Research was conducted to develop gene probes to Baculovirus penaei (BP) from a cloning of BP genomic DNA and to test the applications of the gene probes. Five non-homologous probes representing >20% of the total BP genome were tested in dot blot and in situ hybridization assays. Four different dot blot hybridization assay systems were evaluated. Results obtained from radioactive labeling of the probes indicate that the probes are specific for BP DNA, while the results obtained from the three non-radioactive labeling systems suggest problems with non-specific reactivity and sample preparation for these methods. The results also indicated that in situ hybridization is presently the optimum assaying system for BP gene probes. The BP gene probes were used with an in situ hybridization assay to gather initial data concerning the mode of transmission for BP.

Thesis (Ph.D.)--Pacific Graduate School of Psychology, 1993. / Source: Dissertation Abstracts International, Volume: 54-12, Section: B, page: 6464.

Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / Raymond R. R. Rowland / Infectious cDNA clones of PRRSV make it possible to construct marker viruses for the study of virus replication and pathogenesis. The nonstructural protein 2 (nsp2) of porcine reproductive and respiratory syndrome virus (PRRSV) is the single largest protein produced during virus replication. The cDNA of the pCMV-129 infectious PRRSV clone was modified by creating unique Mlu I and SgrA I restrictions sites at nucleotide (nt) positions 3,219 and 3,614, respectively: both located within the C-terminal region of nsp2. cDNAs coding for oligo- and polypeptide tags, including FLAG, enhanced green fluorescent protein (EGFP) and firefly luciferase were inserted into the newly created restriction sites. The results showed that only the EGFP-containing genomes were properly expressed and produced virus. EGFP fluorescence, but not EGFP immunoreactivity, was lost during passage of recombinant EGFP viruses in culture. Sequencing of a fluorescence-negative EGFP virus showed that the EGFP remained intact, except for the appearance of mutations that may affect chromophore formation. The results show that nsp2 can be a site for the expression of foreign proteins. Removal of the region between Mlu I and SgrA I sites resulted in a virus that contained a 131 amino acid deletion. The deleted region was replaced with EGFP or an eight amino acid influenza hemagglutanin (HA) tag. Recombinant viruses were used to infect pigs. Gross and micro-histopathology showed reduced pathogenesis when compared to the parent wild-type virus. The 131 amino acid peptide, when expressed as a recombinant protein and coated onto enzyme linked immunosorbent assay (ELISA) plates, was recognized by sera from pigs infected with wild-type virus, but not the deletion mutants. The results from this study show that nsp2 is a potential target for the development of marker vaccines that can differentiate infected from vaccinated animals (DIVA) and for virus attenuation.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Gary A. Anderson / Foot-and-mouth disease (FMD) is a highly infectious viral disease of cattle, pigs, sheep, goats, buffalo, and artiodactyl wildlife species. Foot-and-mouth disease virus is endemic and periodic devastating epidemics have occurred and caused heavy economic losses in Iraq for a long time. The first official cases of FMD were recorded in 1937, while the first record of a specific FMD serotype in Iraq was serotype A in 1952. Other serotypes have been reported since then; serotypes O, SAT-1 and Asia1 were recorded in 1957, 1962, and 1975, respectively. Veterinary Services in Iraq has been severely weakened over the past two decades, and its infrastructure has been devastated as a consequence of previous political conflicts, wars and international sanctions. The breakdown of Veterinary Services led to the disruption of disease control strategies, collapse of disease surveillance and monitoring, and weakening of response systems. The destruction of the Al-Dora FMD laboratories for diagnosis and vaccine production by the United Nation in 1996, and the restrictions placed on the importation of vaccines have strongly affected the FMD control program. A severe epidemic of FMD occurred in Iraq in 1998, affecting 2.5 million ruminants and causing heavy losses in newly born animals. It is estimated to have killed about 550,000 animals. The outbreak was due to the serotype O1 Middle East strain which has affected large and small ruminants. In 2009, Iraq was severely affected by new serotype A (subtype A Iran 05). The major efforts of Veterinary Services in Iraq have been directed towards control of FMD by vaccination strategies. Two types of vaccine have been used, trivalent vaccine (O, A 22, and Asia 1) for cattle and buffalo and monovalent vaccine (O Manisa) for sheep and goats. Vaccination has been implemented once yearly on a voluntary basis. Sometimes other limited control measures have accompanied vaccination, which include quarantine, movement control, focused vaccination, disinfection, and public awareness programs. The FMD control program in Iraq has been confronted by many challenges: deficits in FMD surveillance and emergency preparedness, limited diagnostic capabilities, difficulties in restricting animal movement, and lack and irregular supply of appropriate vaccines.

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.

Endocrine regulation of expression of GnRH receptors is an important step in the control of reproduction. During the early follicular phase of the estrous cycle in the ewe, GnRH receptor expression increases in preparation for the preovulatory surge of LH. The studies described herein were designed to further elucidate the hormonal interactions controlling GnRH receptor expression. In long-term ovariectomized ewes, neither removal of progesterone, nor the presence of estradiol affected the expression of GnRH receptors. However, in ewes ovariectomized during the luteal phase of the estrous cycle and immediately implanted with progesterone and estradiol for 48 hours, low levels of estradiol for 24 hours were required to increase GnRH receptor mRNA following the removal of progesterone. In ovariectomized ewes following hypothalamic-pituitary disconnection, low levels of estradiol and pulsatile GnRH were required to increase GnRH receptor expression within 24 hours of treatment initiation. These results suggest an interaction between estradiol and GnRH is involved in increasing GnRH receptor expression during the periovulatory period. How progesterone, estradiol and, GnRH interact to increase GnRH receptors is unknown, but a possible candidate involved in mediating these interactions may be the cell specific transcription factor, steroidogenic factor-1 (SF-1). SF-1 mRNA increased within 24 hours of treatment of ewes with prostaglandin F₂(α) compared to ewes in the luteal phase of the estrous cycle. This suggests that progesterone may have an inhibitory effect on SF-1 mRNA. SF-1 mRNA was similar between ovariectomized ewes and ovariectomized ewes following hypothalamic-pituitary disconnection treated with estradiol and GnRH. Treatment with estradiol or GnRH alone did not increase SF-1 mRNA. The results of these experiments suggest that progesterone removal as well as the presence of estradiol and GnRH are required to increase GnRH receptor expression during the early follicular phase in the ewe. Further, the transcription factor, SF-1 may be involved in mediating the effects of these hormones on GnRH receptor expression.

The competence of lymnaeids as intermediate hosts of Fasciola hepatica has not been studied in southern Louisiana and furthermore the taxonomic status of these snails is still controversial among malacologists. There are different opinions as to whether Fossaria (Bakerilymnaea) cubensis or Lymnaea (Pseudosuccinea) columella acts as the main intermediate host in enzootic areas of Louisiana. In the present study, the suitability of those two snails as hosts for F. hepatica from a single source in Louisiana was compared with four other species of lymnaeids from different geographic areas and the study was undertaken with evaluation of 'vector competence'. Groups of young snails were exposed to three different densities of miracidia, 1,5, and 10, in 0.7ml of spring water in a Beem('(REGTM)) capsule during 3 hrs at room temperature. They were checked for infection at day 14 postexposure; then they were dissected or fixed at different times. Percentages, locations of intramolluscan stages and intensity of infection were recorded, as well as the variations in length of the incubation period. Histopathological studies were also done. The results show that L. (P.) columella from Louisiana is a more suitable host (48.6% became infected) than F. (B.) cubensis (19.2% became infected). The percentages of the positive controls becoming infected were: 68.9% for L. (P.) columella from Michigan, and 32.2% for L. gedrosiana from Iran. The study of histological sections did not show any significant differences in the pathological manifestations in the infected snails. The negative controls were: L. natalensis from Senegal, Stagnicola elodes from Michigan, and Stagnicola attenuata from Hidalgo, Mexico, an enzootic area of fascioliasis. The latter species was studied for the first time and its internal anatomy was compared with S. elodes. Although the results show that F. (B.) cubensis and L. (P.) columella have some degree of competence as intermediate hosts of F. hepatica under laboratory conditions, field observations on the habitat of these snails in southern Louisiana and the characteristic management of cattle, indicate that the former snail is more important as an intermediate host in this enzootic area than is the latter / acase@tulane.edu

Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2009. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3245. Adviser: Robert Knox. Includes bibliographical references. Available on microfilm from Pro Quest Information and Learning.

Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2009. / Source: Dissertation Abstracts International, Volume: 70-06, Section: B, page: 3355. Adviser: Wanda M. Haschek-Hock. Includes bibliographical references. Available on microfilm from Pro Quest Information and Learning.

Two experiments were conducted to determine the effects of grain processing, grain sources, protein sources, and bST treatments on lactation performance, ovarian activity and nutrient digestion in early lactation dairy cows. In experiment 1, forty-eight dairy cows were divided into six groups and fed diets containing 37 to 39% grain as either steam-flaked sorghum (SFS, 360 g/L), steam-flaked corn (SFC, 360 g/L), or steam-rolled corn (SRC, 490 g/L) with one of two protein sources, soybean meal (SBM) or Prolak® for 70 days. Main effects for grain and protein source were not different for DMI, milk yield, 3.5% FCM yield or feed efficiency. Although efficiency of feed conversion into FCM did not differ among treatments, cows fed Prolak® required 10% less NEL for every Mcal of NEL converted into body weight or FCM. Milk protein yield was increased by both flaked gain and Prolak®. Feeding flaked grain increased milk production by 1.5 kg/d compared with rolled grain. Increasing the ratio of RDS/RDP to more than 2.4 increased milk yield by 2.7 kg/d (39.0 vs 36.3 kg/d). Plasma NEFA levels did not differ among treatments and grain and protein source had variable effects on PUN. Compared with other grains, SFC increased digestibilities of DM and OM, and flaked grains increased digestibility of starch. Cows fed SFS had the lowest crude protein digestibility, but was highest in NDF. In experiment 2, thirty-two postpartum (5 DIM)Holstein cows were assigned to one of four treatments in a 2 x 2 factorial design for 90 days. Treatments were: steam-flaked sorghum diet with bST (SFS-B); steam-flaked sorghum diet without bST (SFS-N); steam-rolled corn diet with bST (SRC-B); and steam-rolled corn diet without bST (SRC-N). Cows receiving bST during the first 45 days of treatment had lower DMI and DMI as % of BW. Milk yield was increased by bST 3.1 kg/d and 1.8 kg/d during the 45 d and 90 d periods, respectively, resulting in greater feed efficiency for bST-treated cows. Grain processing did not affect DMI, milk yield and FCM yield. Neither composition nor yields of milk components were influenced by treatments. Compared to the SRC diet, SFS increased plasma glucose 5% and plasma insulin 19%. Grain processing and bST had no effect on either BHBA or hepatic triglycerides. PUN was reduced by bST, but NEFA was increased. Compared with SRC, SFS increased progesterone during the first two postpartum estrous cycles and cows receiving bST had significantly larger CL. Flaked sorghum compared with SRC increased digestibility of DM, OM, and starch, and neither grain affected digestibilities of CP, NDF or ADF. Treatments with bST did not affect any nutrient digestibilities.