transition from a mainly liquid diet to a solid diet of concentrates and forages is important considering the small size and development of the calf's rumen. However, the optimal amount of hay or other high-fiber ingredients in the diet of recently weaned calves is not well defined. The objective of this trial was to determine the effects of feeding 0, 5, or 10% chopped grass hay (6.5% crude protein, 64.6% neutral detergent fiber) with a textured, high-starch starter (20.5% crude protein, 38.4% starch, 14.1% neutral detergent fiber; protein pellet, whole corn, and oats) on performance and digestion in Holstein steer calves between 2 and 4 mo of age. Forty-eight calves (initial body weight = 90.7 ± 2.15 kg) were housed in group pens (4 per pen) and fed diets and water for ad libitum intake. Feed offered and refused was measured daily. Calf body weight, hip width, and body condition score (1-5 scale) were measured initially and at 28 and 56 d. Fecal samples were collected from the pen floor with care not to sample bedding material and composited by pen during d 5-9, 26-30, and 47-51 to estimate apparent total-tract digestibility using acid-insoluble ash. Data were analyzed as a completely randomized design with repeated measures and pen as the experimental unit. As grass hay increased, dry matter intake (kg/d) decreased linearly but tended to change quadratically. Dry matter intake as a % of body weight changed quadratically with increasing hay, increasing from 0 to 5%, and decreasing from 5 to 10% hay. Average daily gain (1.15, 1.12, and 0.95 kg/d), feed efficiency (0.336, 0.319, and 0.309 kg of average daily gain/kg of dry matter intake), and hip width change (4.6, 4.7, and 4.1) decreased linearly with 0, 5, and 10% hay, respectively. Overall estimates of dry matter, organic matter, neutral detergent fiber, acid detergent fiber, and crude protein digestibility changed quadratically with hay, increasing from 0 to 5%, and decreasing from 5 to 10% hay, whereas digestibility of starch, sugar, and fat decreased linearly with increasing hay. Digestibility of dry matter, organic matter, neutral detergent fiber, and acid detergent fiber increased from 2 to 3 mo of age, and decreased from 3 to 4 mo of age. Digestibility of starch decreased linearly and digestibility of fat and crude protein increased linearly with age. Digestive capacity did not appear mature by 4 mo of age. Feeding 5% chopped hay supported optimal digestion and growth in calves 2 to 4 mo of age, which agrees with previously published research.

17 Comparing Natural Vegetation and Food Plot Preference against Supplemental Protein Pellets in Captive White-tailed Deer (Odocoileus virginianus) and Exotic Species

Abstract Supplemental feed is the most expensive input in the captive wildlife industries. This is due to operations utilizing high-energy pellets as supplemental feed. Low fence operations often utilize food plots with high quality vegetation to minimize cost and increase forage availability for wildlife. The objective of this study was to determine forage preference of animals in captivity, and determine the most cost effective method of supplemental feeding. Seven food plots covering 25 acres contained one of three treatments. The treatments were: a commercial blend of soybeans; a commercial blend of soybeans, sunflowers, and milo; native/unplanted. Three utilization cages were set up to inhibit wildlife access to samples within each food plot to act as a control. Height of vegetative samples, inside and outside of the utilization cages, was collected on days 30, 60, and 90 after planting. The 30-d sample showed a preference of native/unplanted vegetation over the commercial treatments, P &amp;lt; 0.05. However, samples taken on days 60 and 90 showed the preference shifted toward the commercial blends, P &amp;lt; 0.05. This browsing preference indicates stage of maturity had an impact on plant selection. Regardless of sample date, the commercial blends showed a difference of P &amp;lt; 0.01, selecting the soybean blend more frequently. With an increased selection of the forages in the food plots, there was a decrease in cost of purchasing feed pellets with a total savings of approximately $5,500.00 from April 5-July 5, 2019. These results suggest that white-tailed deer and exotic species in captivity prefer forages compared to pelleted feed. Supplemental feeding programs that include food plots could allow for natural feeding tendencies, while being more cost effective for operations in the captive wildlife industries.

16 Comparing Natural Vegetation and Food Plot Preference against Supplemental Protein Pellets in Captive White-tailed Deer (Odocoileus virginianus) and Exotic Species

Abstract Supplemental feed is the most expensive input in the captive wildlife industries. This is due to operations utilizing high-energy pellets as supplemental feed. Low fence operations often utilize food plots with high quality vegetation to minimize cost and increase forage availability for wildlife. The objective of this study was to determine forage preference of animals in captivity, and determine the most cost effective method of supplemental feeding. Seven food plots covering 25 acres contained one of three treatments. The treatments were: a commercial blend of soybeans; a commercial blend of soybeans, sunflowers, and milo; native/unplanted. Three utilization cages were set up to inhibit wildlife access to samples within each food plot to act as a control. Height of vegetative samples, inside and outside of the utilization cages, was collected on days 30, 60, and 90 after planting. The 30-day sample showed a preference of native/unplanted vegetation over the commercial treatments, P &amp;lt; 0.05. However, samples taken on days 60 and 90 showed the preference shifted toward the commercial blends, P &amp;lt; 0.05. This browsing preference indicates stage of maturity had an impact on plant selection. Regardless of sample date, the commercial blends showed a difference of P &amp;lt; 0.01, selecting the soybean blend more frequently. With an increased selection of the forages in the food plots, there was a decrease in cost of purchasing feed pellets with a total savings of approximately $5,500.00 from April 5-July 5, 2019. These results suggest that white-tailed deer and exotic species in captivity prefer forages compared to pelleted feed. Supplemental feeding programs that include food plots could allow for natural feeding tendencies, while being more cost effective for operations in the captive wildlife industries.

Optimization of a Colorimetric Assay to Determine Lactate Dehydrogenase B Activity Using Design of Experiments.

Lactate dehydrogenase B (LDH-B) is overexpressed in lung and breast cancer, and it has been considered as a potential target to treat these types of cancer. Herein, we propose a straightforward incomplete factorial (IF) design composed of 12 combinations of two reaction buffers, three pH values, three salt (NaCl) concentrations, and three incubation times, which we called IF-BPST (Buffer/pH/Salt/Time), for the optimization of a colorimetric LDH-B assay in a final volume of 100 µL using 96-well plates. The assay is based on the absorbance change at ~570 nm and the color change of the reaction mixture due to the release of NADH that reacts with nitroblue tetrazolium (NBT) and phenazine methosulfate (PMS), resulting in the formation of a blue-purple formazan. The results obtained using the IF-BPST were comparable with those obtained by response surface methodology. Our work revealed that the NBT/PMS assay with some modifications can be used to measure the activity of LDH-B and other dehydrogenases in a high-throughput screening format at the early stages of drug discovery. LDH-B containing lysates cannot be assayed directly, however, due to the sensitivity of the method toward detergents. Thus, we suggest precipitating the proteins in the lysates to remove the interfering detergents, and then to dissolve the protein pellet in a suitable buffer and carry out the assay.

High-yield skeletal muscle protein recovery from TRIzol after RNA and DNA extraction.

Extraction of DNA, RNA and protein from the same sample would allow for direct comparison of genomic, transcriptomic and proteomic information. Commercially available kits exhibit poor protein yield and the TRIzol® reagent produces a protein pellet that is extremely difficult to solubilize. In response to these limitations, this study presents an optimized method for the extraction of protein from the organic phase of TRIzol that allows for higher yield recovery of skeletal muscle protein compared with direct homogenization in a common protein lysis buffer. The presented method is inexpensive, simple and fast, requires no additional treatment of the protein pellet for dissolution, and is compatible with downstream western blot applications.

Pelletizing of hydrochar biofuels with organic binders

In this study, the effect of four organic binders (protein, starch, lignin, and molasses) on woody biomass pellets was investigated. Properties of binder based pellets were evaluated for energy consumption, tensile strength, equilibrium moisture content, and combustion performance. Results indicate that the energy consumption of all binder based pellets was less than that of hydrochar pellets (45.38 J/g). In addition, starch and protein pellets exhibited better mechanical strength when binder content is 20% (3.07 and 3.40 MPa, respectively). Scanning electron microscope results indicate that starch and protein combined closely with hydrochar. Pellets with starch as the binder (20%) exhibited the highest equilibrium moisture content. Regarding combustion performance, low ignition temperature (Ti) was observed for all binder based pellets.

Supplementation of rangeland primiparous Bos indicus x Bos taurus beef heifers during lactation. 1. Effects on dam milk production and liveweight, bull calf growth, live carcass characteristics and metabolic hormone concentrations

The practice of feeding replacement-breeding bulls on high energy diets after weaning to meet liveweight (LW) and carcass expectations between 18 and 24 months of age negatively affects reproductive potential. This experiment reports upon the effects of an alternative management strategy aimed at improving calfhood nutrition in rangeland-reared bulls to enhance LW and live carcass characteristics at 2 years. Following artificial insemination (AI cohort; n = 26), or natural mating, subsequent to the addition of bulls at 39 d post-AI (NM cohort; n = 36), primiparous Santa Gertrudis heifers grazing rangeland pastures with bull calf progeny were allocated at parturition to receive either nil supplement (control; CON) or provided with unrestricted access to a pelleted vegetable protein meal-based supplement containing 35% CP (SUPP) until weaning at 199 ± (SD) 26 d. The mean estimated pellet consumption by SUPP heifers during lactation was 2.6 ± (SEM) 0.5 kg DM daily. Grazing diet quality measurements indicated nutritional restriction of CON heifers until at least 115 d of lactation. This was confirmed by lower blood urea nitrogen concentrations at 88 d (P < 0.001) and greater mean NEFA (P < 0.001) concentrations. Rainfall during mid-lactation subsequently improved grazing diet quality; thus the CON heifers experienced moderate nutritional restriction across lactation, but sufficient to reduce milk yield by 1.6 kg/d (P < 0.001) and maternal LW at weaning by 18.4 kg (P < 0.001). Bulls reared by SUPP heifers were 17.5 kg heavier at weaning (P = 0.001) and had elevated IGF-I and leptin concentrations between 4 and 4.5 months of age (P < 0.05). Effects on metabolic hormones during calfhood were cohort specific, with greater concentrations of IGF-I confined to AISUPP bulls and NMSUPP bulls demonstrating greater concentrations of leptin. Bulls were amalgamated at weaning and grazed common pastures without supplementation until the experiment concluded at 675 d. Pre-weaning plane of nutrition did not affect the LW, carcass fat depth, IGF-I or leptin concentrations of bulls after weaning. Mean eye muscle area (EMA) was greater in SUPP compared to CON bulls (68.5 ± 0.9 cm2vs 65.2 ± 0.9 cm2; P < 0.05) and AISUPP bulls tended to have greater EMA (P = 0.06) than AICON bulls from 495 d of age. Thus when primiparous heifers experience moderate nutritional restriction during lactation, supplementation may have persistent effects upon increasing carcass muscle characteristics of bull progeny.