Title: Effect of protein (amino acid) ingestion on muscle protein synthesis following resistance exercise. Purpose: The main objective of this thesis is to verify the three basic factors of the amount, type and timing of protein intake based on scientific studies and literature, to provide the most objective and accurate information and procedure on the methodology of nutrition and supplementation associated with the intake of protein / amino acids after strength training and how it all affects muscle synthesis. Summery: The theoretical part of the thesis, discusses the factors affecting muscle protein synthesis, which stimulate growth and tissue regeneration, based on optimal stress response. Logically, it starts from general, i.e. the explanation of terms such as muscle tissue, the stimulation of muscle tissue and its manifestations and changes, nutrition factors and muscle stimulation, the mechanism of dietary factors (proteins/amino acids), specificity of protein/ amino acids in their application to answer the three key issues, which are summarized in the section named scientific studies, which focuses on the effect of intake of protein/ amino acids, in relationship to the efficiency of protein synthesis after strength training. The section summarizes, in detail, the questions of timing,...

A betaína é um trimetil derivado do aminoácido glicina. Os seus principais efeitos fisiológicos são atuar como um osmólito e como doador de radicais metil. Especulase que a betaína possa contribuir para a síntese de creatina no músculo esquelético pelo fornecimento de grupos metil, resultante da conversão de betaína em dimetilglicina, para a remetilação de homocisteína em metionina. Os efeitos da suplementação de creatina sobre o desempenho são conhecidos e relacionam-se principalmente ao aumento na ressíntese de fosforilcreatina (PCR). Autores de estudos recentes têm atribuído seus resultados positivos em relação ao aumento de força muscular a um possível efeito da betaína sobre as concentrações de PCR. Essa variável, entretanto, não foi avaliada, de maneira que os mecanismos responsáveis pelo aumento de força advindo da suplementação de betaína ainda são inexplorados em humanos. Diante disso, este estudo teve como objetivo investigar os efeitos da suplementação de betaína, combinada ou não com a suplementação de creatina, sobre as concentrações intramusculares de PCR, e a produção de força e potência muscular em indivíduos não treinados em força. Além disso, as respostas fisiológicas e ergogênicas da suplementação de betaína e creatina foram comparadas e avaliados os possíveis efeitos aditivos desses suplementos. Foi conduzido um estudo duplo-cego, randomizado, controlado por placebo. Trinta e quatro sujeitos foram divididos em quatro grupos: Betaína (BET; 2 g/dia), Creatina (CR; 20 g/dia), Betaína + Creatina (BET + CR; 2 + 20 g/dia) e Placebo (PL). No período basal (PRÉ) e após 10 dias de suplementação (PÓS), os indivíduos submeteram-se a avaliações do consumo alimentar e da composição corporal, a testes de força e potência muscular e à quantificação intramuscular de PCR. Após a intervenção, as concentrações intramusculares de PCR foram maiores nos grupos CR e BET + CR, quando comparados ao grupo PL (p = 0,004 e p = 0,006, respectivamente). Não houve diferenças significativas entre os grupos BET e PL (p = 0,78) e CR e BET + CR (p = 0,99). Os grupos CR e BET + CR apresentaram maior produção de potência muscular no exercício de agachamento, quando comparados ao grupo PL (p = 0,003 e p = 0,041, respectivamente). Resultados similares foram encontrados para o exercício de supino. Os grupos CR e BET + CR também demonstraram aumento significativo de força muscular (teste de 1-RM) do teste PRÉ para o teste PÓS nos exercícios de supino e agachamento (CR: p = 0,027 e p 0,0001; BET + CR: p = 0,03 e p 0,0001 para membros superiores e inferiores, respectivamente). Não houve diferenças significativas para os testes de força e de potência muscular entre os grupos BET e PL e os grupos CR e BET + CR. Também não houve diferença significativa entre os grupos para a composição corporal. O consumo alimentar permaneceu inalterado ao longo do estudo. Os resultados permitem concluir que a suplementação de betaína, combinada ou não com a suplementação de creatina, não aumenta o conteúdo intramuscular de PCR e não afeta o desempenho de força e de potência muscular / Betaine is a trimethyl derivative of the amino acid glycine. The main physiological functions of betaine are to act as an organic osmolyte and as a donor of methyl radicals. It is speculated that betaine may contribute to the synthesis of creatine in skeletal muscle through the donation of a methyl group, resulting from the conversion of betaine to dimethylglycine, to homocysteine to form methionine. The effects of creatine supplementation on performance are well known and are related primarily to an increase in fosforilcreatina resynthesis (PCR). Authors of recent studies have attributed its positive results regarding the increase of muscle strength to a possible effect of betaine on the concentrations of PCR. However, this variable was not assessed, so that the mechanisms responsible for the increase in muscle strength coming from betaine supplementation in humans are still unexplored. In light of this, the aim of this study was to investigate the effect of betaine supplementation combined or not with creatine supplementation on muscle PCR content, muscle strength and power output in non-resistance trained subjects. Additionally, we compared the ergogenic and physiological responses to betaine versus creatine supplementation. Finally, we also tested the possible additive effects of creatine and betaine supplementation. A randomized, double-blind, placebo-controlled study was conducted. Thirty and four subjects were assigned into four groups: Betaine (BET; 2 g/day), Creatine (CR; 20 g/day), Betaine + Creatine (BET + CR; 2 + 20 g/day) or Placebo (PL). At baseline (PRE) and after 10 days of supplementation (POST) body composition, food intake, muscle strength and power and muscle PCR were assessed. The CR and BET + CR groups presented greater increase in muscle PCR content than PL (p = 0.004 and p = 0.006, respectively). PCR content was comparable between BET versus PL (p = 0.78) and CR versus BET + CR (p = 0.99). CR and BET + CR presented greater muscle power output than PL in the squat exercise following supplementation (p = 0.003 and p = 0.041, respectively). Similarly, bench press average power was significantly greater for the CR-supplemented groups. CR and BET + CR groups also showed significant pre- to post-test increase in 1-RM squat and bench press (CR: p = 0.027 and p 0.0001; BET + CR: p = 0.03 and p 0.0001 for upper- and lower-body assessments, respectively). No significant differences for 1-RM strength and power were observed between BET versus PL and CR versus BET + CR. Body composition did not differ between the groups. Dietary intake was unchanged throughout the study. Thus, we concluded that betaine supplementation does not augment muscle PCR content and betaine supplementation combined or not with creatine supplementation does not affect strength and power performance in non-resistance trained subjects

The effect of post-weaning supplementation of April-born Polypay and White Dorper lambs grazing alfalfa/orchardgrass pasture during two grazing seasons from June to September was studied. One hundred seventy Polypay and 133 White Dorper lambs were randomly allotted to supplemented (2% BW daily) and unsupplemented groups. It was discovered that Polypay lambs weighed more than White Dorpers at wearning, when the grazing season began, and at the end of the 80 (Year 1) and 85 (Year 2) grazing season (P < 0.01). Polypay lambs gained faster (P < 0.01) than White Dorpers. Supplemented lambs gained faster (P < 0.01) than unsupplemented and Polypays had a greater response to supplementation (P < 0.01) than White Dorpers. Polypays had higher (P < 0.01) fecal egg counts, an indicator of Haemonchus contortus infestation. Supplementation did not have any consistent effect on reducing Haemonchus contortus infestation. Differences in forage characteristics of Polypay and White Dorper lambs did appear. Differences in alfalfa and orchardgrass dry matter, neutral detergent fiber, acid detergent fiber, and crude protein availability were measured by subtracting enter from exit availabilities. The largest decrease of alfalfa components from exit to enter was found with unsupplemented Polypays. No consistent effect was found for orchardgrass components.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Tiruvoor G. Nagaraja / Copper, as copper sulfate, is often supplemented at elevated concentrations in swine diets, particularly in piglets, to promote growth. Growth promotional effects of copper are believed to be similar to that of antibiotics in that gut microbial flora is altered to reduce loss of nutrients and suppress pathogens. Bacteria exposed to copper may acquire resistance, and in Enterococcus faecium and E. faecalis, resistance is conferred by a plasmid-borne transferable copper resistance (tcrB) gene. The plasmid also carries macrolide [erm(B)] and glycopeptide (vanA) antibiotics resistance genes. The objectives of the research were to 1) determine the prevalence of tcrB gene in fecal enterococci of piglets in relation to normal (16.5 ppm) and elevated level (125 ppm) of copper supplementation, 2) determine the relationship of tcrB gene and susceptibilities to copper, erythromycin, and vancomycin, and 3) determine the transferability of tcrB gene in enterococci by conjugation. Weaned piglets, housed in pens, fed normal (16.5 ppm; control) or elevated level of copper (125 ppm) were used. Fecal samples were collected weekly for isolation of enterococci. Isolates were speciated by multiplex PCR and sodA gene sequence analysis. The prevalence of tcrB-positive enterococcal isolates was higher (P < 0.05) in the copper supplemented group than the control group. The prevalence of tcrB was affected by sampling days (P < 0.05) with a significant treatment and sampling time interaction (P < 0.05). The tcrB positive isolates were either E. faecium or E. faecalis, and majority of isolates was E. faecium. The mean MIC of copper for tcrB-positive isolates (21.1 mM) was higher (P < 0.001) compared to tcrB-negative isolates (6.1 mM). All isolates were resistant to erythromycin, tetracyclines and susceptible to vancomycin. The transferability of the tcrB gene from tcrB-positive strains to tcrB-negative strains was demonstrated by conjugation. The potential link between tcrB and antibiotic resistance genes and the propensity of enterococci to transfer tcrB to other strains suggests the possibility that copper supplementation may exert selection pressure for antibiotic resistance. The positive association between copper supplementation and prevalence of tcrB gene has important implications for antimicrobial resistance and food safety, which warrants further investigation.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: This thesis aims to determine whether supplementation with a grape-seed derived antioxidant, oligomeric proanthocyanidin (PCO) or the glutathione precursor, N-acetylcysteine (NAC) may prove beneficial as treatment for exercise induced muscle damage (EIMD) in athletes. In this double-blind cohort study, 21 healthy, uninjured male rugby-players in mid-season training phase, aged between 18 and 25 years were randomly divided into three treatment groups. Participants received 210 mg PCO, NAC or placebo treatment for 9 consecutive days. The study comprised a 6-day wash-out period (protocol days: -12 to -7), followed by a 6-day supplement loading period (protocol days: -6 to -1) a plyometric exercise intervention (protocol day 0) and continued supplementation for 2 days (protocol days: 1 to 2). The exercise intervention comprised 15 sets of 10 near maximal, vertical plyometric squat jumps. Blood samples and delayed onset of muscle soreness (DOMS) scores were collected on protocol days: -6, 0, 1 and 2. Assessments included serum creatine kinase (CK) activity, oxygen radical absorbance capacity (ORAC), malondialdehyde (MDA) and soluble vascular cell adhesion molecule-1 (sVCAM-1) concentrations over time as well as a differential circulating leukocyte count (neutrophils, lymphocytes, monocytes, eosinophils and basophils). Data analysis of CK activity revealed no significant differences between groups. However, PCO treatment prevented a significant peak in the CK response at 24 h (as seen in the placebo and NAC groups) when compared to baseline, pre and post readings (p<0.05). NAC supplementation significantly improved serum ORAC after the exercise intervention. By 48 h, serum ORAC had improved significantly from readings taken immediately post exercise (p<0.05) only in the NAC group. For all groups, absolute neutrophil counts peaked at 6 h post exercise from baseline or pre readings (p<0.05). In both NAC and placebo treated groups, neutrophil counts had decreased significantly in circulation by 24 h post exercise from the 6 h time-point (p<0.05). However, neutrophil counts only reached significantly lower levels by 48 h post exercise (p<0.05) in the group supplemented with PCO. The monocyte count also peaked significantly at 6 h post exercise when compared with other time-points before and after the exercise intervention (p<0.05) in all treatment groups. Neither antioxidant treatment significantly altered the responses of other leukocyte sub-populations, MDA or sVCAM-1 concentrations where main effects of plyometric exercise was evident. Although not statistically significant, a trend toward diminished sVCAM-1 expression with either antioxidant supplementation was apparent. These findings suggest that PCO supplementation (210mg/d) which includes a 7 day loading period may diminish plyometric EIMD by limiting (but not completely inhibiting) the neutrophil response. Secondary muscle damage may be prevented by partially blunting neutrophil infiltration, rather than only quenching free radicals released during the neutrophil oxidative burst. Furthermore, the finding that NAC supplementation improves serum ORAC only after exercise may provide added benefit when administered in combination with PCO. / AFRIKAANSE OPSOMMING: Hierde tesis is daarop gerig om vas te stel of aanvulling met ‘n druifsaadekstrak (DSE) gederiveerde antioksidant: pro-antosianiedoliese oligomeer (PSO), of die glutathione voorloopermolekule, N-asetielsistien (NAS) voordelig beskou kan word as behandeling vir atlete onderhewig aan spierskade veroorsaak deur oefening. Gedurende hierdie dubbelblinde kohort studie is 21 gesonde, manlike rugbyspelers sonder beserings tussen die ouderdom van 18 en 25 jaar in middel-seison fase ewekansig in drie behandelingsgroepe verdeel. Deelneemers het elk 210 mg PSO, NAS of placebo-aanvulling geneem vir nege agtereenvolgende dae. Die studie het bestaan uit ‘n 6-dag uitwasperiode (protokoldae: -12 tot -7), as ook ‘n 6-dag aanvullings periode (protokoldae: -6 tot -1), gevolg deur ‘n pliometriese oefeningsintervensie (protokol dag 0) en verdere aanvulling tot en met 2 dae na die oefening (protokol dae: 1 tot 2). Die oefeningsintervensie het 15 stelle van 10 naastenby maksimale, vertikale pliometriese hurkspronge behels. Bloedmonsters en vertraagde aanvang spierseerheid (VAS) tellings is op protokoldae: -6, 0, 1 en 2 geneem. Analiese het serum kreatien kinase (KK) aktiwiteit, suurstof radikaal absorpsie kapasiteit (SRAK), Malondialdahied (MDA) en oplospare vaskulêresel adhesie molekule-1 (oVAM-1) konsentrasie bepalings asook ‘n differentiële sirkulerende leukosiet seltelling ingesluit. KK aktiewiteit het geen merkwaardige verskil tussen groepe getoon nie. PSO aanvulling het wel gelei tot die voorkoming van ‘n merkwaardige piek in die KK response soos in die placebo en NAC behandelde groepe bevind is by die 24 h tydspunt in vergelyking met basislyn-, voor- en na-oefeningslesings (p<0.05). NAS het ‘n merkwaardige verbetering in serum SRAK getoon, maar eers teen 48 h na oefening. Slegs die NAS behandelde groep het op hierdie tydspunt ‘n betekenisvolle verbetering in SRAK getoon in vergelyking met lesings direk na oefening (p<0.05). Vir alle groepe is ‘n betekenisvolle toename in absolute neutrophiltellings waargeneem 6 h na oefening in vergelyking met basislyn- en vooroefeningslesings (p<0.05). Beide NAS en placebo-behandelde groepe het ‘n betekenisvolle afname in neutrophiltellings teen 24 h na oefening getoon in vergelyking met die 6 h tydspunt (p<0.05) maar met die PSO-behandelde groep word hierde afname eers teen 48 h waargeneem (p<0.05). Monosiettellings het in alle groepe 6 h na oefening ‘n betekinsvolle piek getoon (p<0.05). Waar slegs die hoofeffek van die pliometriese oefening betekenisvol was, het nie een van die twee antioksidant aanvullings ‘n merkwaardige verandering aan die respons van ander leukosiet sub-populasies, MDA of oVAM-1 konsentrasies getoon nie. Al kon statistiese beduidenheid nie bewys word nie, wil dit blyk dat ‘n verminderde oVAM-1 uitdrukking onstaan het in die geval van beide antioksidant-behandelde groepe. Tesame stel hierdie bevindinge voor dat PSO toediening (210mg/d) insluitende ‘n 7-dag aanvullingsperiode die vermoë verleen om die neutrophielrespons gedeeltelik te onderdruk (sonder om dit heeltemal te inhibeer) en sodoende spierskade verminder. Dus word verdere spierskade moontlik verlaag deur die voorkoming van neutrophil weefsel infiltrasie eerder as verwydering van reaktiewe spesies wat vrygestel word tydens oefening. Die bevinding dat NAS aanvulling serum SRAK eers na oefening merkwaardig verbeter, kan as voordelig beskou word, veral wanneer toegedien in samewerking met PSO om verdere spierskade te voorkom en herstelling vinniger te bewerkstellig.

These studies determined the effects of sodium chloride supplementation on serum and sweat sodium concentration, cardiovascular function, and physical and cognitive performance. Sweat sodium losses, alone, caused a significant decline in serum sodium concentration (-6.4±1.6 mEq/L, p=0.001) during 3h cycling in the heat in endurance-trained athletes with high sweat sodium losses. However, sodium chloride supplementation matching sweat sodium losses (NA; 5.9±1.5g NaCl/h) maintained serum sodium concentration. Post-exercise maximal cycling power declined and was significantly lower than pre-exercise in placebo (PL; p=0.012), but power was not significantly different in NA (p=0.057). Pre- to post-exercise response time during a Stroop Test improved in NA (p=0.009), while there was no change in PL (p=0.597). Post-exercise postural sway was less in NA vs. PL (p=0.044). Three days of sodium chloride supplementation (~15 g NaCl/d) resulted in a significant increase in plasma volume in healthy untrained males at rest (5.9±7.6 %) and during exercise at 60%VO₂peak (8.6±5.2 %) compared to PL. During NA, stroke volume was 10% higher during exercise vs. PL (139±27 vs. 126±24 ml/beat, respectively, p=0.004). Cardiac output was 8% higher in NA during exercise vs. PL (21.0±3.1 vs. 19.4±2.6 L/min, respectively, p=0.013). Mean arterial pressure during exercise was not different in NA vs. PL (p=0.548) as total peripheral resistance decreased (p=0.027) with the increased cardiac output. Sweat sodium concentration was 9% higher in NA vs. PL during exercise in the heat (70.4±19.5 vs. 64.5±21.7 mEq/L, p=0.044). In summary, serum sodium concentration declines when high sweat sodium losses are not replaced while hydration status is maintained. Acute sodium chloride supplementation during exercise which matches sodium losses maintains serum sodium concentration. This maintenance of serum sodium concentration results in both physical and cognitive benefits compared to when serum sodium concentration declines. Chronic intake of sodium chloride for 3 days increases plasma volume in healthy untrained men and improves cardiovascular function, as both stroke volume and cardiac output are increased, while oxygen consumption and blood pressure are unchanged. Therefore, acute and chronic sodium supplementation positively alters fluid and sodium balance which results in beneficial effects on physical and cognitive performance and cardiovascular function during exercise. / text

Background - The first 1000 days of life (from conception to two years of age) is a critical period of nutritional vulnerability, affecting lifelong health. Iron deficiency (ID) and iron deficiency anaemia (IDA) are considered major public health problems that adversely affect development and growth, impair immunity, and increase morbidity and mortality in infants. ID and IDA in sub-Saharan Africa can be attributed to poor dietary, socioeconomic and disease conditions. One of the major obstacles in determining the prevalence of ID, using serum ferritin (SF) as marker of iron status, is that it not only reflects the amount of iron that is stored in the body, but also functions as an acute phase reactant that is raised in the presence of infection or inflammation. Aim - We conducted a re-analysis of the International Research on Infant Supplementation (IRIS) study’s baseline data to determine a more accurate estimation of the ID prevalence in apparently healthy four to thirteen-month-old infants from rural KwaZulu-Natal while accounting for the effect of chronic and acute inflammation on SF. Study design and methods - A cross-sectional analysis was performed on the baseline data (192 infants) of the IRIS study that was conducted in 2000. Infants’ haemoglobin (Hb), SF, C-reactive protein (CRP) and alpha-1 glycoprotein (AGP) concentrations were interpreted to determine the prevalence of ID. Literature of the past four years served as a guide to compare the ID prevalence obtained from four methods that account for the influence of inflammation on SF concentrations, to a reference method that does not take inflammation into consideration, and to what was reported in the original IRIS study. Weight and recumbent length measurements were converted to z-scores to interpret subjects’ anthropometric nutritional status. Results - A high prevalence of inflammation (52.6%) was present, with 11.5% of the subjects being in the incubation, 17.2% in the early convalescent, and 24% in the late convalescent phase of inflammation. SF was significantly associated with both CRP (ß = 0.200; P = 0.005) and AGP (ß = 0.223; P = 0.002) when adjusting for gender and age. The IRIS study reported an ID prevalence of 18.3%, whereas the results of this study ranged from 17.2 to 52.1%. We derived an IDA prevalence that ranged from 12 to 24.5% according to the different methods. The prevalence of stunting [length-for-age Z-score <-2SD] was 12.5%; while 25.1% of infants were overweight/obese [weight-for-length z-score >2SD]. Conclusion - A double burden of malnutrition was evident from the high prevalence of both overweight and ID, together with inflammation. The disconcertingly large variance in ID prevalence observed between the different methods that were employed highlights that iron supplementation interventions to treat anaemia must be based upon accurate estimates of IDA prevalence, otherwise they pose an increased risk of adverse effects to susceptible, iron-replete, but anaemic infants. Given the detrimental consequences of ID, it is imperative that governments, health care providers and parents must act to prevent or treat ID and IDA among vulnerable infants. / MSc (Dietetics), North-West University, Potchefstroom Campus, 2014

The series of experiments that comprises this thesis consider the use of prebiotics and probiotics to improve success in European lobster Homarus gammarus culture. Research was conducted at the National Lobster Hatchery and The University of Plymouth, UK between April 2007 and March 2010. Great potential was revealed for the application of biotics as dietary supplements in the culture of larval and juvenile H. gammarus. Specific biotics, namely; prebiotic mannon oligosaccharide (MOS) and probiotic Sanolife® (Bacillus spp.) were revealed to increase growth and survival during the culture of early lobster life stages. Lobsters that showed enhanced development and/or survival were found to possess improved feed conversion, immune status and/or stress tolerance. Furthermore it was found that biotic fed lobsters displayed shifts in gastrointestinal (GI) microbiology and enhanced physical GI structures, which may account for improved feed conversion and consequently growth. The work presented thus demonstrates that by making positive changes in GI structure and bacterial community composition combined with influencing immune status culture success can be positively enhanced through the biotic supplementation of diets. However, it must be considered that the outcome of dietary biotic supplementation is heavily influenced by the situation in question.

The American shad Alosa sapidissima has experienced severe declines throughout its native range due to habitat degradation, fragmentation, and over-fishing. Hatchery supplementation is often used for stock restoration, but the effects of supplementation on population structure and genetic diversity are rarely assessed. This study employed molecular markers to evaluate how supplementation of the James River American shad population with Pamunkey River origin larvae since 1994 has impacted genetic diversity and population structure. Population genetic parameters of other major Chesapeake Bay tributaries (Susquehanna, Rappahannock, Potomac, and Nanticoke) also were characterized. Prior to stocking, the James and Pamunkey populations exhibited subtle genetic differentiation, which was absent among post-supplementation samples, presumably due to the stockings. A similar situation was observed among other shad populations of Chesapeake Bay tributaries which were subtly differentiated in the 1990s but lacked any credible among-population differentiation among contemporary samples. Genetic diversity of the James River shad population was high prior to stocking, and remained high throughout years of intensive supplementation, yet the current population decline suggests that the James River shad population still has not recovered. Despite harvest curtailment, elimination of the ocean intercept fishery, and widespread supplementation efforts, Chesapeake Bay tributary American shad populations are collectively at their lowest levels in recorded history. Therefore, success of other restoration goals such as creation of fish passage in James River was investigated in a concurrent radio telemetry study to assess passage at Bosher's Dam fishway. Ninety-four American shad were radio-tagged on the spawning grounds below Bosher's Dam. Approximately one-half of the tagged shad were detected at the escapement receiver within 24 hours after tagging, and the average residence times of remaining shad were approximately one week. No tagged shad were detected above Bosher's Dam. These results imply that restricted passage through Bosher's Dam fishway may be an important factor in the failure of James River American shad to recover. Therefore, improving passage at migratory barriers such as Bosher's Dam, in conjunction with a continued Bay-wide fishing moratorium, may be more beneficial to shad restoration efforts in James and other Chesapeake Bay tributaries than continued supplementation.

Master of Science / Department of Animal Sciences and Industry / John Jaeger / Cattle grazing dormant native range (< 7% crude protein; CP) require supplementation of additional protein to sustain body weight (BW) and body condition score (BCS). Daily delivery of these supplements is an economic burden to cattle producers faced with challenging economic circumstances. Supplementing cows infrequently (as little as once/week) has produced equivalent BW and BCS changes compared to daily delivery. Dried distiller’s grains with solubles (DDGS) provides more ruminally-undegradable protein (RUP; 50-60%) compared to traditional oilseed-meal supplements (i.e. soybean meal) that are >50% ruminally-degradable protein (RDP). Therefore, our objective was to evaluate the effects of supplementation frequency on performance, reproductive success, eating behavior, and subsequent calf performance of spring-calving cows supplemented with DDGS. No differences in ending BW (P = 0.69) and BCS (P = 0.49), or changes in BW and BCS over the supplementation period (P = 0.82 and 0.70, respectively) were observed among cows supplemented every d, every 3 d, or every 6 d. Calf BW at birth, weaning weight (WW), and average daily gain (ADG) were similar among treatments (P = 0.19, 0.12, and 0.10, respectively). First-service conception rate (FSCR) and final pregnancy rate (PR) were also not affected by supplementation frequency (P = 0.62 and 0.76, respectively). The development of replacement heifers is a large expense for cow-calf producers. Improved breeding and heifer development strategies aimed at ensuring the success of replacement females have been developed but reproductive failure still remains a problem. The stress associated with breeding and handling procedures may decrease reproductive success. Therefore, the objective was to determine if intramuscular administration of flunixin meglumine (1.1 mg/kg BW) 14 days post-breeding would improve FSCR and PR in non-transported replacement heifers. Under the conditions of our study, flunixin meglumine did not improve (P = 0.87) first service conception rate above that of control heifers (41.2% and 42.3%, respectively). Final pregnancy rate also was not different between treatments and averaged 81.8% (P = 0.40).