The inclusion of precision dairy farming technologies in dairy operations is an area of increasing research and industry direction. Machine vision based systems are suitable for the dairy environment as they do not inhibit workflow, are capable of continuous operation, and can be fully automated. The research of this dissertation developed and tested 3 machine vision based precision dairy farming technologies tailored to the latest generation of RGB+D cameras. The first system focused on testing various imaging approaches for the potential use of machine vision for automated dairy cow feed intake monitoring. The second system focused on monitoring the gradual change in body condition score (BCS) for 116 cows over a nearly 7 month period. Several proposed automated BCS systems have been previously developed by researchers, but none have monitored the gradual change in BCS for a duration of this magnitude. These gradual changes infer a great deal of beneficial and immediate information on the health condition of every individual cow being monitored. The third system focused on automated dairy cow feature detection using Haar cascade classifiers to detect anatomical features. These features included the tailhead, hips, and rear regions of the cow body. The features chosen were done so in order to aid machine vision applications in determining if and where a cow is present in an image or video frame. Once the cow has been detected, it must then be automatically identified in order to keep the system fully automated, which was also studied in a machine vision based approach in this research as a complimentary aspect to incorporate along with cow detection. Such systems have the potential to catch poor health conditions developing early on, aid in balancing the diet of the individual cow, and help farm management to better facilitate resources, monetary and otherwise, in an appropriate and efficient manner. Several different applications of this research are also discussed along with future directions for research, including the potential for additional automated precision dairy farming technologies, integrating many of these technologies into a unified system, and the use of alternative, potentially more robust machine vision cameras.

The objective of this research was to investigate the effects of slow release urea on N metabolism in cattle. The ruminal behavior of Optigen®II and the effect of basal diet on the in situ degradability of urea and Optigen®II were evaluated. The effect of slow release urea and its interaction with degradable intake protein (DIP) level in the diet on N retention and excretion was evaluated utilizing 8 Holstein steers in a 4 x 4 Latin square experiment. In addition, the effect of slow release urea and DIP level on ruminal and systemic urea kinetics was evaluated using stable isotope techniques with 8 Holstein steers in a 4 x 4 Latin square experiment. Finally, slow release urea was evaluated under a practical beef production setting. The performance of slow release urea was compared to regular feed grade urea in a 42 day receiving study (288 Angus cross steers) as well as a 70 day growing study (240 Angus cross steers). High forage diets increased the ruminal degradation rate of both urea and slow release urea an increased the extent of degradation of slow release urea when compared to high concentrate diets. Lower DIP concentrations in the diet reduced systemic urea production, ruminal ammonia and plasma urea concentrations and urinary urea excretion under most circumstances but also led to a reduction in N retention, reduced diet digestibility, lower feed intake, lower growth rate and decreased feed efficiency. High DIP intakes increased N retention, growth rate, diet digestibility and improved feed efficiency but also lead to increased excretion on urea N in the urine. Slow release urea improved N retention and efficiency of N retention in high DIP diets when compared to urea and generally reduced plasma urea and ruminal ammonia concentrations. Compared to urea, slow release urea did not significantly improve the production of receiving cattle. However Optigen®II improved the feed efficiency when compared to urea on high concentrate diets but reduced feed efficiency on high forage diets.

Developing solutions to the metabolic stress experienced by cows during the transition period is very important because it can negatively influence lactational performance. The objectives were to: 1) compare physiological changes through body weight (BW) and concentrations of non-esterified fatty acids (NEFA) and β-hydroxybutyric acid (BHBA) and 2) evaluate feed intake, milk production, and energy balance (EB) of cows fed brown midrib corn silage (BMRCS)-based diets when compared with conventional corn silage (CCS)-based diets during the transition. At 4 wk prior to parturition, 40 dry multiparous Holstein cows were randomly assigned treatments. The treatment groups consisted of 2 close-up transition diets (CCS-based and BMRCS-based diet) offered to 2 groups of 20 cows each beginning at 4 wk prepartum. After calving, 10 cows from each prepartum group were individually fed one of four dietary treatments. The four dietary treatments postpartum were defined as follows: 1) CC = CCS-based close-up diet + CCS-based lactation diet; 2) CB = CCS-based close-up diet + BMRCS-based lactation diet; 3) BB = BMRCS-based close-up diet + BMRCS-based lactation diet; 4) BC = BMRCS-based close-up diet + CCS-based lactation diet. Cows were sampled weekly for feed intake, and feed composition was taken monthly. After calving, milk yields were recorded daily and milk components were analyzed monthly. Body weights were taken twice per week on wk -4, -2, 1, 2, 3, 4, 8, 12, 16, and 20. Blood serum was sampled 3 times per week from wk -4 through 4 and then on wk 6, 8, 14, and 20. Rumen fluid was sampled on wk -4, 4, 8, 14, and 20. Feeding BMRCS-based diets during the transition did have a positive influence on dry matter intake, milk production, and energy balance. Interestingly, feeding BMRCS-based diets only during the close-up period and feeding a CCS-based diet during the lactation had similar positive effects as feeding a BMRCS-based diet through the dry period and during the lactation. This finding is meaningful because producers, especially in the Intermountain West, have experienced BMR crop yields that have been less than that of conventional crop yields and may be unwilling to utilize BMRCS in dairy rations. However, if feeding a BMRCS-based diet for a limited amount of time is beneficial, producers could be more willing to utilize this silage hybrid as an important transition period management tool.

My dissertation hypothesis is that bovine trophoblast cells express cell-surface and secreted non-classical major histocompatibility complex class I (MHC-Ib) proteins which inhibit NK cells and other leukocytes by binding to inhibitory receptors (e.g., LILRB1, LILRB2, KIR2DL4, and/or CD94/NKG2A). Extremely polymorphic and ubiquitously expressed classical MHC class I (MHC-Ia) proteins, which present foreign antigenic peptides to CD8+ T lymphocytes, are involved in acceptance or rejection of tissue grafts. Non-classical MHC class I (MHC-Ib) glycoproteins, such as Human Leukocyte Antigen-G (HLA-G) and murine Qa-2, are important modulators of the maternal immune system during pregnancy. MHC-Ib proteins are: (a) oligomorphic or monomorphic, (b) expressed in specific tissues under specific condtions, and (c) produced as surface and/or soluble isoforms due to alternative splicing. Third trimester-bovine trophoblast cells express both MHC-Ia and MHC-Ib proteins. The MHC-Ib proteins expressed by trophoblast cells during the third trimester of pregnancy are encoded by four bovine leukocyte antigen (BoLA) loci: BoLA-NC1, BoLA-NC2, BoLA-NC3, and BoLA-NC4. Two MHC-Ia (N*01701 and N*01802) and three MHC-Ib (NC1*00501, NC3*00101 and NC4*00201) proteins showed cell-surface expression in transfection studies performed in murine P815 and human K562 cells. Two additional isoforms, NC1*00401 and NC2*00102, were not detected on the surface of these cells. Nevertheless, both class Ia proteins, N*01701 and N*01802, and five class Ib proteins, NC1*00401, NC1*00501, NC2*00102, NC3*00101, and NC4*00201, were detected in crude cell lysates on Western blots. Precipitation of proteins from culture supernatants showed that cell-surface MHC-Ia (N*01701 and N*01802) and MHC-Ib proteins (NC1*00501, NC3*00101, and NC4*00201) are shed from the surface of these cells into the media. The mechanism of shedding of these proteins is, however, not known. Monoclonal antibodies W6/32, IL-A88, H1A, H6A, H11A, H58A, and PT-85A recognized surface MHC-I isoforms with varying affinity. We were able to develop a sandwich enzyme-linked immunosorbent assay (ELISA) using either H1A or IL-A88 antibody as the capture antibody and the W6/32 antibody for detection. We produced monoclonal antibodies against cattle NC1*00501 and NC3*00101 proteins. One monoclonal antibody generated against BoLA-NC3*00101 was highly specific. Unfortunately, due to failure to clone the NC3*00101- hybridoma, we no longer have an infinite source of this monoclonal antibody for NC3*00101. We eluted peptides from NC3*00101-transfected MHC-null K562 cells and identified peptides using liquid chromatography-mass spectrum (LC-MS) analysis. Analysis of peptide binding data using the SAS Proc mixed statistical program, suggested that the peptide EVTNQLVVL is a potential peptide ligand, which can be used to make tetramers for enumeration of antigen-specific leukocytes.

Toxaphene is reported to cause defects in the collagen of fish. Chronic exposure to toxhaphene weakens the backbone of fish by decreasing the amount of collagen and usually increasing the amount of calcium in the bone which results in a more brittle and fragile bone. We investigated the possible direct action of toxaphene on collagen synthesis by exposing vertebral and swim bladder organ cultures obtained from unexposed rainbow trout (Salmo gairdneri) fingerlings to the same lot of toxaphene found to cause this defect in vivio. Collagen produced by these organ cultures was measured by: (1) total 3H-proline incorporated into the matrix; (2) 3H-proline released during collagenase digestion of acid-precipitated protein; (3) 3H-hydroxyproline extracted from the acid hydrolysate, and (4) tritiated water produced during the hydroxylation of 4-3H-proline. At a relatively high concentration of toxaphene (2.4 mM) these indices of collagen production were reduced, but this was probably caused by a decrease in tissue viability rather than by a direct affect on collagen synthesis. At 240 μM cellular protein synthesis was reduced. Generally no effects were found at toxaphene concentrations below 240 μM. From these studies it was concluded that toxaphene does not have a direct inhibitory effect upon collagen production at the tissue level. For comparison to the in vivo were measured. All three of these parameters were significantly reduced in comparison to controls (α = 0.05) in those fish exposed to the highest concentration of toxaphene (200 ng/1). fish exposed to 150 ng/l toxaphene also had reduced prolyl hydroxylase activity. These results indicated that vertebral prolyl hydroxylase activity may be a sensitive indicator of toxaphene exposure in fish, and inhibition of that enzyme may be involved in the mechanism of toxaphene-induced collagen effects.

Mastitis is one of the most common and expensive dairy cattle diseases. Mastitis prevention and management are key factors in herd health and improved milk quality. One objective of this research was to evaluate management solutions to maintain a low somatic cell count, based on survey responses from Kentucky dairy producers. Because hyperkeratosis may increase mastitis incidence, another objective of this research was to examine changes in teat end hyperkeratosis in a herd transitioning from a standard pulsation milking system to an individual quarter pulsation milking system. The last objective of this research was to evaluate technologies that monitored rumination time, neck activity, reticulorumen temperature, and milk yield as potential mastitis detection devices.

Poor estrus detection can limit the reproductive performance of a dairy herd. One objective of this research was to evaluate an alternative method to traditional estrus detection in the form of automated monitoring technologies. To accomplish this, the first study considered the ability of automatically monitored parameters (activity, number of steps, lying bouts, lying time, feeding time, rumination time, and temperature) to detect estrus. A second study compared automated activity monitoring to timed artificial insemination as reproductive management strategies on commercial herds. The other objective of this research was to evaluate the economic potential of automated estrus detection technologies. This was accomplished by creating and evaluating a farm specific decision support tool to determine the net present value of adopting an automated estrus detection technology.

Precision dairy farming technologies provide a variety of functions to dairy farmers. Little is known about dairy producer perception of these technologies. A study was performed to understand dairy producer perception of parameters monitored by precision dairy farming technologies. Calving has potential to be predicted using these same parameters and technologies. A second study was performed using two commercially marketed technologies in calving prediction. In order for these technologies to generate accurate and useful information for dairy farm use, they must accurately quantify these parameters. The final study evaluated the accuracy of five commercially marketed technologies in monitoring feeding, rumination, and lying behaviors.

Bovine somatotropin (bST) is a protein synthesized at the base of the brain and released by the pituitary gland into the circulatory system. BST is transported by the circulatory system and absorbed only by cells of target organs that possess cell surface receptors for the protein (11, 41). Its effect is initiated via a protein receptor initiation and cyclic AMP cascade . This effect on the cell continues well past the degradation of the bST molecule. BST is likely transported into the cell, where it is degraded . Its constituent blocks, amino acids, are used to synthesize new proteins or converted to other metabolites such as sugars (1).

Enterovirus D68 (EV-D68) virus has become more prevalent over the last 15 to 20 years. EV-D68 attacks the respiratory system and can cause severe disease in individuals who have underlying respiratory problems. There have also been reports of individuals with EV-D68 showing signs of neurological system problems and acute flaccid paralysis. Because of the increase in patients with EV-D68 and also the potential for neurological disease, an animal model is needed to study the disease and to evaluate experimental therapies for EV-D68 infection. To develop the animal model, 4-week old AG129 mice that lack alpha and beta interferon receptors, making them immunosuppressed, were used. The mice were infected with EV-D68 by the intranasal route to allow infection of the lungs. On day-3 post-infection the mice were euthanized and lungs were removed and homogenized for collection of virus. The newly collected virus was then used to infect another set of mice. This procedure was repeated 30 times. As passage number increased so did the amount of virus that was collected from the lungs of mice. The virus titer increased 320-fold between mouse passage 0 to 30. At the end of the thirtieth passage, multiple tissues (lungs, liver, kidney, spleen, blood, brain, spinal cord and leg muscle) were collected from infected mice over several days and titered to demonstrate how quickly the virus spread to various tissues within the mouse. The virus replicated most rapidly in the lungs and remained in the lungs longer than the other tissues evaluated. However, large quantities of virus were found in all tissues evaluated. Finally, several experimental antiviral compounds were evaluated: rupintrivir, pleconaril, ribavirin, enviroxime and guanidine, all of which showed therapeutic potential in cell culture. In the animal model rupintrivir, pleconaril, ribavirin and enviroxime did not show any therapeutic effect. Only guanidine reduced the amount of virus that was found in the lungs as well as in whole blood.