The Effect of the Species Source of Muscle and/or Digestive Enzymes on the Utilization of Fish Protein Hydrolysates as a Dietary Protein Source in First Feed for Larval Walleye (Sander vitreus).

Fish protein hydrolysates used in larval diets have been prepared from a variety of fish species, with different enzymes used to hydrolyze the protein. This study's objectives were to determine the effect of the dietary inclusion of fish muscle hydrolysates obtained from species-specific muscle/enzymes-versus hydrolysates produced from muscle/enzymes of a different species-on the growth performance, survival, skeletal development, intestinal peptide uptake, and muscle-free amino acid (FAA) composition of larval Walleye (Sander vitreus). Eight protein products were obtained for this study, comprising an unhydrolyzed and hydrolyzed product from each combination of muscle/enzymes from Walleye and Nile tilapia (Oreochromis niloticus). Four diets were produced, and the dietary protein was provided in a 50/50 ratio of unhydrolyzed and hydrolyzed protein from the respective muscle/enzyme combination. Four groups were fed one of the corresponding formulated diets, and two groups of larvae, fed a commercial starter diet and Artemia, respectively, served as reference groups. Larval Walleye fed the diet containing protein produced with the species-specific muscle and enzymes had a significantly higher weight after the study-30% higher than any other group. A significant interaction effect between muscle and enzyme sources on the growth of Walleye larvae was observed. The species-specific combination also led to a significant increase in postprandial FAA and indispensable amino acid concentrations in muscle. No significant differences were observed between the hydrolysate-fed groups in survival, deformity occurrence, or peptide uptake. Each hydrolysate-based diet significantly reduced skeletal deformities and survival compared to the commercial diet. The results of this study suggest that species-specific muscles and enzymes produce a more optimal dietary protein source for larval fish than non-species-specific products. Further research should focus on improving the physical properties of the formulated diets to reduce possible leaching of hydrolyzed protein and improve the survival of fish larvae.

A Novel Approach in the Development of Larval Largemouth Bass Micropterus salmoides Diets Using Largemouth Bass Muscle Hydrolysates as the Protein Source.

This study's objectives were to determine the effect of Largemouth Bass (LMB) muscle hydrolysates obtained using same-species digestive enzymes and the degree of LMB muscle hydrolysis when included in the first feeds of growth performance and survival, skeletal development, intestinal peptide uptake, and muscle-free amino acid composition of larval LMB. LMB muscle was mixed with digestive enzymes from adult LMB, and hydrolyzed for 1.5, 3, and 6 h, respectively. Five diets were produced, the intact diet containing non-hydrolyzed muscle and four diets with 37% muscle hydrolysate inclusion. Those diets were characterized by their level of each hydrolysate (presented as a ratio of 1.5, 3, and 6 Ts hydrolysates): 1:1:1, 1:3:6, 1:3:1, 6:3:1 for diets A, B, C, and D, respectively. To account for gut development, one group of larval LMB was fed a weekly series of diets B, C, and D to provide an increasing molecular weight profile throughout development. This group was compared against others that received either; (1) diets D, C, and B; (2) diet A; or (3) intact diet. The initial inclusion of the hydrolysates significantly improved the total length of the larval LMB; however, neither the hydrolysate inclusion nor the series of dietary molecular weight profiles improved the overall growth of larval LMB. The inclusion of hydrolysates significantly decreased the occurrence of skeletal deformities. The degree of hydrolysis did not have a significant effect on the parameters measured, except for intestinal peptide uptake, which was increased in the group that received the most hydrolyzed diet at the final time of sampling. The lack of overall growth improvement suggests that while the hydrolysates improve the initial growth performance, further research is necessary to determine the optimal molecular weight profile, hydrolysate inclusion level, and physical properties of feeds for larval LMB.

Seaweed proteins are nutritionally valuable components in the human diet.

The global population is expected to reach 11 billion people by the year 2100 and will require sustainable sources of dietary protein. Most dietary protein originates from animal and terrestrial plant agriculture, which leads to deforestation, water pollution, and greenhouse gas emissions. Discovering alternative protein sources that are nutritionally adequate for the human diet without harmful environmental effects is imperative. Seaweeds are a promising option as they produce abundant protein with a low carbon footprint. Experimental evidence shows that seaweeds contain high concentrations of the essential amino acids (EAAs) necessary for human consumption, but seaweeds have yet to be evaluated with standardized metrics to compare their nutritional value to other protein sources. In this technical note, independent literature describing the EAA content and protein digestibility of 3 commonly consumed species of seaweeds was evaluated alongside traditional protein sources using a novel hybrid protein quality (HPQ) metric. HPQ is derived from the protein digestibility-corrected amino acid score and digestibility indispensable amino acid score but includes modifications to address the lack of in vivo digestibility data for seaweeds. Seaweed proteins are similar in quality to common plant protein sources such as peas, soy, and tree nuts. Furthermore, seaweed proteins from different species have complementary EAA profiles and can be mixed to form protein blends that are nutritionally on par with animal products such as milk and whey. Thus, seaweeds may be viable protein sources with a reduced footprint that provide beneficial ecosystem services.

Fish muscle hydrolysate obtained using largemouth bass Micropterus salmoides digestive enzymes improves largemouth bass performance in its larval stages.

The present study utilized digestives tracts from adult largemouth bass (LMB) to hydrolyze Bighead carp muscle and obtain an optimal profile of muscle protein hydrolysates that would be easily assimilated within the primitive digestive tract of larval LMB. Specifically, muscle protein source was digested for the larva using the fully developed digestive system of the same species. The objectives of this study were: 1) to develop an optimal in vitro methodology for carp muscle hydrolysis using LMB endogenous digestive enzymes, and 2) to evaluate the effect of dietary inclusion of the carp muscle protein hydrolysate on LMB growth, survival, occurrence of skeletal deformities, and whole-body free amino acid composition. The study found that the in vitro hydrolysis method using carp intact muscle and LMB digestive tracts incubated at both acid and alkaline pH (to mimic digestive process of LMB) yielded a wide range of low molecular weight fractions (peptides), as opposed to the non-hydrolyzed muscle protein or muscle treated only with acid pH or alkaline pH without enzymes from LMB digestive tracts, which were comprised of large molecular weight fractions (polypeptides above 150 kDa). Overall, the dietary inclusion of the carp muscle hydrolysate improved growth performance of larval LMB in terms of final average weight, weight gain, DGC, SGR, and body length after 21 days of feeding compared to fish that received the diet based on non-hydrolyzed carp muscle. The study also found that hydrolysate-based feed significantly reduced skeletal deformities. The positive growth performance presented by fish in the hydrolysate-fed group possibly resulted from matching the specific requirements of the larvae with respect to their digestive organ development, levels of digestive enzymes present in the gut, and nutritional requirements.

The effects of age, sex, and hot carcass weight on cooked lamb flavor and off-flavor in four muscle cuts.

The present study used 48 lambs originating from three different locations in the Western United States (16 lambs per location; 8 ewes and 8 wethers per location). Each consisting of similar breed composition (Suffolk cross) that were selected to represent weight by age at harvest treatments: light weight carcasses at 5 mo (LW5, 31.81 ± 1.88 kg), light weight carcasses at 12 mo (LW12, 35.09 ± 4.45 kg), heavy weight carcasses at 12 mo (HW12, 57.89 ± 4.70 kg) with different carcass weight compositions. Older heavy weight lambs (HW12) had greater (P ≤ 0.01) hot carcass weight, ribeye area, backfat and body wall thickness, and yield grade compared with light weight lamb carcasses (LW5 and LW12). The longissimus thoracis longissimus thoracis (LT) from older lamb carcasses (LW12 and HW12) had a greater (P ≤ 0.01) total lipid percentage compared with younger lamb carcasses (LW5). Across harvest weight and age treatments, wether carcasses had greater (P ≤ 0.05) total lipid percentage compared with ewe carcasses. Slice shear force values were greater (P ≤ 0.01) for both the LT and semimembranosus from older lambs (LW12 and HW12) compared with LW5 lambs, with no differences between ewes and wethers. Lamb flavor intensity was greater (P ≤ 0.05) for the LT of LW12 lambs and tended (P = 0.08) to be greater for HW12 lambs, compared with the LT from LW5 lambs. The off-flavor intensity of the LT was greater (P ≤ 0.01) for older lambs (LW12 and HW12) compared with LW5 lambs. Interestingly, the lamb flavor and off-flavor intensity scores of the ground shoulder exhibited a treatment × sex interaction. Lamb flavor intensity of LW12 lamb was greater (P ≤ 0.05) from ewes compared with wethers, whereas wethers had a greater (P ≤ 0.05) lamb flavor intensity compared with ewes for HW12 lambs, and LW12 ewe lambs had a greater (P ≤ 0.05) off-flavor intensity compared with all other treatment × sex treatment combinations. Overall, lambs in the present study possessed a mild lamb flavor, typically with greater lamb flavor and off-flavor intensities for older animals; while slice shear force and LT lipid percentage increased as animal age increased at the time of harvest.

Proteomic profile and morphological characteristics of skeletal muscle from the fast- and slow-growing yellow perch (Perca flavescens).

The objective of the present study was to compare skeletal muscle proteomic profiles, histochemical characteristics, and expression levels of myogenic regulatory factors (MRFs) between fast- versus slow-growing yellow perch Perca flavescens and identify the proteins/peptides that might play a crucial role in the muscle growth dynamic. Yellow perch were nursed in ponds for 6 weeks from larval stage and cultured in two meter diameter tanks thereafter. The fingerlings were graded to select the top 10% and bottom 10% fish which represented fast- and slow-growing groups (31 yellow perch per each group). Our statistical analyses showed 18 proteins that had different staining intensities between fast- and slow-growing yellow perch. From those proteins 10 showed higher expression in slow-growers, and 8 demonstrated higher expression in fast-growers. Fast-growing yellow perch with a greater body weight was influenced by both the muscle fiber hypertrophy and mosaic hyperplasia compared to slow-growing fish. These hyperplastic and hypertrophic growth in fast-grower were associated with not only metabolic enzymes, including creatine kinase, glycogen phosphorylase, and aldolase, but also myoD and myogenin as MRFs. Overall, the results of the present study contribute to the identification of different expression patterns of gene products in fast- and slow-growing fish associated with their muscle growth.

Effect of energy source and level, animal age, and sex on the flavor profile of sheep meat.

The effects of dietary energy source, energy level, sheep age, and sheep sex on flavor and off-flavor intensity were evaluated. Consumer panelists, with previous lamb-eating experience, assessed lamb flavor and off-flavor intensity on a 100-point, end-anchored scale (0 = very mild to 100 = very intense), with off-flavor being defined as anything different than lamb flavor. Lamb longissimus thoracis (LT) and whole, boneless ground shoulder (GS) formed into patties were evaluated. Trial 1 was a randomized complete block design with a 3 × 2 factorial arrangement of treatments. Sheep age (ewe lambs, yearling ewes, and mature ewes; n = 16/age) and ad libitum access to diets [alfalfa pellets (AP) or whole-shelled corn (WSC100)] were treatments. The LT from mature ewes had a greater (P ≤ 0.02) off-flavor intensity when compared with yearling ewes and ewe lambs. Ground shoulder from sheep raised on AP had a greater lamb flavor (P ≤ 0.04) and off-flavor (P ≤ 0.04) intensity than GS from sheep consuming WSC100. Trial 2 was a randomized complete block design with a 3 × 2 × 2 factorial arrangement of treatments. Three dietary treatments [AP, WSC100, and restricted intake of whole-shelled corn to 85% of ad libitum (WSC85)], lamb sex (ewes and wethers; n = 48/sex), and lamb age [short fed, 177 ± 16.6 d of age and 93 ± 20.5 d on feed (DOF); long fed, 294 ± 7.0 d of age and 219 ± 3.8 DOF]. Flavor intensity of the LT was greater (P ≤ 0.05) from lambs offered AP when compared with lambs offered WSC85, whereas lamb flavor of the LT from lambs fed WSC100 was intermediate and not different from the lamb flavor of the LT of lambs fed AP or WSC85. The LT of long-fed lambs had a greater (P ≤ 0.01) lamb flavor and off-flavor intensity when compared with short-fed lambs. Lambs offered AP resulted in a GS with greater lamb flavor intensity (P ≤ 0.01) when compared with lambs offered WSC85 and WSC100, with no diet influence on GS off-flavor intensity. Long-fed lambs produced GS with a greater lamb flavor (P ≤ 0.01) and off-flavor (P ≤ 0.01) intensity when compared with GS from short-fed lambs. Results from the two trials indicate lamb flavor and off-flavor intensity were greater from sheep offered a high-forage (AP) diet when compared with a high-concentrate (WSC) diet. Lamb flavor intensity increased as age of the sheep at harvest increased, suggesting dietary management and associated age-related effects at harvest will influence consumer perception of lamb flavor.

Transcriptomic response to soybean meal-based diets as the first formulated feed in juvenile yellow perch (Perca flavescens).

With increasing levels of fish meal (FM) protein in aquafeeds being replaced with soybean meal (SBM) protein, understanding the molecular mechanisms involved in response to alternative diets has become a critical concern. Thus, the goal of this study was to examine transcriptional differences in the intestine of juvenile yellow perch through RNA-sequencing (RNA-seq), after their initial introduction to a formulated diet with 75% SBM protein inclusion for 61 days, compared to those fed a traditional FM-based diet. Transcriptomic analysis revealed a concise set of differentially expressed genes in juveniles fed the SBM-based diet, the majority of which were intrinsic to the cholesterol biosynthesis pathway. Analysis of total body lipid and cholesterol levels were also investigated, with no between-treatment differences detected. Results of this study demonstrate that in response to SBM-based diets, yellow perch juveniles up-regulate the cholesterol biosynthesis pathway in order to maintain homeostasis. These findings suggest that the upregulation of the cholesterol biosynthesis pathway may negatively impact fish growth due to its large energy expenditure, and future studies are warranted.

Potassium carbonate improves fresh pork quality characteristics.

Meat enhancement strategies like sodium tripolyphosphate (STP) are used to improve fresh meat quality attributes like color, water-holding capacity, and tenderness. However, alternatives are necessary because of reduced consumer acceptance of STP. One alternative is potassium carbonate (K2CO3). A study was conducted to evaluate K2CO3's impact on fresh, boneless, center-cut pork loins enhanced with one of five treatments: a negative control, positive control (0.3% STP), and three concentrations of K2CO3 (0.1, 0.3, and 0.5%). Loins were cut into chops, stored under simulated retail display, and analyzed for color (L*, a*, b*), pH, cook loss, and tenderness. For each quality characteristic measured, the 0.3% and 0.5% K2CO3 maintained redness (a*), decreased yellowness (b*), reduced cooking loss, and maintained tenderness compared to STP. SDS-PAGE analysis further determined that both K2CO3 and STP extracted myosin heavy chain. Combined, these data suggest that K2CO3 may function as an alternative to STP in the fresh pork industry provided microbial safety and shelf-life are appropriately controlled.

Assessment of chicken breast meat quality after freeze/thaw abuse using magnetic resonance imaging techniques.

BACKGROUND: Freezing/thawing meat can result in quality losses as a result of the formation, melting and reformation of ice. These changes in water state can result in alterations in texture, water holding and other key quality attributes. It was hypothesized that magnetic resonance imaging (MRI) could quantify changes in mobility and localization of water as a function of freezing/thawing, which could be correlated with quality measurements. RESULTS: Drip loss increased significantly for unbrined samples by over 100% after each freeze/thaw cycle (1.5% to 3.3% to 5.3% drip loss). Brine uptake decreased 50% after 2 cycles (from 53% to 28% mass uptake). Drip loss for brined samples increased after 2 cycles; other attributes were not significantly affected. MRI showed brined samples had less change in both proton density and T2 distributions. High-field nuclear magnetic resonance (NMR) imaging showed greater change in T2 distributions. CONCLUSION: As freeze/thaw damage increased, meat quality was reduced in both brined and unbrined chicken breasts, with more prominent changes in unbrined meat. These decreases in quality correlated with changes, albeit small, in water mobility and localization as measured by MRI. High-field NMR micro-imaging showed more dramatic changes in T2 distributions in unbrined samples. These MRI techniques are shown to be useful in the assessment of meat quality after freeze/thaw abuse. © 2018 Society of Chemical Industry.

Effect of embryonic thermal manipulation on heat shock protein 70 expression and immune system development in Pekin duck embryos.

Two experiments were conducted to study the effects of thermal manipulation on heat shock protein 70 (HSP70) transcription and immune cell characteristics' in Pekin duck embryos. In experiments I and II, fertile duck eggs were incubated at either a standard (S; 37°C) or high (H; 38.0°C) temperature from embryonic day (ED) 1 to 10. At ED11, half the eggs incubated at 37.5°C or 38.0°C were either kept at 37.5°C (SS) or 38.0°C (HH) respectively, or moved to the opposite incubator/temperature of either 38.0°C (SH) or 37.5°C (HS). Beginning on ED 21, all eggs (embryos) were incubated at 37.5°C. In experiments I and II, the mean egg shell temperature (EST) of eggs incubated at 38.0°C was higher than that from 37.5°C at ED10 (P < 0.01). In both experiments, the yolk free wet embryo body weight (YFWEBW) of embryos from the HS treatment was increased while that in the HH treatments was decreased compared with the SS treatment at ED25 (P < 0.01). In experiment II, embryos from the HH treatment had increased HSP70 mRNA compared with the SS treatment in the liver, thymus, and bursa (P < 0.05). The thymus and bursa from embryos in the SH treatment had increased MHCI mRNA when compared with the SS treatment (P < 0.05). The thymus from HH and SH treatments had increased MHCII mRNA while the bursa from the HH and SH treatments had decreased MHCII mRNA when compared with the SS treatment (P < 0.05). The spleen and bursa from the HH treatment had increased IL-6, IFNγ, and IL-10 mRNA when compared with the SS treatment (P < 0.05). The thymus and spleen from embryos in the HH treatment had increased CD8+/CD4+ ratios and CD4+CD25+ cell percentages compared with the SS treatment (P < 0.05). In conclusion, embryos incubated at 0.5°C higher than the standard incubation temperature from ED1 to 21 had increased HSP70 mRNA, MHCI, and inflammatory cytokines, and the immune response was skewed toward cell-mediated immunity.

Onset of white striping and progression into wooden breast as defined by myopathic changes underlying Pectoralis major growth. Estimation of growth parameters as predictors for stage of myopathy progression.

The broiler industry has incurred significant economic losses due to two muscle myopathies, white striping (WS) and wooden breast (WB), affecting the Pectoralis major (P. major) of commercial broilers. The present study documented macroscopic changes occurring with age/growth in the P. major and P. minor muscles of commercial broilers from day 2 through day 46 (n = 27/day). Distinct myopathic aberrations observed in both breast muscles corresponded to the onset of WB. These distinct morphological changes were used as determinants in developing a ranking system, defining the ontogeny of WB as the following four stages: (1) WS, (2) petechial epimysium haemorrhages, (3) intramuscular haemorrhages and (4) ischaemia. A cumulative logit proportional odds model was used to relate the rank probabilities with the following growth parameters: body weight, P. major and P. minor weight/yield/length/width/depth. The best-fit model included P. major length/width/depth, P. minor width, P. major and P. minor yield as predictors for rank. Increasing P. major depth, P. minor width and P. major yield increased the odds of falling into higher ranks (more severe myopathy). Conversely, increasing P. major length, P. major width and P. minor yield increased the odds of falling into smaller ranks (less severe myopathy). This study describes the macroscopic changes associated with WB ontogeny in the development of a ranking system and the contribution of growth parameters in the determination of rank (WB severity). Results suggest that physical measurements inherent to selection for high-yielding broiler genotypes are contributing to the occurrence and severity of WS and WB.

Reverse Stability Kinetics of Meat Pigment Oxidation in Aqueous Extract from Fresh Beef.

The use of kinetic models is an evolving approach to describing quality changes in foods during processes, including storage. Previous studies indicate that the oxidation rate of myoglobin is accelerated under frozen storage conditions, a phenomenon termed reverse stability. The goal of this study was to develop a model for meat pigment oxidation to incorporate the phenomenon of reverse stability. In this investigation, the model system was an aqueous extract from beef which was stored under a range of temperatures, both unfrozen and frozen. The kinetic analysis showed that in unfrozen solutions, the temperature dependence of oxidation rate followed Arrhenius kinetics. However, under in frozen solutions the rate of oxidation increased with decreasing temperature until reaching a local maximum around -20 °C. The addition of NaCl to the model system increased oxidation rates at all temperatures, even above the initial freezing temperature. This observation suggests that this reaction is dependent on the ionic strength of the solution as well as temperature. The mechanism of this deviant kinetic behavior is not fully understood, but this study shows that the interplay of temperature and composition on the rate of oxidation of meat pigments is complicated and may involve multiple mechanisms. PRACTICAL APPLICATION: A better understanding of the kinetics of quality loss in a meat system allows for a re-examination of the current recommendations for frozen storage. The deviant kinetic behavior observed in this study indicates that the relationship between quality loss and temperature in a frozen food is not as simple as once thought. Product-specific recommendations could be implemented in the future that would allow for a decrease in energy consumption without a significant loss of quality.

Temperature-dependent Henry's Law constants of 4-alkyl branched-chain fatty acids and 3-methylindole in an oil-air matrix and analysis of volatiles in lamb fat using selected ion flow tube mass spectrometry.

RATIONALE: 4-Alkyl branched-chain fatty acids and 3-methylindole are characteristic flavor compounds associated with sheep meat. Determining their partitioning behavior between the gas and condensed phase and ultimately developing a correlation between the compound's headspace concentration and sensory descriptive grouping are important for high-throughput characterization and grading classification. METHODS: The headspace concentrations of 3-methylindole, 4-methyloctanoic acid, 4-ethyl-octanoic acid, and 4-methylnonanoic acid above corn-oil-based standard solutions and lamb fat samples were measured using selected ion flow tube-mass spectrometry (SIFT-MS). The standard solutions were equilibrated at 80, 100, 110 and 125°C while the fat samples were equilibrated at 125°C. Statistical evaluation, linear and polynomial regression analyses were performed to establish the compound-specific and temperature-dependent Henry's Law constants, enthalpy (ΔH) and entropy (ΔS) of phase changes. RESULTS: The Henry's Law constants (kHcp ) were calculated from the regression analysis with a high degree of confidence (p < 0.05) and linearity (r2  > 0.99). The kHcp increased with increase in equilibrium temperature. The empirical calculation of ΔH and ΔS at different temperatures confirmed the temperature-dependence of the Henry's Law constants. The headspace concentrations of the lamb-flavor compounds were determined above actual lamb fat samples and the corresponding condensed-phase concentrations were successfully derived. CONCLUSIONS: The temperature-dependent Henry's Law constants, ΔH, and ΔS of phase changes for 3-methylindole, 4-methyloctanoic acid, 4-ethyloctanoic acid, and 4-methylnonanoic acid in an air-oil matrix were empirically derived. The effectiveness of SIFT-MS for the direct, real-time, and rapid determination of key flavor compounds in lamb fat samples was established.

Gas-phase chemical ionization of 4-alkyl branched-chain carboxylic acids and 3-methylindole using H3 O+ , NO+ , and O2+ ions.

RATIONALE: 4-Methyloctanoic acid, 4-ethyloctanoic acid, 4-methylnonanoic acid, and 3-methylindole are primary contributors to the distinctive aroma and flavor of lamb meat. The reactions of H3 O+ , NO+ , and O2+ with these compounds, and identification of the product ions and their distribution, are fundamental to their characterization and rapid, real-time trace analysis using selected ion flow tube mass spectrometry (SIFT-MS). METHODS: The chemical ionization of pure standards of 4-ethyloctanoic acid, 4-methyloctanoic acid, 4-ethylnonanoic acid, and 3-methylindole was carried out using the H3 O+ , NO+ , and O2+ reagent ions of a V200™ SIFT mass spectrometer. Kinetic data were calculated using the Langevin collision rate with parameterized trajectory equations. Identification of product ions, distribution, and interferences was performed by further evaluation of the pertinent ion-molecule reaction mechanisms, careful spectral analyses, and molecular mass-molecular structure pairing. RESULTS: The collisional capture rate constants of the reaction of the precursor ions H3 O+ , NO+ , and O2+ , their extended hydrates and the analytes, which were assumed to occur at or near the collisional rate, were all of the order of 10-9  cm3 molecule s-1 - typical for bimolecular ion-molecule reactions. Positive identification of the primary and secondary product ions, fragmented ionic species, and potential ion conflicts and interferences, from each reagent ion channel, was determined for each compound. CONCLUSIONS: We have established the ion chemistry involved in the ionization of the 4-alkyl branched-chain fatty acids and 3-methylindole using the precursor ions, H3 O+ , NO+ , and O2+ in SIFT-MS. The ion-molecular chemistry and the associated kinetics serve as a fundamental basis for the accurate characterization of these compounds by SIFT-MS.

Degenerative primer design and gene sequencing validation for select turkey genes.

We successfully designed and validated degenerative primers for turkey genes MUC2, RPS13, TBP and TFF2 based on chicken sequences in order to use gene transcription analysis to evaluate (quantify) the mucin transcription to probiotic supplementation in turkeys. Primers were designed for the genes MUC2, TFF2, RPS13 and TBP using a degenerative primer design method based on the available Gallus gallus sequences. All primer sets, which produced a single PCR amplicon of the expected sizes, were cloned into the TOPO(®) vector and then transformed into TOP 10(®) competent cells. Plasmid DNA isolation was performed on the TOP10(®) cell culture and sent for sequencing. Sequences were analyzed using NCBI BLAST. All genes sequenced had over 90% homology with both the chicken and predicted turkey sequences. The sequences were used to design new 100% homologous primer sets for the genes of interest.

Identification, purification and characterization of furfural transforming enzymes from Clostridium beijerinckii NCIMB 8052.

Generation of microbial inhibitory compounds such as furfural and 5-hydroxymethylfurfural (HMF) is a formidable roadblock to fermentation of lignocellulose-derived sugars to butanol. Bioabatement offers a cost effective strategy to circumvent this challenge. Although Clostridium beijerinckii NCIMB 8052 can transform 2-3 g/L of furfural and HMF to their less toxic alcohols, higher concentrations present in biomass hydrolysates are intractable to microbial transformation. To delineate the mechanism by which C. beijerinckii detoxifies furfural and HMF, an aldo/keto reductase (AKR) and a short-chain dehydrogenase/reductase (SDR) found to be over-expressed in furfural-challenged cultures of C. beijerinckii were cloned and over-expressed in Escherichia coli Rosetta-gami™ B(DE3)pLysS, and purified by histidine tag-assisted immobilized metal affinity chromatography. Protein gel analysis showed that the molecular weights of purified AKR and SDR are close to the predicted values of 37 kDa and 27 kDa, respectively. While AKR has apparent Km and Vmax values of 32.4 mM and 254.2 mM s(-1) respectively, using furfural as substrate, SDR showed lower Km (26.4 mM) and Vmax (22.6 mM s(-1)) values on the same substrate. However, AKR showed 7.1-fold higher specific activity on furfural than SDR. Further, both AKR and SDR were found to be active on HMF, benzaldehyde, and butyraldehyde. Both enzymes require NADPH as a cofactor for aldehydes reduction. Based on these results, it is proposed that AKR and SDR are involved in the biotransformation of furfural and HMF by C. beijerinckii.

Functional proteomic analysis predicts beef tenderness and the tenderness differential.

Inconsistent tenderness is one of the most detrimental factors of meat quality. Functional proteomics was used to associate electrophoretic bands from the myofibrillar muscle fraction to meat tenderness in an effort to gain understanding of the mechanisms controlling tenderness. The myofibrillar muscle fraction of the Longissimus dorsi from 22 Angus cross steers was analyzed by SDS-PAGE and linearly regressed to Warner-Bratzler shear values. Six significant electrophoretic bands were characterized by electrophoretic and statistical analysis and sequenced by nano-LC-MS/MS. The protein(s)/peptide(s) identified in these bands encompass a wide array of cellular pathways related to structural, metabolic, chaperone, and developmental functions. This study begins to assemble information that has been reported separately into a more complete picture that will lead to the establishment of a coherent mechanism accounting for meat tenderness.

Proteomic analysis of proteins associated with body mass and length in yellow perch, Perca flavescens.

The goal of commercial yellow perch aquaculture is to increase muscle mass which leads to increased profitability. The accumulation and degradation of muscle-specific gene products underlies the variability in body mass (BM) and length observed in pond-cultured yellow perch. Our objective was to apply a combination of statistical and proteomic technologies to identify intact and/or proteolytic fragments of muscle specific gene products involved in muscle growth in yellow perch. Seventy yellow perch randomly selected at 10, 12, 16, 20, and 26 wk of age were euthanized; BM and length were measured and a muscle sample taken. Muscle proteins were resolved using 5-20% gradient SDS-PAGE, stained with SYPRO Ruby and analyzed using TotalLab software. Data were analyzed using stepwise multiple regression with the dependent variables, BM and length and proportional OD of each band in a sample as a potential regressor. Eight bands associated with BM (R(2) = 0.84) and nine bands with length (R(2) = 0.85) were detected. Protein sequencing by nano-LC/MS/MS identified 20 proteins/peptides associated with BM and length. These results contribute the identification of gene products and/or proteolytic fragments associated with muscle growth in yellow perch.