Adenoid reservoir for pathogenic biofilm bacteria.

Biofilms of pathogenic bacteria are present on the middle ear mucosa of children with chronic otitis media (COM) and may contribute to the persistence of pathogens and the recalcitrance of COM to antibiotic treatment. Controlled studies indicate that adenoidectomy is effective in the treatment of COM, suggesting that the adenoids may act as a reservoir for COM pathogens. To investigate the bacterial community in the adenoid, samples were obtained from 35 children undergoing adenoidectomy for chronic OM or obstructive sleep apnea. We used a novel, culture-independent molecular diagnostic methodology, followed by confocal microscopy, to investigate the in situ distribution and organization of pathogens in the adenoids to determine whether pathogenic bacteria exhibited criteria characteristic of biofilms. The Ibis T5000 Universal Biosensor System was used to interrogate the extent of the microbial diversity within adenoid biopsy specimens. Using a suite of 16 broad-range bacterial primers, we demonstrated that adenoids from both diagnostic groups were colonized with polymicrobial biofilms. Haemophilus influenzae was present in more adenoids from the COM group (P = 0.005), but there was no significant difference between the two patient groups for Streptococcus pneumoniae or Staphylococcus aureus. Fluorescence in situ hybridization, lectin binding, and the use of antibodies specific for host epithelial cells demonstrated that pathogens were aggregated, surrounded by a carbohydrate matrix, and localized on and within the epithelial cell surface, which is consistent with criteria for bacterial biofilms.

Proteomic assessment of the acute phase of dystrophin deficiency in mdx mice.

Duchenne muscular dystrophy (DMD) is caused by the absence of a functional dystrophin protein and is modeled by the mdx mouse. The mdx mouse suffers an early necrotic bout in the hind limb muscles lasting from approximately 4 to 7 weeks. The purpose of this investigation was to determine the extent to which dystrophin deficiency changed the proteome very early in the disease process. In order to accomplish this, proteins from gastrocnemius from 6-week-old C57 (n = 6) and mdx (n = 6) mice were labeled with fluorescent dye and subjected to two-dimensional differential in-gel electrophoresis (2D-DIGE). Resulting differentially expressed spots were excised and protein identity determined via MALDI-TOF followed by database searching using MASCOT. Proteins of the immediate energy system and glycolysis were generally down-regulated in mdx mice compared to C57 mice. Conversely, expression of proteins involved in the Kreb's cycle and electron transport chain were increased in dystrophin-deficient muscle compared to control. Expression of cytoskeletal components, including tubulins, vimentin, and collagen, were increased in mdx mice compared to C57 mice. Importantly, these changes are occurring at only 6 weeks of age and are caused by acute dystrophin deficiency rather than more chronic injury. These data may provide insight regarding early pathologic changes occurring in dystrophin-deficient skeletal muscle.

Characterizing membrane phospholipid hydrolysis of pork loins throughout three aging periods.

Three chops from 20 pork carcasses were aged for 1, 8, and 21 days. Electrospray ionization-tandem mass spectrometry was used to comprehensively analyze profiles of phospholipids from each sample (n = 60). Total phospholipid quantity decreased 4-folds (P < .01) from 1 to 21 days of aging in pork loins. Phosphatidylinositol (PI) and phosphatidylserine (PS) increased by 30% and 73%, respectively, from 1 to 21 days of aging in pork loins (P < .01). This increase was mainly due to relative percentage increase from PI 38:4 (18:0-20:4) and PS 36:2 (18:0-18:2; P < .01). The results also showed that the relative percentage of lysophosphatidylcholine increased by 35% after short term aging (8d), and phosphatidic acid increased by 10-folds after extended aging (21d; P < .01). These results documented that phospholipids undergo enzymatic hydrolysis during aging, but also indicated that lipid species containing 18:2 or 20:4 within PI and PS were slightly more resistant to enzymatic hydrolysis compared with the other phospholipids.

Effects of breed, sex and halothane genotype on fatty acid composition of triacylglycerols and phospholipids in pork longissimus muscle.

The objective of this study was to estimate the effects of breed, sex, and halothane (HAL-1843) genotype on fatty acid composition of triacylglycerols (TAG) and phospholipids (PL) extracted from porcine longissimus muscle (LM). Purebred Yorkshire (n = 131), Duroc (n = 136), Hampshire (n = 49), Spotted (n = 35), Chester White (n = 74), Poland China (n = 51), Berkshire (n = 169) and Landrace (n = 82) pigs (n = 727; 427 barrows and 300 gilts) from the 1994 and 2001 National Barrow Show Sire Progeny Tests were used. For statistical analyses, a mixed model was used that included fixed effects of breed, sex, HAL-1843(TM) genotype, year, slaughter date within each year, interaction of breed x sex and random effects of sire and dam within breed. Breeds and sex were significantly associated with the percentages of the majority fatty acids in TAG. Duroc pigs had greater total saturated fatty acids (SFA) and lower total monounsaturated fatty acids (MUFA) (p < 0.05) contents than did pigs of all other breeds except Berkshire (p > 0.05). The concentration of total polyunsaturated fatty acids (PUFA) was the greatest in Hampshire pigs (p < 0.05). The content of total SFA was greater (p < 0.01), whereas the concentrations of total MUFA and PUFA were lower (p < 0.01) in barrows than those in gilts. The contents of major SFA in PL did not differ significantly among pigs from different breeds and sex groups. However, breed and sex significantly affected the concentrations of major MUFA and PUFA in PL and strong negative correlation between the total contents of MUFA and PUFA in PL was observed in the current study. Chester White pigs had greater total MUFA and lower total PUFA contents (p < 0.05) in PL than did pigs of all other breeds except Spotted (p > 0.05). In contrast to breed and sex effects, the concentrations of fatty acids in PL were more affected by HAL-1843 genotype than those in TAG. The content of C16:0, a major SFA in PL, differed significantly in pigs with different HAL-1843 genotypes. In conclusion, these results suggest that breed and sex are important sources of the variations for fatty acid composition of TAG and PL in LM.

Progress in reducing the pale, soft and exudative (PSE) problem in pork and poultry meat.

Research in the area of the pale, soft and exudative (PSE) pork and poultry meat is reviewed in this article with an emphasis on genetic, biochemical and metabolic factors contributing to the problem. Over the past five decades, there has been much more work in the pork meat area where a few genetic markers have been identified, and are currently used to remove susceptible animals from the herd. Some of the markers are linked to aberrant calcium regulation in the early postmortem muscle. The poultry industry is still not at the point of using genetic marker(s); however, some recent work has revealed several potential markers. The review also discusses environmental factors such as antemortem stress and early postmortem processing practices (e.g. chilling rate) that can influence the development and severity of the PSE phenomenon. Some of these factors are known to cause protein denaturation at the early stage of postmortem and directly contribute to poor water-holding capacity and inferior texture in fresh meat and later in processed products. A newer hypothesis suggesting that variation in protein oxidation, in response to antemortem stress and early postmortem tissue environment, can contribute to development of PSE pork is also discussed. Finally, a few recommendations for future work are proposed.

Porcine epidemic diarrhea virus reduces feed efficiency in nursery pigs.

Porcine epidemic diarrhea virus (PEDV) infects enterocytes and in nursery pigs, results in diarrhea, anorexia, and reduced performance. Therefore, the objective of this study was to determine how PEDV infection influenced growth performance and repartitioning of amino acids and energy in nursery pigs. A total of 32 barrows and gilts, approximately 1 wk post-wean (BW = 8.46 ± 0.50 kg), and naïve for PEDV were obtained, weighed, and allotted based on sex and BW to one of two treatments: 1) Control, PEDV naïve and 2) PEDV-inoculated (PEDV) with eight pens of two pigs each per treatment. On day post-inoculation (dpi) 0, PEDV pigs were inoculated via intragastric gavage with PEDV isolate (USA/Iowa/18984/2013). Pig and feeder weights were recorded at dpi -7, 0, 5, and 20 in order to calculate ADG, ADFI, and G:F. Eight pigs per treatment were euthanized on dpi 5 and 20, and tissues and blood were collected. At dpi 5, all PEDV pigs were PCR positive for PEDV in feces. Overall, PEDV pigs tended (P < 0.10) to increase ADFI, which resulted in reduced (P < 0.05) feed efficiency. At dpi 5, PEDV pigs had reduced (P < 0.05) villus height and increased (P < 0.05) stem cell proliferation in the jejunum compared with Control pigs. Pigs inoculated with PEDV had increased (P < 0.05) serum haptoglobin and increased insulin-to-glucose ratios compared with Control pigs at dpi 5. Markers of muscle proteolysis were not different (P > 0.05) between treatments within dpi; however, at dpi 5, 20S proteasome activity was increased (P < 0.05) in longissimus dorsi of PEDV pigs compared with Control pigs. Liver and jejunum gluconeogenic enzyme activities were not different (P > 0.05) between treatments within dpi. Overall, PEDV-inoculated pigs did recover the absorptive capacity that was reduced during PEDV infection by increasing proliferation of intestinal stem cells. However, the energy and nutrients needed to recover the epithelium may be originating from available luminal nutrients instead of muscle proteolysis and gluconeogenesis. This study provides insight into the effects of an enteric coronavirus on postabsorptive metabolism in nursery pigs.

Predicting aged pork quality using a portable Raman device.

The utility of Raman spectroscopic signatures of fresh pork loin (1 d & 15 d postmortem) in predicting fresh pork tenderness and slice shear force (SSF) was determined. Partial least square models showed that sensory tenderness and SSF are weakly correlated (R2 = 0.2). Raman spectral data were collected in 6 s using a portable Raman spectrometer (RS). A PLS regression model was developed to predict quantitatively the tenderness scores and SSF values from Raman spectral data, with very limited success. It was discovered that the prediction accuracies for day 15 post mortem samples are significantly greater than that for day 1 postmortem samples. Classification models were developed to predict tenderness at two ends of sensory quality as "poor" vs. "good". The accuracies of classification into different quality categories (1st to 4th percentile) are also greater for the day 15 postmortem samples for sensory tenderness (93.5% vs 76.3%) and SSF (92.8% vs 76.1%). RS has the potential to become a rapid on-line screening tool for the pork producers to quickly select meats with superior quality and/or cull poor quality to meet market demand/expectations.

Effects of heat stress and insulin sensitizers on pig adipose tissue.

Heat stress (HS) negatively impacts several swine production variables, including carcass fat quality and quantity. Pigs reared in HS have more adipose tissue than energetically predicted, explainable, in part, by HS-induced hyperinsulinemia. Study objectives were to evaluate insulin's role in altering fat characteristics during HS via feeding insulin-sensitizing compounds. Forty crossbred barrows (113 ± 9 kg BW) were randomly assigned to one of five environment by diet treatments: 1) thermoneutral (TN) fed ad libitum (TNAL), 2) TN and pair-fed (TNPF), 3) HS fed ad libitum (HSAL), 4) HS fed ad libitum with sterculic oil (SO) supplementation (HSSO; 13 g/d), and 5) HS fed ad libitum with dietary chromium (Cr) supplementation (HSCr; 0.5 mg/d; Kemin Industries, Des Moines, IA). The study consisted of three experimental periods (P). During P0 (2 d), all pigs were exposed to TN conditions (23 ± 3 °C, 68 ± 10% RH) and fed ad libitum. During P1 (7 d), all pigs received their respective dietary supplements, were maintained in TN conditions, and fed ad libitum. During P2 (21 d), HSAL, HSSO, and HSCr pigs were fed ad libitum and exposed to cyclical HS conditions (28 to 33 °C, 58 ± 10% RH). The TNAL and TNPF pigs remained in TN conditions and were fed ad libitum or pair-fed to their HSAL counterparts. Rectal temperature (TR), respiration rate (RR), and skin temperature (TS) were obtained daily at 0600 and 1800 h. At 1800 h, HS exposed pigs had increased TR, RR, and TS relative to TNAL controls (1.13 °C, 48 bpm, and 3.51 °C, respectively; P < 0.01). During wk 2 and 3 of P2, HSSO pigs had increased 1800 h TR relative to HSAL and HSCr (~0.40 and ~0.42 °C, respectively; P ≤ 0.05). Heat stress decreased ADFI and ADG compared to TNAL pigs (2.24 vs. 3.28 and 0.63 vs. 1.09 kg/d, respectively; P < 0.01) and neither variable was affected by SO or Cr supplementation. Heat stress increased or tended to increase moisture content of abdominal (7.7 vs. 5.9%; P = 0.07) and inner s.c. (11.4 vs. 9.8%; P < 0.05) adipose depots compared to TNAL controls. Interestingly, TNPF pigs also had increased adipose tissue moisture content and this was most pronounced in the outer s.c. depot (15.0 vs. 12.2%; P < 0.01) compared to TNAL pigs. Heat stress had little or no effect on fatty acid composition of abdominal, inner, and outer s.c. adipose tissue depots. In summary, the negative effects of HS on fat quality do not appear to be fatty acid composition related, but may be explained by increased adipose tissue moisture content.

New frontiers in understanding drip loss in pork: recent insights on the role of postmortem muscle biochemistry.

The aim of this review was to summarize some of the most important and novel hypotheses regarding the development of water-holding capacity in pork. Among the key factors that determine water-holding capacity are rate and extent of pH decline and the structure of the muscle and the muscle cell itself. However, there have been numerous reviews on the importance of pH yet relatively few on the concept of how muscle cell structure influences the ability of the postmortem cell to maintain water/moisture. Recent advances in the ability of scientists to monitor the movement of water within the cell and within the muscle and to monitor protein degradation have led to new hypotheses regarding the development of drip or purge from fresh meat. One of the more novel and promising hypotheses that has surfaced recently is that alterations in the architecture of the cell may govern the retention/loss of moisture from the muscle cell and ultimately the muscle. The degradation of cytoskeletal and intermediate filament proteins (like desmin) early postmortem may release constraints within the cell that limit the space available for water/fluid that has been forced from the myofibril during rigour. Conversely, degradation of some membrane proteins (like integrin) could actually contribute to the formation of drip channels and thus might actually improve the ability of moisture to 'escape' from the muscle cell. Thus, water-holding capacity in fresh meat is associated with specific changes that occur in early postmortem muscle.

Influence of feed efficiency classification and growing and finishing diet type on meat tenderness attributes of beef steers.

Enhancing feed efficiency (FE) in cattle continues to be an important goal in the beef industry. Previous research suggests improving FE may alter calpain system activity, potentially having negative effects on meat tenderness. The objective of this study was to assess the potential influence of beef cattle FE on postmortem meat tenderness. During the growing phase, 181 crossbred steers were fed for 76 d at the University of Missouri on a whole shell corn-based diet (MU-Corn; = 90) or a roughage-based diet (MU-Rough; = 91). Within diet, steers were classified for FE based on residual feed intake (RFI) calculations. Within each growing phase diet, the 12 most feed efficient (HFE; average RFI -3.33 ± 0.77) and 12 least feed efficient (LFE; average RFI 2.90 ± 0.94) steers (48 steers total) were selected and shipped to Iowa State University for the finishing phase. Steers were fed in pens with GrowSafe bunks and equally assigned to a cracked corn-based finishing diet (ISU-Corn) or a byproduct-based finishing diet (ISU-Byp) for the 87-d finishing phase. After a 24-h chill, rib sections were collected from all carcasses and aged for 2 or 14 d prior to analysis of calpastatin activity and calpain 1 autolysis (d 2), troponin-T degradation (d 2 and 14), proximate composition (d 2), and Warner-Bratzler shear force (WBSF; d 14). Data were analyzed as a 2 × 2 × 2 factorial, with steer or steak as the experimental unit ( = 6 per treatment combination). There were no 3- or 2-way interactions ( ≥ 0.12) noted for any data, with the exception of steak lipid content, which tended ( = 0.08) to be affected by FE × Iowa State University diet. Steaks from LFE steers had greater lipid content compared with steaks from HFE steers within ISU-Byp, with no differences due to FE within ISU-Corn. Growing diet did not affect calpastatin activity, calpain 1 autolysis, or troponin T degradation ( ≥ 0.12); however, MU-Rough had greater WBSF than MU-Corn ( = 0.05). Day-2 calpastatin activity tended ( = 0.10) to be greater in steaks from HFE vs. LFE steers, although no differences due to FE classification were observed for calpain 1 protein, troponin-T degradation, or WBSF ( ≥ 0.13). Finishing diet did not affect calpastatin activity, calpain 1 autolysis, d-2 troponin-T degradation, or WBSF ( ≥ 0.24); however, d-14 troponin-T degradation was greater in ISU-Corn than in ISU-Byp ( = 0.005). In this study, using phenotypic extremes for FE revealed a tendency for greater calpastatin activity only in highly feed-efficient steers; however, these data indicate that high-fiber diets may negatively impact meat tenderness, and further work is needed to clarify the influence of diet type on parameters of meat tenderness.

Proteomic features linked to tenderness of aged pork loins.

There is considerable evidence that the protein component of fresh pork makes a major contribution to tenderness. In particular, the proteomic profile can be linked to postmortem events including pH decline, tissue oxidation, and protein degradation. The objectives for this study were to determine differences in sarcoplasmic proteomes that contribute to tenderness variation in aged pork longissimus dorsi muscles (LM). A defined set of pork loins selected to be similar in pH, color, and lipid yet different in tenderness were used. Pork loins were assigned to tenderness groups based on their star probe values; a high star probe group (HSP; n=12 mean star probe 7.75 kg) and low star probe group (LPS; n=12 star probe 4.95 kg) Samples were selected for proteomic experiments based on star probe values, and selected samples were within specified ranges for ultimate pH (5.54-5.86), marbling score (1.0-3.0), and percent total lipid (1.61-3.37%). Two-dimensional difference in gel electrophoresis (2D-DIGE) and mass spectrometry were used to examine sarcoplasmic protein abundance and potential modifications. Proteins spots that were significantly different across groups were selected for identification. Results from 2D-DIGE showed that HSP samples had significantly more abundant metabolic, stress response, and regulatory proteins in the sarcoplasmic fraction compared with LSP samples. The stress response protein peroxiredoxin-2 was more abundant in HSP samples as determined by 2D-DIGE ( ≤ 0.01; 2 spots) and western blot assay ( = 0.02). Low star probe samples showed significantly more degradation of the structural protein desmin in 2D-DIGE ( < 0.01) and western blot assay ( < 0.01). These results demonstrate that extreme proteolytic differences influenced measured tenderness of LSP and HSP samples and that soluble desmin and peroxiredoxin-2 may be used as biomarkers to differentiate between tough and tender aged pork products.

Temporal proteomic response to acute heat stress in the porcine muscle sarcoplasm.

Heat stress (HS) is an important topic in the swine industry, costing hundreds of millions of dollars in economic losses annually, figures that could easily rise in light of global climate change. Muscle biology during HS is particularly important given skeletal muscle's large proportion to the body and its ultimate conversion to meat. Here we report the proteomic changes that occur during acute HS (37°C and 40% relative humidity) lasting 2, 4, or 6 h in the muscle sarcoplasm of growing pigs in comparison with 6 h of thermal neutral (TN; 21°C and 70% relative humidity) conditions ( = 8 per treatment). The red and white areas of the semitendinosus muscle were used to compare the differential effects of HS on oxidative or glycolytic muscles. The results support the hypothesis of proteomic profile differences between the acute HS and TN groups. Altered abundance ( < 0.05) of several proteins occurred in as little as 2 h of HS, affecting metabolism, cell structure, and chaperone, antioxidant, and proteolytic activity. We determined that the muscle HS response is both fiber type and time specific. Overall, more differences were observed in the red semitendinosus than in the white semitendinosus, although the time point at which differences were observed varied. These data show that as little as 2 h of HS has measurable effects on muscle proteins, indicating that acute HS has the potential to impair muscle function and growth.

Postmortem protein degradation is a key contributor to fresh pork loin tenderness.

The objective of this study was to determine factors that influence tenderness independent of variation in pH, color, or marbling. To achieve the objective, 2 sample groups were chosen from a population of 159 pork loins aged 11 to 16 d. Predetermined ranges (ultimate pH, 5.54 to 5.86; marbling score, 1.0 to 3.0; percent total lipid, 1.61 to 3.37%) were defined for inclusion of individual loins in the study. The pork loins with the greatest ( = 12) and least ( = 12) Instron star probe values were assigned to 2 classification groups. The high star probe group had an average star probe that was 2.8 kg greater than the low star probe group (7.75 vs. 4.95 kg). Pork quality and sensory characteristics of pH, subjective and instrumental color values, cook loss, sensory tenderness, chewiness, juiciness, pork flavor, and off flavor were determined on fresh, never frozen pork chops. Lipid content, sarcomere length, myosin heavy-chain profile, and calpain autolysis were determined. Degradation of troponin-T, desmin, filamin, and titin were evaluated on the protein extracts from each sample. Pork loin pH, subjective color scores, Minolta L values, sarcomere length, and myosin heavy-chain composition were not different across groups. Chops from the low star probe group had a significantly greater marbling score (2.3 vs. 1.9) and lipid content (2.61 vs. 2.23%). Calpain-1 was completely autolyzed in both high and low star probe samples, demonstrating that calpain-1 potentially had been active in all samples. Low star probe whole-muscle protein extracts had more troponin-T ( < 0.01), desmin ( < 0.01), and filamin degradation ( < 0.01) than high star probe samples. Both classification groups showed degradation of titin. Remarkably, some high star probe samples still had observable intact bands of titin on SDS-PAGE gels. These results demonstrate that significant variation in instrumental tenderness is observed within a moderate pH range. Lipid content and proteolysis both appear to contribute to this variation.

A systematic review and meta-analysis of tenderness metrics in control groups used in comparative nutrition experiments.

This review reports the pork quality attributes, Warner-Bratzler Shear Force, Slice Shear Force, Star Probe, pH, marbling, color (Minolta L*/L or Hunter L*/L), and sensory tenderness evaluation, in control groups used in comparative nutrition experiments over the past 20 yr. The original aim of this study was to evaluate if changes in pork quality based on the above metrics occurred over time. To address this question, it was anticipated that data may come from 3 sources with decreasing relevance: representative retail pork surveys, representative post-harvest carcass surveys, and control groups from comparative nutrition experiments. To identify the study population, a review of studies reported in Centre for Agricultural Biosciences International Abstracts (Web of Knowledge; 1994-2014) was conducted. Two national level surveys of retail pork and 146 relevant nutritional experiments studies, with 228 control groups, were identified by the search. It was not possible to conduct a meta-analysis of the retail pork surveys based on only 2 time points. For the comparative studies, a random effects meta-analysis was conducted with year as a covariate to assess the impact of time on the outcome. In the absence of modifiers, there was no evidence of meaningful change in the mean Warner-Bratzler Shear Force, pH, color, marbling, or sensory scores over the study period. There was evidence of substantial between-study heterogeneity in the characteristics of control pigs used over the years for Warner-Bratzler Shear Force and measures of color. The absence of publicly-available representative surveys of pork quality meant the changes in pork quality over time were not clear. If changes in pork quality have occurred, the data suggest that pigs used as controls in experiments may have become less representative of commercial pigs over time and the translatability of study findings from nutrition experiments might be reduced over time. Alternately, if commercial pigs have not changed, then control pigs reflect this. The study does not address if control groups in other experimental intervention studies had similar tenderness patterns as reported here for nutritional interventions. A large amount of potentially available data was excluded from the analysis due to incomplete reporting in the original study reports.

Identification of potential serum biomarkers to predict feed efficiency in young pigs.

Identification of biomarkers for feed efficiency in livestock will aid in the efficient production of high-quality protein to meet the demands of a growing population. The overall objective of this research was to identify biomarkers in serum for swine feed efficiency and to discover pathways affected by divergent selection for residual feed intake (RFI). Serum was collected from young pigs (between 35 and 42 d of age) from 2 lines of pigs that have been genetically selected to be either more efficient (low-RFI) or less efficient (high-RFI). After blood collection, during finishing, pigs from each line were placed on either a low-energy/high-fiber diet or a traditional high-energy/low-fiber diet to test for any diet effects on RFI. Subsets of 6 pigs per line within each diet were used in 3 independent experiments. Pigs with extreme RFI phenotypes from the low-energy/high-fiber diet were used to confirm the results from the first 2 comparisons. Two-dimensional difference in gel electrophoresis and mass spectrometry were used to identify proteins with different abundances between RFI line or finishing diet. Three proteins had consistent and significant ( < 0.05) RFI line differences for both diets: gelsolin, vitronectin, and serine protease inhibitor A3 (serpinA3). Abundance of gelsolin, a protein with roles in actin filament assembly and immune response, was greater in the more efficient low-RFI pigs (9 to 39%). Vitronectin was also more abundant in the low-RFI pigs (39 to 56%) and has known roles in blood homeostasis and may regulate adiposity. SerpinA3 is a member of a very large family of proteins referred to as serine protease inhibitors. A total of 14 spots that were more abundant in the low-RFI line, some at least twice as abundant, were identified as serpinA3. Multiple isoforms of serpinA3 have been reported (serpinA3-1 to serpinA3-4 in pigs and serpinA3-1 to serpinA3-8 in cattle) with serpinA3 having many different functions dependent on isoform. Gelsolin, vitronectin, and serpinA3 are 3 proteins that may play direct and important biological roles in the pathways that control RFI and, ultimately, feed efficiency through energy utilization and homeostasis. These data demonstrate that serum proteins can be a useful source of potential biomarkers for feed efficiency and provide information on pathways with distinct expression patterns between animals that differ in feed efficiency.

Associations between two gene markers and indicator traits affecting fresh and dry-cured ham processing quality.

The study objective was to evaluate the effects of mutations in two genes, PRKAG3 and CAST, on fresh and dry-cured processing characteristics. Processing and curing followed normal commercial procedures. A 100 g sample of the semimembranosus was excised and frozen for later DNA harvest. Genotypes for CAST, and PRKAG3 were determined after DNA extraction from the frozen samples. The PRKAG3 marker had no affect on dry-cured ham processing characteristics. The CAST gene marker was a significant source (P<0.05) of variation for cured ham moisture content and tended to be a significant source (P<0.10) for yield, ham weight loss, salt content and Minolta color change. The data demonstrate that the CAST 11 genotype is associated with greater processing yields. Alternatively, selection or sorting for the CAST 22 genotype should produce cured hams that exhibit more efficient moisture loss and, as a result, require less processing time and contain greater salt content.

Influence of pH on rheological properties of porcine myofibrillar protein during heat induced gelation.

Texture of meat products is dependent on the gelation characteristics of myofibrillar protein. Gaining an understanding of the gelation mechanism of meat gel systems is beneficial for the development of processed meat products as well as maintaining quality in meat products. The aim of this study was to investigate the impact of pH (5.6, 6.0, 6.5, and 7.0) on heat-induced gelation properties of myofibrillar proteins from porcine semimembranosus muscle. Dynamic rheological measurements were taken as the temperature increased by 1°C/min from 20 to 85°C, followed by a holding phase at 85°C for 3min to ensure complete gelation and during a subsequent cooling where the temperature dropped from 85 to 5°C at a rate of 5°C/min. Storage modulus (G') increased as gel formation occurred, but decreased after reaching the temperature of myosin denaturation (52°C) until approximately 60°C when the gel strength increased again. This resulted in a peak and subsequent depression in the data. This depression in the curve was more pronounced with increasing pH. Results indicate protein denaturation and gel formation are pH dependent. Furthermore, rate of gelation appears to influence water-holding capacity.

Impact of supplementing vitamin C for 56, 90, or 127 days on growth performance and carcass characteristics of steers fed a 0.31 or 0.59% sulfur diet.

The objective was to examine differential timing of vitamin C (VC) supplementation during the finishing period (for the first 56, 90, or 127 d) on performance, VC, and glutathione (GSH) concentrations and carcass traits of steers receiving a 0.31 or 0.59% S diet. Angus steers (n = 42) were stratified to pens by initial BW (304 ± 13 kg) and GeneMax marbling score (4.3 ± 0.12), and pens were randomly assigned to 1 of 7 treatments (6 steers/pen and 1 pen/treatment), including a high-S (HS; 0.59% S) control (HS CON), HS CON + 10 g VC∙steer(-1)∙d(-1) for the first 56 d of the finishing period, HS CON + 10 g VC∙steer(-1)∙d(-1) for the first 90 d of the finishing period, HS CON + 10 g VC∙steer(-1)∙d(-1) for the entire 127-d finishing period (HS VC127), low-S (LS; 0.31% S) diet + 10 g VC∙steer(-1)∙d(-1) for the first 56 d of the finishing period (LS VC56), LS diet + 10 g VC∙steer(-1)∙d(-1) for the first 90 d of the finishing period, or LS diet + 10 g VC∙steer(-1)∙d(-1) for the entire 127-d finishing period. Jugular blood and ultrasound measures were taken from all steers before feeding on d 0, 56, 90, and 127, and liver biopsies and ruminal hydrogen sulfide measurements were collected on d 121 or 122. Steers (n = 40) were harvested on d 127, and carcass data were collected. Data were analyzed by ANOVA as a completely randomized design with the fixed effect of treatment. Because individual intake data were collected, steer was the experimental unit. Final BW and ADG were greater (P ≤ 0.03) and DMI tended (P = 0.09) to be greater in the LS steers compared to HS steers, but G:F did not differ (P = 0.41) by treatment. A treatment × time effect (P = 0.04) for DMI was noted, likely due to lesser DMI between d 91 and 127 for all treatments except the HS VC127 and LS VC56. Plasma VC concentrations of LS steers were less (P = 0.05) than the HS steers. Total (P = 0.06) and reduced (P = 0.03) plasma GSH were greater in HS steers supplemented with VC than the HS CON, but liver GSH were not different due to S or VC (P ≥ 0.13). The ratio of oxidized to reduced liver GSH was greater (P < 0.01) in HS CON than HS steers supplemented with VC. Marbling score, LM area, KPH, and quality grade were not different (P ≥ 0.19) due to diet, but LS steers had greater (P = 0.05) back fat than HS steers. In conclusion, steers fed a HS diet had poorer live performance and unexpectedly greater plasma ascorbate concentrations than the LS-fed steers. Interestingly, increasing days of VC supplementation across the HS diets increased GSH indices, suggesting that although HS diets may negatively affect antioxidant capacity of cattle, supplementing VC may help correct this.

Effects of duration of vitamin C supplementation during the finishing period on postmortem protein degradation, tenderness, and meat color of the longissimus muscle of calf-fed steers consuming a 0.31 or 0.59% sulfur diet.

High-S (HS) diets have been identified as a causative agent in the development of oxidative stress in cattle, which in postmortem muscle can negatively alter meat quality. Vitamin C (VC) is a potent antioxidant produced endogenously by cattle; however, exogenous supplementation of VC may be useful when HS diets are fed to cattle. The objective of this study was to examine the impact of duration of VC supplementation, for the first 56, 90, or 127 d, during the finishing period on meat color and tenderness of the longissimus thoracis (LT) collected from calf-fed steers consuming a 0.31 or 0.59% S diet. Angus steers ( n= 42) were stratified to pens by initial BW (304 ± 13 kg) and GeneMax marbling score (4.3 ± 0.12), and each pen was randomly assigned to 1 of 7 treatments (6 steers/pen, 1 pen/treatment), including HS (0.59% S, a combination of dried distillers grains plus solubles and sodium sulfate) control (HS CON), HS CON + 10 g VC·steer·(-1)d(-1) for the first 56 d (HS VC56), 90 d (HS VC90), or 127 d (HS VC127), low S (LS; 0.31% S) + 10 g VC·steer·(-1)d(-1) for the first 56 d (LS VC56), 90 d (LS VC90), or 127 d (LS VC127). Steers were harvested (n = 40) and, after a 24-h chill, rib sections (LT) were collected. pH was determined on each rib section before division into 3 sections for determination of 1) 7-d retail display and color and Warner-Bratzler shear force (WBSF), 2) 14-d WBSF determination, and 3) protein degradation and collagen content (2 d postmortem). Data were analyzed by ANOVA as a completely randomized design, with the fixed effect of treatment. Individual feed intake was recorded, and steer was the experimental unit. The HS steers had a greater and lesser percent of the 80- and 76-kDa subunits of calpain-1 (P ≤ 0.05), respectively, and tended to have less (P = 0.08) troponin T degradation (d2), and more (P = 0.02) collagen than LS steers. Increasing days of VC supplementation decreased (P = 0.05) the percentage of the 80 kDa subunit of calpain-1 in HS steers but actually increased it in LS steers (P= 0.003). Supplementing VC, regardless of dietary S, did not affect meat collagen, WBSF, or color (P ≥ 0.12). a* and b* values were greater (P ≤ 0.05) in the LS treatments compared to the HS treatments. Increasing the days of VC supplementation to steers fed a HS diet appears to alleviate the negative effects of the HS diet on calpain-1 but has no effect on muscle tenderness or meat color.

Investigation of the efficacy of albumin removal procedures on porcine serum proteome profile.

Improving the ability to predict livestock performance using biomarkers will provide a benefit for livestock genetic evaluation and improvement. The most practical biological sample to screen for development of biomarkers is serum due to the ease of collection. However, protein profiles in serum are complex and dynamic. Strategies are needed to manage variation in serum proteins used for biomarker identification. Albumin is the most abundant protein in serum, comprising over 50% of the overall protein content, and has historically been depleted from serum before biomarker identification. The objective of this study was to investigate the use of gel-based proteomic techniques to evaluate the need for porcine albumin depletion in biomarker identification. Albumin is known to bind many proteins in the blood, thus potential biomarkers could be removed during albumin depletion. Using two-dimensional difference in gel electrophoresis (2D-DIGE), we show whole serum can be used for biomarker discovery. The data obtained show that albumin removal methods are effective for porcine sera. Over 85% of the protein spots resolved on at least half of the gels were changed in abundance between whole and albumin depleted sera. Of the 204 protein spots significantly altered in abundance, 59 were changed over 400%. However, albumin removal also altered the serum proteome in an unpredictable manner; in the depleted sera, 86 protein spots were increased in abundance and 118 were decreased. Furthermore, the abundance of 59.4% of the protein spots in the albumin depleted samples had a larger standard error than whole sera. However, the resolution of albumin in 2D-DIGE analysis of whole sera permitted the detection and quantification of substantial numbers of proteins. Thus, it is proposed that whole serum can be used in a gel-based proteomics system for the identification of porcine biomarkers.