Scientists have developed techniques like ovum pick up (OPU) and follicular ablation as a large source of oocytes for creating IVP bovine embryos. These techniques have allowed for more efficient dissemination of valuable female genetics compared to traditional artificial insemination or embryo flushing. IVP embryos have lower embryo development rates and quality, leading to lower pregnancy rates. Nerve growth factor-beta (NGF), however, has been previously shown to improve 48-hour cleavage rates and the number of hatching/ hatched blastocysts out of total presumptive zygotes. We hypothesize that NGF will improve IVP embryo development by positively influencing cleavage and blastocyst rates. The first two experiments' objectives were to determine the effect of recombinant bovine (60 or 90% purity) and human NGF (97% purity) supplementation during in vitro fertilization on 24- and 48-hour cleavage and day 8 blastocyst development rates. The objective of the third experiment was to assess the effect of the supplementation of bovine NGF (90% purity) on heat shocked and non-heat shocked in vitro-matured cumulus-oocyte complexes, assessing cleavage rates at 48 and 72 hours post insemination and blastocyst development rates. The results of experiment 1 show there were no differences between any of the three treatment groups (bNGF60, hNGF95, and control) for 24 hour (P = 0.66) or 48 hour (P = 0.33) embryonic cleavage rates. Additionally, there were no differences between treatments in the total percentage of blastocysts per oocyte (P = 0.91) or the percentage of blastocysts per cleaved embryo (P = 0.32). The results of experiment 2 also showed no differences between any of the three treatment groups (bNGF90, hNGF95, and control) for 24 hour (P = 0.16) or 48 hour (P = 0.18) embryonic cleavage rates. Additionally, there were no differences between treatments in the total percentage of blastocysts per oocyte (P = 0.42) or the percentage of blastocysts per cleaved embryo (P = 0.57). In the 3rd experiment, there was not a significant effect of treatment (P ≤ 0.05) at all stages of embryonic development assessed. On the contrary, in the third experiment, non-heat stressed NGF treatment had an interestingly detrimental effect on early cleavage rates of embryos compared to the non-treated control embryos. These results showed that NGF could not improve in vitro embryonic development rates in standard conditions; however, this negative impact of NGF on early cleavage was not observed in heat-shocked embryos. Suggesting that there could be a protectant factor in NGF that warrants further investigation. / Master of Science / Nerve growth factor (NGF) was initially thought to only play a role in nerve cell development, but research has since shown an influence on female reproduction in cattle. NGF and its receptors have been identified in the follicular fluid and reproductive cell types of females, contributing to egg maturation. Previous data on NGF supplementation with IVP embryos, which took place during the summer, showed that NGF positively affected in vitro-produced embryo development when added to fertilization media, specifically on cleavage rates (division without growth, must be two cells or greater) and blastocyst development. The actual role of NGF on embryo development is still unclear. Therefore, replication of this study is essential. First, we added either recombinant human nerve growth factor (90% pure) or bovine nerve growth factor (60% or 90% pure) to the IVF medium. The goal was to determine if NGF would have the same effects on cleavage rates as bovine purified NGF when supplemented during the fertilization stage, as well as to decide if protein purity and species affected how NGF influenced embryo development rates. For the second part of the study, we heat-shocked oocytes during maturation in a "hot incubator" and supplemented them with bovine 90% NGF. This was done to mimic the summer month heat stress that may have occurred in the abstract data. Our objective was to determine if NGF could mitigate the detrimental heat shock during development and potentially improve embryo developmenNerve growth factor (NGF) was initially thought to only play a role in nerve cell development, but research has since shown an influence on female reproduction in cattle. NGF and its receptors have been identified in the follicular fluid and reproductive cell types of females, contributing to egg maturation. Previous data on NGF supplementation with IVP embryos, which took place during the summer, showed that NGF positively affected in vitro-produced embryo development when added to fertilization media, specifically on cleavage rates (division without growth, must be two cells or greater) and blastocyst development. The actual role of NGF on embryo development is still unclear. Therefore, replication of this study is essential. First, we added either recombinant human nerve growth factor (90% pure) or bovine nerve growth factor (60% or 90% pure) to the IVF medium. The goal was to determine if NGF would have the same effects on cleavage rates as bovine purified NGF when supplemented during the fertilization stage, as well as to decide if protein purity and species affected how NGF influenced embryo development rates. For the second part of the study, we heat-shocked oocytes during maturation in a "hot incubator" and supplemented them with bovine 90% NGF. This was done to mimic the summer month heat stress that may have occurred in the abstract data. Our objective was to determine if NGF could mitigate the detrimental heat shock during development and potentially improve embryo development rates under these stressful conditions. The results of all experiments indicated that NGF could not influence development rates. positively. On the contrary, in the third experiment, non-heat stressed NGF treatment had an interestingly detrimental effect on early cleavage rates of embryos when compared to non-treated control embryos. This negative impact of NGF on early cleavage was not observed in heat-shocked embryos pointing to a possible protectant factor in NGF that needs further investigation.t rates under these stressful conditions. The results of all experiments indicated that NGF could not influence development rates. positively. On the contrary, in the third experiment, non-heat stressed NGF treatment had an interestingly detrimental effect on early cleavage rates of embryos when compared to non-treated control embryos. This negative impact of NGF on early cleavage was not observed in heat-shocked embryos pointing to a possible protectant factor in NGF that needs further investigation.

Return to estrus following the postpartum interval to achieve pregnancy success on time is a considerable obstacle for beef females. Chromium supplementation increases available glucose and insulin sensitivity within cells. Two experiments were conducted to investigate the effects of supplementing Chromium propionate (CrP) during the peripartum until weaning on productive and reproductive performance in Bos taurus beef cows. In Exp. 1, 62 Angus-based beef cows were stratified by predicted calving date, body weight (BW) and randomly assigned to one of two treatments: 1) CON, (n=30) supplementation of corn gluten, soy hull pellet feed (50:50) with a mineral pack at 1 kg -1hd-1d; or 2) TRT, (n=32) supplementation at 1 kg -1hd-1d of corn gluten, soy hull pellet feed (50:50) with a mineral pack containing 1.4 g of Chromium Propionate (KemTRACE® Chromium 0.4%, Kemin Industries, Inc., Des Moines, IA). Cows remained on a single pasture equipped with SmartFeed trailers for individual supplement intake (SmartFeed®, C-lock Inc., Rapid City, SD). The experiment lasted 98 days, starting 63 days pre-breeding to 35 post-fixed-time artificial insemination (TAI). Ovarian ultrasonography was performed on days -10, -3, 0 (TAI Day), and 7 to determine the diameter of the largest follicle and corpus luteum (CL) volume. Age, days postpartum (DPP), initial and final BW, and supplement intake were similar (P>0.05) between treatments. However, TRT cows had a larger follicle (P=0.028) on d 0, increased CL volume (P=0.038), and increased (P=0.0213) circulating progesterone (P4) on day 7. In Exp. 2, 953 beef cows across nine locations were assigned to one of two treatments: 1) CON, supplementation of a mineral product at 113 g -1hd -1d (n=464 cows; 16 experimental units); or 2) TRT, supplementation of mineral product at 113 g -1hd -1d containing 1.4 g of CrP n=489 cows; 16 experimental units). Supplementation started approximately 37 days pre-calving and continued until weaning for 345 days. Age, DPP, d-10 body condition score (BCS), initial and final BW, BCS, calf birth and weaning weight, and mineral disappearance were similar (P>0.05) between treatments. However, CrP cows tended (P=0.081) to have greater estrus expression (68.3 and 60.2 ± 3.1 %, for CrP and CON, respectively) and greater (P=0.045) TAI pregnancy rates (55.2% vs. 49.9% ± 2, for CrP and CON, respectively). We conclude that supplementation of CrP to beef cows during the peripartum through weaning did not affect BW or BCS, but increased ovulatory follicle diameter, estrus expression, CL volume, and P4 concentration, and one or more of these positive effects of CrP likely contributed to the improvement in TAI pregnancy rate. / Master of Science / Reproductive efficiency in beef cow-calf operations relies heavily on pregnancy success as early in the breeding season as possible. Several factors influence the return to cyclicity after calving, including days post-calving, body condition, and nutritional status of the dam. Chromium is an essential trace mineral that can be added to cattle diets to promote glucose uptake by cells and mitigate the negative impacts of the stressful postpartum period. Two studies were designed to evaluate the effect of chromium propionate supplementation to beef cattle during the pre-calving through weaning period on reproductive parameters and overall cow performance. In experiment one, 62 cows were assigned to two treatment groups: control (feed containing a standard mineral pack) and treatment (feed containing 1.4g of chromium propionate supplementation). The second experiment included 953 cows assigned to one of two treatment groups: control (supplementation of free choice mineral product) and treatment (supplementation of free choice mineral containing 1.4 grams of chromium propionate). Both groups in experiment two had the same target feed intake. In both experiments, beef females were exposed to estrous synchronization and timed artificial insemination and were exposed to a bull for natural service in the next estrous cycle. Cows receiving the chromium supplementation did not experience any changes in body condition score, weight change, or average daily gain in either experiment. However, reproductive performance was affected by chromium-supplemented cows had an increased dominant follicle size on the day of timed-artificial insemination (TAI), larger corpus luteum volume 7 days post artificial insemination, increased progesterone concentrations, and increased progesterone to corpus luteum ratio in experiment 1. Supplementation of Cr in experiment 2 tended to increase estrus expression and showed an increase in the pregnancy rate of TAI. Calf performance was not impacted by chromium propionate supplementation. These results indicate that chromium propionate supplementation could benefit the reproductive performance of beef cows.

Bovine lactoferrin (BLF) and bovine transferrin (BTF) are major-iron transport and regulation proteins found in bovine whey. BLF and BTF must interact with the eukaryotic cell surface to mediate their biological function of iron delivery and cellular functions of inflammatory and immunological modulation. As common components of the eukaryotic cell surface, gangliosides were used for affinity purification of BLF and BTF. Bovine gangliosides were isolated from fresh buttermilk and covalently immobilized onto controlled-pore glass beads (66 μg/g beads). After the matrix was loaded with whey protein (WPI or WPC), lactoferrin was eluted with 1 M NaCl and lll identified by N-terminal protein sequencing. Pretreated whey isolate (1 % wt/vol) showed the highest lactoferrin purity with 40% among protein sources, and whey protein isolate (10% wt/vol) showed the highest recovery with 105%. Bovine transferrin was eluted with sodium phosphate buffers at pH 7 after the immobilized matrix was loaded with a 2% (wt/vol) whey solution. The ganglioside column resulted in a 74.2% recovery of BTF from whey, and the BTF was enriched to 61% purity after Mono-Q chromatography. Bovine transferrin was identified by SDS-PAGE analysis, Western analysis, and isoelectrofocusing. In conclusion, immobilized gangliosides can be used to purify BLF and BTF from bovine whey.

Thesis (M. Sc.)--University of Alberta, 2009. / Title from pdf file main screen (viewed on Dec. 22, 2009). "A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Master of Science in Animal Science, Department of Agricultural, Food and Nutritional Science, University of Alberta." Includes bibliographical references.

Orientador: Roberto Sartori Filho / Banca: José Buratini Júnior / Banca: Pietro Sampaio Baruselli / Resumo: Objetivou-se avaliar o efeito da condição corporal e/ou da alta ingestão de matéria seca (AIMS) na produção in vivo de embriões de fêmeas bovinas superovuladas. Em um primeiro experimento, 14 vacas Simental x Nelore não-lactantes com elevado escore de condição corporal (ECC) foram divididas em grupos de Manutenção=M ou alta ingestão de matéria seca=AIMS. As vacas do grupo AIMS receberam dieta com 180% da manutenção, entre 7 dias antes do início da superovulação (SOV) e o final das aplicações de FSH. O grupo M recebeu dieta de manutenção. O número de folículos recrutados e ovulados não diferiu entre os grupos (P>0,10). Entretanto, os números de estruturas totais e embriões viáveis colhidos foram maiores no grupo M (P<0,05). Em um segundo experimento avaliou-se a influência do ECC associado ou não da AIMS na produção embrionária em 36 novilhas Nelore. AIMS ocorreu por 14 dias antes do início da SOV. Após colheita, os embriões viáveis foram congelados para posterior cultivo até eclosão. Não houve diferença entre os grupos na população folicular ao início da SOV, na resposta superestimulatória ou superovulatória, nem no número ou qualidade dos embriões colhidos. As novilhas com <ECC apresentaram maiores concentrações séricas de IGF-I, entretanto as com >ECC tiveram insulina mais alta. Os embriões dos animais com >ECC apresentaram diâmetro, taxas de eclosão e expressão relativa de mRNA do gene BAX superiores após cultivo do que os coletados no grupo com <ECC. Os resultados conflitantes entre os experimentos e com dados da literatura sugerem haver diferentes respostas à alimentação dependendo da raça ou estado nutricional na produção embrionária. / Abstract: The aim of this study was to investigate the effect of body condition and/or high dry matter intake (flushing) on in vivo embryo production in superovulated female cattle. A first experiment used 14 non-lactating Nelore x Simmental cows with a high body condition score (BCS) divided into Maintenance=M or Flushing=F groups. Seven days prior to onset of superovulation (SOV) until the last day of treatment with FSH, group F cows were fed a diet to achieve 180% of maintenance. Group M cows were fed a maintenance diet. The number of recruited or ovulated follicles did not differ between groups (P>0.10). However, the total number of embryos/ova and the number of viable embryos recovered were greater in the M group (P<0.05). A second study investigated whether differences in BCS, associated or not with nutritional flushing, influence the embryo production in 36 Nelore heifers. Nutritional flushing was conducted during 14 days prior to the onset of SOV. After recovery, viable embryos were frozen to be subsequently cultured until hatching. There was no difference among groups for follicle population at onset of SOV, superstimulatory or superovulatory responses, nor number or quality of recovered embryos. Heifers with <BCS had grater blood concentrations of IGF-I, whereas heifers with >BCS had grater insulin. Embryos collected from >BCS heifers had greater diameter, hatching rates and relative expression of the BAX gene mRNA than the ones recovered from <BCS heifers. The conflicting results between the experiments and also in relation to data from the literature suggest that there are different responses to diets depending on animal breed or nutritional status on embryo production. / Mestre

Call number: LD2668 .T4 1966 G883 / Master of Science

Master of Science / Department of Clinical Sciences / David E. Anderson / Lameness in cattle is a common cause of pain however there are no approved cattle analgesic drugs. Flunixin meglumine, the only non-steroidal anti-inflammatory drug approved for use in adult dairy cattle, is labeled for pyrexia associated with bovine respiratory disease, endotoxemia, acute mastitis and associated inflammation. There is currently a lack of objective data regarding the analgesic efficacy of flunixin meglumine in cattle. The objectives of this study were to characterize an amphotericin B-induced lameness model and to ascertain the analgesic effects of flunixin meglumine using multimodal assessment. We hypothesized that flunixin meglumine would provide analgesia as evidenced by increased activity levels as well as increased exerted force and contact area on the affected limb in flunixin treated steers. Amphotericin B-induced synovitis arthritis was induced in the distal interphalangeal joint of 10 dairy steers. The cattle were randomly allocated between a treatment and a control group. The treatment steers received flunixin meglumine at the time of arthritis induction and at 12 hours post-induction. Accelerometric, gait, pressure mat, vital parameter and plasma cortisol data were gathered in the pre and post-induction phases. The data were analyzed using linear mixed models with treatment and time designated as fixed effects. Induction of amphotericin B arthritis produced a moderate, transient lameness. Control steers were more than twice as likely to be lame as flunixin meglumine treated steers using visual lameness assessment (92.2% ± 8.1 versus 40.7% ± 2.5) (P<0.03). Flunixin meglumine treated steers placed significantly greater force and contact area on the affected foot. Control steers also placed significantly greater force, impulse and contact area on the paired claw as compared to control steers. Flunixin treated steers spent considerably less time in recumbency than their control counterparts, particularly in the immediate post-induction time period. This is one of the first studies to document the character of an amphotericin B-induced synovitis arthritis model in cattle as well as to document analgesic efficacy of a nonsteroidal anti-inflammatory drug in an induced lameness model. Flunixin meglumine was efficacious in providing analgesia in an amphotericin B-induced lameness model in dairy steers.

Die vorliegende Arbeit beschäftigt sich mit der Identifikation neuer immunologischer Indikatoren (Biomarker) für den allgemeinen Gesundheitszustand von Hochleistungsmilchkühen. Diese Biomarker sollen möglichst einfach und schnell mittels eines Stalltests nachweisbar sein, weshalb die gelösten Proteine in der Milch im Fokus standen. Die neuen Biomarker sollten nicht nur Mastitis, sondern vor allem auch Entzündungen außerhalb des Euters anzeigen können. Zu Beginn sollte das Gesamtspektrum an Immunkomponenten erfasst werden, weshalb zunächst auf Proteinexpressionsebene angesetzt wurde. Das schloss die Analyse von vorhandenen Immunzellpopulationen in Blut- und Milchproben ein, um einen Überblick über potentielle Produzenten der immunologischen Indikatoren zu erhalten. Es konnte erstmals Cluster of Differentiation (CD) 25 (alpha-Kette des Interleukin-2-Rezeptors, IL2R) auf bovinen polymorphnukleären, neutrophilen Granulozyten (PMN) aus peripherem Blut nachgewiesen werden. Die Expression (mittlere Fluoreszenzintensität, MFI) von CD25 stieg dabei mit dem Schweregrad der entzündlichen Erkrankung an. Die Ergebnisse konnten auf Transkript- wie auch auf Proteinexpressionsebene bestätigt werden. Gleiche Tendenzen waren auch für Milchzellen erkennbar. In der statistischen Analyse zeigte CD25 auf PMN im peripheren Blut ein hohes Abgrenzungsvermögen für erkrankte Kühe. Die Messung von CD25 auf PMN könnte somit zur Bestimmung des allgemeinen Gesundheitszustandes von Hochleistungsmilchkühen genutzt werden.

No description available.

The bovine papilloma virus type 2 (BPV-2) L2 open reading frame (ORF) was cloned into a λ pL promoter expression vector. This clone was shown to express a fusion protein which comprised 75% of the BPV-2 ORF linked to the first 13 N-terminal amino acids of the λ cIl gene product. Antisera was generated against this fusion protein and subsequently used to identify the L2 gene product as a 64,000 dalton protein in BPV-2 virions. It was also demonstrated that the L2 viral protein was present in full caps ids, but only in very limited amounts in empty caps ids. Densitometer analysis indicated that the L2 protein comprised only 8% of the total L1 + L2 "Coomassie blue stainable" protein in full capsids. The antisera was also used to demonstrate that the BPV-2 L2 gene product is antigenically related to the BPV-1 L2 gene product. Finally, an attempt was made to determine the location of the L2 gene product within the capsid structure. Hemagglutination inhibition and enzyme-llnked-immunosorbent- assay data both indicate that the L2 protein is exposed on the surface of the capsid. Immune electron microscopy data was inconclusive in determining the location of the L2 gene product.