Immunogenicity in dogs of three recombinant antigens (TSA, LeIF and LmSTI1) potential vaccine candidates for canine visceral leishmaniasis

Control of canine visceral leishmaniasis (VL) remains a difficult and serious problem mostly because there is no reliable and effective vaccine available to prevent this disease. A mixture of three recombinant leishmanial antigens (TSA, LeIF and LmSTI1) encoded by three genes highly conserved in the Leishmania genus have been shown to induce excellent protection against infection in both murine and simian models of cutaneous leishmaniasis. A human clinical trial with these antigens is currently underway. Because of the high degree of conservation, these antigens might be useful vaccine candidates for VL as well. In the present study, using the dog model of the visceral disease, we evaluated the immunogenicity of these three antigens formulated with two different adjuvants, MPL-SE and AdjuPrime. The results were compared with a whole parasite vaccine formulated with BCG as the adjuvant. In order to investigate if sensitization with the recombinant antigens would result in recognition of the corresponding native parasite antigens upon infection, the animals were exposed for four weeks after the termination of the immunization protocol with the recombinant antigens to a low number of L. chagasi promastigotes, an etiological agent of VL. Immune response was evaluated by quantitative ELISA in the animal sera before and after exposure to the viable parasites. Both antigen specific IgG1 and IgG2 antibody levels were measured. Immunization of dogs with the recombinant antigens formulated in either MPL-SE or AdjuPrime resulted in high antibody levels particularly to LmSTI1. In addition, this immunization although to low levels, resulted in the development of antibody response to the whole parasite lysate. Importantly, experimental exposure with low numbers of culture forms of L. chagasi promastigotes caused a clear boost in the immune response to both the recombinant antigens and the corresponding native molecules. The boost response was predominantly of the IgG2 isotype in animals primed with the recombinant antigens plus MPL-SE. In contrast, animals primed with the recombinant antigens formulated in AdjuPrime as well as animals vaccinated with crude antigen preparation responded with mixed IgG1/IgG2 isotypes. These results point to the possible use of this antigen cocktail formulated with the adjuvant MPL-SE in efficacy field trials against canine VL.     

Effect of field pea level on intake, digestion, microbial efficiency, ruminal fermentation, and in situ disappearance in beef steers fed forage-based diets

Four ruminally and duodenally cannulated crossbred beef steers (397+/-55 kg initial BW) were used in a 4 x 4 Latin square to evaluate the effects of increasing level of field pea supplementation on intake, digestion, microbial efficiency, ruminal fermentation, and in situ disappearance in steers fed moderate-quality (8.0% CP, DM basis) grass hay. Basal diets, offered ad libitum twice daily, consisted of chopped (15.2-cm screen) grass hay. Supplements were 0, 0.81, 1.62, and 2.43 kg (DM basis) per steer daily of rolled field pea (23.4% CP, DM basis) offered in equal proportions twice daily. Steers were adapted to diets on d 1 to 9; on d 10 to 14, DMI were measured. Field pea and grass hay were incubated in situ, beginning on d 10, for 0, 2, 4, 8, 12, 16, 24, 36, 48, 72, and 96 h. Ruminal fluid was collected and pH recorded at -2, 0, 2, 4, 6, 8, 10, and 12 h after feeding on d 13. Duodenal samples were taken for three consecutive days beginning on d 10 in a manner that allowed for a collection to take place every other hour over a 24-h period. Linear, quadratic, and cubic contrasts were used to evaluate the effects of increasing field pea level. Total DMI and OMI increased quadratically (P = 0.09), whereas forage DMI decreased quadratically (P = 0.09) with increasing field pea supplementation. There was a cubic effect (P < 0.001) for ruminal pH. Ruminal (P = 0.02) and apparent total-tract (P = 0.09) NDF disappearance decreased linearly with increasing field pea supplementation. Total ruminal VFA concentrations responded cubically (P = 0.008). Bacterial N flow (P = 0.002) and true ruminal N disappearance (P = 0.003) increased linearly, and apparent total-tract N disappearance increased quadratically (P = 0.09) with increasing field pea supplementation. No treatment effects were observed for ruminal DM fill (P = 0.82), true ruminal OM disappearance (P = 0.38), apparent intestinal OM digestion (P = 0.50), ruminal ADF disappearance (P = 0.17), apparent total-tract ADF disappearance (P = 0.35), or in situ DM disappearance of forage (P = 0.33). Because of effects on forage intake and ruminal pH, field peas seem to act like cereal grain supplements when used as supplements for forage-based diets. Supplementing field peas seems to effectively increase OM and N intakes of moderate-quality grass hay diets.     

Results of single-session bilateral triple pelvic osteotomy with an eight-hole iliac bone plate in dogs: 95 cases (1996-1999)

OBJECTIVE: To evaluate feasibility of single-session bilateral triple pelvic osteotomy with 8-hole iliac bone plates in dogs with bilateral hip dysplasia. DESIGN: Retrospective study. ANIMALS: 95 dogs with bilateral hip dysplasia. PROCEDURE: Medical records were reviewed, and information was obtained on signalment; body weight; angles of subluxation and reduction prior to surgery; durations of surgery and hospitalization; postoperative mobility; severity of lameness, radiographic grade of hip dysplasia, Norberg angle, and femoral head coverage before and after surgery; time required for radiographic evidence of iliac healing; change in pelvic diameter; implant integrity; and complications. RESULTS: Mean age at the time of surgery was 10.8 months, and mean weight was 35.2 kg (77.4 lb). Prior to surgery, mean angles of subluxation were 2.2 degrees on the right and 2.6 degrees on the left; mean angles of reduction were 25.9 degrees on the right and 27.3 degrees on the left. Mean surgical time was 95 minutes. All but 1 dog were able to walk on their own by the fourth day after surgery. Mean hospitalization time was 7.5 days. Clinical signs of lameness and radiographic grade of hip dysplasia were significantly improved during follow-up examinations. Mean time for radiographic iliac healing was 8 weeks. None of the plates and only 7 of the 1,520 (0.5%) screws loosened after surgery. Nineteen dogs had complications, but all complications were minor. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that single-session bilateral triple pelvic osteotomy with 8-hole iliac bone plates is effective for treatment of dogs with bilateral hip dysplasia.     

Technique for equine cervical articular process joint injection

Degenerative changes and osteochondrosis of articular processes are common sources of stiffness or pain in the equine cervical spine. Temporary relief of the clinical signs related to these maladies may be achieved by injecting corticosteroids into the joint. This is routinely done by ultrasound-guided needle placement. The cervical articular processes and joint form an easily identifiable sonographic landmark, a step-like echogenic surface described as having the appearance of a "chair."     

QUANTIFICATION OF LEFT VENTRICULAR MASS USING CARDIAC MAGNETIC RESONANCE IMAGING COMPARED WITH ECHOCARDIOGRAPHY IN DOMESTIC CATS

The hypotheses were that cardiac magnetic resonance imaging (cMRI) would accurately determine LV mass in domestic cats and would do so more accurately than echocardiography (ECHO). ECHO was performed on seven sedated cats. LV mass was calculated using the truncated ellipse formula from a right parasternal long-axis view. T1 weighted gradient echo cMRI was acquired from anesthetized cats during multiple phases of the cardiac cycle. Short-axis images were obtained by acquiring 3 mm thick contiguous slices perpendicular to the cardiac long axis. LV mass was determined using Simpson's rule. Endocardial and epicardial borders were traced on each slice at end-systole, end-diastole, and mid-cycle and the difference in areas was myocardial area. Myocardial area was multiplied by slice thickness to calculate myocardial volume. Total (summated) myocardial volume was multiplied by myocardial density (1.05) to obtain LV mass at three measured phases of the cardiac cycle. Cats were euthanized and the LV was dissected and weighed to determine true mass. CMRI at end-systole most accurately quantified LV mass and was more accurate than echocardiography (P = 0.0078). Actual LV mass ranged from 6.5 to 10.5 g (mean = 8.5 g, SD = 1.6 g) compared with MRI LV mass at end-systole, which ranged from 6.7 to 11.1 g (mean = 8.7 g, SD = 1.7 g) and echocardiographic LV mass at enddiastole, which ranged from 5.2 to 9.1 g (mean= 7.1 g, SD = 1.8 g). Inter- and intraobserver variability for cMRI was 2%. CMRI obtained at end-systole accurately and reliably quantifies LV mass in domestic cats. It is more accurate than the echocardiographic method used in this study.     

Nonsteroidal anti-inflammatory drug-induced duodenal ulceration and perforation in a mature rottweiler

A mature male rottweiler was evaluated for acute collapse and abdominal pain. The history consisted of concurrent administration of meloxicam and aspirin. On exploratory laparotomy, a large perforated ulcer was discovered in the proximal duodenum, with secondary peritonitis. The pathogenesis of NSAID-induced gastrointestinal ulceration and the supposed safety of COX-2 selective agents are discussed.     

Caprine serum fraction immunomodulator as supplemental treatment of lower respiratory disease in the horse

Suppurative lower airway disease is a common debilitating disease in performance horses and, while rarely fatal, is often recalcitrant to conventional therapy. A variety of treatments have been used to combat this condition and we conducted two types of studies to determine if caprine serum fraction--immunomodulator (CSFI), a nonspecific immunomodulator, improved recovery from lower respiratory disease. Two dose response studies were performed to ascertain the efficacy of CSFI. Horses were maintained daily on conventional antibiotic therapy. Respiratory tract exudate, nasal discharge, dyspnoea, chest auscultation and cough frequency were monitored weekly. One hundred percent of the horses treated with 2 i.m. injections of either 60 or 120 mg CSFI one week apart showed significant improvement with each weekly evaluation and were fully recovered by week 3. Horses treated with 15 or 30 mg CSFI did not differ significantly from the control group. Only 10% of the control horses responded to conventional antibiotic therapy. An expanded field trial utilising 80 horses diagnosed with lower respiratory disease and housed at 4 equine clinics was conducted. Thirty-five percent of the 40 control horses, treated solely by conventional antibiotic therapies, recovered while 75% of the horses treated with a supplemental administration of 60 mg CSFI as described above recovered. The combined data from these studies showed that CSFI was able to promote an overall recovery from lower respiratory disease of 86%.     

Collection and analysis of peritoneal fluid from healthy llamas and alpacas

OBJECTIVE: To describe a technique for abdominocentesis in camelids and report peritoneal fluid biochemical and cytologic findings from healthy llamas and alpacas. DESIGN: Prospective study. Animals-17 adult llamas and 5 adult alpacas. PROCEDURES: Right paracostal abdominocentesis was performed. Peritoneal fluid was collected by gravity flow into tubes containing potassium-EDTA for cell count and cytologic evaluation and lithium heparin for biochemical analysis. Blood samples were collected via jugular venipuncture into heparinized tubes at the same time. Cytologic components were quantified. Fluid pH and concentrations of total carbon dioxide, sodium, potassium, chloride, lactate, and glucose were compared between peritoneal fluid and venous blood. RESULTS: All but 3 camelids had peritoneal fluid cell counts of < 3,000 nucleated cells/microL, with < 2,000 neutrophils/microL and < 1,040 large mononuclear cells/microL. All but 1 had peritoneal fluid protein concentrations of > or = 2.5 g/dL. Peritoneal fluid of camelids generally contained slightly less glucose, lactate, and sodium and roughly equal concentrations of potassium and chloride as venous blood. CONCLUSIONS AND CLINICAL RELEVANCE: Peritoneal fluid was collected safely from healthy camelids. Compared with blood, peritoneal fluid usually had a low cell count and protein concentration, but some individuals had higher values. Electrolyte concentrations resembled those found in blood. High cell counts and protein concentrations found in peritoneal fluid of some healthy camelids may overlap with values found in diseased camelids, complicating interpretation of peritoneal fluid values.