Bovine mastitis, or inflammation of the mammary gland, has substantial economic and animal welfare implications. A genetic basis for mastitis resistance traits is recognized and can be used to guide selective breeding programs. The discovery of regions of the genome associated with mastitis resistance, and knowledge of the underlying molecular mechanisms responsible, can facilitate development of efficient mastitis control and therapeutic strategies. The objectives of this dissertation research were to identify sites of genetic variation associated with mastitis resistance, and to define the contributions of the milk-secreting epithelial cells to mammary gland immune responses and mastitis resistance. Twenty seven regions of the bovine genome potentially involved in mastitis resistance were identified in Holstein dairy cattle. Additionally, this research demonstrates a role of bovine mammary epithelial cells in mastitis resistance, and provides guidance for the use of an in vitro model for mastitis studies. Primary bovine mammary epithelial cells from mastitis-resistant cows have differential expression of 42 inflammatory genes compared with cells from mastitis-susceptible cows, highlighting the importance of epithelial cells in mastitis resistance. Bovine mammary epithelial cells display both similarities and differences in pro-inflammatory gene expression compared to fibroblasts, and their expression of inflammatory genes is influenced by administration of the enzyme phospholipase A2. The growth potential of milk-derived bovine mammary epithelial cells in vitro can be extended, facilitating their use in mastitis studies, by transfection with a viral protein. Collectively, this research contributes to current knowledge on bovine mastitis resistance and in vitro models.

A recombinat Chinese Hamster Ovary (rCHO) cell line designated as CHO SEAP was utilized in this investigation to optimize protein production. Two bench top stirred-tank bioreactors, namely a pitched-blade and a packed-bed basket bioreactor, were utilized for a comparative study to determine which bioreactor would produce the best results in terms of protein production. The objective of this research project was to provide basic data that shows cells cultured in a packed-bed basket bioreactor in perfusion mode will generate more protein product than cells in batch mode suspension culture with a pitched-blade bioreactor. The packed-bed bioreactor creates a homeostatic environment similar to the environment found in vivo, where waste products are constantly removed and fresh nutrients are replenished. Closed batch cultures do not provide a homeostatic environment. In batch culture systems, nutrients are depleted and waste products accumulate. The results from this experiment could help investigators involved in protein and/or vaccine production facilities select the appropriate bioreactor and mode of operation to optimize cell productivity for generation of a specific protein product. CHO cells have been used for the production of vaccines, recombinant therapeutic proteins, and monoclonal antibodies, and these cells are now the cell line of choice in the biopharmaceutical industry. Traditional vaccine production methods in egg embryos are slow and outdated, whereas roller bottle-based cell culture techniques are time consuming and have limited scalability. These limitations justify the need for development of stirred tank bioreactors. Cells cultured in a packed-bed bioreactor are not exposed to hydrodynamic forces, as is the case with pitched-blade bioreactors, allowing for maximum growth and protein expression. This mode of operation involves the constant removal of media depleted of nutrients and the addition of fresh media with more nutrients to keep the cells growing. Long run times decrease the constant need for re-seeding cells and re-establishing seed cultures, thus, reducing setup time and labor dramatically. Secreted products are automatically separated from cells in perfusion, eliminating filtration and membrane fouling. A detailed description of both modes of operation are discussed in this thesis.

A live birth from a somatic cell nuclear transfer (SCNT) embryo represents a small percentage of donor cells that survived the reprogramming gauntlet. The inability to reprogram histone modifications in the donor cell line could add to the reprogramming deficiencies associated with SCNT. The effects of two histone modifications associated with transcriptional activation (H3K4m3 and H4K16ac) and two histone modifications associated with repressing transcription (H3K9m2 and H3K27me3) were evaluated in the context of their association to three genes known to contribute to maintaining totipotency: Nanog, Oct4, and Sox2. A µChIP assay was utilized using antibodies specific for each histone modification followed by real time PCR (qPCR) analysis to quantify the percentage of each gene associated with each particular histone modification. Gene expression analysis was followed by immunofluorescence and protein analysis. Results of these analyses suggest that gene association to certain histone modifications did not accurately predict gene expression in bovine blastocyst embryos. Of the three genes studied, only Oct4 expression differed significantly between in vitro fertilized (IVF; control) and SCNT blastocysts. Protein levels detected through immunofluorescence correlated directly with the gene expression analysis. Nanog and Sox2 expression profiles of IVF and SCNT bovine blastocysts are similar, yet the histone modification profiles associated with all three genes differ significantly. Altered expression levels in developmentally important genes will likely result in abnormal activity of the associated cellular pathway. Aberrant histone modifications, along with abnormal Oct4 expression, may contribute to the low percentage of SCNT embryos that result in live offspring.

Members of the genus Peromyscus, White-footed Mice, are known to occur in nearly all habitats of North America. Because of their numerical abundance and widespread distribution, these mice are extensively used in the study of the dynamics of animal populations. Thus, information perteining to the local distruibution and activities of these mice is of value to the more comprehensive studies that are being carried out in the field of zoological research.

Mastitis, an infection of the mammary gland, is the most common and expensive animal health problem for the dairy industry and affects every dairy farm to some degree. This disease complex is painful for dairy cows, increases the on-farm use of antibiotics, presents a threat to milk quality and is a waste of time, money and milk production. Each year, the dairy industry loses as much as a billion dollars to mastitis. Many cows will experience mastitis at least once during a lactation cycle and some animals will develop recurring mastitis episodes in a single mammary quarter. These mastitic quarters can be difficult to manage during the lactation cycle. Cessation of production in the quarter while continuing to milk the other three can be a beneficial management decision in this scenario. However, the current methods available for cessation of lactation in a single quarter are limited. This study investigated the use of casein hydrolysate as a non-antibiotic option for causing cessation of lactation in a quarter. From this preliminary study we were able to apply our results to another aspect of mastitis prevention, which is the routine use of intramammary antibiotics at the end of the lactation cycle. This management practice is known as dry treatment and is a standard practice in the dairy industry with many years of proven efficacy against clearing infections present at the end of the lactation cycle. Increasing pressure from consumers to decrease antibiotic use in food production animals has caused this practice to come under scrutiny. This secondary study investigated the use of casein hydrolysates as a nonantibiotic alternative to standard antibiotic dry cow treatment. Overall, these studies demonstrated that casein hydrolysate has some efficacy in inducing mammary involution of a single quarter mid-lactation and also potentially as an alternative dry cow treatment. None of the animals treated in these studies displayed any symptoms of pain or discomfort, and all treated quarters resumed milk production after the next calving. Additionally, all antimicrobial milk tests on treated animals were negative. Casein hydrolysates may be a useful management tool for milk quality and animal health within the dairy industry.

A computer program was developed at Utah State University (USU) to aid in obtaining a more complete individual performance record keeping system for beef cattle in Utah. Some computer programs for beef cattle records presently exist but a program was needed that was readily available to the USU animal science extension and resident staff. The program was written in FORTRAN for use on the Burrogh 6700 computer located at the Utah State University Computer Center. It was designed to read input data for individual animals, perform various calculations (i. e. days of age, adjusted weaning weight and weaning weight ratio), print out the input data and results of the calculations for each animal as well as the average adjusted weight for each sex group (heifer, bull, steer). The computer program will manipulate weights in either the English or metric system and will convert weights from the English to metric system if desired. A unique feature of the program is the ranking of animals from highest to lowest based on the weaning weight ratio with accompanying animal number. The records can be evaluated to identify potential animals to use as replacements and those to be culled. The input data are collected on the ranch by a cooperative arrangement between the ranch operator and the USU Extension Staff. The ranch operator collects the preliminary data such as: birth date, tag number, tattoo number, dam, age of dam, and sire, and records it on the beef cattle performance input record. The extension specialist weights, gives a conformation score and records the information for each calf on the input record. The beef cattle performance input record is arranged in the same order as the data card is key punched thus facilitating the punching of the data cards. The staff can change from using the desk calculator to the use of the computer to improve efficiency and flexibility output as well as having more time available to spend with the public teaching that maintaining accurate records can help improve their herds for production and for inventory control. This can help the beef cattle industry to improve quality and type of beef animal produced in Utah and should improve the potential efficiency and profit. The rancher can transfer his records, with minor modifications, to one of the existing national computer programming organizations if desired. This computer program with or without modification has application for current research and university teaching. The computer program was designated for use with beef cattle, but could be modified to use for any class of livestock. This program is not an end in itself but is a foundation from which to build an improved record keeping system in Utah which could improve the production and quality of the beef cattle industry.

Influenza viruses cause between 3 and 5 million cases of respiratory infection each year and are responsible for between 250 and 500 thousand deaths. There are principally two avenues for the treatment and prevention of influenza. They are vaccination and antiviral regimens. Prevention of infection is largely accomplished through vaccination. While vaccines remain the preferred method for controlling the spread of influenza, treatment with antiviral drugs is important for treatment of severe infections that are caused by viruses that are different from the vaccination strains. The two major classes of antiviral drugs for influenza treatment are the adamantanes and the neuraminidase inhibitors. While most viruses have become resistant to the adamantanes, the neuraminidase inhibitors remain the primary choice for treatment of infections. Oseltamivir is the most important of the neuraminidase inhibitors. Data from an experiment run at Utah State University displayed a characteristic that is reflected in other published data. Oseltamivir, which has been shown to be effective against influenza virus strains in vitro, is unable to sufficiently protect mice from lethal infections. The focus of the present research was to identify viral differences that might explain for this discrepancy. Four viral strains were chosen that display differing susceptibilities to oseltamivir in mice. The viruses used were Influenza A/Duck/MN/1525/81 (H5N1), A/Victoria/3/75 (H3N2), A/NWS/33 (H1N1) and A/California/04/2009 (Pandemic H1N1). Oseltamivir was unable to protect mice that were infected with the A/Duck/MN/1525/81 and A/Victoria/3/75 viruses. These two viruses, along with the A/NWS/33 and the A/California/04/2009 viruses, were compared in vitro using virus replication kinetics, neuraminidase inhibition assays, and antiviral assays in cell culture. The viruses were studied in vivo by comparing survival, weight loss, lung scores and weights, lung virus titers, complete blood counts, cytokine assays, and histopathology. A second in vivo experiment was run to determine the effects of oseltamivir on survival, weight loss, lung scores and weights, lung virus titers, and histopathology. The two in vivo experiments summarized in this study confirmed previous data since oseltamivir was unable to protect mice infected with the influenza A/Duck and A/Victoria viruses. Overall, the virus infections behaved remarkably similar. The most interesting difference was that the A/Duck/MN/1525/81 and A/Victoria/3/75 viruses were able to induce more severe histopathological damage in mouse lungs earlier in the infection. The ability to cause severe disease more quickly might explain why the A/Duck/MN/1525/81 and A/Victoria/3/75 viruses remain lethal, despite oseltamivir treatment.

The composition of the gut microbiome can be affected by environmental factors, such as diet. The Western dietary pattern is associated with microbiome dysbiosis and adverse health outcomes, including obesity and metabolic disorders. The objective of this study was to examine the effect of gut microbiota from lean or obese human donors on metabolism and weight gain in recipient mice fed one of three basal diets: 1) the standard AIN93G diet, which promotes rodent health; 2) the total Western diet (TWD), which mimics the American dietary pattern and promotes inflammation-associated colorectal carcinogenesis; and 3) a 45% high fat diet-induced obesity (DIO) diet, which promotes excessive weight gain and symptoms of metabolic syndrome. We hypothesized that fecal microbiome transfer (FMT)from obese human donors would lead to an obese phenotype with symptoms of metabolic syndrome in recipient mice, and that consumption of TWD or DIO diets would further exacerbate the metabolic syndrome phenotype. The experiment design consisted of two main factors: body type of the human donor (obese or lean) and experimental diet (AIN, DIO or TWD), which was fed to mice for 22 weeks. Prior to FMT, the resident gut microbiome in mice was depleted using an established broad spectrum antibiotic/antifugal oral dosing regimen. Interestingly, human donor body type did not significantly affect final body weight or body composition in recipient

The manufacture of cheddar cheese is greatly dependent on bacterial growth for acid production. The quality of cheese depends upon the type and extent of microbial activity. An important function of the lactic fermenting bacteria is the production of acid resulting from cellular metabolism. If little or no acid is production of acid resulting from cellular metabolism. If little or no acid is produced the resulting cheese will have an inferior body, flavor, and texture and may even cause the cheese to be used as grinders. The major causes for inhibited lactic bacterial growth are poor starter handling procedures, antibiotic in mild coming from cows treated for mastitis, quaternary ammonium compounds used in plant sanitation, and bacteriophage. The occurrence of bacteriophage contamination and the increased use of quaternary ammonium compounds and antibiotics have caused a serious problem in cheese manufacturing. Purpose of project. The purpose of this project is to determine if cheddar cheese, comparable to normal cheese, can be made from slow or non-acid milk with the use of added lactic acid. Lactic acid will be used for the purpose of substituting for the acid that is normally produced by bacterial metabolism in the manufacture of cheddar cheese.

Heifers should represent the greatest genetic potential within a dairy herd. To maximize this potential, heifers must be inseminated to proven sires, a practice requiring management changes on many dairies. Holstein heifers (n=115) were allotted to one of three groups to compare alternatives to daily estrous detection that may facilitate the management of a program of artificial insemination for heifers. Group 1 heifers (Controls) received no treatment but were observed twice daily for signs of estrus. Group 2 heifers (2X-PGF) were synchronized with two injections of prostaglandin F2 alpha given 11 days apart and observed for estrus. Group 3 heifers (MGA+PGF) were synchronized with 9 days of melengestrol acetate feeding and an injection of prostaglandin F2 alpha 14 days after the last feeding and observed for estrus. Comparisons were made on the effectiveness of each treatment in estrous response and pregnancy rates and on the management requirements and economics associated with each program. Estrous response was significantly higher for the 2X-PGF group, while synchronized pregnancy and first-service conception rates did not differ. On an annual basis, the MGA+PGF group was calculated to require three-quarters of the labor input as compared to the 2X-PGF group and less than a fifth of the labor required for the Control group. A program of daily estrous detection was calculated to have an economic advantage over synchronization the frequency estrous synchronization programs was scheduled four times per year. was increased to six times per when When year, synchronization had a slight economic advantage over daily estrous detection when animal numbers were low and labor costs were high. The economic advantage of daily estrous detection is reduced when synchronization is performed more frequently throughout the year.