In vitro study of neutrophil apoptosis in dogs

In the present study, to investigate the apoptosis of the polymorphonuclear neutrophil (PMN) from healthy dogs, we carried out TUNEL assay and DNA analysis by electrophoresis on dog PMNs. The TUNEL assay indicated that apoptotic PMNs in dogs were 0.15+/-5% before incubation, 0.3+/-5% at 4h incubation, 1+/-6% at 8h, 9+/-4% at 12h and 28+/-5% at 24h, respectively. The ladder formation was much more clearly observed in DNA from PMNs after 24h incubation at 37 degrees C than that before incubation. The results in this study indicated that healthy dog PMNs undergo apoptosis spontaneously within hours to days, and that the apoptosis of PMNs might be related to the high turnover of these circulating cells in dogs.     

On ancestors of dog breeds with focus on Weimaraner hunting dogs

Paternally inherited Y chromosomal markers and maternally inherited mitochondrial (mt) DNA sequences were investigated in 27 dog breeds (Canis familiaris), of which the Weimaraner hunting dog was studied in greater detail. Altogether, nine potentially polymorphic markers of the Y chromosome were examined as well as parts of the canine mt genome (1947 base pairs) in 111 male dogs and four wolves for comparison. Twenty Y chromosomal and fifty-nine mitochondrial DNA (mtDNA) haplotypes were identified in the canine breeds and wolves. In 34 Weimaraners, four distinct Y chromosomal haplotypes were observed as well as three mtDNA types thus reflecting at least four male and three female ancestors for the current population in Germany. Tracing patri- and matrilineages, several entries in the Weimaraner stud book cannot be reconciled with the male-only, Y chromosomal neither the female-only, mt inheritance patterns, respectively. The investigated breeds represent 9 of 10 groups defined by the Fédération Cynologique Internationale (FCI). The level of Y chromosomal and especially mtDNA diversity was immense considering the relatively small number of individuals investigated per breed. Unique haplotypes were found only in a few breeds and the wolf. Other haplotypes were shared among several breeds, also across different FCI groups, suggesting that these canine breeds had common male and female ancestors.     

Thyroid and immunologic status of dogs with perianal fistula

Lymphocyte-proliferation responses, absolute lymphocyte counts, and thyrotropin-stimulation responses were determined in 33 dogs with perianal fistula; serum immunoglobulin values also were determined in 15 of the 33 dogs. Lymphocytes were stimulated with concanavalin A, pokeweed mitogen, and phytohemagglutinin and were cultured with medium containing normal pooled canine serum or fresh patient's autologous serum. Initially, lymphocytes from 9 dogs (27.3%) had depressed stimulation responses to greater than or equal to 1 phytomitogen, and 4 of the 9 dogs had absolute lymphopenia. One month after recovery in these 9 dogs, lymphocytes from 4 dogs (66.7%) had normal proliferation responses. Of immunoglobulin determinations in 15 dogs, serum IgA values were 32 to 185 mg/dl (mean, 69 +/- 10 mg/dl) and were low in 2 dogs (13%), and serum IgM values were 48 to 610 mg/dl (mean, 263 to 46 mg/dl) and were high in 8 dogs (53%). Serum IgG values were 1,050 to 3,220 mg/dl (mean, 2,339 +/- 165 mg/dl) and were high in 10 dogs (71%). After thyrotropin stimulation, 1 dog was considered hypothyroid. Neither pathogenesis nor prognosis of canine perianal fistula was clarified via immunoglobulin concentrations or absolute lymphocyte counts. Based on lymphocyte-proliferation assays, suppression of cell-mediated immunity was probably a result of perianal fistula, rather than a cause of the fistula.     

Vaginal resection and anastomosis for treatment of vestibulovaginal stenosis in 4 dogs with recurrent urinary tract infections

CASE DESCRIPTION-4 dogs were evaluated because of recurrent urinary tract infections. CLINICAL FINDINGS-All dogs had recurrent urinary tract infections and similar clinical signs; 3 dogs had urinary incontinence. Digital vaginal examination revealed vestibulovaginal stenosis in all dogs, which was confirmed by results of contrast vaginourethrography. From image measurements, the vestibulovaginal ratio (ratio of the height of the vestibulovaginal junction to the maximum height of the vagina on a lateral vaginourethrogram) was calculated for each dog. Three dogs had severe stenosis (vestibulovaginal ratio, < 0.20; severe stenosis is defined as a vestibulovaginal ratio < 0.20), whereas the fourth dog had moderate stenosis (vestibulovaginal ratio, 0.24; ratio range for moderate stenosis is 0.20 to 0.25). TREATMENT AND OUTCOME-All dogs were anesthetized for surgical correction of the vestibulovaginal stenosis. Vaginal resection and anastomosis of the stenosis was performed in all 4 dogs, with 1 dog also undergoing episioplasty. Complete resolution of clinical signs was apparent in 3 dogs; 1 dog had postoperative complications including pollakiuria and stranguria, which resulted in rectal and vaginal prolapse. This dog underwent ovariohysterectomy, after which clinical signs resolved. All dogs had resolution of urinary tract infections at the time of follow-up (6 to 8 months after surgery). CLINICAL RELEVANCE-Resection and anastomosis may resolve recurrent urinary tract infections in dogs with severe or moderate vestibulovaginal stenosis. Episiotomy was not necessary for success of surgical treatment, and overall, that procedure increased morbidity, the severity of intraoperative hemorrhage, and duration of surgery.     

In vitro responses of distal airways in horses with recurrent airway obstruction

Distal airway segments (ID, 3 to 4 mm; length, 5 mm) from 2 groups of horses were isolated and suspended in tissue baths filled with Krebs solution, aerated with 5% CO2 in oxygen and maintained at 37 degrees C. Responses to exogenous acetylcholine, isoproterenol, or electrical field stimulation were compared. Control horses (n = 30) had no history of recurrent airway obstruction, whereas principal horses (n = 15) had recurrent airway obstruction and were studied during an acute episode of airway obstruction. Although the distal airways contracted in response to the cumulative half-logarithmic addition of acetylcholine (10(-10) M to 10(-3) M) in both groups, bronchi obtained from principals were less sensitive to acetylcholine than were bronchi obtained from controls. Tetdrodotoxin-sensitive electrical field stimulation-induced contractions were observed in both groups of airways, but the tension achieved in principal bronchi was less than in controls. All electrical field stimulation-induced contractions were abolished by atropine, indicating that the only excitatory innervation of equine distal airway is through the parasympathetic system. To examine the effect of isoproterenol and determine inhibitory innervation, bronchi were precontracted with histamine. Electrical field stimulation did not cause relaxation of precontracted bronchi in either group, thus indicating that distal airways lack inhibitory innervation. Isoproterenol caused similar, dose-dependent relaxation in both groups.     

In vitro antifungal susceptibility of Malassezia pachydermatis from dogs with and without skin lesions

Canine Malassezia dermatitis is frequently treated with systemic ketoconazole (KTZ) and itraconazole (ITZ). However, no information is available on the antifungal susceptibility to azoles and allilamine of Malassezia pachydermatis isolates from dogs with or without skin lesions. The present study was designed to evaluate the in vitro antifungal susceptibility of M. pachydermatis strains from dogs with or without skin lesions to KTZ, ITZ, miconazole (MICO), fluconazole (FLZ), posaconazole (POS), voriconazole (VOR) and terbinafine (TER) using the Clinical and Laboratory Standards Institute reference Broth Microdilution Method (CLSI M27-A2). The association between the susceptibility to antifungal compounds and the origin of M. pachydermatis, from skin with or without lesions has been also assessed. A total of 62 M. pachydermatis strains from healthy dogs (i.e., Group A=30) or with skin lesions (i.e., Group B=32) were tested. ITZ, KTZ and POS showed the highest activity against M. pachydermatis strains, whereas MICO TER and FLZ the lowest. A higher number of Malassezia resistant strains were registered among isolates from Group B than those from Group A. This study indicates that M. pachydermatis strains were susceptible to ITZ, KTZ, and POS. However, dogs with lesions may harbour strains with low susceptibility to antifungal agents and displaying cross-resistance phenomena to azole. The antifungal therapy in Malassezia infections requires careful appraisal of choice of drugs especially in cases of unresponsiveness to antifungal treatment or recurrent infections.     

Lymphoscintigraphy in healthy dogs and dogs with experimentally created thoracic duct abnormalities.

Lymphoscintigraphic evaluation of the thoracic duct (TD) was performed in 10 healthy and 12 dogs with experimentally created TD abnormalities (6 dogs with TD lacerations and 6 dogs with cranial vena ligations). Complete imaging took 4 hours and caused no adverse effects or complications. Lymphoscintigraphy of healthy dogs failed to image the TD; however, background activity in the abdomen and thorax, and radioactivity in the kidneys, bladder, liver, and heart were noticed. Lacerations and transections of the TD were experimentally created in 6 dogs to ascertain whether TD rupture could be detected with lymphoscintigraphy. Lymphoscintigraphy was performed within 48 hours of creating the TD defect. There was no significant difference in the scintigraphic pattern of healthy dogs and those with experimentally created TD defects. Ligation of the cranial vena cava was performed in 6 dogs; 3 dogs developed chylothorax. In those 3 dogs, diffuse radioactivity was imaged in the thorax and was compatible with thoracic lymphangiectasia. In one of these dogs, linear activity consistent with the TD and localized regions of radioactivity cranial to the heart (compatible with the mediastinal lymph nodes) were noticed. Lymphoscintigraphic findings in these dogs correlated with lymphangiographic findings.     

In vitro fluoroquinolone susceptibility of Pseudomonas aeruginosa isolates from dogs with ulcerative keratitis

OBJECTIVE: To determine the in vitro fluoroquinolone susceptibility profiles of Pseudomonas aeruginosa isolates from dogs with ulcerative keratitis. Animals-27 dogs with P. aeruginosa-associated ulcerative keratitis. PROCEDURES: P. aeruginosa isolates from dogs with ulcerative keratitis were collected during a 3-year period. Isolates were tested by use of the disk diffusion method for their susceptibility to 7 fluoroquinolones that are available as commercial ophthalmic preparations. The antimicrobials included second- (ciprofloxacin, ofloxacin, norfloxacin, and lomefloxacin), third- (levofloxacin), and fourth-generation (gatifloxacin and moxifloxacin) fluoroquinolones. Isolates were designated as susceptible, intermediate, or resistant to the various antimicrobials. The percentage of susceptible isolates was compared among individual fluoroquinolones and among fluoroquinolone generations. RESULTS: None of the dogs had received topical or systemic fluoroquinolone treatment prior to referral. Twenty-seven P. aeruginosa isolates were collected during the study period. In vitro, bacterial resistance to the tested fluoroquinolones was infrequently identified (24/ 27 isolates were susceptible to all fluoroquinolones evaluated); susceptibility percentages ranged from 88.9% to 100% for individual antimicrobials. There were no significant differences among isolate susceptibilities to the individual antimicrobials or among generations of fluoroquinolones. CONCLUSIONS AND CLINICAL RELEVANCE: On the basis of these in vitro data, none of the 7 evaluated fluoroquinolones (individually or collectively by generation) appeared to offer a clinically important advantage in the treatment of P. aeruginosa-associated ulcerative keratitis in dogs. Among the P. aeruginosa isolates collected from dogs with ulcerative keratitis in this study, the likelihood of susceptibility to the fluoroquinolones evaluated was high.     

In vitro characteristics of normal and dystrophic skeletal muscle from dogs

Explants were prepared from skeletal muscle tissue from 5 nondystrophic pups and from 5 pups with X-linked muscular dystrophy; pups were 2 to 17 weeks old. A serial reexplant method resulted in optimal cell density with minimal fibroblast growth. Cultures were examined daily by use of phase-contrast microscopy; differentiated (post-fusion) cultures were examined by electron microscopy. Moderate nuclear pleomorphism and cell clustering were observed in cultures of normal and dystrophic muscle cells. Cultures were maintained to 27 days after plating. Minimal myofilament synthesis was observed in multinucleate cells from nondystrophic and dystrophic pups, but spontaneous contraction of myotubes was not observed during this period. Differences in growth, fusion, or differentiation of myogenic cells into multinucleate cells and myotubes were not found between dystrophic and normal muscle.     

Leukoencephalitis and vasculitis with perivascular demyelination in a Weimaraner dog

Aseptic and noninfectious diseases of the central nervous system are being recognized with increasing frequency. After multiple episodes of neurologic illness, this 7-year-old Weimaraner dog was euthanatized and submitted for postmortem examination. Lesions in the central nervous system were found mainly in the white matter of the cerebral cortex and cervical spinal cord and represented acute and more chronic injury. Necrotizing vasculitis with fibrinoid change and a marked neutrophilic infiltrate dominated the acute lesions. More chronic changes consisted of perivascular demyelination and accumulation of foamy macrophages with positive staining for myelin. An immune-complex (Arthus-type) vasculitis is suspected.     

Electrocardiographic abnormalities associated with tetanus in two dogs

Bradycardia, sinus arrest, and second-degree atrioventricular block developed in 2 dogs with tetanus. Clinical signs attributable to bradycardia were not apparent. Administration of atropine resulted in resolution of the arrhythmias. Both dogs responded well to supportive treatment; the bradycardia resolved within 4 days of onset without specific treatment. Tetanus should be included in the differential diagnosis when increased neuromuscular excitability and bradycardia are evident, as is found in toxicity with acetylcholinesterase inhibitors, increased intracranial pressure, and other neurologic disorders.     

Assessment of neutrophil function in dogs with primary ciliary dyskinesia

Compared with neutrophils from healthy dogs, neutrophils from 2 dogs with primary ciliary dyskinesia had increased distance of random migration, but fewer of the neutrophils migrated. The affected dogs had an increase in the numbers of Staphylococcus aureus phagocytized. Lymphocyte blastogenesis in the affected dogs in response to standard mitogens was considered to be normal.     

Acid-base and electrolyte abnormalities in dogs with gastrointestinal foreign bodies

Gastrointestinal foreign bodies occur commonly in dogs. The objective of the study was to describe the acid-base and electrolyte abnormalities identified in dogs with gastrointestinal foreign bodies and determine if these abnormalities are related to the site or type of foreign body present. Medical records of 138 dogs were reviewed, and information on signalment, initial venous electrolyte and acid-base values, surgical findings, relevant historical information, imaging modalities used, cost of hospital visit, intra- or postoperative complications, and survival was obtained. The site of the foreign body was recorded in 94.9% of cases and the most common site was the stomach (50%), followed by the jejunum (27.5%). The foreign bodies were linear in 36.2% of cases. The most common electrolyte and acid-base abnormalities regardless of the site or type of foreign body were hypochloremia (51.2%), metabolic alkalosis (45.2%), hypokalemia (25%), and hyponatremia (20.5%). No significant association was found between electrolyte or acid-base abnormalities and the site of foreign body. Linear, as opposed to discrete, foreign bodies were more likely to be associated with a low serum sodium concentration (odds ratio, 0.85; 95% confidence interval, 0.75-0.95). Hyperlactatemia (> 2.4 mmol/L) was seen in 40.5% of dogs. A wide variety of electrolyte and acid-base derangements are found in dogs with gastrointestinal foreign bodies. Hypochloremia and metabolic alkalosis are common in these dogs. Hypochloremic, hypokalemic metabolic alkalosis is seen with both proximal and distal gastrointestinal foreign bodies.     

Thyroid hormone abnormalities and outcome in dogs with non-thyroidal illness

OBJECTIVES: To document thyroid hormone abnormalities in dogs with non-thyroidal illness and identify markers of prognostic value. METHODS: Circulating total and free thyroxine, total triiodothyronine and thyrotropin concentrations were measured in 196 dogs with non-thyroidal illness. Clinical signs, previous medications and outcome were recorded in each case. Data were analysed to determine endocrine prognostic factors, and to document the prevalence of thyroid hormone abnormalities. RESULTS: Total triiodothyronine, and total and free thyroxine concentrations were decreased in 75.9, 34.7 and 4.5 per cent of cases, respectively. Dogs which were euthanased had significantly decreased total triiodothyronine, and total and free thyroxine concentrations compared with those which made a full recovery. Total triiodothyronine concentrations were significantly lower in dogs that were euthanased compared with those which made a partial recovery. CLINICAL SIGNIFICANCE: Thyroid hormone concentrations may be used as prognostic indicators in dogs with non-thyroidal illness. Low triiodothyronine syndrome may be more common in dogs than previously recognised.