Cryprosporidium parvum is a coccidian protozoan that colonizes epithelial cells lining respiratory and digestive tracts of animals and humans. Cryptosporidiosis is a well-recognized zoonotic disease infecting primarily neonates and immunocompromised hosts, including human immunodeficiency virus-infected patients. Clinical disease is manifested as a chronic diarrheal illness that is self-limiting in immunocompetent hosts and prolonged and often life-threatening in hosts with compromised immune systems.The lack of a suitable small animal model for screening anti-cryptosporidial drugs and for examining the pathogenicity and immunobiology of chronic cryptosporidiosis was the impetus for this research effort. The objectives of the present study were three-fold: to characterize chronic Cryptosporidium parvum infections in dexamethasone-immunosuppressed mice; evaluate the effects of Cryprosporidium parvum and dexamethasone on B and T lymphocyte proliferation; and determine the effects of the immunomodulator dehydroepiandrosterone on oocyst shedding intensities of mice infected with Cryptosporidium parvum. Adult C57BL/6N mice were immunosuppressed with the synthetic glucocorticoid dexamethasone, then infected with Cryprosporidium parvum (106 oocysts/mouse) investigated for their ability to sustain a four-month chronic infection. Dexamethasone was administered intraperitoneally (125 Jlg/mouse/day) or orally (8 Jlg/ml) in the drinking water ad libitum. Infection chronicity was characterized by evaluating mouse monality, oocyst excretion in the feces, tissue distribution of the parasite, and parasite-induced pathology. A progressive infection with Cryptosporidium parvum occurred in mice immunosuppressed intraperitoneally and orally as long as dexameth sone was administered. Mice receiving dexamethasone given intraperitoneally had a shoner prepatent period and a more consistent, although cyclic, oocyst shedding pattern when compared with mice given dexamethasone orally. Mice given dexamethasone orally exhibited a delayed prepatent period, with a steady increase in oocyst shedding. All mice receiving dexamethasone orally died within three months following oocyst inoculation. Clinical signs included dehydration, icterus, and reduction in spleen and body weights. Clinical signs were more abrupt in mice receiving oral dexamethasone. Parasite colonization involved the entire intestinal tract, including the pyloric ring and Peyer's patches, but was the heaviest in the terminal ileum. Parasites were present in the lungs, gallbladder, and pancreatic ducts. Pathologic abnormalities were isolated to the terminal small intestine and included blunting and fusion of intestinal villi and crypt hyperplasia. Cryptosporidium parvum and dexamethasone administered in vivo reduced B and T lymphocyte responses to the mitogens lipopolysaccharide and concanavalin A. Dehydroepiandrosterone and dehydroepiandrosterone-sulfate resulted in no significant reductions in cryptosporidial activity as determined by oocyst shedding in the feces.

Thesis (Ph. D.)--Rutgers University, 2008. / "Graduate Program in Animal Sciences." Includes bibliographical references (p. 138-165).

Master of Science / Animal Sciences and Industry / Evan C. Titgemeyer / Six ruminally-cannulated Holstein steers (202 ± 15 kg) were used to study the effects of ruminal ammonia loading on whole-body lysine utilization. Steers were housed in metabolism crates and used in a 6 × 6 Latin square design. All steers received 2.52 kg of DM/d of a diet (10.1% CP) containing 82% soybean hulls, 8% wheat straw, 5% cane molasses, and 5% vitamins and minerals. Ten g/d of urea was infused continuously into the rumen of all steers to ensure adequate ruminal ammonia concentrations; concurrently, steers were ruminally infused continuously with 200 g/d acetic acid, 200 g/d propionic acid, and 50 g/d of butyric acid and abomasally infused with 300 g/d of glucose continuously to increase energy supply without increasing microbial protein supply. Steers were also abomasally infused continuously with an excess of all essential amino acids except lysine to ensure that lysine was the only limiting amino acid. Treatments were arranged as a 3 × 2 factorial with 3 additional levels of urea (0, 40, or 80 g/d) continuously infused ruminally to induce ammonia loading and 2 levels of lysine (0 or 6 g/d) continuously infused abomasally. Treatments did not affect fecal N output (P = 0.37). Lysine supplementation decreased (P < 0.01) urinary N excretion from 51.9 g/d to 44.3 g/d, increased (P < 0.01) retained N from 24.4 to 33.3 g/d, and tended (P = 0.09) to reduce plasma urea-N. Urea infusions linearly increased retained N (26.7, 28.8, and 31.1 g/d; P = 0.05) and also linearly increased (P < 0.01) urinary N excretion (31.8, 48.1, and 64.4 g/d), urinary urea (21.9, 37.7, and 54.3 g/d), urinary ammonia (1.1, 1.4, and 1.9 g/d), and plasma urea (2.7, 4.0, and 5.1 mM) for 0, 40, and 80 g urea/d, respectively. Assuming that retained protein is 6.25 × retained N and contains 6.4% lysine, the incremental efficiencies of infused lysine utilization were 51, 59, and 69% for steers receiving 0, 40, and 80 g/d of urea, respectively, suggesting that the ruminal ammonia loads might improve the efficiency of lysine utilization; this is supported by the observed increases in whole-body protein deposition in response to ammonia loading of our steers that were, by design, lysine deficient.

Doctor of Philosophy / Department of Grain Science and Industry / Praveen Vadlani / Whole stillage and thin stillage from the ethanol production process were evaluated as substrate sources for the production of [beta]-carotenes using Sporobolomyces Roseus (ATCC 28988). This product has the potential to be used as a novel feed ingredient for poultry, swine, or cattle diets. [Beta]-carotenes have been supplemented in animal diets to improve animal health, enhance meat color and quality and increase vitamin A concentrations in milk and meat. Microbial fermentations involving growth and product kinetics were performed in 500 mL baffled shake flasks and in a 5 L fermentation bioreactor. Media optimization was conducted in shake flasks to evaluate two carbon sources: glucose and glycerol, and two nitrogen sources: ammonium sulfate and urea. Final [beta]-carotene concentration of 272.57±4.34 [mu]g [beta]-carotene/g biomass was found to be highest for the whole stillage, with 10 g/L added glucose and 10 g/L nitrogen added through ammonium sulfate supplementation. Glycerol addition yielded no significant increase (P<.05) in [beta]-carotene yield, while urea addition significantly decreased (P<.05) the final [beta]-carotene concentrations. The resulting fermented product can be blended with regular feed using either whole stillage as a dry feed ingredient or thin stillage as a liquid feed additive. The fermentation of whole stillage significantly influenced the physical and flow properties of the material. Even though there was a significant decrease (P<0.05) in bulk density and increase (P<0.05) in tapped density between DDGS and fermented whole stillage, there was a less pronounced difference between the whole stillage and fermented whole stillage. The fermentation of whole stillage significantly influenced the physical and flow properties of the material. This showed that the fermentation process and resulting nutritional profile had a significant effect on the resulting fermented whole stillage. A 50 L bioreactor was specifically designed to evaluate the scalability of the process and to perform subsequent feed production trails. Pilot scale feed pelleting runs were conducted and the resultant product was put in environmental chambers to determine if [beta]-carotene concentration was reduced as a result of storage. There was a significant decrease (P<0.05) in [beta]-carotene levels after pelleting and after 28 d of storage at elevated temperature and humidity. These decreases were consistent with previous research.

Master of Science / Department of Animal Sciences and Industry / Karol E. Fike / Trace mineral supplementation is necessary for proper reproductive success. Little research has evaluated the effect of an injectable trace mineral product, in conjunction with a dietary mineral supplementation program, on reproduction. This thesis includes two separate studies evaluating the use of an injectable trace mineral product, in addition to a dietary mineral program, on the reproductive success of yearling bulls and heifers. In the first study, we hypothesized that when dietary trace mineral needs are met, administration of an injectable trace mineral product to developing beef bulls would cause a short-term increase in circulating trace mineral concentrations, but not alter semen quality nor ability to pass a breeding soundness examination. Trace mineral treatment did not affect scrotal circumferences and BW of bulls throughout the trial (P [equal to or greater than] 0.20). Trace Mineral bulls had greater (P [equal to or less than] 0.0001) trace mineral concentrations at 8 h post-treatment than Control bulls. Semen trace mineral concentrations on d 42 and 91 were similar (P [equal to or greater than] 0.52) between treatments. Sperm parameters improved (P [equal to or less than] 0.003) from d 42 to 91, but did not differ (P [equal to or greater than] 0.06) between treatments. A similar (P = 0.94) percentage of Trace Mineral (67%) and Control (68%) bulls passed a BSE 91 d post-treatment. In the present study, supplemental trace mineral injection was successful at raising circulating trace mineral levels, but did not alter semen trace mineral levels nor improve semen quality. In the second study we hypothesized that when dietary trace mineral needs are met, the use of an injectable trace mineral product in developing heifers would not affect pregnancy rates at single service fixed-time artificial insemination (FTAI). Trace Mineral heifers had greater (P = 0.02) pregnancy rates (51.28%) than Control heifers (25.58%). The percentage of Trace Mineral (30.77%) and Control heifers (47.50%) that displayed estrous behavior prior to FTAI as indicated by a red estrous detection patch was not different (P = 0.13) between treatments. In the present study, despite dietary trace mineral requirements being met, use of an injectable trace mineral injection improved pregnancy rates following FTAI, but did not affect estrous behavior.

Master of Science / Grain Science and Industry / Cassandra K. Jones / A total of 7 experiments were conducted to evaluate the effects of particle size and thermal processing on swine growth performance or to develop improved analytical methods for particle size prediction. First, 5 experiments utilized 596 nursery pigs to assess how corn particle size and pelleting affected nursery pig growth performance and feed preference. The improvements from reducing particle size were mixed among experiments, potentially because pigs preferred to consume more coarsely ground corn in both mash (P < 0.05; 79.3 vs. 20.7%) and pelleted diets (P < 0.05; 58.2 vs. 31.8%) diets. Pelleting diets led to a reduction in feed disappearance, which tended to improve feed efficiency in nursery pigs (P < 0.05; 0.61 vs. 0.64 for pigs fed mash vs. pelleted diets in Exp. 1). Next, a total of 270 finishing pigs were utilized to determine the effects of long-term conditioning or extrusion of low energy feedstuffs on finishing pig nutrient digestibility, growth performance and carcass characteristics. Treatments included the same basal diet processed as: 1) non-processed mash, 2) pelleted with 45 s conditioner retention time, 3) pelleted with 90 s conditioner retention time, or 4) extruded. Thermal processing, regardless of type, improved daily gain and feed efficiency (P < 0.05), but did not affect feed intake (P > 0.10). Extruded diets tended to improve feed efficiency compared to pelleted diets (P < 0.10). However, pigs fed thermally-processed diets had greater jowl iodine value compared to those fed mash diets (P < 0.05). Finally, 420 samples were used to determine the impact of top sieve size, grain type, technician, and flow agent on the ability of a 3-sieve analytical method to accurately predict the mean particle size determined by a standardized 12-sieve method. The experiment was a 3 × 2 × 2 × 3 factorial with 3 technicians, 2 sieve sizes (U.S. No. 12 vs. 16 sieve as the top sieve), 2 flow agent levels (0 vs. 0.5 g), and 3 grain types (corn, sorghum, or wheat). Linear regression was used to develop individual equations to predict the mean particle size for each of the 3-sieve methods compared to the standard 12-sieve method recognized as ASAE S319.4, and the GLIMMIX procedure of SAS was used to evaluate the impact of main effects and interactions on predication accuracy. All interactions were removed from the model due to insignificance (P > 0.10). Technician, screen size and flow agent did not affect (P > 0.10) the accuracy of the prediction equations. Grain was the only main effect of significance (P < 0.05), where the prediction equation overestimated the particle size of wheat by approximately 15 µm and underestimated the particle size of corn by approximately 12 µm. While statistically significant, these variations were deemed to be sufficiently accurate for the 3-sieve method, and that separate equations for each grain type were not warranted to retain the simplicity of the method. In summary, technician, sieve size, grain type, and the use of flow agent did not greatly affect the accuracy of the 3-sieve particle size analytical method, so the original method was concluded to be accurate and the preferred method.

Doctor of Philosophy / Animal Sciences and Industry / Joel M. DeRouchey / Eight experiments using a total of 2,964 finishing pigs and 2,947 feed, phytase, or premix samples were used to determine the effects of: 1) wheat source, particle size and feed form on finishing pig performance; 2) feed form feeding strategies; 3) fine generation from pellets during feed manufacturing and delivery, and 4) thermal stability and shelf life of phytase products. Exp. 1 and 2 evaluated wheat sources, particle size, and diet form for finishing pigs. Fine gound hard red winter wheat fed in meal form improved G:F and nutrient digestibility, whereas wheat ground from ~700 to 250 µ in pelleted diets did not influence growth or carcass traits. Feeding hard red winter wheat improved ADG and ADFI compared with feeding soft white winter wheat. In Exp. 3, pellet feeding regimens were used to evaluate finishing pig performance and stomach morphology. Feeding pelleted diets improved G:F but increased stomach ulceration and pig removals; however, rotating pellets and meal diets provided an intermediate G:F response with fewer stomach ulcers and pig removals. Experiments 4 to 6 investigated fines formation during pelleted feed manufacturing and delivery. Pellet quality worsened as pellets were transported through the feed mill post pelleting and during delivery. Unloading speed or feed line location had little effect on pellet quality. There were significant differences between the fines and pellet nutrient profiles as noted by the increased concentration of ADF, crude fiber, Ca, ether extract, and starch in the fines and decreased CP and P when compared to pellets. In Exp. 7 and 8, the thermal stability and shelf life of 4 commercial phytase products was determined. Increasing conditioning temperatures decreased phytase stability regardless of product. Phytase activity was affected by storage duration, temperature, product form, and phytase source. Pure products stored between 15 and 22˚C were the most stable and premixes were affected by longer storage times and higher temperatures.

Doctor of Philosophy / Department of Animal Sciences and Industry / Terry Houser / One experiment was conducted to determine the collagen and adipocyte characteristics in pork belly fat with different iodine values (IV) and if these factors contribute to belly firmness. An additional two experiments were conducted to create an objective method to score bacon distortion during cooking and to determine how IV and cooking method contribute to bacon distortion. Experiment 1 sorted pork bellies (n=72) into three IV categories: High 76.5 g/100g, Intermediate 70.5 g/100g, and Low 64.9 g/100g. Belly characteristics and firmness were measured before processing into bacon. After processing, 3 bacon slices were selected from the belly and analyzed for histochemistry and collagen analysis. No differences were observed between belly characteristics, while High IV bellies showed softer bellies. Adipocyte characteristics remained unchanged between IV groups. High IV bellies showed greater amounts of collagen. Experiment 2 cooked bacon slices (n=585) on three different appliances (griddle, microwave, and oven) and scored the resulting distortion using a subjective scale. Raw and cooked bacon characteristics were measured to determine which response variables contributing to distortion. Bacon slices were removed from 6 different locations within each belly sampled. Two distortion measurements were created to objectively describe distortion response (crest frequency and bacon distortion index. Subjective distortion scores, crest frequency, bacon distortion index, and raw and cooked bacon characteristics were shown to change between locations of the belly. Accuracy of predictive equations developed to predict distortion scores were low. Experiment 3 evaluated how IV interacts with cooking methodology to influence cooking characteristics, fat quality and distortion of bacon. Bacon slices (n=300) were organized into two IV categories, Low (61.52 to 65.54 g/100g) and High (78.83 to 85.34 g/100g) and cooked using three different appliances (oven, microwave, and griddle). Bacon from the Low IV group had the greatest amount of fat. Cooking bacon on a griddle showed the greatest distortion scores, while the oven produced bacon with the lowest distortion scores. Bacon with higher IV produced bacon with increased distortion scores. Bacon from the High IV group showed smaller cooked dimensions than the Low IV bacon. Neither cooking method nor IV level affected the cooked fatty acid composition.

Master of Science / Department of Animal Sciences and Industry / James S. Drouillard / Hops β-acid extract was fed to 80 heifers (389 ± 23.6 kg initial BW) to assess impact on feedlot performance and ruminal fermentation. Heifers were randomly assigned to individual pens and fed once daily for 147 d. Treatments were a control (no additive); 33 mg monensin (Elanco Animal Health, Greenfield, IN)/kg diet DM; and 10, 25, 50 mg β-acid extract of hops (DSM Nutritional Products, France)/kg diet DM. Ruminal fluid was collected on d 44 and 86 by rumenocentesis for analyses of VFA, lactate, and NH[subscript]3 concentrations. Cattle were harvested at a commercial abattoir on d 147. Hops β-acids decreased propionate (P = 0.01) concentrations and increased caproate (P = 0.05), A:P (P = 0.04), and ammonia concentrations (P = 0.03) compared to monensin. Growth performance of heifers fed β-acid or monensin was not different than that of heifers fed the control diet. Additionally, two in vitro studies were conducted to evaluate effects of hops β-acid extract on starch fermentation by mixed microbial populations from the bovine rumen. In trial 1, 2 treatments were assigned in triplicate to fermentation bottle, fitted with Ankom[superscript]RF1 Gas Production System modules (Ankom[superscript]RF Technology, Macedon, NY) using starch as substrate (Difco Soluble Starch; Dickinson and Company, Sparks, MD) and either 0 or 33 mg hops β-acid extract (10.99% active hops beadlet; DSM Nutritional Products, France)/kg substrate. Gas production was measured over 30 h. Terminal pH, IVDMD, and VFA and lactate were measured after 30 h of fermentation. Gas production increased in response to β-acid (P ≤ 0.05). Terminal pH, IVDMD, VFA, and lactate were unaffected by addition of β-acid extract (P ≤ 0.05). In trial 2, pH, VFA concentrations, and IVDMD were measured at 6-h intervals during a 30-h incubation period using 36 fermentation tubes. There was no effect of hops β-acid on in vitro fermentation (P > 0.05). In conclusion, under the conditions of these experiments, hops β- acid extracts hops had little impact on feedlot performance, though there are indications of an impact on ruminal fermentation.

Master of Science / Department of Animal Sciences and Industry / Teresa Douthit / Joann Kouba / Gonadotropin-releasing hormone (GnRH-I), as well as its receptor, GnRHR-I, once thought to be localized solely to the hypothalamus and anterior pituitary, have since been detected in the testis of numerous mammals. Another isoform of GnRH, GnRH-II, has been isolated from the testis of numerous mammals and binds a specific receptor, GnRHR-II. Our objective was to establish whether GnRH-I and GnRH-II, along with their specific receptors, are produced and present in the equine testis. Testicular tissue was collected from colts < 2 yr (n = 5) and stallions ≥ 2 yr (n = 10) of age during routine castrations. Total RNA extracted from testicular tissue was reverse transcribed and cDNA was subjected to conventional PCR using gene specific primers for GnRH-I, GnRHR-I, GnRH-II, and GnRHR-II. Protein was extracted and subjected to dot blot and Western blot using antibodies directed against GnRH-I, GnRH-II, GnRHR-I, or GnRHR-II. Transcripts for both ligands and receptors were detected in all testes. Product identity was confirmed by sequencing, which also clarified that unusual band sizes were the result of alternative splicing of GnRHR-II, and the retention of an intron in the GnRH-II mRNA was discovered. Prepro-GnRH-I and prepro-GnRH-II protein was detected in all stallion testes via dot blot technique. On Western blots, testicular samples from colts (n = 4) had 3-fold greater GnRHR-I levels compared to stallions (n = 7; P < 0.022). Conversely, there was a tendency for GnRHR-II protein to be greater in tissue collected from stallions compared to colts (P < 0.0756). Finally semen was collected from mature stallions (9 to 18 yr; n = 4) and purified using a discontinuous gradient. By utilizing immunocytochemistry, GnRHR-II was localized to the connecting piece of mature stallion spermatozoa. This is the first report identifying GnRH-I and -II and their receptors in the equine testis and GnRHR-II on mature stallion spermatozoa. These decapeptide hormones may act via autocrine and/or paracrine signaling to affect steroidogenesis and spermatogenesis in the stallion testis.