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- Author or Editor: Lisle W. George x
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SUMMARY
The efficacy of an ophthalmic ointment containing benzathine cloxacillin for treatment of infectious bovine keratoconjunctivitis was determined in 2 experiments. In the first experiment, Holstein calves (n = 6/group) were inoculated with Moraxella bovis and treated on postinoculation days 3 and 6 with either topically applied benzathine cloxacillin (250 mg/eye) or long-acting oxytetracycline formulation (20 mg/kg of body weight, im). A third group of inoculated calves remained untreated as controls. For the second experiment, 4 groups of calves (n = 6/group) were inoculated and treated on postinoculation days 3 and 6 with 50, 125, 250, or 375 mg of benzathine cloxacillin; a fifth untreated group served as controls. Ocular specimens were obtained for microbiologie culture, and eyes were observed and assigned a clinical score daily. Eyes were photographed on alternate days. Ulcer surface area was measured, using a planimeter.
In experiment 1, the week-2 ulcer surface area measurements for both groups of treated calves were smaller than those for controls. There was a greater frequency of M bovis isolation from the ocular secretions of controls than from those of benzathine cloxacillin-treated calves during postinoculation weeks 2 and 3. The number of M bovis isolations from the benzathine cloxacillin- and oxytetracycline-treated calves was not significantly different at any sample collection interval. On week 3, the scores of the benzathine cloxacillin-treated calves were smaller than those of controls.
In experiment 2, calves of the 250- and 375-mg groups had smaller ulcer surface area measurements than did controls on week 2. By week 3, calves of the 375-mg group had smaller scores than did controls. During postinoculation weeks 1 through 3, M bovis was isolated less frequently from the ocular secretions of calves of the 250- and 375-mg groups than from those of the control calves.
Abstract
Objective—To characterize serum biochemical abnormalities in goats with uroliths.
Design—Retrospective case-control series.
Animals—107 male goats with uroliths and 94 male goats with various nonrenal diseases (controls).
Procedures—For male goats, results of serum biochemical analyses collected from 1992 through 2003 were retrieved from computerized records, as were signalment, clinical diagnoses, and discharge status. Results of analyses for BUN, creatinine, phosphorus, calcium, Na, K, Cl, total CO2, anion gap, and glucose were compared between goats with uroliths and control goats.
Results—Goats with uroliths had higher mean BUN, creatinine, total CO2, K, and glucose concentrations and lower mean phosphorus, Na, and Cl concentrations than control goats, with no difference in mean calcium concentration and anion gap. Goats with uroliths had higher frequency of azotemia, hypophosphatemia, hypochloridemia, and increased total CO2 and lower frequency of decreased total CO2 than control goats. Urolithiasis occurred more frequently in castrated males than in sexually intact males and in dwarf African breeds than in other breeds.
Conclusions and Clinical Relevance—Goats with uroliths often had hypophosphatemia at admission. Hypochloridemic metabolic alkalosis was the most common acid-base disorder. Rupture in the urinary tract system was associated with increased prevalence of hyponatremia and hyperkalemia. Clinicians should be aware of these abnormalities when determining fluid therapy.
SUMMARY
Disposition kinetics of cloxacillin were examined in calves after topical administration of benzathine cloxacillin and single iv administration of sodium cloxacillin, and the susceptibility of 17 field isolates of Moraxella bovis was measured. For the IV pharmacokinetic phase, sodium cloxacillin was administered at dosage of 10 mg/kg of body weight to male Holstein calves (n = 6, weighing 146 to 170 kg), and serum concentration of cloxacillin was measured thereafter for 10 hours.
For the ocular pharmacokinetic phase, 6 calves were given either of 4 benzathine cloxacillin topical formulations consisting of 50-, 125-, 250-, or 375-mg doses. Treatment was repeated every 10 days until all 4 benzathine cloxacillin dosages were tested in the same 6 calves. Blood and tears were collected for 72 hours after each benzathine cloxacillin formulation was administered, and the concentration of cloxacillin in each specimen was measured, using a bioassay.
The minimal inhibitory concentration of cloxacillin for 17 field isolates of M bovis was determined by use of an agar pour-plate dilution assay.
After single iv administration of sodium cloxacillin, its half-life, body clearance, and volume of distribution were 19.5 ± 12.8 minutes, 18.3 ± 2.2 ml/min·kg, and 496 ± 290 ml/kg, respectively. After topical administration of benzathine cloxacillin, cloxacillin concentration in lacrimal fluid peaked between 30 and 45 minutes and ranged between 963 μg/ml and 3,256 μg/ml for the 125- and 375-mg doses, respectively. There was no detectable cloxacillin activity in the lacrimal fluid of any calf by 36 hours after topical administration of benzathine cloxacillin, and cloxacillin was not detected in the serum at any time. The mean lacrimal fluid cloxacillin concentration for the 4 groups during the first 8 hours was not significantly different; however, by 12 hours, the cloxacillin concentration in tears from calves of the 250- and 375-mg groups was significantly (P < 0.05) greater than that in calves of the 50- and 125-mg groups.
Cloxacillin concentration ≥ 3.13 μg/ml was maintained for a significantly (P < 0.05) longer time after treatment, using the 375-mg dose, compared with the 50-mg dose of benzathine cloxacillin. The minimal inhibitory concentration of cloxacillin for 1 isolate was 6.25 μg/ml, but was ≤ 3.13 μg/ml for 16 other M bovis isolates.
Abstract
Objectives—To compare stability, antigenicity, and aggregation characteristics of Moraxella bovis cytolysins among isolates from geographically diverse areas.
Study Population—8 isolates of M bovis.
Procedure—Filter-sterilized broth culture supernatants of M bovis were concentrated, diafiltered, and chromatographed. The endotoxin and cytolysin activities in samples were measured. Chromatographed cytolysins of M bovis were examined by immunoblotting. Hemolytic and leukotoxic activities were measured from samples collected at each step of purification and before and after storage. Hemolysis was measured directly by use of washed bovine erythrocyte targets. Leukotoxicity was measured by use of a 51Cr release assay.
Results—Cytolysin was retained by a filter with 100-kd nominal molecular weight limit. Hemolytic activity, leukotoxic activity, and endotoxin were eluted together in void volume of a gel-filtration column (molecular mass exclusion limit = 4 × 107 d). Gel-column chromatographed diafiltered retentate had the greatest specific cytolytic activity and the highest endotoxinto- protein ratio. Frozen diafiltered retentate(–80°C, 4 months) was cytolytic after thawing. Immunoblots of gel-column chromatographed cytolysin contained 4 proteins with molecular masses between 90 and 68 kd. Fractions with high lytic activities also had additional protein bands with molecular masses of 98 and 63 kd. Immunoblots of gel-column chromatographed diafiltered retentate revealed proteins with molecular masses between 90 and 68 kd.
Conclusions and Clinical Relevance—Diafiltered M bovis cytolysin is aggregated with endotoxin. Antigenicity and cytolytic activities in diafiltered retentate are conserved among M bovis isolates. Diafiltration could be useful for bulk semipurification of M bovis cytolysin. Cytolysin-enriched vaccines of M bovis could be contaminated by endotoxin. ( Am J Vet Res 2004;65:977–983)
Abstract
Objective—To identify the Moraxella bovis cytotoxin gene.
Procedure—Hemolytic and nonhemolytic strains of M bovis were compared by use of western blotting to identify proteins unique to hemolytic strains. Oligonucleotide primers, designed on the basis of amino acid sequences of 2 tryptic peptides derived from 1 such protein and conserved regions of the C and B genes from members of the repeats in the structural toxin (RTX) family of bacterial toxins, were used to amplify cytotoxin-specific genes from M bovis genomic DNA. Recombinant proteins were expressed, and antisera against these proteins were produced in rabbits.
Results—Several proteins ranging in molecular mass from 55 to 75 kd were unique to the hemolytic strain. An open reading frame encoding a 927-amino acid protein with a predicted molecular mass of 98.8 kd was amplified from M bovis genomic DNA. The deduced amino acid sequence encoded by this open reading frame was homologous to RTX toxins. Antisera against the recombinant carboxy terminus encoded by this open reading frame neutralized hemolytic and cytolytic activities of native M bovis cytotoxin.
Conclusions and Clinical Relevance—A gene was identified in M bovis that encodes a protein with sequence homology to other RTX toxins. Results of cytotoxin neutralization assays support the hypothesis that M bovis cytotoxin is encoded by this gene and belongs in the RTX family of bacterial exoproteins. Identification of this gene and expression of recombinant cytotoxin could facilitate the development of improved vaccines against infectious bovine keratoconjunctivitis. (Am J Vet Res 2001;62:1222–1228)
Abstract
Objective—To determine the immunogenicity of a Moraxella bovis cytolysin-enriched vaccine for prevention of infectious bovine keratoconjunctivitis (IBK).
Animals—104 mixed-breed beef calves ranging between 4 and 8 months of age.
Procedure—Vaccines were prepared by the diafiltration of broth culture supernatant from hemolytic M bovisor or sterile media. The diafiltered retentate was combined with Quil A adjuvant. Calves were randomly assigned to receive either the cytolysin vaccine (n = 35) or, as controls, adjuvant (35) or saline (0.9% NaCl) solution (34). Eyes of all calves were examined weekly for signs of IBK for 15 weeks. Calves that developed severe IBK were treated SC with florfenicol.
Results—Cytolysin vaccine contained 4 proteins with molecular masses ranging between 65 and 90 kd. Cytolysin-vaccinated calves had fewer instances of IBK than control calves. The time of onset of corneal lesions in cytolysin-vaccinated calves that developed IBK was delayed, compared with that of calves in either control group. The cytolysin-Quil A vaccine contained endotoxin, but calves did not have clinical signs of illness after vaccination.
Conclusions and Clinical Relevance—Calves that were vaccinated with a cytolysin-enriched vaccine had some resistance to IBK. Vaccines containing concentrated diafiltered M bovis cytolysin could protect beef calves against IBK. (Am J Vet Res 2005;66:136–142)
Abstract
Objective—To explore the use of urethral endoscopy and laser lithotripsy in the diagnosis and management of urolithiasis in goats and pot-bellied pigs.
Design—Prospective clinical study.
Animals—16 male goats and 6 male pot-bellied pigs with dysuria.
Procedure—Abdominal ultrasonography and urethral endoscopy were performed on all 22 animals. Endoscopic-guided holmium:yttrium-aluminum-garnet laser lithotripsy was performed in 3 goats and 2 pot-bellied pigs.
Results—Urolithiasis was identified in 15 goats and 5 pot-bellied pigs. Primary urinary bladder paralysis and cystitis were identified in the remaining pot-bellied pig and goat. Mean bladder diameters of obstructed small- and large-breed goats were 7 and 9.5 cm, respectively. The mean bladder diameter of obstructed pot-bellied pigs was 9.5 cm. Five of 20 animals with obstructive urolithiasis had severe urethral necrosis or stricture formation at the time of urethroscopy. All of these animals were euthanatized within 6 months because of persistent dysuria. When used, laser lithotripsy successfully fractured the distally located obstructing stones in the 3 goats and 2 pot-bellied pigs.
Conclusions and Clinical Relevance—Urethral endoscopy is useful for evaluating urethral patency in goats and pot-bellied pigs. Examination of the urethral mucosa following relief of urethral obstructions aids in the assessment of the long-term prognosis for urethral stricture. Urethral endoscopy also expands the therapeutic options for management of urolithiasis by providing a route for conducting laser lithotripsy. Laser lithotripsy proved to be safe and effective for clearing distally located calculi refractory to removal by traditional urethral flushing. Lithotripsy application is restricted to calculi lodged in the urethra. (J Am Vet Med Assoc 2002; 220:1831–1834)
Abstract
Objective—To evaluate the clinical efficacy of a single injection of tulathromycin, compared with saline (0.9% NaCl) solution-treated control calves, for treatment of induced infectious bovine keratoconjunctivitis in calves.
Design—Clinical trial.
Animals—30 Holstein bull calves ranging from 5 to 6 months old and 75 to 200 kg (165 to 440 lb) with no history of Moraxella bovis infections, no history of M bovis vaccination, and negative results for M bovis on 3 consecutive ocular bacterial cultures.
Procedures—Both eyes of each calf were infected with 1 X 1010 colony-forming units of piliated M bovis for 3 consecutive days prior to the trial. On day 0, ocular lesion scores were determined for each calf and the calves were weighed and assigned to a treatment (2.5 mg/kg [1.14 mg/lb] of body weight, SC) or control group according to a stratified random allocation based on weight and lesion score. Eyes were stained with fluorescein and photographed daily to record healing. Eyes were evaluated bacteriologically for M bovis on days 0 to 6 and at 3-day intervals thereafter.
Results—Median time to ulcer resolution in calves treated with tulathromycin was 9.1 days. More than 50% of control calves still had ulcers at the end of the trial (21 days). Moraxella sp was isolated less often from the eyes of treated calves than from the control calves. By day 10, the treated calves had lower ocular lesion scores than control calves.
Conclusions and Clinical Relevance—A single dose of tulathromycin (SC) was an effective treatment of calves with experimentally induced infectious bovine keratoconjunctivitis. The long serum half-life of tulathromycin, along with the results of this trial, suggests that tulathromycin may be a rational choice as a single-injection treatment for infectious bovine keratoconjunctivitis.