Porcine adenovirus as a delivery system for swine vaccines and immunotherapeutics

Porcine adenovirus (PAdV) has many qualities which make it an ideal choice for use as a delivery vector in swine. It is a low grade pathogen, present almost world-wide in a number of serotypes varying in their virulence and tissue tropism, which may allow for serotype specific vaccine targeting. PAdV is species specific having only been isolated from swine, reducing the possibility of its spread to other animals or man following administration. When engineered to contain a foreign gene, recombinant PAdV (rPAdV) can be grown to high titres in tissue culture cells making it cheap to produce. Knowledge of the complete nucleotide sequence of the PAdV genome has enabled rationally directed insertions of foreign genes which remain stably inserted in the genome and can be expressed at high levels following delivery to the target host. Importantly, recombinant PAdV can be administered by injection or by the oral route in feed or drinking water. We have delivered a range of antigens and immunomodulatory molecules to commercially available pigs using rPAdV and found it to be a very effective delivery system. Significantly, recombinant PAdV serotype 3 is highly effective as a delivery vehicle even when administered in the face of high levels of artificially induced serotype specific neutralising antibody to the vector.     

Biological responses to porcine respiratory and reproductive syndrome virus in pigs of two genetic populations

One hundred pigs from the NE Index Line (NEI) and 100 Hampshire-Duroc cross pigs (HD) were inoculated intranasally with porcine respiratory and reproductive syndrome virus (PRRSV 97-7895 strain) at 26 d of age to determine whether genetic variation in response to PRRSV exists. An uninfected littermate to each infected pig served as a control. Pigs were from 163 dams and 83 sires. Body weight and rectal temperature were recorded, and blood samples were drawn from each pig on d 0 before inoculation and on d 4, 7, and 14 after inoculation. Pigs were sacrificed on d 14. Lung and bronchial lymph nodes were collected, placed in optimal cutting temperature compound, and frozen at -80 degrees C. The presence of PRRSV in serum and in lung tissue and bronchial lymph nodes was determined by isolation in cell culture. The presence of antibodies in serum collected on d 14 was determined by a commercial ELISA test. Lung tissue was examined microscopically and scored for incidence and severity of lesions (score of 1 to 3; 1 = no or few lesions, and 3 = severe interstitial pneumonia). Data were analyzed with a mixed model that included random sire and dam effects. The interaction of line x treatment was significant (P < 0.001) for weight change and rectal temperature. Un-infected HD pigs gained 0.67 kg more from d 0 to 14 and averaged 0.32 degrees C higher rectal temperature than uninfected NEI pigs (P < 0.001), whereas infected NEI pigs gained 0.34 kg more and had -0.54 degrees C lower temperature than infected HD pigs (P < 0.001). Viremic titer (cell culture infectious dose 50%/mL) was greater (P < 0.05) in HD than NEI at d 4 (10(4.52) vs. 10(4.22)), 7 (10(4.47) vs. 10(3.99)), and 14 (10(3.49) vs. 10(3.23)). Viral titer loads in lung (P = 0.11) and bronchial lymph nodes tended (P = 0.07) to be greater in HD than NEI pigs. Antibody signal-to-positive (S/P) ELISA ratios in infected pigs ranged from 0.18 to 3.38, and 88% had levels > or = 0.40, which is the positive threshold for this ELISA. The S/P range in uninfected pigs was 0 to 1.11, and 99% had levels < or = 0.40. Mean S/P ratio for infected pigs was 0.23 units higher in HD than in NEI (P < 0.001). The HD pigs had a greater incidence of interstitial pneumonia and 0.65 higher mean lesion scores than NEI pigs (P < 0.001). In summary, responses of pigs of the two lines to infection with PRRSV differed, indicating that underlying genetic variation existed.     

Thermographic Eye Temperature as an Index to Body Temperature in Ponies

Infrared thermography (IRT) is a passive, remote, and noninvasive method of measuring surface temperatures. Select surface locations, such as the eye, could indicate body temperature. To investigate whether thermographic eye temperatures were associated with body temperatures and could be used to detect febrile ponies, we measured IRT eye temperatures, rectal temperatures, and implanted thermal microchip temperatures from 24 male ponies daily for 3 consecutive days. Information regarding distance of the pony from the IRT device, presence of direct sunlight during the measurement period, and ambient temperature were also collected. A multivariate linear regression analysis indicated that 60.41% of the variance in IRT was accounted for by rectal temperature, sunlight, and distance between the camera and the pony, with rectal temperature being the major contributor to variance (46.23%). Using a rectal temperature of >38.6°C (101.5°F) as the indicator of febrile status, sensitivity and specificity of the IRT device used to detect the febrile ponies were found to be 74.6% and 92.3%, respectively, when using the maximum IRT eye temperature per pony per day. In conclusion, IRT eye temperature could be a preliminary screening tool to determine whether a more time-consuming, labor-intensive, and invasive method (e.g., rectal temperature) is warranted for fever validation.     

Differential response of cull cow muscles to the hypertrophic actions of ractopamine-hydrogen chloride

Ractopamine-HCl (RAC) is a beta-adrenergic agonist with variable effects on cattle performance and carcass variables. Cull cows fed RAC (200 mg . head(-1) . d(-1)) demonstrate an increased size of type I and II muscle fibers that does not translate into a larger ribeye area. The objective of this study was to examine the dose-dependent effects of RAC on cull cow muscle morphometrics. Eighty-eight cull beef cows representing 2 breed types (n = 44 each) were fed 0, 100, 200, and 300 mg . head(-1) . d(-1) of RAC for the last 28 d of a 54-d feeding period. On d 54, cows were slaughtered, and samples of the LM and semimembranosus muscle (SM) from 16 randomly selected carcasses (n = 4 per treatment) were taken for measurement of beta (2)-adrenergic receptors and type I, IIA, and IIX myosin heavy chain (MyHC) gene expression. Twenty-four hours postmortem, LM, SM, infraspinatus (INF), and vastus lateralis samples from 40 randomly selected carcasses (n = 10 per treatment) were obtained and frozen for immunohistochemical analysis. Muscle fiber cross-sectional area and diameter, MyHC isoform expression, and fiber-associated nuclei numbers were measured. Ractopamine dosage exhibited differential effects on muscle morphometrics and MyHC gene expression. Muscle fiber cross-sectional area and diameter were increased (P < 0.05) by RAC in INF type I and IIA fibers and SM type IIA fibers. Ractopamine increased (P < 0.05) MyHC type IIX mRNA and tended to increase (P < 0.10) beta(2)-adrenergic receptors in the SM; a change in mRNA abundance was not detected for either gene in the LM. Treatment with RAC decreased (P < 0.05) fiber-associated nuclei numbers in the INF, vastus lateralis, and LM but did not affect (P > 0.05) MyHC or beta-adrenergic receptor expression. These results indicate that cull cow feeding programs may consider supplementing RAC as a means of adding value to cuts within the chuck, such as the INF.     

Osteoma of the oral and maxillofacial regions in cats: 7 cases (1999-2009)

Objective-To describe clinical features of oral and maxillofacial osteomas in cats. Design-Retrospective case series. Animals-7 cats with oral or maxillofacial osteoma or both. Procedures-Medical records were reviewed for information on signalment, history, clinical signs, physical examination findings, diagnostic imaging findings, results of serum biochemical analyses and histologic testing, surgical procedures performed, and perioperative complications. Outcome was determined on the basis of follow-up telephone interviews of owners. Results-Cats ranged from 1 to 23 years of age. Clinical signs were observed in 5 cats and were attributed to the presence of the mass. Diagnostic imaging (radiography and computed tomography) and histologic examination confirmed the diagnosis of osteoma. Three cats were euthanatized; 1 cat was treated by mandibulectomy, 1 was treated by maxillectomy, and 2 were treated by debulking. At the time of follow-up at least 1 year after surgery, all 4 treated cats were alive, with owners reporting an acceptable quality of life. Conclusions and Clinical Relevance-Osteoma of the oral and maxillofacial regions is an uncommon tumor in cats. Most cats are examined during an advanced stage of the disease, when treatment options may be limited. Although osteoma is a benign tumor, the recommendation is to perform a clinical evaluation, diagnostic imaging, biopsy, and treatment early in the disease process, when less invasive surgical approaches may be feasible.     

Dose and release pattern of anabolic implants affects growth of finishing beef steers across days on feed

Four experiments evaluated the effect of implant dose and release pattern on performance and carcass traits of crossbred beef steers. In Exp. 1, steers (4 to 7 pens/treatment; initial BW = 315 kg) were fed an average of 174 d. Treatments were 1) no implant (NI); 2) Revalor-S [120 mg of trenbolone acetate (TBA) and 24 mg of estradiol 17β (E(2)); REV-S]; 3) Revalor-IS followed by REV-S (cumulatively 200 mg of TBA and 40 mg of E(2); reimplanted at 68 to 74 d; REV-IS/S); and 4) Revalor-XS (200 mg of TBA and 40 mg of E(2); REV-X). Carcass-adjusted final BW was greater (P < 0.05) for REV-X and REV-IS/S than for REV-S (610, 609, and 598 kg, respectively). Daily DMI did not differ (P > 0.10) among the 3 implants, but carcass-adjusted G:F was greater (P < 0.05) for REV-X and REV-IS/S than for REV-S (0.197 and 0.195 vs. 0.188). Both HCW and LM area were greater (P < 0.05) for REV-X and REV-IS/S than for REV-S. Marbling scores were greatest (P < 0.05) for REV-S and least (P < 0.05) for REV-IS/S; REV-X was intermediate to NI and REV-IS/S. In Exp. 2, steers (10 pens/treatment; initial BW = 391 kg) were fed 131 d, with treatments of REV-S, REV-IS/S (reimplanted at 44 to 47 d), and REV-X. Carcass-adjusted final BW (598 kg), ADG (1.6 kg), DMI (9.4 kg), G:F (0.17), and HCW did not differ (P > 0.10) among treatments. The percentage of Choice was less (P < 0.05) and percentage of Select greater (P < 0.05) for REV-IS/S than for REV-S and REV-X. In Exp. 3, steers (10 pens/treatment; initial BW = 277 kg) were fed 197 d and received either REV-IS/S (reimplanted at 90 to 103 d) or REV-X. Carcass-adjusted final BW (625 vs. 633 kg) and ADG (1.81 vs. 1.76 kg) were greater (P < 0.05) for REV-X-implanted steers. Daily DMI did not differ, but G:F tended (P < 0.10) to be increased and HCW was greater (P < 0.05) for REV-X than for REV-IS/S. In Exp. 4, steers (8 pens/treatment; initial BW = 238 kg) were fed 243 d and received either REV-IS/S (reimplanted at 68 to 71 d) or REV-X. Carcass-adjusted final BW (612 kg), ADG (1.54 kg), DMI (7.55), and G:F (0.21) did not differ (P > 0.10) for REV-IS/S and REV-X-implanted steers. Carcass traits did not differ among implants, but the percentage of Choice carcasses was greater (P < 0.05) and percentage of Select was less (P < 0.05) for REV-X than for REV-IS/S. These data indicate that when TBA/E(2) dose is equal, the altered release rate of REV-X can improve performance and quality grade, but these effects depend on duration of the feeding period and timing of initial and terminal implants.     

Neutralization kinetics of bovine viral diarrhea virus by hyperimmune serum: one or multi-hit mechanism

The kinetics of bovine viral diarrhea (BVD) virus neutralization (VN) by hyperimmune serum followed first order kinetics at low dilutions (1:8 and 1:16) of hyperimmune bovine serum. A lag phase in the VN curve occurred when the serum was further diluted. Addition of rabbit anti-bovine IgG, but not anti-IgM, significantly increased the degree of VN after BVD virus was reacted with further diluted (1:256 dilution) anti-BVD bovine hyperimmune serum. Incubation of virus-hyperimmune serum (1:64 dilution) at 4 degrees C for 0 to 60 min, followed by incubation at 37 degrees C indicated VN occurred as a two-stage process: a binding (temperature-independent) phase that was followed by a triggering (temperature-dependent) phase. The data favor the thesis that neutralization of BVD virus occurs by a multi-hit mechanism and requires combination of at least two molecules of antibody with each virus.     

Acute myeloid leukemia with multilineage dysplasia in an alpaca

An 18-year-old female alpaca was presented to the Colorado State University Veterinary Teaching Hospital for chronic ill thrift over a 1-year period. Six weeks previously, an infected left mandibular cheek tooth was removed by oral extraction. On physical examination the patient was cachectic, lethargic, and weak. Abnormalities on the CBC included neutropenia, thrombocytosis, and severe nonregenerative, macrocytic, hypochromic anemia. Dysplastic nucleated erythrocytes and micromegakaryocytes were observed on the peripheral blood smear. Neutrophils, bands, and metamyelocytes appeared markedly toxic. Numerous blasts containing variable numbers of fine azurophilic granules were also observed. Based on their morphology, the cells were interpreted to be progranulocytes and myeloblasts, and a presumptive diagnosis of acute myeloid leukemia (AML) was made. The blast cells accounted for 60% of the nucleated cell population on bone marrow aspirates, further supporting a diagnosis of AML with multilineage dysplasia. Post mortem examination showed infiltration of the neoplastic cells into spleen, liver, kidney, and lymph nodes. Based on histologic findings, the morphologic diagnoses were disseminated myeloid neoplasia, chronic regionally extensive tooth root abscess, and membranous glomerulonephritis. The neoplastic cells were CD172a-positive on flow cytometry, chloroacetate esterase-positive by cytochemistry, and myeloperoxidase-positive by immunohistochemistry, confirming myeloid origin. To our knowledge, this is the first case of AML with multilineage dysplasia in an alpaca, with only one other case of myelodysplasia described previously in this species.     

Transition From Sagebrush Steppe to Annual Grass (Bromus tectorum): Influence on Belowground Carbon and Nitrogen

Vegetation changes associated with climate shifts and anthropogenic disturbance have major impacts on biogeochemical cycling. Much of the interior western United States currently is dominated by sagebrush (Artemisia tridentata Nutt.) ecosystems. At low to intermediate elevations, sagebrush ecosystems increasingly are influenced by cheatgrass (Bromus tectorum L.) invasion. Little currently is known about the distribution of belowground organic carbon (OC) on these changing landscapes, how annual grass invasion affects OC pools, or the role that nitrogen (N) plays in carbon (C) retention. As part of a Joint Fire Sciences-funded project called the Sagebrush Treatment Evaluation Project (SageSTEP), we quantified the depth distribution of soil OC and N at seven sites experiencing cheatgrass invasion. We sampled plots that retained sagebrush, but represented a continuum of cheatgrass invasion into the understory. Eighty-four soil cores were taken using a mechanically driven diamond-tipped core drill to a depth of 90 cm, or until bedrock or a restrictive layer was encountered. Samples were taken in 15-cm increments, and soil, rocks, and roots were analyzed for OC and total N. We determined that cheatgrass influences the vertical distribution of OC and N within the soil profile and might result in decreased soil OC content below 60  cm. We also found that OC and total N associated with coarse fragments accounted for at least 10% of belowground pools. This emphasizes the need for researchers to quantify nutrients in deep soil horizons and coarse fragments.