VTechWorks Repository :: Browsing by Author "Johnson, Sally E."

Browsing by Author "Johnson, Sally E."

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Additive effects among uterine paracrine factors in promoting bovine trophoblast cell proliferation
Xie, Ming (Virginia Tech, 2014-06-10)
Several uterine-derived paracrine factors have been implicated as critical regulators of conceptus development in cattle, but it remains unclear how these factors work together to establish and maintain pregnancies. The primary objectives of this work were to establish if cooperative interactions between epidermal growth factor (EGF), fibroblast growth factor-2 (FGF2) and insulin-like growth factor-1 (IGF1) promote bovine trophoblast cell proliferation, and to decipher the intracellular signaling mechanisms employed by these growth factors to regulate cell proliferation. Pilot studies established effective concentrations for each growth factor on a bovine trophoblast cell line (CT1). The first set of studies examined how each factor worked individually or in conjunction with each other to impact CT1 proliferation. Mitotic index (percentage of EdU-positive nuclei after a 45 min challenge) was increased (P<0.05) by supplementation with 10 ng/ml EGF, 10 ng/ml FGF2, or 50 ng/ml IGF1 when compared with non-treated controls. In addition, a greater increase (P<0.05) was detected when all three factors were supplemented together. A follow-up study determined that supplementation of any two growth factors could not replicate the cooperative effect noted when all three factors were provided. A second set of studies was undertaken to examine how mitogen-activated protein kinase (MAPK) and phosphoinositide 3-kinase/AKT (PI3K/AKT) signaling systems mediate the independent and cooperative effects of these paracrine factors. Both EGF and IGF1 transiently activated mitogen-activated protein kinase3/1 (MAPK3/1) in CT1 cells as determined by Western Blot analysis. By contrast, FGF2 did not affect MAPK3/1 phosphorylation status, but increased AKT phosphorylation status. Neither EGF nor IGF1 impacted AKT activity. Supplementation with a pharmacological inhibitor of MAPK3/1 (PD98059) prevented EGF-, IGF1-, and FGF2-dependent increases in CT1 cell proliferation. This inhibitor also completely abolished the increases in cell proliferation observed when all three factors were supplemented together. Supplementation with a pharmacological inhibitor of AKT (Wortmannin) reduced FGF2-stimulated CT1 proliferation, but did not impact EGF- and IGF1 effects. The AKT inhibitor partially attenuated the cooperative effects of all three factors on CT1 cell proliferation. A final study examined how the combination of EGF, FGF2, and IGF1 affect bovine embryo development. In vitro produced bovine blastocysts were cultured either with the combination of growth factors or vehicle only from day 8 to day 12 post-in vitro fertilization (IVF). The combination of EGF, FGF2, and IGF1 increased (P<0.05) the percentage of hatched blastocysts and outgrowth formation versus controls. Increased (P<0.05) diameters were detected in blastocysts treated with the combination of three growth factors on day 12 post-IVF when compared to controls. Treatment with the combination of EGF, FGF2, and also IGF1 increased (P<0.05) the change of diameter from day 8 to 12 post-IVF. In summary, these observations provide evidence that cooperative interactions of uterine-derived factors promote trophoblast proliferation and conceptus development in ways that may promote the establishment and maintenance of pregnancy in cattle. The mechanisms utilized for these activities remain unresolved, but MAPK3/1 and PI3K/AKT signaling systems appear to play integral roles in some of these processes.
Biochemical Basis of Fresh Ham Color Development
Stufft, Kristen Marie (Virginia Tech, 2015-09-14)
Commercial hams display variation in color uniformity across the cut surface, especially the semimembranosus (SM) muscle. This variation in fresh ham color, or two-toning, persists through further processing and contributes to production of a less desirable end product. In an attempt to understand the underlying source of this color variation, we evaluated the differences in muscle fiber-type composition and glycolytic metabolism in the SM muscle of fresh hams differing in color uniformity. Fifty-eight fresh SM muscles, ranging in color, were selected at 24 h postmortem and each partitioned into four distinct regions and three color classes based on color uniformity in the caudal region. The L* (lightness) values were greatest and a* (redness) values lowest in the most caudal portions of the muscle. The caudal portion also exhibited the lowest ultimate pH (P < 0.0001), lowest myoglobin (P < 0.05), greatest glycolytic potential (GP) (P < 0.0001) and the lowest myosin heavy chain type I isoform (P < 0.0001) abundance of all regions in 'normal' colored hams. After segregating based on L* values, the caudal region had identical pH, GP, LDH, and MyHC-I, despite significant differences in L* (P < 0.0001). These data show the most caudal aspects of the SM are indeed more prone to adverse postmortem metabolism and suggest that inherent differences in muscles of the ham may make some areas of the ham more vulnerable to temperature abuse during harvesting.
A Carnitine-Containing Product Improves Aspects of Post-Exercise Recovery in Adult Horses
Johnson, Sally E.; Barshick, Madison R.; Gonzalez, Madison L.; Riley, Julia Wells; Pelletier, Megan E.; Castanho, Beatriz C.; Ealy, Elayna N. (MDPI, 2023-02-14)
Strenuous exercise can cause tissue damage, leading to an extended recovery period. To counteract delayed post-exercise recovery, a commercial product containing L-carnitine (AID) was tested in adult horses performing consecutive exercise tests to exhaustion. Fit Thoroughbreds were administered an oral bolus of placebo (CON) or AID prior to performing an exercise test to exhaustion (D1). The heart rate (HR) and fetlock kinematics were captured throughout the exercise test. Blood was collected before, 10 min and 1, 4 and 6 h relative to exercise for the quantification of cytokine (IL1β, IL8, IL10, TNFa) gene expression and lactate concentration. Horses performed a second exercise test 48 h later (D2), with all biochemical and physiological measures repeated. The results demonstrate that the horses receiving AID retained a greater (p < 0.05) amount of flexion in the front fetlock on D2 than the horses given CON. The horses presented a reduced (p < 0.05) rate of HR decline on D2 compared to that on D1. The expression of IL1β, IL8 and IL10 increased at 1 h post-exercise on D1 and returned to baseline by 6 h; the cytokine expression pattern was not duplicated on D2. These results provide evidence of disrupted cytokine expression, HR recovery and joint mobility in response to consecutive bouts of exhaustive exercise. Importantly, AID may accelerate recovery through an undetermined mechanism.
Conversion of equine umbilical cord matrix mesenchymal stem cells to the trophectoderm lineage using the Yamanaka reprogramming factors
Reinholt, Brad M. (Virginia Tech, 2015-07-21)
Induced pluripotent stem (iPS) cells that possess embryonic stem (ES) cell-like properties are generated through the use of the Yamanaka transcription factors, OCT4, SOX2, KLF4, and MYC (OSKM). Advanced transgene delivery methods utilizing non-integrating viruses for transduction of target cells has provided new opportunities for regenerative medicine in humans and other species. We sought to use this technology to generate equine iPS cells to address challenges in equine regenerative medicine. Umbilical cord matrix mesenchymal stromal cells (MSC) were transduced with the non-integrating Sendai virus encoding for the OSKM transcription factors. The cells initially were cultured on mouse embryonic feeder cells supplemented with LIF (10 ng/mL) and FGF2 (4 ng/mL). Transduction generated 21 initial colonies. Of these, four survived beyond 20 passages. The transduced equine cells morphologically resembled ES cells and expressed cell surface antigens indicative of ES cells. Molecular evaluation revealed the cells maintained expression of endogenous OSKM while the exogenous OSK transgenes were extinguished, but MYC was maintained. The transduced equine cells did not express the ES marker NANOG, but did express the trophectoderm markers CDX2 and TFAP2A. Both OCT4 and CDX2 were colocalized to the nucleus. The transduced equine cells were termed equine induced trophoblast (iTr) cells. Culture of the iTr cell in suspension resulted in formation of blastocyst-like spheres rather than solid cell aggregates indicative of ES and iPS cells. The iTr cells were transitioned to a feeder free monolayer culture. Transformation of the iTr cells to the spherical arrangement stimulated expression of genes that mark differentiation of trophoblast cells and up-regulated 250 transcripts over the monolayer arrangement. The iTr monolayer arrangement up-regulated 50 transcripts compared to the spherical arrangement. The iTr spheres respond to BMP4, EGF, and FGF2 by phosphorylating signal transduction proteins. Addition of BMP4, EGF, or FGF2 in combined pairs was able to alter TFAP2A, NEU1, and SLC35A1 expression. The generation of iTr cells by transduction of the Yamanaka reprogramming factors is not unique to equine cells. However, this report marks the generation of the first equine trophoblast cell line capable of recapitulating early equine trophoblast development. The new iTr line could prove valuable in gaining greater understanding of equine trophectoderm development.
Defining the role of mitochondria in fresh meat quality development
Matarneh, Sulaiman K. (Virginia Tech, 2017-07-12)
During postmortem metabolism, hydrogen ions accumulate in the muscle and gradually lower the pH from 7.2 to an ultimate pH near 5.6. The ultimate pH of meat is widely valued as an indicator of fresh meat quality as it directly affects the quality characteristics of color, texture, and water holding capacity. Therefore, our research was conducted to identify the processes responsible for determining ultimate pH. Pigs harboring the AMPKï¿½•3R200Q mutation produce meat with extremely low ultimate pH (pH ~ 5.3) that is detrimental to quality. This phenomenon is often attributed to a greater glycogen content in muscle from the mutant pigs compared to wild-type pigs. However, our research indicated that greater glycolytic flux in muscle from these pigs causes a lower ultimate pH rather than greater tissue glycogen deposition. On the other hand, however, AMPKï¿½•3R200Q pigs contain more mitochondria and retain greater oxidative capacity. Hence, we hypothesized that mitochondria may contribute to the lower ultimate pH in muscle of these pigs. To test our hypothesis, isolated mitochondria were incorporated into an in vitro system the mimics postmortem glycolysis. Mitochondria enhanced glycolytic flux and pH decline in the in vitro system similar to that of AMPKï¿½•3R200Q pigs. After a series of experiments, we found that the causative agent for enhanced glycolytic flux is a soluble mitochondrial protein. In other experiments, mitochondrial F1-ATPase was found to be responsible for the majority of this effect, principally through promoting greater ATP hydrolysis at lower pH values, thereby allowing for greater flux through glycolysis. These data suggest that variations in ultimate pH may be more thoroughly explained and predicted by the abundance of mitochondria. Broiler pectoralis major muscle, which is a highly glycolytic muscle, possesses high ultimate pH (pH ~ 5.9) compared to pork and beef. We postulated that rapid carcass chilling reduces the flux through glycolysis, thereby causing premature termination of postmortem metabolism. Yet, chilling was only partially responsible for the high ultimate pH of pectoralis major muscle. However, we showed that pectoralis major of broiler chicken exhibits lower phosphofructokinase-1 activity compared to porcine longissimus lumborum muscle, which limits the flux through glycolysis.
Determining muscle plasticity and meat quality development of low-input extended fed market-ready steers
Wicks, Jordan C.; Wivell, Alexis L.; Beline, Mariane; Zumbaugh, Morgan D.; Bodmer, Jocelyn S.; Yen, Con-Ning; Johnson-Schuster, Chantal; Wilson, Thomas B.; Greiner, Scott P.; Johnson, Sally E.; Shi, Tim H.; Silva, Saulo Luz; Gerrard, David E. (Oxford University Press, 2024-05-02)
In March 2020, the World Health Organization declared COVID-19 a pandemic, which ultimately led to many meat processors temporarily shutting down or reducing processing capacity. This backlog in processing capacity forced many feedlots to retain cattle for longer periods of time and assume the risk of major market fluctuations. The aim of this study was to understand how a dietary insult affects meat quality and muscle metabolism in market-ready steers (590 kg). Sixteen market-ready (590 kg) commercial Angus crossbred steers were subjected to a maintenance diet of either forage or grain for 60 d. Longissimus lumborum (LL) muscle samples were collected immediately postmortem and processed for characteristics reflecting the underlying muscle fiber type and energy state of the tissue. Despite cattle being subjected to a 60-d feeding period, there were no detectable differences (P > 0.05) in carcass characteristics, color of lean, or ultimate pH (pHu). Moreover, our data show that muscle plasticity is rather resilient, as reflected by lack of significance (P > 0.05) in oxidative and glycolytic enzymes, myosin heavy chain isoforms (MyHC), myoglobin, and mitochondrial DNA (mtDNA) contents. These data show that market-ready steers are capable of withstanding a low-input feeding strategy up to 60 d without dramatically impacting underlying muscle characteristics and meat quality development.
Diminished satellite cell fusion and S6K1 expression in myotubes derived from skeletal muscle of low birth weight neonatal pigs
Chen, Ying; Zhu, Haibo; McCauley, Sydney R.; Zhao, Lidan; Johnson, Sally E.; Rhoads, Robert P.; El-Kadi, Samer Wassim (The American Physiological Society, 2017-02-09)
Low birth weight (LBWT) is consistently associated with impaired postnatal muscle growth in mammals. Satellite cell (SC)‐mediated myonuclear incorporation precedes protein accumulation in the early stages of postnatal muscle development and growth. The objective of this study was to investigate proliferation and differentiation of SCs and the regulation of protein synthesis signaling in response to insulin‐like growth factor (IGF)‐I stimulation in SC‐derived myotubes of LBWT neonatal pigs. SCs isolated from Longissimus dorsi muscle of LBWT and NBWT pigs (3‐d‐old, n = 8) were cultured and induced to proliferate and differentiate to myotubes in vitro. On day 3 of differentiation, myotubes were fasted in serum‐free media for 3 h and treated with human recombinant R³‐insulin‐like growth factor‐I (rh IGF‐I) at 0, 25, and 50 ng × mL⁻¹ for 30 min. There was no difference in proliferation rates of SCs from LBWT and NBWT pigs. However, LBWT SC fusion was 15% lower (P ≤ 0.05) without a difference in MyoD or myogenin mRNA expression in comparison with NBWT pigs, suggesting SCs are not intrinsically different between the two groups. IGF‐Ι stimulation at physiological concentrations activated downstream effectors of mTOR similarly in myotubes from LBWT and NBWT pigs. However, abundance of ribosomal protein S6 kinase 1(S6K1) was lower in myotubes of LBWT compared to their NBWT siblings (P ≤ 0.05). These results indicate that the modest reduction in SC fusion and S6K1 expression are not the major contributors to the impaired postnatal muscle growth of LBWT pigs.
Downregulated Translation Initiation Signaling Predisposes Low-Birth-Weight Neonatal Pigs to Slower Rates of Muscle Protein Synthesis
Chen, Y.; McCauley, Sydney R.; Johnson, Sally E.; Rhoads, Robert P.; El-Kadi, Samer Wassim (Frontiers, 2017-07-11)
Low-birth-weight (LBWT) neonates experience restricted muscle growth in their perinatal life. Our aim was to investigate the mechanisms that contribute to slower skeletal muscle growth of LBWT neonatal pigs. Twenty-four 1-day old male LBWT (816 ± 55 g) and normal-birth-weight (NBWT; 1,642 ± 55 g) littermates (n = 12) were euthanized to collect blood and longissimus dorsi (LD) muscle subsamples. Plasma glucose, insulin, and insulin-like growth factor-I (IGF-I) were lower in LBWT compared with NBWT pigs. Muscle IGF-I mRNA expression were lower in LBWT than NBWT pigs. However, IGF-I receptor mRNA and protein abundance was greater in LD of LBWT pigs. Abundance of myostatin and its receptors, and abundance and phosphorylation of smad3 were lower in LBWT LD by comparison with NBWT LD. Abundance of eukaryotic initiation factor (eIF) 4E binding protein 1 and mitogen-activated protein kinase-interacting kinases was lower in muscle of LBWT pigs compared with NBWT siblings, while eIF4E abundance and phosphorylation did not differ between the two groups. Furthermore, phosphorylation of ribosomal protein S6 kinase 1 (S6K1) was less in LBWT muscle, possibly due to lower eIF3e abundance. In addition, abundance and phosphorylation of eIF4G was reduced in LBWT pigs by comparison with NBWT littermates, suggesting translation initiation complex formation is compromised in muscle of LBWT pigs. In conclusion, diminished S6K1 activation and translation initiation signaling are likely the major contributors to impaired muscle growth in LBWT neonatal pigs. The upregulated IGF-I R expression and downregulated myostatin signaling seem to be compensatory responses for the reduction in protein synthesis signaling.
Effect of amino acids and vitamin D3 on performance and biological responses in poultry
Wen, Jinlei (Virginia Tech, 2018-06-08)
As productive performance is improved by breed selection, amino acid requirements may change to support this higher performance in poultry. The first objective of this dissertation was to update the valine and tryptophan requirement of small-framed laying hens and the lysine requirement of young broilers using empirical dose-response methods. The tryptophan requirement was estimated as 155.8 mg/d for egg mass, 153.2 mg/d for egg production and 140.4 mg/d for feed conversion ratio using a linear broken line model. For valine, the requirement was highest for egg mass, 597.3 mg/d, followed by egg production, 591.9 mg/d and feed conversion ratio (FCR), 500.5 mg/d. The lysine requirement of young chicks was estimated by conducting four short term experiments from 1 to 3, 3 to 5, 5 to 8 and 8 to 11 days of age, respectively. The lysine requirement from 1 to 3, 3 to 5 and 5 to 8 days of age were not able to be estimated as no dose response was observed on growth performance most likely due to an overestimation of the lysine requirement. Digestible lysine requirement from 8 to 11 days of age was 1.057%, 1.050% and 1.016% based on body weight gain, FCR and pectoralis major weight using a linear broken line model, respectively. In addition to determining amino acid requirements, research was conducted to develop a new bacterial protein meal for use in laying hens diets. The data suggested that diets containing 7.5% of the bacterial protein meal was able to at least maintain egg production in laying hens, but 15% bacterial protein meal resulted in reduced performance. The second objective of this dissertation was to investigate the effects of various concentrations of dietary vitamin D3 on pullet and laying hen performance, eggshell quality and bone health in laying hens. Pullets/hens were randomly assigned to five dietary treatments containing vitamin D3 from 1,681 to 68,348 IU/kg diet from day of hatch until 68 weeks of age. These data suggested that dietary vitamin D3 fed at 68,348 IU/kg resulted in reduced egg production, but vitamin D from 8,348 to 35,014 IU/kg diet maintained egg production, increased egg vitamin D content in a dose dependent manner, and generally increased both eggshell quality and pullet and hen bone mineral status.
The Effect of Phytonutrient Supplementation on Pig Growth
Seligman, Lexie Camille (Virginia Tech, 2024-08-26)
As the global population continues to increase, the demand for livestock production also rises. This has led to growing interest in efficient approaches to enhance animal growth and development. Phytonutrients are considered a promising alternative to synthetic compounds to improve animal growth performance. The objective of the study was to examine the effects of varying doses of phytonutrient supplementation on pig growth and metabolism. Thirty-two crossbred pigs (approximately 2 months of age, 17.7 kg± 0.82 kg) were randomly assigned to one of four phytonutrient groups (Control: 0 ppm; 1: 62.5 ppm; 2: 125 ppm; 3: 250 ppm) fed ad libitum for 28 days. Body weight was recorded on days -5, -1, 0, 7, 14, 21, and 28. Feed intake was recorded daily, and body composition measured by dual x-ray absorptiometry (DEXA) was obtained on days -1, 14, and 28. Pigs increased (P<0.0001) in body weight, irrespective of treatment (P≤0.0535). Control pigs had greater average daily gain (ADG) when compared with other diets (P<0.001). Treatment 3 had the highest (P<0.05) feed:gain when compared with other treatments. Lean percentage of body weight decreased (P<0.0001) while fat mass increased (P<0.0001) over the duration of the study. Overall, supplementation of this phytonutrient blend did not show significant improvement to the growth of the pigs.
The effects of Bacillus subtilis, dietary calcium and coccidial vaccines in broilers experiencing naturally occurring necrotic enteritis
Calvert, Alamanda J. (Virginia Tech, 2022-06-03)
Necrotic enteritis (NE) is a disease that negatively affects broiler performance and has increasingly become a problem with the reduced use of antibiotics. There is increased interest to utilize alternative methods including coccidial vaccines and direct fed microbials (DFM) to reduce or prevent NE. Additionally, it has been demonstrated that reduced dietary Ca can reduce NE mortality. The objective of the first study was to examine the effects of Bacillus subtilis (DFM) in combination with attenuated (AV) and non-attenuated (NV) coccidial vaccines in broilers with NE. The objective of the second study was to determine the effects of DFM, antibiotic (BMD) or negative control (NC) with standard dietary Ca (0.9%) or below standard Ca (0.75%, 0.6%) in broilers with NE. To induce a natural NE occurrence, birds were vaccinated with the appropriate coccidial vaccine and then placed on reused litter from a previous flock exhibiting NE lesions and mortality. In the first trial DFM reduced BW and BW gain (P ≤ 0.05) during the starter phase (0 to 14 d). The addition of a DFM in the AV group resulted in decreased (P ≤ 0.05) BW gain and BW at 42 d. The DFM in the NV group resulted in improved (P ≤ 0.05) mortality corrected feed conversion ratio from 0 to 42 d. Overall, with a natural NE occurrence, the effect of DFM was variable depending on what type of coccidial preventative was utilized and was beneficial when used with NV. In the second trial BW and BW gain were increased in broilers fed 0.60% dietary Ca and DFM compared to 0.90% Ca and NC (P ≤ 0.05). Broilers fed 0.60% Ca had reduced NE mortality compared to broilers fed 0.90% Ca (P ≤ 0.05). Reducing dietary Ca to below industry standards was able to lessen the severity of NE though the addition of DFM was needed to improve BW. Overall, with a natural NE occurrence, the DFM was beneficial when used with NV and in diets with 0.6% dietary Ca.
Effects of beta-hydroxy beta-methylbutyrate (HMB) supplementation on gluteus medius muscle fiber composition and muscle performance in adult Thoroughbred horses exercising to fatigue on a high-speed treadmill
Busse Esser, Nicolas Ignacio (Virginia Tech, 2021-09-16)
Consumption of β-hydroxy β-methylbutyrate (HBM), a leucine metabolite, alters muscle composition and metabolism leading to strength and agility improvements in human athletes. To determine if HMB affects athletic performance and muscle function in horses, Thoroughbred geldings were fed a control (CON; n=5) or HMB (n=6) supplement (30 mg/kg/day) for 6 weeks prior to completing a standardized exercise test (SET). Gluteus medius (GM) muscle samples were obtained before the SET for fiber-typing and venous blood was collected before and immediately upon completion of the SET for lactate measurements. Heart rate (HR), biceps femoris (BF) and semitendinosus (ST) surface electromyograms, and fore- and hindlimb metacarpophalangeal joint angles were captured for the duration of the SET. Results demonstrate that HMB supplementation increased (P < 0.05) the percentage of type IIA muscle fibers in the GM with a corresponding decrease (P < 0.05) in type IIX fibers. The percentage of type I fibers was unaffected by diet. Supplementation with HMB did not result in any significant effects on performance, muscle function or biomechanical properties by comparison to CON. Increasing treadmill speed resulted in an increase (P < 0.05) in stride length and maximal extension angle of the fore fetlock, and a shortening (P < 0.05) of the stance phase of the gait cycle. Integrated EMG (iEMG) increased (P < 0.05) with increasing treadmill speeds for both the BF and ST, with the BF exhibiting greater iEMG values than the ST. In summary, HMB increased the percentage of type IIA fibers which did not translate into immediate, improved athletic performance
Effects of early weaning on finishing feed efficiency, marbling development and retail product quality of beef steers
Smith, Jason Kaleb (Virginia Tech, 2015-09-03)
Beef cattle producers continuously search for nutritional management options that provide flexibility to production scenarios. Due to its positive effects on maternal productivity, early weaning is one such alternative strategy that has received considerable interest. To better understand the effects of early weaning on calf productivity, an extensive literature review and three experiments were conducted to evaluate the effects of early weaning on finishing feed efficiency, marbling development and retail product quality of beef steers. In experiment I, which included 90 Angus-sired steers from four calving seasons, early weaning followed by a short ad libitum concentrate-feeding and pasture-backgrounding phase reduced finishing residual feed intake (RFI) by 7 % (P < 0.0001) and increased carcass marbling score (MS) by 10 % (P < 0.01) when compared to conventionally weaned (CW) contemporaries. Similar effects were observed in experiment II, which included 28 Angus and Simmental-sired steers, as early weaning reduced RFI (P < 0.01) and increased carcass MS (P < 0.01). Lung mass of early weaned (EW) steers was greater than their CW contemporaries (P < 0.05), and was inversely related to RFI (R2 = 0.17; P < 0.05). Finishing treatments in this experiment included a high corn ration and an alternative low corn ration that iso-calorically replaced 50 % of the DM from corn with dried corn gluten feed. Iso-caloric replacement of corn reduced lung mass (P < 0.01), and when combined with the observed increase of EW steers suggests that lung development is affected by dietary energy type at various stages of growth. In experiment III, objective analyses of ribeye steaks obtained from steers included in experiment two revealed that early weaning increased cross-sectional muscle fiber area by 28 % (P < 0.001) and tended to increase (P = 0.08) Warner-Bratzler shear force by 36 %. Nonetheless, these effects were not great enough to alter un-trained consumer perception of texture (P ≥ 0.65), juiciness (P ≥ 0.55), flavor (P ≥ 0.25) or overall acceptability (P ≥ 0.34). Collectively, these results indicate that early weaning enhances finishing feed efficiency and carcass marbling without affecting un-trained consumer sensory perception.
The effects of furosemide on equine skeletal muscle satellite cell myogenesis and metabolism in vitro
Helsel, Patricia J. (Virginia Tech, 2020-01-29)
Thoroughbred racehorses undergo strenuous exercise which often leads to the occurrence of exercise-induced pulmonary hemorrhage (EIPH), in which capillaries rupture within the alveoli in the lungs causing bleeding. Severe cases of EIPH lead to epistaxis and may result in fatality. Presently, the loop diuretic furosemide is the only medication approved to mitigate the effects of EIPH. Often regarded in the racing industry as "performance enhancing" due to 4% weight loss ensued by its diuretic effect, it is unknown what effects furosemide may have on muscle recovery. Therefore, the objective of this study was to determine the effects various doses of furosemide may have on equine satellite cell (eqSC) myogenesis and metabolism. Mitotic index was increased (P<0.05) as a result of treatment with 100 µg/mL furosemide, a 10-fold pharmacological dose, in comparison to vehicle, but was not different (P>0.05) compared to the physiological dose of 10 µg/mL furosemide. Average cell number decreased (P<0.05) in the excess furosemide group compared to all other groups. Pax7 expression did not differ (P>0.05) between groups. Expression of the differentiation transcription factor myogenin, and embryonic sarcomeric myosin heavy chain decreased (P<0.05) when cells were treated with 100 µg/mL furosemide. Fusion index and myotube area decreased (P<0.05) as a result of treatment with excess furosemide. Glycogen concentration in myotubes was lower (P<0.05) following treatment with 100 µg/mL furosemide, while IGF-1 was unsuccessful in rescuing the effects of furosemide. Excess furosemide decreased expression of muscle creatine kinase while increasing expression of phosphoglucomutase 1, glycogen synthase 1, and glycogen branching enzyme 1 (P<0.05). Excess furosemide decreased basal oxygen consumption rate (OCR) and increased OCR after addition of oligomycin (P<0.05). Excess furosemide did not affect myotube glycolysis rates in vitro. In conclusion, furosemide inhibits muscle differentiation and oxidative metabolism in eqSCs.
The effects of hyaluronic acid and exercise on equine skeletal muscle
Gregg, Savannah Renee (Virginia Tech, 2023-08-18)
Unaccustomed, strenuous exercise can cause skeletal muscle damage that subsequently induces an acute inflammatory response in the tissue which is marked by an infiltration of leukocytes into the damaged muscle. To try and suppress the initial pro-inflammatory response in skeletal muscle of horses performing a single exercise stress test, a commercial sodium hyaluronate (HA) treatment was administered and tested for anti-inflammatory properties. Unfit, adult Thoroughbreds were intravenously injected three times with HA or received no injection at all (CON) over a 3-week period before performing a single submaximal exercise test. Gluteal muscle biopsies were collected before and 1 h after the completion of exercise for RNA-Seq and staining. The results indicated that HA treatment in horses down regulated genes associated with lymphocyte activation and cytokine production (Il17RA, OSCAR, LYL1, TLR1, TLR2, TLR8, TLR10) but did not irreversibly down regulate these genes with the addition of exercise. Exercise as a stressor did cause an acute inflammatory response in muscle which was seen through global expression of macrophage and neutrophil surface markers (NCF2, ELANE, CD168I). These results determine that HA treatment does act as an anti-inflammatory in equine skeletal muscle but does not possess prolonged effects with the initiation of inflammation.
Effects of mid-gestational L-citrulline supplementation to twin-bearing ewes on umbilical blood flow, placental development, and lamb production traits
Kott, Michelle Lynn (Virginia Tech, 2021-01-11)
The interaction between the embryo and fetus with the maternal environment can have both short- and long-term consequences on health and development after birth. In some cases, these changes may be detrimental to the individual, but in other cases these developmental changes may be beneficial and manipulated to produce desired effects. Our interest is to use this concept of fetal programming to improve skeletal muscle development and meat production in ruminants. To achieve this, we targeted the period of gestation when fetal muscle fiber formation occurs. Primary muscle fibers form during embryonic development, and it is this small number of primary muscle fibers that will serve as templates for secondary fiber formation that occurs in the fetus during mid-gestation. Supplementing amino acids that influence blood flow within the reproductive tract is one potential way to provide fetuses with added nutrients during gestation, and this supplementation strategy may be especially useful when the maternal diet is compromised. L-citrulline was chosen for this work because of its long half-life in maternal circulation. This work utilized twin-bearing ewes with a moderate dietary energy restriction to assess the effects of mid-gestational L-citrulline supplementation on umbilical blood flow, placental function, neonatal lamb size, and lamb performance. We hypothesize that i.v. administration of L-citrulline will increase uterine and placental blood flow in gestating ewes and this will improve fetal growth, development, and overall postnatal performance. Blood flow parameters were not influenced by treatment (P>0.05). Circulating levels of progesterone and pregnancy-specific protein B (PSPB) were used as indicators of placental function and were unaffected by treatment administration (P>0.05). A treatment by time interaction was detected in both analyses, but no differences between treatments were detected within any time points. There was no effect of treatment on lamb weights or survival to weaning (P>0.05). Lamb sex effects are absent with the exception that body weights were greater in ewe lambs (P>0.05). There was no effect of treatment on any carcass traits or visceral organ weights assessed, though there was an effect of sex on dressing percentage and pancreas weight with wethers having a greater dressing percentage and heavier pancreases per kg body weight than that of ewes (P<0.05). In summary, contrary to our hypothesis L-citrulline supplementation to pregnant ewes under a minor to moderate metabolic challenge had no impact on blood flow and provided no programming benefit to the lambs.
Effects of mid-gestational l-citrulline supplementation to twin-bearing ewes on umbilical blood flow, placental development, and lamb production traits
Kott, Michelle L.; Pancini, Stefania; Speckhart, Savannah L.; Kimble, Lauren N.; White, Robin R.; Stewart, Jamie L.; Johnson, Sally E.; Ealy, Alan D. (Oxford University Press, 2021-07)
The objective of the study was to examine how l-citrulline supplementation to ewes during mid-gestation influences placental activity, placental blood flow, lamb body weight, and carcass characteristics. Two studies were completed. A pharmacokinetic study to compare circulating plasma amino acid concentrations after a single intravenous injection of 155 µmol/kg BW l-citrulline or after an isonitrogenous amount of l-alanine (control; 465 µmol/kg BW). Increases (P < 0.05) in circulating citrulline concentrations were detected for 8 h after l-citrulline injection versus the control. Similarly, increases (P < 0.05) in circulating arginine concentrations were detected for 24 h after l-citrulline treatment. The second study used 12 ewes with twin pregnancies. Daily intravenous injections of either l-citrulline or l-alanine were administered for 39 d from d 42-45 to 81-84 of gestation. Ewes were limit-fed at 85% daily energy requirements during the injection period. A decrease (P < 0.0001) in body weight was observed in both treatment groups during this period. No treatment differences were observed in circulating pregnancy-specific protein B concentrations or placental blood flow during the treatment and post-treatment gestational period. No treatment differences were observed in lamb survival nor in lamb birth, weaning and slaughter weights. Treatment did not influence lamb carcass composition or organ weights. However, there was a tendency (P = 0.10) for an increase in antral follicle numbers in ovaries from ewe lambs derived from ewes treated with l-citrulline. In summary, a daily l-citrulline injection increased both circulating citrulline and arginine concentrations in ewes, but daily l-citrulline injections during mid-gestation did not produce any detectable changes in placental activity and blood flow, neonatal and postnatal lamb development, and lamb carcass composition at slaughter. In conclusion, no benefits in placental function and lamb development were observed after providing l-citrulline during mid-gestation in ewes exposed to a mild energy restriction, but there was an indication that follicle numbers in ewe lambs were positively influenced by l-citrulline treatment during fetal development.
Effects of nutrient restriction on the metabolic profile of Bos indicus-influenced and B. taurus suckled beef cows
Fontes, Pedro L. P.; Oosthuizen, N.; Ciriaco, F. M.; Sanford, C. D.; Canal, L. B.; Cooke, Reinaldo F.; Pohler, K. G.; Henry, D. D.; Mercadante, Vitor R. G.; Ealy, Alan D.; Johnson, Sally E.; DiLorenzo, N.; Lamb, G. C. (2021-03)
Recent research from our group demonstrated that Bos indicus-influenced suckled beef cows had greater resilience to withstand nutrient restriction and establish pregnancy compared with B. taurus cows exposed to the same conditions. To further understand these findings, differences in metabolic profile between these same B. indicus-influenced and B. taurus females were explored. Suckled beef cows (n = 134) were enrolled in a completely randomized design with a 2 x 2 factorial arrangement of treatments. On day - 21, Angus (AN; Bos taurus) and Brangus (BN; B. indicus-influenced) cows were randomly assigned to 1) a diet that met daily energy maintenance requirements (MAINT), or 2) a diet that restricted intake to 70% of the daily energy maintenance requirements (RESTR). Cows were exposed to an estrus synchronization protocol and received an embryo 7 d after ovulation was pharmacologically induced on day 0. Blood samples were collected on days - 21 and 19 to determine circulating concentrations of non-esterified fatty acids (NEFA), beta-hydroxybutyrate (BHB), insulin, glucose, and IGF-1. Pregnancy status after embryo transfer was determined on day 28. As a consequence of the proposed diets, cows in the RESTR diet had less body condition score (BCS) on day 19 (P = 0.008) across breed types. Moreover, BCS change from day - 21 to 19 was included as independent covariate into subsequent analyses, allowing for the comparison of breed types under an equivalent level of body reserve mobilization. A breed x diet interaction was observed for plasma insulin (P = 0.03) and IGF-1 (P = 0.04) on day 19, where AN-RESTR cows had less plasma concentrations on day 19 compared with AN-MAINT cows. Diets did not impact (P > 0.10) plasma insulin and IGF-1 concentrations in BN cows. No diet or breed effects were observed in circulating concentrations of NEFA, BHB, and glucose (P > 0.10). Across breed types and nutritional treatment, there was positive linear effect (P = 0.04) of plasma concentrations of insulin and IGF-1 on the probability of pregnancy to fixed-time embryo transfer. In summary, the negative impacts of nutrient restriction on the somatotropic axis, independently of body tissue mobilization, were heightened in Bos taurus females compared with B. indicusinfluenced cohorts, which corroborate with the differences observed in fertility between these subspecies.
Effects of Nutritional Manipulation on Poultry under Normal and Stressful Conditions
Sarsour, Albaraa Hisham (Virginia Tech, 2021-11-01)
Genetic selection for improved performance has necessitated the frequent update of amino acid requirements to support this improvement. The first objective of this dissertation was to investigate the tryptophan requirement of laying hens in peak production and the lysine and sulfur amino acid requirements of broiler chicks under a phase feeding scenario using different models. Digestible tryptophan requirements were estimated to be 137 for egg production, 133 for egg mass, and 133 for feed efficiency using the linear broken-line model. The lysine and sulfur amino acid requirement were estimated by conducting 3 experiments within the starter phase from 2 to 5, 5 to 8, and 8 to 11 days of age. The linear broken line estimate for digestible lysine was 1.22, 1.17 and 1.16% for BWG and 1.31, 1.21, and 1.14% for FCR in experiments 1, 2, and 3, respectively. The linear broken line estimate for sulfur amino acids was 0.82, 0.81 and 0.94% for BWG and 0.82, 0.80, and 0.90 for FCR in experiments 4, 5, and 6, respectively. The lysine requirements decreased from 2 to 5 to 8 to 11 days, however the sulfur amino acid requirements increased during that same period. This could be due to other roles that sulfur amino acids play in the body other than growth. The second objective of this dissertation was to investigate the effects of a direct fed microbial (DFM) on broilers exposed to a cyclic heat stress in 2 consecutive experiments. In this experiment, the heat stress treatment reduced body weight gain and lean tissue accretion from 0 to 35 d in both experiments. In Experiment 2, when the litter was reused BWG was increased by 36 g/bird with supplementation of DFM. Ileal digestibility at 28 d (2 hr post HS) was improved with DFM supplementation in both experiments. Serum FITC-d increased with HS at both 28 and 35 d. Serum FITC-d was generally decreased with DFM at 28 d but the response was inconsistent at 35 d. Overall, the results suggest that HS reduced broiler performance and DFM treatment improved intestinal permeability and nutrient digestibility responses to HS in both experiments but did not improve performance until built up litter was used in Experiment 2. The last objective of this dissertation was to investigate the effects of sulfur amino acids (SAA) on broilers exposed to a cyclic heat stress. As expected, HS reduced BWG and worsened FCR. The supplementation of SAA had no effect on live performance. At 28 d of age, supplementation of SAA to birds exposed to HS resulted in reduced intestinal permeability. The interaction was lost at 31 d, but HS still increased intestinal permeability (P ≤ 0.05). Potential oxidative damage was reduced by increased SAA supplementation as indicated by an increase in the reduced glutathione to oxidized glutathione ratio. These data suggest that intestinal permeability is compromised acutely to at least three days of heat exposure before the bird can adjust, but oxidative damage is more chronic building over the entire 7 d HS period. SAA might have some protective effect on both intestinal permeability and oxidative stress responses to HS.
The effects of tributyrin and butyrate on equine skeletal muscle
Gonzalez, Madison Louise (Virginia Tech, 2022-09-02)
In the equine industry, there is a need for supplements that can improve performance and muscle recovery. Tributyrin and butyrate affect satellite cells and oxidative metabolism in other species. To assess the effects of tributyrin on equine muscle repair, Thoroughbred horses were supplemented tributyrin, and a submaximal exercise test was performed. RNA isolated from the gluteal muscle of horses supplemented with tributyrin had increased myogenin mRNA. Satellite cells isolated from supplemented horses had a higher percentage of proliferating cell nuclear antigen immunopositive cells, indicating tributyrin primed satellite cells to activate. Another experiment was performed to test the effects of tributyrin supplementation on equine muscle metabolism. Horses were fed tributyrin for 30 days while partaking in light exercise training. After the supplementation period, horses performed an exhaustive exercise test. Tributyrin supplementation did not affect performance or measures of oxidative metabolism in the muscle. To measure the effects of butyrate on equine muscle metabolism, Thoroughbred horses were supplemented butyrate for 30 days. At the end of supplementation gluteal muscle from butyrate fed horses had a higher percentage of type IIA fibers. Tributyrin supplementation demonstrated positive effects on satellite cell activation, but failed to increase oxidative metabolism measures. Butyrate did statistically increase the percentage of type IIA fibers, but not oxidative enzyme activity and the modest increase seen would likely not effect performance. Higher doses and longer supplementation of butyrate and tributyrin should be investigated to see if a metabolic shift beneficial to racehorses can be achieved. Furthermore, future research should focus on tributyrin's effects on satellite cells and how supplementation can improve muscle recovery in racehorses.

Browsing by Author "Johnson, Sally E."

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