The primary lines of defense in preventing any exotic species from becoming endangered or extinct should be the protection of their habitat and from poaching. For wildlife conservationists, these primary measures are often not feasible, and thus cannot be the only means to prevent extinction. The objective of this research was to identify a semen extender that contained little to no egg yolk and could still effectively freeze domestic cat epididymal spermatozoa, for the purpose to sort spermatozoa into X and Y, using flow cytometry. In the first experiment we compared a TesT (Tes + Tris) 20% egg yolk extender (control) to a modified human sperm preservation medium (HSPM) and a Tris citrate extender that contained bovine serum albumin (TCBSA) to compare their effectiveness of preserving feline epididymal spermatozoa in relation to control spermatozoa. Results from this one experiment showed no significant difference among any of the treatments when measuring membrane integrity or acrosomal status. There was significant difference when comparing the pre-cool motility value between the control and the human sperm preservation medium extender (HSPM), but this difference was not detected in either of the post-thaw evaluations. In the second experiment, an attempt was made to determine what concentration could egg yolk be reduced to, and still exhibit cryoprotection. Three egg yolk concentrations (10%, 5% and 2%) were evaluated. It was determined that even at the lowest concentration, 2% egg yolk was not significantly different from the 10% or the 5% when comparing motility, membrane integrity and acrosomal status. In the third series of experiments, a TesT 2% egg yolk extender was evaluated to determine if exposure of the spermatozoa, before and after freezing, to 0nM, 1nM and 5nM of pentoxifylline (a motility stimulant) would have an effect on the post-thaw parameters measured. It was determined that there was no significant in post-thaw sperm motility, membrane integrity or acrosomal status when the epididymal sperm were exposed to the pentoxifylline both before and after freezing. In the last series of experiments TesT 2% egg yolk extender was compared with the BioXCell® and Biolife® extenders (neither containing egg yolk, and predominantly used for cool liquid sperm storage) on their effectiveness to maintain feline epididymal spermatozoa at 4°C for 72 hours. Although no significant difference was noted, the TesT 2% egg yolk and the BioXCell® extenders showed the most promising results for cooled liquid storage up to 72 hours. These experiments have shown that viable epididymal cat sperm can be collected from the epididymides of castrated toms, cryopreserved in extenders that contain little to no egg yolk and thawed, resulting in acceptable post-thaw values sufficient enough for IVF, possibly for AI and most certainly for ICSI. Finally, sperm were frozen in the TesT extender with and without egg yolk and frozen from room temperature and after cooling to 4°C. The TesT without egg yolk, frozen from room temperature, consistently had lower sperm motility and membrane integrity then sperm frozen in one or both of the extenders frozen after cooling to 4°C. There was no difference between the two TesT extenders frozen after cooling to 4°C for any of the parameters measured.

This study was designed to determine whether kefir accentuates the positive health benefits assessed by measures in fitness and/or body composition, as a measure of cardiovascular disease risk as well as the biomarker c-reactive protein. Sixty-seven adult males and females aged 18-35 years were assigned to one of four groups: 1) endurance training + control beverage, 2) endurance training + kefir beverage, 3) active control + control beverage or 4) active control + kefir beverage. The exercise groups completed 15 weeks of structured endurance training while the active control groups maintained their usual exercise routine. Baseline physiological and exercise measurements were collected pre-intervention and post-intervention. Blood and saliva samples were analyzed for C-reactive protein, tumor necrosis factor-alpha and secretory immunoglobulin A. Instances of sickness and illness throughout the intervention were also examined. Additionally, each group was assigned to either a kefir or a calorie/macronutrient matched placebo beverage that was consumed twice per week. The endurance training protocol was effective as demonstrated by a significant improvement (p<0.05) in the 1.5-mile times of the endurance training groups. The endurance training groups also experienced a significantly higher (p<0.05) number of sicknesses than the active control groups. There were no significant interactions among groups with respect to physiological outcome variables with the exception of serum C-reactive protein. The endurance training group receiving a control beverage demonstrated a significant increase (p<0.05) in C-reactive protein following the 15 week training program while the endurance training group receiving a kefir beverage had no significant change in C-reactive protein. Although the reported sickness was not significantly different between the endurance training group + kefir beverage and the endurance training group + control beverage, kefir supplementation may have been a factor in attenuating the increase in C-reactive protein that was observed over the course of the intervention period. This preliminary study suggests that kefir may be involved in improving the risk profile for cardiovascular disease as defined by C-reactive protein.

Active immunisation against GnRH inhibits reproductive function by inducing a hypogonadotropic condition associated with gonadal atrophy. Despite economic, ethical and environmental advantages of GnRH immunisation in cattle over conventional castration methods, the technology has not yet been commercially adopted. Primarily because of the requirement for numerous booster vaccinations because of the reversibility of physiological effects, the commercial efficacy of immunocastration is currently poor. However, neonatal GnRH immunisation in sheep can result in a permanent suppression of reproduction (Brown et al., 1994; 1995; Clarke et al., 1998). These findings and a study in pigs (Molenaar et al., 1993) indicate that, the hypothalamic/pituitary gland unit (HPU) may be particularly susceptible to GnRH antibodies during a specific window of development in the pre-pubertal animal, but no long-term studies in cattle have been conducted. Therefore the primary objective of this project was to determine the effect of neonatal immunisation against GnRH in cattle. Beef cross bull (n=9; Chapter 3) and heifer calves (n=9; Chapter 4) were vaccinated against a newly developed (Pfizer®) GnRH construct vaccine at -2, 6 and 13 weeks of age. Nine calves of each sex served as negative controls, receiving saline injections only. The GnRH vaccine had proved effective (Dr. A.R. Peters, personnel communication 2000) in inducing immune responses and reducing variation between animals in unpublished industrial studies, compared to earlier vaccines, and hence was reasoned to be capable of raising GnRH antibodies despite the relative immaturity of the neonatal immune system. Following vaccination, circulating GnRH antibodies and reproductive hormones, such as FSH (Chapters 3 and 4), testosterone (Chapter 3), progesterone (to assess onset of puberty) and oestradiol (Chapter 4) were measured and additional intensive serial bleeds were carried out to assess LH parameters up to and beyond puberty (puberty defined by testes circumference in bulls). Gonadal (antral follicles and testes growth) and accessory gland development was quantified throughout the trial using ultrasound scanning. Sexual behaviour (Chapter 3) was studied from 38 weeks of age, while an assessment of sperm quality (Chapter 3), and anabolic response to vaccination was also performed post-mortem (Chapters 3 and 4). GnRH immunisation in neonatal calves did not permanently impair reproduction. A temporary suppression in reproductive function was evident through the disruption of pituitary gland function, as indicated by a reduction of LH pulse amplitude and mean plasma LH concentrations (Chapters 3 and 4). In addition, a reduction in medium- sized follicle numbers, testes growth, plasma testosterone concentration, vesicular gland length and juvenile aggression occurred. Some beneficial anabolic effects were observed e.g., carcass composition grades. Changes all occurred subsequent to increased GnRH antibody titres in immunised cattle. Despite some evidence of prolonged effects on LH amplitude and circulating testosterone after anti-GnRH titres had dissipated, all inhibited parameters, except carcass quality, returned to levels comparable to control animals by 72 weeks of age. No treatment effects on FSH concentrations, large follicle numbers, reproductive tracts (post mortem) or peri- and post-pubertal behaviours were observed following treatment. Sperm morphological abnormalities tended to be more prevalent in GnRH immunised bulls. A significant increase in GnRH antibody titres occurred at -23 weeks of age (Chapter 4), this may have been a rebound in antibody titre, possibly caused by an anti-idiotype immune response (antibody response to GnRH antibodies), or due to significant maturational changes in immune function at this time causing a delayed response to vaccination. Alternatively a novel "auto-immune" response may have been detected, which if confirmed/repeatable might be incorporated into an immunisation protocol to act as a "self-booster". However, no previous reports of such an event have been published and further investigation is urgently required. A more prolonged or permanent suppression of reproductive function may be possible following an earlier, greater and more sustained elevation of antibody titres during the neonatal period. Further development of GnRH vaccines and/or protocols (prime-boost, cytokine modulation vaccines, concomitant passive and active immunisation and pregnant cow GnRH vaccination), and studies of performance and GnRH antibody mechanism(s) of action in cattle are required. Chapters 3 and 4 provide a comprehensive study on pubertal development and neonatal GnRH vaccination, thus contributing significantly to knowledge in these fields. Currently, the vaccine used in this trial may be used to delay puberty in older calves or transiently suppress reproductive function to aid management. The economical viability of animal production systems such as beef and lamb are closely related to rates of reproduction. The Fec B gene in ewes increases ovulation rate and litter size, possibly through the development of precocious follicles, which can switch their primary dependence from FSH to LH. As a result, more follicles are selected to continue growth to an ovulatory size. The precise mechanisms by which these processes occur have recently been shown to involve oocyte follicle interactions (see section 1.1.5). Follicle development is modulated by GHIIGF and inhibin, however attempts to increase follicular development and ovulation through active inhibin immunisation alone have been variable and hence not commercially attractive. To develop successful protocols to induce twin ovulations in cows· and ewes, without superovulation, a clearer, more details understanding of follicullogenesis is required. The objective of the current study was to better understand these mechanisms through investigating interactions of GH/GF and inhibin in the ovary, follicle development, steroidogenesis, and receptor populations using an anoestrous sheep model. Spring born Mule x Charolais ewe lambs were actively immunised (n=8) against porcine inhibin α-C 1-26 peptide conjugated to KLH in NUFCA (primary and 3 boosters (NUFA», while 8 served as negative controls. Seven days following the final booster, the ewes were subdivided to give four groups: (1) controls + saline (n=4); (2) controls + rbGH (4ml s.c; 1mg. mr1; n=4); (3) inhibin immunised + saline (n=4); and (4) immunised + rbGH (n=4). Recombinant bovine growth hormone (rbGH) was given (Lm.) for 6 days. On day 4 GnRH (Receptal®; 1 ml) was injected s.c, to all animals to initiate the beginning of a new follicular wave. Blood samples were collected fortnightly to measure inhibin antibody titres, IGF-I, FSH and steroids. On the seventh day ensuing slaughter serum antibodies and ovaries were harvested. Left ovaries were intended for ISH (mRNA for P450arom) and/or immunohistochemical analysis. Follicles from right ovaries were dissected out, counted, measured and cultured in M199 at 37°C for 2 hours. Culture media was then assayed for oestradiol. Follicle shells were stored at -180°C for LH receptor binding studies. This work reports on the influence of different treatments on follicle populations. All immunised animals produced antibodies, which bound to 1251-inhibin. Using ANOVA to compare treatments it was observed that, Inhibin immunisation significantly (P<O.05) increased the number of follicles >3.5mm in diameter, but did not affect the smaller <3.5mm population. In contrast, rbGH administration led to a significant (P<O.05) elevation of follicles <3.5mm, without increasing the >3.5mm follicle numbers. These findings are in agreement with previous research. The molecular studies of left ovaries are not presented herein as due to time constraints the work was not completed and is currently on going. In conclusion, additional results of this study are required to meet the objectives of the experiment. Further research is required on dominant follicle selection if superovulatory programmes in both livestock and humans are to be more precisely controlled and readily accepted.

The available supply of domestic goat meat has not matched the increased demand for goat meat. High cost of production is a concern of goat producers, with feed being a major factor in input expenses. Increasing slaughter weight of kid meat goats would increase the available goat meat, but requires added nutrition beyond that obtained from typical forage based systems for goat production. Savannah bucklings (n=31) and Boer bucklings (n=28) were stratified by weight and breed and were randomly assigned a treatment of 0 (T1), 15 (T2), 30 (T3), or 45 (T4) percent dried distillers grain with solubles (DDGS). One goat from each pen was harvested on day 0 (H1), and every 21 days (H2, H3, H4) so that equal numbers of goats from each breed were sacrificed each harvest time. Bucklings and feed refusal were weighed weekly. Data was analyzed for ANOVA using Proc Mixed for fixed effects of treatment, harvest time and breed. There were no significant interactions for any traits measured. Breed did not affect (P>0.05) live performance, carcass traits, or cutability. Average daily gains (ADG) tended to linearly decrease with inclusion of DDGS, but significant difference were only observed in the second 21 days with T4 goats having the lowest (P<0.05) ADG. Treatment had no effect on feed efficiency. Goats in H4 had the highest (P<0.05) 1 and 3-hour temperatures and goats in H1 had the lowest (P<0.05) 1 and 3-hour pH values. The H4 carcasses had the largest ribeye areas and heaviest weights for most primal cuts. Carcasses and most primal cut weights of T4 goats were lighter (P<0.05) than those of goats in T1 and T2. Percentage of primal cuts in relation to the cold carcass did not differ (P>0.05) for treatments, but were influenced by harvest time. Warner-Bratzler shear force did not differ (P>0.05) for treatments and harvest time. The level and length of time feeding DDGS can affect goat carcass characteristics. This study found no differences in live traits, carcass characteristics, or meat from Boer- and Savannah-cross buckling kid goats.

Distillers dried grains with solubles (DDGS) is a common byproduct of the ethanol industry and is used in animal feeds. Carotenoids (xanthophylls) are already present in the human eye, and increasing the amount of carotenoids in the eye can help prevent eye diseases. The purpose of this research was to confirm that adding DDGS to standard corn and soybean meal hen diets may increase the amount of lutein available in egg yolks. An experiment was conducted with Hy-Line W-36 hens to evaluate the effects of DDGS in corn-soybean meal diets. Three hundred fifteen hens were fed one of seven treatment diets with five replications of nine hens per replicate in a completely randomized design. This was a 56-d trial. The treatment diets were: 1) Control (no DDGS), 2) 10% DDGS processed with heat treatment (DDGS+H), 3) 10% DDGS processed without heat treatment (DDGS-H), 4) 20% DDGS+H, 5) 20% DDGS-H, 6) 30% DDGS+H, and 7) 30% DDGS-H. Average daily feed intake, feed efficiency, egg specific gravity, egg mass, yolk color, and Haugh units were determined on three consecutive days at the end of each 28-d period. The eggs collected on the last three days of each 28-day period were stored either at room temperature or under refrigeration. Half of the stored eggs were broken out after three days of storage while the other half were broken out on day seven of storage, and measurements were collected. Throughout the trial, there was no effect of dietary treatment on average daily feed intake, feed efficiency, hen day production, egg weight, specific gravity, or hen weight. At the end of both 28-d periods, yolk redness (a*) was increased in eggs from hens fed DDGS-H or DDGS+H. Yolk yellowness (b*) was increased in hens fed diets with 20% of either DDGS+H or DDGS-H at the end of the second 28-d period. Storage method did affect egg quality. Eggs stored in refrigeration were higher in quality. The inclusion of any level of DDGS in hen diets did not affect hen egg production or egg quality but did increase yolk redness (a*) and yellowness (b*) which could be an indicator of increased lutein content.

Seventy-one Holstein calves were used to evaluate the effects of milk replacer (MR) feeding management alone or in combination with a multivitamin-electrolyte supplement on growth and mitigation of heat stress. Milk replacer treatments consisted of Land OLakes Herdmaker Supreme (20% CP, 20% fat) and Land OLakes Warm Front (27% CP, 10% fat). Calves received either 0 or 20 ml Palamountains Calf Boost® in MR once daily. Calves were offered treatments beginning on day 4. Calves on 27% CP : 10% fat MR were fed 2.72kg MR twice daily for the first three weeks of life, and 3.86kg twice daily until weaning. Beginning on day 42, MR feeding was reduced to 1 time per day to decrease MR intake by 50%. On day 49 calves were weaned. Water and calf starter (20% CP) were offered ad libitum beginning on day 4. Body weight, hip height, wither height, hip width, and body length were recorded weekly, and grain and water intakes were measured twice daily. Blood was collected on days 14, 28, 42, and 56 for analysis of plasma urea nitrogen (PUN), glucose, and â-hydroxybutyrate (BHBA), as well as rumen fluid for analysis of volatile fatty acids (VFA) and pH. There was a main effect of MR, with calves fed 27% CP: 10% fat MR showing greater body weights and increased hip height, wither height, and body length (P<0.05). Calves fed 27% CP: 10% fat MR consumed less grain than 20% CP: 20% fat MR calves (P<0.05) until the end of week 7, but showed no difference at week 8). Calves fed 27% CP: 10% fat MR had greater PUN concentrations (P < 0.05) than 20% CP: 20% fat. Glucose concentrations decreased (P < 0.05) as calves aged. There was no treatment effect (P > 0.05) on plasma BHBA or VFA concentrations; however, concentrations increased (P < 0.05) as calves aged. No effects of treatment or time were observed (P > 0.05) for rumen pH. These data indicate that MR feeding management may improve growth performance in neonatal dairy calves, but multivitamin mineral supplements may not provide any additional benefit.

Histone proteins are proteins found in eukaryotic cells that associate with DNA to form the most basic structural unit of chromatin, called nucleosomes. The post-translational modifications of histones regulate developmental competence in bovine oocytes and early embryos. The difference in developmental competence between in vitro matured oocytes and in vivo matured oocytes was used to investigate the accumulation of transcripts for histone methyltransferases (HMTs) during oocyte maturation. Methyltransferases ASH1L, EHMT2, SUV39H1 and KDM6B were selected as genes of interest. Transvaginal ultrasound-guided aspiration (TUGA) was used to collect immature and in vivo matured bovine cumulus-oocyte complexes (COCs). Immature COCs collected via TUGA were randomly assigned to either the immature or the in vitro mature treatments. Transcriptome analysis was performed in COCs, oocytes, and cumulus cells. Results showed no differences in transcriptome levels between immature and in vivo treatments, suggesting that there are no major accumulations of transcripts for HMTs during the antral phase of oocyte maturation in vivo. Higher accumulations of transcripts for the EHMT2 and ASH1L genes were found in the in vitro maturation Treatment for COCs and oocytes (p = 0.005 and p = 0.001, respectively). Immunocytochemistry was used to investigate the consequences of this increase in transcripts accumulation for HMTs during in vitro maturation of oocytes. Methylation levels of lysine 9 in histone 3 measured in both oocytes at the metaphase II stage and early embryos showed that the increase in the accumulation of transcripts coding for HMTs during in vitro maturation correlates with a decrease in the level of methylation of lysine 9 in histone 3 in oocytes at the metaphase II stage, as well as a decrease in the levels of methylation of lysine 9 in histone 3 in the blastomeres of early cleaving embryos. The decrease in the levels of tri-methylation of lysine 9 in histone 3 potentially affect the capacity of the oocyte and early embryo to silence gene and stabilize heterochromatic regions and potentially compromise the developmental potential of the embryo.

Two hundred forty beef calves (BW = 217 ± 20.6 kg) were used to evaluate performance, blood metabolites, and rumen development from feeding bermudagrass or ryegrass and rye baleage. Calves were stratified by BW, sex, and breed and assigned to one of 12 paddocks (0.40 ha each) with 4 treatment diets and fed for a 60 d backgrounding period. Diets included: early boot stage bermudagrass hay, (BERH); early boot stage ryegrass and rye baleage (ERRG); late bloom stage ryegrass and rye baleage, (LRRG); and early boot stage bermudagrass baleage, (BERB). Calves on BERH, LRRG, and BERB had free choice access to a 35% CP (as fed basis) liquid supplement. Body weights and rectal temperatures were collected on d -1, 0, 29, 30, 60, and 61 for comparison of BW, BW gain, ADG, and body temperature. Ruminal fluid and blood samples were collected for analysis of pH, NH3, VFA, PUN, and glucose from calves (n = 5 and 10/paddock, respectively) on d 0, 30, and 60. There was a treatment by day interaction (P < 0.01) for BW, temperature, PUN and ruminal pH. Body weights were heavier (P < 0.05) for LRRG compared with BERB and BERH, and heavier (P = 0.01) for ERRG compared with BERB on d 60, respectively. Body temperatures declined (P <0.01) from day 0 to 60. Plasma urea nitrogen was lowest (P <0.01) LRRG on d 30 compared with BERB and BERH; whereas, LRRG has the lowest (P <0.01) PUN on d 60 compared with the remaining treatments. Ruminal pH was lowest (P <0.01) for BERH and LRRG compared with ERRG on d 30, and highest (P <0.01) for ERRG on d 60 compared with BERB, respectively. Body weight gain and ADG were greater (P < 0.01) during the 60 d backgrounding period for calves fed ERRG and LRRG. A treatment effect existed for glucose where concentrations in the ERRG and BERH fed calves were greater (P <0.05) compared with the LRRG and BERB fed calves. A day effect for NH3 and glucose existed where concentrations decreased (P<0.01) from d 0 to 30 among all treatments. A treatment by day interaction existed (P =0.05) for butyrate where levels were greater for BERH and LRRG on d 30 compared with LRRG on d 60. Main effect of treatment (P <0.01) was observed for acetate and propionate, where BERB and LRRG had the lowest concentrations compared with ERRG and BERH. Performance of backgrounded calves fed ryegrass and rye baleage with or without supplementation, based on harvest stage, was improved over feeding bermudagrass hay with supplementation.

Variation in cryotolerance exists between embryos from different animal breed, species and management conditions. Reduced tolerance to chilling and cryotolerance of oocytes and embryos has been associated with greater cytoplasmic lipids (Kim et al., 2001; Seidel, 2006). Previous studies in the cow have demonstrated nutrition-induced modification of follicular components. Trans-10, cis-12 conjugated linoleic acid (CLA) was identified as a potent inhibitor of milk fat synthesis in lactating cows (Baumgard et al., 2000) and inclusion of CLA in bovine embryo culture medium improved post-thaw embryo survival (Pereira et al., 2007). Dietary supplementation of cows with CLA could alter oocyte fatty acid metabolism, oocyte lipid composition and embryo cryotolerance, and responses may be different between Bos indicus and Bos taurus breeds of cattle. Therefore, a series of experiments were conducted to evaluate effects of dietary CLA supplementation of cows on (1) milk fat depression in lactating Holstein cows, (2) follicle and oocyte production and lipid content of oocytes from Brahman and Holstein cows, (3) mRNA expression of genes involved in lipid metabolism in oocytes from Brahman and Holstein cows and (4) cryosurvival of in vitro-produced embryos from CLA-supplemented oocyte donor cows. Milk fat was depressed by 10.1% in lactating Holstein cows fed CLA. Follicle, oocyte and embryo production of cows were not influenced by CLA supplementation. Dietary supplementation of cows with CLA before oocyte collection did not influence cryotolerance of in vitro-produced embryos or expression of genes in oocytes involved in lipid metabolism. Lipid content of oocytes was not influenced by CLA supplementation. The ovarian response to dietary CLA was similar among Brahman, Holstein and crossbred beef cows. The highly regulated mechanisms involved in fatty acid uptake by ovarian components may help explain the lack of ovarian response to dietary CLA in the current study.

The main problem addressed in animal ethics is on what grounds and to what extent we owe animals moral consideration. I argue that many animals deserve direct moral consideration in virtue of their agency, selfhood and autonomy. I start by providing an account of agency and selfhood that admits of degrees, from minimal to complex, among animal species that is supported by current research on consciousness and the mental capacities of animals. I posit that agency and selfhood are morally valuable as they allow for subjective mental experiences that matter to conscious individuals. I then develop a view of autonomy that corresponds to my view of agency and selfhood, whereby the degree to which an individual is self-aware indicates the degree to which that being is autonomous. I argue that autonomy not only consists in the rational and reflective capacities of humans, but also at a more minimal level where autonomy is simply the ability to make choices. I support this view of autonomy as choice with an account of ???naturalized autonomy??? and explain some of the implications of this view for animals. After considering the views of Peter Singer, Tom Regan and Bernard Rollin on animal ethics, I analyze the flaws in their reasoning and argue that my own view provides a stronger account for the direct moral consideration of animals. This is due to my inclusion of agency, selfhood and autonomy, which these philosophers mainly neglect. I review some current reinterpretations of Kant???s moral arguments that claim animals ought to be considered ends-in-themselves. I present reasons why the inclusion of selfhood would strengthen this claim and further develop my argument for respecting the autonomy of animals. I conclude that a theory of animal ethics based on agency, selfhood and autonomy provides the strongest account for the direct moral consideration of animals, as it is empirically informed and provides a moral middle path between animal welfare and animal rights.