Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

Salmonella Typhimurium ghost cells expressing K88ab, K88ac, K99, and FasA fimbriae of enterotoxigenic Escherichia coli (ETEC) in their envelopes were constructed. The genes encoding the fimbriae were individually cloned into an expression plasmid, pMMP81, carrying the asd gene, which was subsequently electroporated into the Δasd S. Typhimurium mutant. Plasmid pJHLP99, carrying the phiX174 lysis gene E, was also subsequently electroporated into the Salmonella mutant. The presence of the individual fimbriae on the ghost cells was examined by Western blot analysis. Forty BALB/c mice were equally divided into 2 groups of 20 mice each. Group A mice were intramuscularly vaccinated with a mixture of the 4 ghost cells expressing the individual fimbriae. The group B mice were inoculated with sterile phosphate-buffered saline as a control. The antigen-specific serum IgG concentrations were significantly higher in group A than in group B from week 2 until week 6 after inoculation. In addition, the antigen-specific IgA concentrations in fecal samples were significantly higher in group A than in group B at week 2 after inoculation. A large difference between the groups in the number of antigen-specific IgA-secreting cells in the small intestine was observed by immunohistochemical study. Also, the splenic lymphocyte proliferative responses were significantly greater in group A than in the control mice. These results suggest that vaccination with our Salmonella ghost cells can induce both humoral and cell-mediated immune responses and that the increased number of antigen-specific IgA-secreting cells in the small intestine may be correlated with the elevated fecal IgA immune response.     

Deoxynivalenol in chicken feed alters the vaccinal immune response and clinical biochemical serum parameters but not the intestinal and carcass characteristics

This study was conducted to investigate the impacts of deoxynivalenol (DON) feeding either alone or in combination with a microbial feed additive (MFA) on the immune response to a viral vaccine and serum clinical chemical parameters. Forty 1‐day‐old boiler chicks were weighed and randomly divided into four groups, 10 birds in each group: (i) control group fed with basal diet; (ii) DON group fed with basal diet artificially contaminated with 10 mg DON/kg feed; (iii) DON + MFA group fed with basal diet contaminated with 10 mg DON/kg feed and supplemented with 2.5 kg of MFA/ton feed; and (iv) MFA group fed with basal diet supplemented with 2.5 kg of MFA/ton feed. At 35 days of age, birds were slaughtered and blood was collected for investigating the antibody titre against infectious bronchitis virus (IBV) and clinical chemical parameters. The results showed that DON reduced (p = 0.032) the titre against IBV, decreased (p = 0.005) the level of alanine transaminase (ALT) (4.2 ± 0.5 U/l) compared with control birds (6.4 ± 0.5 U/l), increased (p = 0.002) the serum cholesterol concentration (144 ± 6 mg/dl) compared with their control counterparts (123 ± 5 mg/dl) and increased (p = 0.074) the amount of circulating triglycerides (62.25 ± 7.50 mg/dl) compared with controls (39.55 ± 4.74). These results indicate that dietary DON altered the humoral immune response to viral vaccine and affected the serum clinical biochemistry. However, DON in combination with MFA did not affect serum IBV titre. Taken together, DON in the feed of broilers produced an impairment of the success of IBV vaccine and affected the health of birds.     

Cardiac evaluation of anesthetized Grevy's zebras (Equus grevyi)

OBJECTIVE: To determine ECG and echocardiographic measurements in healthy anesthetized Grevy's zebras (Equus grevyi). ANIMALS: 20 healthy zebras. PROCEDURES: Auscultation, base-apex ECG, and echocardiography were performed on anesthetized zebras. RESULTS: Low-grade systolic murmurs were detected in the left basilar region in 4 of 20 zebras. Evaluation of ECGs from 19 zebras revealed sinus rhythm with a predominantly negative QRS complex and a mean +/- SD heart rate of 67 +/- 10 beats/min. Echocardiograms of sufficient image quality were obtained for 16 zebras. Interventricular septal thickness in diastole, left ventricular chamber in diastole and systole, left atrial diameter, and left ventricular mass were significantly and moderately correlated with estimated body weight (r values ranged from 0.650 to 0.884). Detectable swirling of blood in the right and sometimes the left ventricles was detected in 9 of 16 zebras, whereas physiologic regurgitation of blood was detected for the aortic valve in 3 zebras, pulmonary valve in 2 zebras, mitral valve in 2 zebras, and tricuspid valve in 1 zebra. CONCLUSIONS AND CLINICAL RELEVANCE: Results of this study provide reference information for use in the cardiac evaluation of anesthetized Grevy's zebras.     

Effects of the cyclooxygenase inhibitor meloxicam on recovery of ischemia-injured equine jejunum

OBJECTIVE: To determine the effect of meloxicam and flunixin meglumine on recovery of ischemia-injured equine jejunum. ANIMALS: 18 horses. PROCEDURES: Horses received butorphanol tartrate; were treated IV with saline (0.9% NaCl) solution (SS; 12 mL; n = 6), flunixin meglumine (1.1 mg/kg; 6), or meloxicam (0.6 mg/kg; 6) 1 hour before ischemia was induced for 2 hours in a portion of jejunum; and were allowed to recover for 18 hours. Flunixin and SS treatments were repeated after 12 hours; all 3 treatments were administered immediately prior to euthanasia. Selected clinical variables, postoperative pain scores, and meloxicam pharmacokinetic data were evaluated. After euthanasia, assessment of epithelial barrier function, histologic evaluation, and western blot analysis of ischemia-injured and control jejunal mucosa samples from the 3 groups were performed. RESULTS: Meloxicam- or flunixin-treated horses had improved postoperative pain scores and clinical variables, compared with SS-treated horses. Recovery of transepithelial barrier function in ischemia-injured jejunum was inhibited by flunixin but permitted similarly by meloxicam and SS treatments. Eighteen hours after cessation of ischemia, numbers of neutrophils in ischemia-injured tissue were higher in horses treated with meloxicam or flunixin than SS. Plasma meloxicam concentrations were similar to those reported previously, but clearance was slower. Changes in expression of proteins associated with inflammatory responses to ischemic injury and with different drug treatments occurred, suggesting cyclooxygenase-independent effects. CONCLUSIONS AND CLINICAL RELEVANCE: Although further assessment is needed, these data have suggested that IV administration of meloxicam may be a useful alternative to flunixin meglumine for postoperative treatment of horses with colic.     

Cerebral Blastomyces dermatitidis infection in a cat

CASE DESCRIPTION: An 8-year-old domestic shorthair cat was evaluated because of signs of depression, circling, and visual deficits. CLINICAL FINDINGS: The cat had no cutaneous lesions, and results of an ophthalmologic examination and thoracic radiography were within reference limits. Computed tomography of the brain revealed a mass lesion involving the right parietal, temporal, and occipital lobes; the mass was in broad-based contact with the skull and smoothly marginated and had strong homogenous enhancement after contrast agent administration. During craniectomy, samples of the mass were collected for cytologic and histopathologic evaluations and microbial culture. A diagnosis of Blastomyces dermatitidis-associated meningoencephalitis with secondary pyogranulomatous inflammation was made. TREATMENT AND OUTCOME: Amphotericin B (0.25 mg/kg [0.11 mg/lb], IV) was administered on alternate days (cumulative dose, 1.75 mg/kg [0.8 mg/lb]). To minimize the risk of nephrotoxicosis, assessments of serum biochemical variables (urea nitrogen and creatinine concentrations) and urinalyses were performed at intervals. The third dose of amphotericin B was postponed 48 hours because the cat became azotemic. The cat subsequently received fluconazole (10 mg/kg [4.5 mg/lb], PO, q 12 h) for 5.5 months. Six months after discontinuation of that treatment, the cat appeared healthy and had no signs of relapse. CLINICAL RELEVANCE: Brain infection with B dermatitidis is typically associated with widespread disseminated disease. The cat of this report had no evidence of systemic disease. Blastomycosis of the CNS should be considered as a differential diagnosis for brain lesions in cats from areas in which B dermatitidis is endemic.     

Disseminated cryptococcosis including osteomyelitis in a horse

A 4-year-old Arab mare was diagnosed with disseminated cryptococcosis, including osteomyelitis of the proximal phalanx of the left hind limb, osteomyelitis with associated soft tissue granuloma of a rib and disseminated, large cryptococcal nodules in the lungs. The lesion in the dorsoproximal aspect of the proximal phalanx had a large area of cortical lysis with spiculated periosteal new bone and extensive soft tissue swelling. The affected rib had a pathological fracture. Cryptococcal osteomyelitis has not been previously reported in horses but should be considered as a differential diagnosis, particularly in endemic regions.     

Characterisation of European varieties of triticale with special emphasis on the ability of plant phytase to improve phytate phosphorus availability to chickens

1. A total of 30 varieties and selection lines of triticale grown under similar conditions were characterised. Thousand grain weight, specific weight, Hagberg falling number and N were 50.2 +/- 5.0 g, 72.4 +/- 2.1 kg/hl, 96 +/- 48 s and 16.1 +/- 0.11 g/kg, respectively. 2. Mean phosphorus (P) concentration was 2.86 +/- 0.31 g/kg, of which 77% was of phytic origin. Mean phytase activity was 1018 +/- 319 phytase units (PU)/kg. A genotypic effect on phytase activity was detected amongst 5 varieties studied out of 30. Potential and real applied viscosities were positively correlated and mean values were 3.53 +/- 0.66 and 2.15 +/- 0.31 ml/g, respectively. 3. The efficacy of plant phytase in improving P availability was assessed in chickens up to 3 weeks of age. Growth performance and bone ash concentration were compared in birds given either a maize (450 g/kg) and soybean meal (230 g/kg) phosphorus deficient diet containing 3.5 g P/kg, this basal diet supplemented with 1 or 2 g P/kg as monocalcium phosphate (MCP) or triticale (450 g/kg) and soybean meal (230 g/kg) diets containing 3.2 to 3.8 g P/kg with no MCP. To achieve graded levels of phytase activity, 4 varieties of triticale, intact or in which phytase was denaturated by heat treatment, were used. Estimated metabolisable energy, protein, amino acids and calcium concentrations were similar in all diets. 4. Phytase activity in the triticale-based diets ranged between 135 and 1390 PU/kg. Growth performance and bone ash were responsive to plant phytase and to MCP. Non-linear models of these responses were adjusted with the best fit for bone ash parameters. The values of 250, 500 and 1000 PU of plant phytase were estimated to be equivalent to 0.46, 0.67 and 0.81 g P as MCP, respectively.     

Investigation and clinical application of a novel axial pattern flap for nasal and facial reconstruction in the dog

OBJECTIVE: To describe the vascular supply to a facial skin flap based at the commissure of the lip in the dog and report on its use in four dogs. STUDY DESIGN: Experimental and prospective clinical study. Animals Five canine cadavers and four client-owned dogs. METHODS: In the cadavers, the ventral aspect of the zygomatic arch, the ventral margin of the caudal mandible and the wing of the atlas were marked as anatomical boundaries of a skin flap that was elevated from the subcutaneous tissues to the level of the medial canthus of the eye. Methylene blue dye and barium sulphate solution were independently infused through a common carotid (three dogs) or facial artery (two dogs) catheter. Distribution of dye throughout the harvested skin was assessed subjectively. After contrast infusion the flap was excised and radiographed. The technique was used to reconstruct large facial or nasal defects in four dogs after tumour or skin lesion excision. RESULTS: Cadaver dissections and contrast studies clearly demonstrated three direct cutaneous arteries, the superior and inferior labial arteries and the angularis oris artery, arborising within the base of the flap. A separate direct cutaneous branch of the angularis oris artery was identified. An arterial plexus was identified within the distal flap, within which this artery communicates with the transverse facial artery and a cutaneous branch of the masseteric artery. Dye infusion caused discolouration of the elevated skin and vasculature within the flap. The flap survived in all clinical cases with marginal distal necrosis in one dog. CONCLUSIONS: The complex facial flap described is perfused by three direct cutaneous arteries and functions reliably in clinical cases.