Animal & Avian Sciences Theses and Dissertations
Permanent URI for this collectionhttp://hdl.handle.net/1903/2741
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Item Quantification of fatty acids over the gametogenic cycle of striped bass (Morone saxatilis) with varying dietary levels of DHA, EPA and AA(2000) Dickey, Lisa Ann; Woods, L. Curry III; Animal and Avian Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md)The purpose of this thesis was to investigate the effects of dietary treatments on the fatty acid composition of striped bass (Marone saxatilis) blood, liver and ovary over the course of the gametogenic cycle. Striped bass were fed experimental diets in which docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and arachidonic acid (AA) were increased in a stepwise fashion progressing from diets 1-4 respectively. A significant difference was seen in the tissues between fish from dietary treatment group 1 when compared to those fish from dietary treatment group 4 suggesting a direct impact on the tissues by the fatty acid composition of the diet. The second purpose of this thesis was to explore the use of a novel method of fatty acid extraction from fish tissues. This method was compared (UMD method) was compared to the well-known Bligh and Dyer method commonly used for fatty acid extraction. The UMD method proved to be superior quantitatively in terms of fatty acid extraction when compared to the Bligh and Dyer method. The UMD is also more cost affective and leaves less room for human error.Item NUTRITIONAL AND ENVIRONMENTAL IMPLICATIONS OF STARCH DEGRADABILITY AND NITROGEN FERTILIZATION OF ORCHARDGRASS SILAGE IN TOTAL MIXED RATIONS FED TO LACTATING COWS(2003) Woodward-Greene, Mary Jennifer; Erdman, Richard; Animal & Avian Sciences; University of Maryland, College Park, Md.; Digital Repository at the University of MarylandThe objectives of this research were to determine the effect of starch and nitrogen (N) availability on microbial protein production and N efficiency, ruminal N efficiency and ammonia, and to assess forage fertilization and grain selection decisions. Diets were incubated in vitro batch culture, and fed in a 6 x 6 Latin square in vivo digestion trial. Total mixed rations (TMR) contained 50:50 forage:concentrate (dry matter (DM) basis) of second-cutting orchardgrass silage fertilized with 200 (OG200) or 400 (OG400) pounds per acre N, plus concentrate mixes using high to low rumen available starches: barley, corn, and milo. TMR crude protein (CP) was 17% and 18% for in vivo, and 20% and 21% for in vitro OG200 and OG400 diets, respectively. Synchronous diets were low:low or high:high rumen starch availability:diet N (corn or milo withOG200, and barley with OG400). No effects on ruminal microbial protein synthesis and flow, N flow, or milk production were observed. DM, organic matter (OM) (P<0.01), N, and neutral detergent fiber (NDF) (P<0.02) total digestibilities increased with synchronous diets. N digestibility was depressed in diets of low:high rumen starch availability:diet N, due to increased hindgut fermentation adding microbial protein to the feces (P<0.001). All OG400 diets had higher fecal N percentage (P<0.001). OG400 had higher ruminal ammonia both in vitro and in vivo (P<0.05), and higher total in vivo volatile fatty acid (VFA) concentration (P<0.001), but rumen pH was stable due to increased recycling of urea. Orchardgrass fertilized at high N can be digested as well as lower N fertilized forages when combined with a rapidly available ruminal starch such as barley, and decrease outputs of fecal DM by up to 401.5 and N by nearly 22 kilograms per year per cow. Crop fertilization and grain selection decisions affect forage composition, rumen fermentation, ration digestibility, and fecal DM and N output.Item DELINEATING THE ROLES OF C. ELEGANS HEME RESPONSIVE GENES HRG-2 AND HRG-3 IN HEME HOMEOSTASIS(2009) Chen, Caiyong; Hamza, Iqbal; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Heme is an essential cofactor for diverse biological processes such as oxygen transport, xenobiotic detoxification, and circadian clock control. Since free heme is hydrophobic and cytotoxic, we hypothesize that within eukaryotic cells, specific trafficking pathways exist for the delivery of heme to different subcellular destinations where hemoproteins reside. To identify molecules that may be involved in heme homeostasis, we conducted a C. elegans microarray experiment on RNA extracted from worms grown at different concentrations of heme in axenic liquid medium. Analysis of the microarrays revealed that the mRNA levels of heme-responsive gene-2 (hrg-2) and hrg-3 increased more than 70 fold when worms were grown at 4 µM compared to 20 µM heme. hrg-2 is expressed in hypodermal tissues in the worm, and the protein localizes to the endoplasmic reticulum and the apical plasma membrane. In vitro hemin agarose pull-down experiments indicate that HRG-2 binds heme. Deletion of hrg-2 in C. elegans leads to reduced growth rate at low heme. Moreover, expression of HRG-2 in hem1δ, a heme-deficient yeast strain, results in growth rescue at submicromolar concentrations of exogenous heme. These results indicate that HRG-2 may either directly participate in heme uptake or facilitate heme delivery to another protein. Unlike hrg-2, hrg-3 is exclusively expressed in the worm intestine under heme deficiency. Following its synthesis, HRG-3 is secreted into the body cavity pseudocoelom. Deletion of hrg-3 results in increased heme levels in the worm intestine, suggesting that HRG-3 may function in intercellular heme transport in C. elegans. To identify the functional network or pathways for HRG-2 and HRG-3, we performed a genome-wide microarray analysis using RNA samples prepared from the worms grown at different concentrations of heme and oxygen. The results showed that a total of 446 genes were transcriptionally altered by heme and/or oxygen. Among them, 41 and 29 genes exhibited similar expression profiles to hrg-2 and hrg-3, respectively. We postulate that these genes may function in conjunction with hrg-2 and hrg-3. Taken together, we have identified two novel heme-responsive genes in metazoa that may play critical roles in modulating organismal heme homeostasis in C. elegans.Item Proteomic Profiling and Label-Free Quantification of Bovine Milk Proteins during Experimentally Induced Escherichia coli Mastitis(2009) Boehmer, Jamie Layne; Peters, Robert R; Bannerman, Douglas D; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Coliform mastitis has been a primary focus of dairy cattle disease research due to staggering affiliated losses, severe systemic complications arising from host inflammatory response to lipopolysaccharide, and the poor response of coliform pathogens to antimicrobials. Reliable biomarkers are needed to evaluate the efficacy of adjunctive therapies for the treatment of inflammation associated with coliform mastitis, and to aid in the approval of new veterinary drugs. The aims of the current analyses were to utilize proteomic methodologies to evaluate protein expression in whey from cows with experimentally induced coliform mastitis, and to employ label-free quantification strategies to estimate changes in relative abundance of proteins identified in milk over the course of clinical infection. Two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ionization time-of-flight (MALDI- TOF) mass spectrometry (MS) resulted in the identification of complement factors, antimicrobial proteins, and acute phase proteins in mastitic milk. Analysis using liquid chromatography (LC) inline with electrospray ionization - quadrupole TOF tandem mass spectrometry (MS/MS) resulted primarily in the identification of abundant whey and casein proteins, and the transient detection of proteins related to host response. Nano-LC- nanospray-MS/MS using a linear ion trap, however, led to the robust discovery of over fifty inflammatory proteins in whey from mastitic milk, including the novel markers kininogen-2 and inter-alpha trypsin inhibitor heavy chain-4. Normalized spectral counts were compared to enzyme-linked immunosorbant assay (ELISA) for select proteins to assess the accuracy of the spectral count data. Similar expression patterns were detected using spectral counts and ELISA. Results indicate that proteomic methodologies can detect biomarkers of coliform mastitis in bovine milk during clinical infections, and that spectral counts are a viable means of evaluating relative changes in protein biomarkers of mastitis, including those for which no antibody currently exists.Item Identification and characterization of a heme responsive element in the hrg-1 promoter(2008) Sinclair, Jason; Hamza, Iqbal; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Despite its biological significance, little is known about how animals sense and respond to heme to maintain homeostasis. C. elegans is a heme auxotroph, which makes it an excellent model to identify and dissect heme homeostasis pathways. Using C. elegans we have identified HRG-1, a vesicular heme transporter that is transcriptionally upregulated when environmental heme is low. The current study seeks to address how hrg-1 is regulated by heme. Here, we show that a putative 23 base pair (bp) heme-responsive element (HRE) and GATA-binding motifs are necessary for heme-dependent regulation of hrg-1. The HRE comprises both enhancer and repressor elements and works in conjunction with ELT-2 to regulate hrg-1 expression. We propose that the HRE could be used as a molecular tool in C. elegans to tightly regulate internal gene expression by modulating environmental heme. Our ultimate goal is to identify the trans-acting factor to eventually create a whole animal sensor for monitoring organismal heme homeostasis.Item QUALITY ASSESSMENT OF ATLANTIC STURGEON (ACIPENSER OXYRINCHUS) SPERMATOZOA UNDER CONDITIONS OF SHORT-TERM STORAGE(2009) Dorsey, Kathryn Michelle; Woods, Curry; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Short-term storage trials were conducted in 2008 and 2009 on Atlantic sturgeon semen obtained from captive males, held at the U.S. Fish and Wildlife Service, Northeast Fish Technology Center, that were hormonally induced to spermiate; and wild males collected during the spawning season from the Hudson River. Samples were stored under refrigeration (4 + 1°C) in treatments consisting of different gaseous environments (oxygen, nitrogen or air) and experimental diluents (Modified Tsvetkova extender, Park & Chapman extender and neat, i.e. undiluted). Analyses of gamete quality were performed on day 0 (pre-treatment), and then every other day for 7 days in 2008 and for 21 days in 2009. Sperm quality parameters evaluated included: viability, motion analysis, curvilinear velocity and cellular ATP levels. Higher gamete quality was maintained when spermatozoa were diluted in the PC extender and stored in the presence of oxygen.Item ASSOCIATION OF SINGLE NUCLEOTIDE POLYMORPHISMS WITH PHENOTYPIC PRODUCTION TRAITS IN BROILER CHICKENS(2009) Liu, Xuan; Porter, Tom E; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)This research investigated the association between SNPs and phenotypic production traits in fat and lean chicken broiler lines. In previous research, eleven SNPs in the promoter regions of four candidate genes were selected. In this study, significant associations were detected between AKR1B10 SNP1 and SDC1 SNP1 and fat yield. SDC1 SNP1 was significantly associated with fat weight. SOD3 SNP2 was associated with breast yield. Five sire-SNP interactions and one sex-SNP interaction were significant. There was a significant interaction between sex and SDC1 SNP3 on muscle-related factor. GPC3 SNP1 interacted with time period on body weight from week 1 to week 9. QTLs on chromosomes 1, 3 and 4 for body fat were refined by incorporating these SNPs into QTL analysis. These genetic markers may be of great value for marker-assisted selection (MAS) for chickens with less abdominal fat as well as genetic markers for body fat accumulation in humans.Item CAN CHOLINE SPARE METHIOININE FROM CATABOLISM IN LACTATING MICE AND DAIRY COWS?(2009) Benoit, Sarah Lee Ann; Erdman, Richard A; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)Several studies have demonstrated that supplementation of rumen-protected choline (RPC) improves milk production in the lactating dairy cow; however, there are an equal number of studies failing to observe production responses. To date, there are only three studies that provide quantitative information in ruminants on the metabolic fates of methyl groups derived from choline and Methionine (Met). This has limited the ability to predict when, and under what conditions, RPC supplementation will be beneficial. The objectives of this thesis were to determine the interaction of choline and Met methyl group metabolism and the extent of methyl group transfer during lactation, and define what role, if any, is there for RPC in remethylation of homocysteine and in the sparing of Met in lactating animals. A preliminary study with lactating mice consuming a low protein basal diet (10%) was conducted. From 11 to 15 d postpartum, stable isotopes of [methyl,2H3] choline and [methyl,2H3] Met replaced the unlabeled choline and Met in the basal diet to measure the metabolic fates of choline and Met including Met remethylation and sources of Met methyl in the mammary gland. Isotopic analysis revealed that the liver is a major site of Met remethylation from dietary choline with a minimum choline methyl group contribution to Met remethylation of 35%. Mammary tissue was not a major site of Met remethylation from dietary choline (< 10% of Met methyl) as measure by Met methyl in mammary tissue and milk casein. However, there was a significant unlabeled source of methyl groups contributing at a minimum of 45% Met remethylation in the mammary tissue, presumably by de novo synthesis. This suggested that in addition to the liver, the mammary gland is an active site of methyl group transactions. In a subsequent study, lactating dairy cows were fed a total mixed ration formulated to meet the nutrient requirements with exception of metabolizable Met that was restricted to 1.49 % of metabolizable protein. Treatments included a Control (basal diet) and RPC supplemented diet where the basal diet was top dressed with 15g/d RPC, diets were fed in a single reversal design with 2 week experimental periods. Stable isotopes of Met, [1-13C] Met, [13CH3] Met, and [methyl-2CH3] choline were continuously infused on d 14 of each period to determine the metabolic fate and methyl transactions of Met methyl as measured in blood and milk casein. Treatment had no effect on milk production or composition. However, plasma free Met from choline transmethylation was shown to act as a significant contributor to casein synthesis. Fractional Met remethylation rates in the control and RPC treatments were 26 and 23%, respectively. Methionine Met methyl loss within the mammary tissue appears to be minimal. Based on casein Met enrichment, upwards of 40% of Met present in casein had undergone transmethylation with choline serving as the ultimate methyl donor. Furthermore, plasma versus casein Met methyl enrichment data suggested that a significant amount of de novo methyl group synthesis occurs in the dairy cow's mammary gland with choline serving as a major methyl donor.Item Effects of barrier perches and stocking density on the behavior, space use, and leg health of the domestic fowl (Gallus gallus domesticus)(2009) Ventura, Beth Ann; Estevez, Inmaculada; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)The objective of this study was to discern whether providing enrichment in the form of barrier perches across a range of densities might improve leg and foot health and promote behavioral expression and more even use of space in broilers. To investigate this, 2,088 day-old broiler chicks were randomly assigned to one of three barrier treatments at one of three densities. Effects on behavior, space use, foot and hock health, tibia fluctuating asymmetry, fear and production were subsequently assessed. Higher densities appeared to compromise broiler welfare, seen by increased tibia length asymmetry, poorer foot and hock health, suppression of activity, increased disturbances, and decreased use of space. Conversely, barrier perches - particularly simple barriers - appeared to improve footpad quality, promote increased perching and activity, decrease aggression and disturbances, and improve use of the central pen space, all without negatively impacting production traits.Item THE FUNCTIONAL REGULATION OF FCRN EXPRESSION AND FCRN-MEDIATED ANTIGEN PRESENTATION(2009) Liu, Xindong; Zhu, Xiaoping; Animal Sciences; Digital Repository at the University of Maryland; University of Maryland (College Park, Md.)The neonatal Fc receptor for IgG (FcRn), a major histocompatibility complex (MHC) class I-related molecule, plays an important role in IgG transport and protection. The transport of IgG across epithelial and endothelial barriers and the IgG homeostasis maintained by FcRn contributes to the effective humoral immunity. Thus, the level of FcRn itself will affect the IgG-associated immune responses. Although FcRn is expressed in a variety of tissues and cell types, the extent to which FcRn expression is regulated by immunological and inflammatory events remains unknown.I showed here that FcRn was up-regulated by the stimulation of inflammatory cytokines or Toll-like receptor ligands in human peripheral blood mononuclear cell (PBMC) and THP-1 cell line. By chromatin immunoprecipitation, I identified three NF-κB binding sites within introns 2 and 4 of the human FcRn gene. These intronic binding sites boost FcRn transcription activities through looping with the promoter region. In contrast, FcRn expression was down-regulated by Th1 cytokine IFN-γ, and the down-regulation of FcRn was not caused by apoptosis or the instability of FcRn mRNA. It has been demonstrated that IFN-γ activated STAT1 bound with GAS sequence in human FcRn promoter, and which blocked the transcriptional machinery. Fc gamma receptors (FcγRs) expressed in macrophages (MФ) and dendritic cells (DCs) can mediate antigen presentation in both MHC class II and MHC class I pathways. We tested here the role for FcRn in antigen presentation of IgG-restricted Immune complexes (ICs). It was observed that the expression of FcRn in MФ, but not in DC enhanced the phagosomal ICs antigen presentation to CD4 T cells. A low pH value in phgosome of MФ facilitated FcRn binding to ICs, stabilizing the antigens and promoting the efficient MHC II-peptide assembly. However, the alkalized phagosomes in DC failed FcRn to enhance the antigen presentation of ICs.