Flea bite hypersensitivity: new aspects on the involvement of mast cells

A study was performed to test the effect of sensitization to flea antigen, followed by exposure to fleas on mast cells (MCs), their subtypes, and IgE+ cells. Biopsies were taken from flea-sensitized dogs (n=28) and non-sensitized dogs (n=5) that had been exposed to fleas. Control groups consisted of flea-sensitized (n=12) and non-sensitized dogs (n=9) that were not exposed to fleas. Biopsies, taken before, 24 and 72 h after local flea exposure, were stained with haematoxylin and eosin (H&E), toluidine blue, a double labelling technique for MC chymase and tryptase and anti-IgE. An intradermal test for flea antigen was performed and serum titres of allergen-specific IgE and IgG were measured. Significantly higher numbers (P<0.001) of double labelled MCs compared to toluidine blue stained MCs were detectable in flea-sensitized dogs independent of flea exposure. In contrast, in non-sensitized dogs, the number of toluidine blue stained MCs and the number of double labelled MCs did not differ. In flea-sensitized dogs after flea exposure the percentage of C-MC was significantly increased at day 1 (P<0.001) and day 3 (P<0.001), whereas the percentage of TC-MCs decreased significantly at day 1 (P<0.001) and day 3 (P<0.05). The percentage of T-MCs decreased (P<0.05 day 0 versus day 1; P<0.05 day 0 versus day 3). No significant difference was detectable after toluidine blue staining and staining for IgE+ cells between the groups nor between the MC density and the number of IgE+ cells. All flea-sensitized dogs had positive skin tests to flea antigen and high serum titres of flea-specific serum IgE and IgG antibodies. In non-sensitized dogs, these results were negative. Our data provide strong evidence for an upregulation of MC proteases during the process of sensitization and a generalized selective release of mast cell tryptase after exposure to the antigen.     

Histologic evaluation of the immediate effects of diamond burr debridement in experimental superficial corneal wounds in dogs

Purpose To evaluate the corneal changes immediately after diamond burr debridement of superficial corneal wounds in dogs. Spontaneous chronic corneal epithelial defects (SCCEDs) are the most common form of canine recurrent corneal ulcers. The diamond burr has been used in the management of corneal lesions in humans since 1983. Recently, it has been successfully used in the treatment of SCCEDs in dogs; however, little has been documented as to its mechanism of action. Methods Five adult female research dogs euthanized for reasons unrelated to the study were included, providing 10 normal eyes. An excimer laser spatula was used for epithelial removal after delineation with an 8 mm punch biopsy trephine. Diamond burr debridement was performed for 30 and 45 s in five eyes each (groups 1 and 2 respectively). The procedure was performed on the ventral half of the experimental defect as well as ventral normal cornea, immediately after euthanasia, and prior to enucleation. Samples were processed routinely for histologic evaluation and stained with periodic acid–Schiff. Results No stromal defects could be identified under light microscopy. In experimental corneal wounds, multi‐focal areas remained covered by the epithelial basement membrane (BM) after diamond burr treatment in both groups (group 1 = 48%±16SD, group 2 = 26%±12SD). Removal of BM on group 2 was significantly higher than group 1 (P < 0.05). Conclusions The diamond burr allows a safe method of debridement and does not create defects beyond the epithelial BM in corneal wounds in normal dogs. Evaluation of the diamond burr debridement in cases of SCCEDs is warranted.     

Clinical findings,bronchoalveolar lavage fluid cytology and matrix metalloproteinase-2 and -9 in canine pulmonary eosinophilia

We characterized clinical and clinicopathological features, and the involvement of gelatinolytic matrix metalloproteinases (MMP-2 and -9) in canine pulmonary eosinophilia (PE). Study material consisted of 20 PE dogs and 16 healthy beagles. All dogs underwent a similar clinical examination and bronchoalveolar lavage (BAL). Analysis for cell count and differential cell count of BAL fluid (BALF), arterial blood gas analysis before and after BAL, and thoracic radiographs before BAL and after treatment were obtained. Twelve dogs were re-evaluated and six relavaged. MMP-2 and MMP-9 in BALF were analysed by zymography, Western immunoblotting and immunocytochemistry.In the PE dogs, BALF, cell count, number and percentage of eosinophils, and numbers of macrophages, lymphocytes, neutrophils, mast cells and epithelial cells were all significantly elevated. Blood eosinophilia was detected in half of the PE dogs. Three PE dogs had mild hypoxaemia. The BAL procedure had an equal effect on PE and healthy dogs' arterial blood gas values. Bronchointerstitial densities were seen in PE dogs' radiographs. Treatment of PE decreased BALF cell count, eosinophil count and percentage and diminished radiographic changes. Gelatinolytic activity was higher in PE dogs' BALF. BALF macrophages and epithelial cells were the principal sources of the MMP-9.     

Pathogenicity and protective activity in pregnant goats of a Brucella melitensis Deltaomp25 deletion mutant

The Brucella melitensis mutant BM 25, which lacks the major 25 kDa outer membrane protein Omp25, has previously been found to be attenuated in the murine brucellosis model. In the present study, the capacity of the Deltaomp25 mutant to colonise and cause abortions in the caprine host was evaluated. The vaccine potential of BM 25 was also investigated in goats. Inoculation of nine pregnant goats in late gestation with the B. melitensis mutant resulted in 0/9 abortions, while the virulent parental strain, B. melitensis 16M, induced 6/6 dams to abort (P<0.001, n=6). BM 25 also colonised fewer adults (P<0.05, n=6) and kids (P<0.01, n=6) than strain 16M. The Deltaomp25 mutant was found capable of transient in vivo colonisation of non-pregnant goats for two weeks post-infection. Owing to the ability of BM 25 to colonise both non-pregnant and pregnant adults without inducing abortions, a vaccine efficacy study was performed. Vaccination of goats prior to breeding with either BM 25 or the current caprine vaccine B. melitensis strain Rev. 1 resulted in 100 per cent protection against abortion following challenge in late gestation with virulent strain 16M (P<0.05, n=7). However, unlike strain Rev. 1, BM 25 does not appear to cause abortions in late gestation based on this study with a small number of animals. The B. melitensis Deltaomp25 mutant, BM 25, may be a safe and efficacious alternative to strain Rev. 1 when dealing with goat herds of mixed age and pregnancy status.     

Collagenolytic activity in bronchoalveolar lavage fluid in canine pulmonary eosinophilia

We studied and characterized the collagenolytic matrix metalloproteinases (MMP-8 and MMP-13) in the pathogenesis of canine pulmonary eosinophilia (PE). Twenty dogs with PE and 16 healthy control dogs underwent similar clinical examination and collection of bronchoalveolar lavage fluid (BALF). Analyses of total cell and differential cell counts and collagen I degradation with and without aminophenyl mercuric acetate (APMA) treatment were performed. Correlations between cell counts and percentage of degraded collagen I in BALF were studied. Collagenase activity detected in BALF was characterized by Western immunoblotting for collagenase-2 (MMP-8) and collagenase-3 (MMP-13), and their cellular location was studied by immunocytochemical means. Collagenolytic activity was significantly increased in cell-free and native BALF of PE dogs compared to healthy controls. APMA treatment had no significant effect on BALF collagenase activity, indicating that collagenolytic activity occurred in diseased BALF in vivo in active form. Western immunoblotting identified the presence of MMP-8 and MMP-13 immunoreactivities, of which the latter was converted to active form. Major immunoreactivity for MMP-8 was observed in macrophages and epithelial cells, and major immunoreactivity for MMP-13 was observed in macrophages. A significant positive correlation was noted between the percentage of degraded collagen I and the counts of eosinophils, macrophages, lymphocytes, and mast cells. These findings suggest that the up-regulation of collagenolysis eventually contributes to pulmonary tissue destruction in canine PE.     

Results of ligation of patent ductus arteriosus in dogs: 201 cases (1969-1988)

Surgical treatment of 201 dogs with patent ductus arteriosus at the College of Veterinary Medicine, The Ohio State University was evaluated retrospectively to determine risk factors for development of surgical complications. During surgery, 15 dogs (7%) died because of hemorrhage associated with ductus dissection (n = 8), pulmonary edema (n = 4), ventricular fibrillation (n = 1), hemorrhage not associated with ductus dissection (n = 1), and cardiac arrest immediately after ductus ligation (n = 1). An additional 8 dogs (4%) died less than 1 month after surgery (total mortality before, during, and immediately after surgery, 11%). Nineteen dogs (9.5%) developed hemorrhage during surgery. Sixteen dogs developed complications other than hemorrhage (pulmonary edema [n = 4], cardiac arrest [n = 4], iatrogenic lung trauma [n = 3], ventricular fibrillation [n = 2], septicemia [n = 2], and recanalized ductus [n = 2]). Correlation was not found between age, sex, body weight, surgical technique (Jackson method vs standard method of dissection), or surgeon level of training and development of hemorrhage during surgery, other complications, or survival less than 5 days. Positive correlation (P less than 0.05) was found between hemorrhage and death within 5 days after surgery. Positive correlation (P less than 0.05) was also found between other complications and death within 5 days after surgery. Nineteen dogs survived surgery, but later died of unrelated causes (mean life span, 57 months); 63 of the dogs were still alive and doing well as of January 1990 (mean life span, 47 months after surgery). Contrary to previous reports, age, body weight, and surgical technique did not affect results.(ABSTRACT TRUNCATED AT 250 WORDS)     

Topical 0.1% tacrolimus for the treatment of discoid lupus erythematosus and pemphigus erythematosus in dogs

Topical 0.1% tacrolimus was used for treatment of localized lesions associated with 10 cases of discoid lupus erythematosus (DLE) and two cases of pemphigus erythematosus (PE) either as a sole therapy (n=2) or as an adjunctive treatment (n=10). Eight of 10 dogs with DLE and both dogs with PE were improved following 8 weeks of topical application. In six of the eight dogs that improved, other medications were discontinued. No adverse effects in clinical or laboratory parameters were noted throughout the study.     

Leishmania infantum-specific IgG, IgG1 and IgG2 antibody responses in healthy and ill dogs from endemic areas. Evolution in the course of infection and after treatment

The expression of IgG, IgG1 and IgG2 specific antibodies for Leishmania infantum was studied in five groups of dogs in Catalonia (Spain): I, 99 asymptomatic dogs (infected and uninfected) from a highly endemic area for leishmaniosis; II, 139 untreated dogs with clinically patent leishmaniosis; III, 11 naturally infected asymptomatic dogs monitored for up to 5 years since they were found seropositive to Leishmania antigen and without treatment; IV, 25 naturally infected dogs with clinically patent leishmaniosis and treated with either meglumine antimoniate and allopurinol or allopurinol alone and V, six experimentally infected dogs, treated with meglumine antimoniate and controlled for 5 years. The levels (ELISA units) of IgG, IgG1 and IgG2 in asymptomatic dogs (group I) were very variable (24+/-33, 32+/-31 and 26+/-31, respectively), and, as expected, lower than in ill dogs (group II) (168+/-34, 84+/-71 and 172+/-31, respectively). In both groups, the correlation between IgG and IgG2 levels (r=0.95, P<0.001 in group I and r=0.63, P<0.001 in group II) was higher than between IgG and IgG1 levels (r=0.01, P>0.05 in group I and r=0.31, P<0.001 in group II). In group III, IgG and IgG2 expression increased during infection, while IgG1 expression remained the same. In dogs of group IV, IgG levels after 1 year of treatment decreased more in responsive (mean values, 163+/-42 before treatment (b.t.) and 100+/-36 after treatment (a.t.)) than in unresponsive dogs (158+/-29 b.t. and 124+/-51 a.t.), especially for IgG1 (94+/-89 b.t. and 20+/-21 a.t. in responsive dogs and 35+/-25 b.t. and 22+/-13 a.t. in unresponsive dogs) rather than for IgG2 (156+/-16 b.t. and 114+/-45 a.t. in responsive and 151+/-11 b.t. and 125+/-36 a.t. in unresponsive dogs). Similar results were observed in the evolution of experimentally infected animals after consecutive and specific treatments. Overall results show the great variation in Leishmania-specific IgG1 expression in asymptomatic and symptomatic dogs, their lack of correlation with that of IgG2 and chemotherapy is more effective in dogs with initially high expression of IgG1.     

Influence of ampicillin, ceftiofur, attenuated live PRRSV vaccine, and reduced dose Streptococcus suis exposure on disease associated with PRRSV and S. suis coinfection

The objective of this research was to evaluate the efficacy of two antimicrobials (ampicillin and ceftiofur), a modified-live porcine reproductive and respiratory syndrome virus (PRRSV) vaccine, and low dose exposure to Streptococcus suis on disease associated with PRRSV/S. suis coinfection. Fifty-six, crossbred, PRRSV-free pigs were weaned at 10-12 days of age and randomly assigned to five treatment groups. All pigs were inoculated with 2ml of 10(6.4) TCID50/ml of high virulence PRRSV isolate VR-2385 intranasally at 29-31 days of age (day 0 of the study) followed 7 days later by intranasal inoculation with 2ml of 10(8.9)colony forming units(CFU)/ml S. suis type 2 isolate ISU VDL #40634/94. Pigs in group 1 (n=10) served as untreated infected positive controls. Pigs in group 2 (n=12) were treated with 5.0 mg/kg ceftiofur hydrochloride intramuscularly (IM) on days 8, 11, and 14. Pigs in group 3 (n=11) were treated with 11 mg/kg ampicillin IM on days 8-10. Pigs in group 4 (n=12) were vaccinated 14 days prior to PRRSV challenge with a commercial modified-live PRRSV vaccine. Pigs in group 5 (n=11) were exposed to a 1:100 dilution of the S. suis challenge inoculum 19 days prior to S. suis challenge. Mortality was 80, 25, 82, 83, and 36% in groups 1-5, respectively. The reduced dose S. suis exposure had some residual virulence, evidenced by S. suis induced meningitis in two pigs after exposure. Treatment with ceftiofur hydrochloride and reduced dose exposure to S. suis were the only treatments which significantly (P<0.05) reduced mortality associated with PRRSV/S. suis coinfection, significantly (P<0.05) reduced recovery of S. suis from tissues at necropsy, and significantly (P<0.05) reduced the severity of gross lung lesions.     

Fracture of the proximal tibial epiphysis and tuberosity in 10 dogs

Ten dogs were presented with fractures of the proximal tibial epiphysis and tuberosity. All dogs had a cranioproximal-caudodistal angulation of the tibial plateau. Six dogs had marked caudal displacement of the proximal tibial epiphysis, five of which had also sustained fractures of the proximal fibula. The estimated mean angle of inclination of the tibial plateau of affected limbs was 45.8 +/- 9.6 degrees, which was significantly greater (P<0.0005) than the estimated mean angle of the normal contralateral limb 26.2 +/- 6.6 degrees. The mean angle of inclination of the tibial plateau of dogs with fibular fractures (n=5) was not significantly different from dogs without fibular fractures (n=5) (P>0.25). Five dogs were treated conservatively and five were treated by three different methods of surgical repair. Surgically treated dogs had significantly greater preoperative tibial plateau angles (P<0.05). All dogs regained full limb usage, regardless of the method of treatment chosen.