Serum and Plasma Vascular Endothelial Growth Factor Levels in Various Cat Populations

Introduction:  Vascular endothelial growth factor (VEGF) is one of the most important mediators of angiogenesis. Elevated levels within tumors, and in serum, plasma, and tumor effusion have been correlated with the development of metastatic disease, recurrence, and poor prognosis in many tumors in humans. Canine VEGF has been sequenced as homologous with the human form, and elevated serum and plasma VEGF have been found in dogs with hemangiosarcoma. Feline VEGF has also been sequenced, and shares homology with the human and canine forms.
Materials and Methods:  Stored serum and plasma samples from normal cats, cats with various neoplasms, and cats with non‐neoplastic disease were evaluated with a commercial ELISA kit (R&D Systems, Minneapolis MN). Samples were run in duplicate, and a standard curve was performed for each plate. The data were analyzed for differences between populations, and between serum and plasma levels in the same patient to determine the optimal sample for evaluating VEGF in cats.
Results:  In seven apparently healthy cats mean plasma VEGF was 95.6 pg/mL. In non‐neoplastic disease (7 cases), mean plasma VEGF was 117.3 pg/mL and mean serum VEGF level was 219.7 pg/mL. In ten tumor‐bearing patients mean plasma VEGF was 247.1 pg/mL, and mean serum VEGF was 322.3 pg/mL. Statistical analysis showed no significant difference between mean serum and plasma VEGF concentrations within each group or between groups (p > 0.05).
Discussion:  Serum and plasma VEGF levels could not be used to distinguish between healthy cats and cats with neoplastic or non‐neoplastic disease.     

Total intravenous anesthesia with ketamine–medetomidine–propofol in horses

Propofol is a potentially useful intravenous anesthetic agent for total intravenous anesthesia (TIVA) in horses. The purpose of this study was to compare the anesthetic and cardiorespiratory effects of TIVA following the administration of propofol alone(P–TIVA) and ketamine–medetomidine–propofol (KM–P–TIVA) in adult horses. The carotid artery was translocated to a subcutaneous position during TIVA with P–TIVA (n = 6) or KM–P–TIVA (n = 6). All horses were premedicated with medetomidine [0.005 mg kg^–1, intravenously (IV)]. Anesthesia was induced with midazolam (0.04 mg kg^–1 IV) and ketamine (2.5 mg kg IV). All horses were orotracheally intubated and breathed 100% oxygen. The KM drug combination (ketamine 40 mg mL^–1 and medetomidine 0.05 mg mL^–1) was infused at a rate of 0.025 mL kg^–1 hour^–1. Subsequently, a loading dose of propofol (0.5 mg kg^–1, bolus IV) was administered to all horses; surgical anesthesia (determined by horse response to incision and surgical manipulation, positive response being purposeful or spontaneous movement of limbs or head) was maintained by varying the propofol infusion rate as needed. Arterial blood pressure and HR were also monitored. Both methods of producing TIVA provided excellent general anesthesia for the surgical procedure. Anesthesia time was 115 ± 17 (mean ± SD) and 112 ± 11 minutes in horses anesthetized with KM–P–TIVA and P–TIVA, respectively. The infusion rate of propofol required to maintain surgical anesthesia with KM–P–TIVA was significantly less than for P–TIVA (mean infusion rate of propofol during anesthesia; KM–P–TIVA 0.15 0.02 P–TIVA 0.23 ± 0.03 mg kg^–1 minute^–1, p = 0.004). Apnea occurred in all horses lasting 1–2 minutes and intermittent positive pressure ventilation was started. Cardiovascular function was maintained during both methods of producing TIVA. There were no differences in the time to standing after the cessation of anesthesia (KM–P–TIVA 62 ± 10 minutes versus P–TIVA 87 ± 36 minutes, p = 0.150). The quality of recovery was good in KM–P–TIVA and satisfactory in P–TIVA. KM–P–TIVA and P–TIVA produced clinically useful general anesthesia with minimum cardiovascular depression. Positive pressure ventilation was required to treat respiratory depression. Respiratory depression and apnea must be considered prior to the use of propofol in the horse.     

Isolation of shiga-toxigenic Escherichia coli O157 from hide surfaces and the oral cavity of finished beef feedlot cattle

OBJECTIVE: To determine whether viable shiga-toxigenic Escherichia coli (STEC) O157 could be isolated from hide surface locations and the oral cavity of finished beef feedlot cattle. DESIGN: Within-animal prevalence distribution survey. ANIMALS: 139 finished cattle in 4 pens in a feedlot in Nebraska; prevalence of fecal STEC O157 shedding ranged from 20 to > 90%. PROCEDURE: Samples were collected from 7 sites from each animal: feces, oral cavity, and 5 hide surface locations (lumbar region, ventral aspect of the neck, ventral abdominal midline [ventrum], dorsal thoracic midline [back], and distal aspect of the left hind limb [hock]). RESULTS: Viable STEC O157 were isolated from the oral cavity or 1 or more hide surfaces of 130 cattle, including 50 fecal isolation-negative cattle. Site-specific prevalence of STEC O157 was 74.8% for oral cavity samples, 73.4% for back samples, 62.6% for neck samples, 60.4% for fecal samples, 54.0% for flank samples, 51.1% for ventrum samples, and 41.0% for hock samples. Only 5 cattle tested negative for STEC O157 at all 7 sites. Multiple correspondence and cluster analyses demonstrated that bacterial culture of feces, oral cavity samples, and back samples detected most cattle with STEC O157. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that viable STEC O157 may be isolated from the oral cavity, multiple hide surfaces, and feces of a high percentage of fed beef cattle and that bacterial culture of feces alone generally underestimates the percentage of fed beef cattle from which STEC O157 can be isolated.     

Effect of different adjuvants on the immune responses of cattle vaccinated with Mycobacterium tuberculosis culture filtrate proteins

The development of improved vaccines for bovine tuberculosis is urgently required as a cost effective solution for control and eventual eradication of tuberculosis in domestic animals. Studies in small animal models of tuberculosis have shown that vaccination with culture filtrate proteins (CFP), prepared from Mycobacterium tuberculosis or M. bovis, can induce cellular immune responses and confer a level of protection against aerogenic challenge with virulent mycobacteria. As a first step in the development of a mycobacterial CFP vaccine for protection of cattle against bovine tuberculosis, the immune responses of cattle vaccinated with short-term culture filtrate proteins (ST-CFP) from M. tuberculosis and formulated with different adjuvants were compared with those vaccinated with bacille Calmette-Guerin (BCG). The adjuvants included dimethyldioctyldecyl ammonium bromide (DDA), diethylaminoethyl (DEAE)-dextran, and ST-CFP adsorbed onto polystyrene beads. Vaccination with ST-CFP/DEAE-dextran induced high levels of interleukin-2 (IL-2) but low levels of interferon-gamma (IFN-gamma) from whole-blood cultures stimulated with M. tuberculosis ST-CFP in comparison with the strong IFN-gamma and IL-2 responses induced after vaccination with BCG. ST-CFP/DEAE-dextran also induced a strong antigen-specific immunoglobulin antibody response with both immunoglobulin G1 (IgG1) and IgG2 isotypes. Vaccination with ST-CFP/beads induced a weak IgG1-biased antibody response but no IFN-gamma or IL-2 response. DDA did not induce significant immune responses in animals vaccinated with ST-CFP. In comparison to the moderate delayed-type hypersensitivity (DTH) responses induced by vaccination with subcutaneous BCG, none of the ST-CFP vaccines induced a significant DTH response to either M. tuberculosis ST-CFP or bovine purified protein derivative (PPD). While the ST-CFP vaccines used in this study have not induced strong antigen-specific cellular immune responses in cattle comparable to those induced by BCG, they are immunogenic in cattle and it may be possible to overcome this problem by using adjuvants that more effectively promote IFN-gamma responses in this species.     

Astaxanthin Normalizes Epigenetic Modifications of Bovine Somatic Cell Cloned Embryos and Decreases the Generation of Lipid Peroxidation

Astaxanthin is an extremely common antioxidant scavenging reactive oxygen species (ROS) and blocking lipid peroxidation. This study was conducted to investigate the effects of astaxanthin supplementation on oocyte maturation, and development of bovine somatic cell nuclear transfer (SCNT) embryos. Cumulus–oocyte complexes were cultured in maturation medium with astaxanthin (0, 0.5, 1.0, or 1.5 mg/l), respectively. We found that 0.5 mg/l astaxanthin supplementation significantly increased the proportion of oocyte maturation. Oocytes cultured in 0.5 mg/l astaxanthin supplementation were used to construct SCNT embryos and further cultured with 0, 0.5, 1.0 or 1.5 mg/l astaxanthin. The results showed that the supplementation of 0.5 mg/l astaxanthin significantly improved the proportions of cleavage and blastulation, as well as the total cell number in blastocysts compared with the control group, yet this influence was not concentration dependent. Chromosomal analyses revealed that more blastomeres showed a normal chromosomal complement in 0.5 mg/l astaxanthin treatment group, which was similar to that in IVF embryos. The methylation levels located on the exon 1 of the imprinted gene H19 and IGF2, pluripotent gene OCT4 were normalized, and global DNA methylation, H3K9 and H4K12 acetylation were also improved significantly, which was comparable to that in vitro fertilization (IVF) embryos. Moreover, we also found that astaxanthin supplementation significantly decreased the level of lipid peroxidation. Our findings showed that the supplementation of 0.5 mg/l astaxanthin to oocyte maturation medium and embryo culture medium improved oocyte maturation, SCNT embryo development, increased chromosomal stability and normalized the epigenetic modifications, as well as inhibited overproduction of lipid peroxidation.