Isolation of Tritrichomonas foetus from cats sampled at a cat clinic, cat shows and a humane society in southern Ontario.

Tritrichomonas foetus is a protozoan parasite that has been associated with chronic diarrhea in cats. This study aimed to determine (i) the prevalence of T foetus shedding in cats from three different populations in southern Ontario, and (ii) associations between the presence of T foetus and potential cat management, health and demographic risk factors. A cross-sectional study was conducted involving 140 cats from a cat clinic in Guelph, 46 cats from a humane society in Guelph and 55 cats from two cat shows. Risk factor information was assessed through a questionnaire. The InPouch TF (feline) culture method was used to determine the presence of T foetus in all samples. Polymerase chain reaction was conducted on all samples positive by the InPouch TF, as well as 132 negative samples. The assays were interpreted in series and the prevalence of T foetus shedding and 95% confidence intervals (CI) were estimated at 0.7% (95% CI: 0.0-3.9%; n = 140) from the cat clinic, 0% (95% CI: 0.0-7.7%; n = 46) from the humane society and 23.6% (95% CI: 13.2-37.0%; n = 55) from the cat shows. 'Attendance at cat shows' was the only variable significant in both the univariable and multivariable analyses (P <0.05). No significant association was found between the presence of T foetus and diarrhea at the time of sampling or having a history of diarrhea in the past 6 months. The prevalence of T foetus was highly variable among populations of cats in southern Ontario, with shedding being most common in show cats.

Migration of cells from the yolk sac to hematopoietic tissues after in utero transplantation of early and mid gestation canine fetuses.

BACKGROUND: In utero hematopoietic cell transplantation offers a means of early intervention for the treatment of diseases before birth. Delivery of cells to the yolk sac is a minimally invasive approach that results in low levels of chimerism. However, there is little information on the optimal doses, timing of delivery, and migration of transplanted cells from the yolk sac into the fetus. METHODS: Varying cell doses of mesenchymal stromal cells or bone marrow mononuclear cells labeled with fluorescent supraparamagnetic iron oxide nanoparticles and a fluorescent intracellular dye, 5- and 6-([(4-chloromethyl)benzoyl]-amino) tetramethylrhodamine, were transplanted under ultrasound guidance to the yolk sacs of day 25 or day 35 canine fetuses. Ex vivo whole body fluorescence imaging and microscopy of tissue sections were correlated with the presence of iron oxide in injected and control fetuses. RESULTS: Day 25 and day 35 recipients showed similar survival rates after injection of cells into yolk sacs, although increased fetal morality was associated with cell doses greater than 10 cells/kg to day 25 fetuses. The fluorescence and iron oxide signals were predominantly localized to the abdominal regions, with no fluorescence visible in yolk sacs. Microscopy of tissues revealed colocalization of fluorophore with iron oxide in donor cells detected in the fetal livers and bone marrow of recipients 7 and 17 days after receiving mesenchymal stromal cells or bone marrow mononuclear cells. CONCLUSIONS: These studies demonstrated that cells injected into the yolk sacs of early gestation canine fetuses migrate to recipient hematopoietic tissues. Thus, yolk sac injection offers a safe and effective approach for engraftment of cells to fetal hematopoietic tissues.

Multiple endocrine diseases in cats: 15 cases (1997-2008).

The objective of this retrospective study was to characterize a population of cats from a tertiary care center diagnosed with multiple endocrine disorders, including the specific disorders and time intervals between diagnosis of each disorder. Medical records of 15 cats diagnosed with more than one endocrine disorder were reviewed. The majority of cats were domestic shorthairs, and the mean age at the time of diagnosis of the first disorder was 10.3 years. The most common combination of disorders was diabetes mellitus and hyperthyroidism. Two cats had concurrent diabetes mellitus and hyperadrenocorticism, one cat had concurrent central diabetes insipidus and diabetes mellitus. A mean of 25.7 months elapsed between diagnoses of the first and second endocrine disorder, but this was variable. This study suggests the occurrence of multiple endocrine disorders is uncommon in cats.

Invasive cutaneous angiomatosis and thrombocytopenia in a cat.

CASE DESCRIPTION: A 9-year-old 6.9-kg (15.18-lb) castrated male Siamese cat was evaluated because of a 3-year history of repeated hemorrhage from the right metacarpal pad. CLINICAL FINDINGS: Physical examination findings were unremarkable except for a 2-mm-diameter erosion of the right metacarpal pad. A CBC revealed marked thrombocytopenia. Serum biochemical analyses, retroviral screening, thoracic radiography, and abdominal ultrasonography revealed no abnormalities. Via ultrasonographic examination, the vasculature in the right metacarpal pad appeared increased, compared with that of the left pad; an aberrant arterial plexus that was confined to the metacarpal pad was identified via arterial angiography. TREATMENT AND OUTCOME: Surgical resection of the metacarpal pad (without digital pad transposition) with primary closure was performed. Histologic evaluation of the pad tissue revealed invasive cutaneous angiomatosis. The incision healed without complications, and limb function was considered normal. Administration of prednisone (2 mg/kg [0.91 mg/lb], PO, q 24 h) was initiated 4 weeks prior to surgery to treat suspected immune-mediated thrombocytopenia and continued afterwards with a tapering dosage. Platelet count was within reference limits 4 months after surgery; at 12 months, there was no evidence of recurrence of abnormal vasculature in the right metacarpal pad region. CLINICAL RELEVANCE: Complete resection of the metacarpal pad (without pad transposition) resulted in successful and well-tolerated treatment of cutaneous angiomatosis of the metacarpal pad of a cat. Recurrence of abnormal vasculature was not evident at a 12-month follow-up examination. Thrombocytopenia is commonly associated with vascular anomalies in humans and may have been a contributing factor in this cat.

Derivation and characterization of canine embryonic stem cell lines with in vitro and in vivo differentiation potential.

Embryonic stem cells (ESCs) represent permanent cell lines that can be maintained in an undifferentiated state. In an environment that induces differentiation, they form derivatives of the three embryonic germ layers: mesoderm, ectoderm, and endoderm. These characteristics give ESCs great potential for both basic research and clinical applications in the areas of regenerative medicine and tissue engineering. The establishment of ESCs from large animals that model human diseases is of significant importance. We describe the derivation of permanent canine cell lines from preimplantation-stage embryos. Similar to human ESCs, canine ESCs expressed OCT3/4, NANOG, SOX2, SSEA-3, SSEA-4, TRA-1-60, TRA-1-81, and alkaline phosphatase, whereas they expressed very low levels of SSEA-1. They maintained a normal karyotype and morphology typical of undifferentiated ESCs after multiple in vitro passages and rounds of cryopreservation. Plating cells in the absence of a feeder layer, either in attachment or suspension culture, resulted in the formation of embryoid bodies and their differentiation to multiple cell types. In vivo, canine ESCs gave rise to teratomas comprising cell types of all three embryonic germ layers. These cells represent the first pluripotent canine ESC lines with both in vitro and in vivo differentiation potential and offer the exciting possibility of testing the efficacy and safety of ESC-based therapies in large animal models of human disease.

Endoscopic evaluation of the gastroduodenal mucosa to determine the safety of short-term concurrent administration of meloxicam and dexamethasone in healthy dogs.

OBJECTIVE: To evaluate the safety, with respect to the development of gastric ulcers and erosions, of concurrent administration of meloxicam and dexamethasone for 3 days to healthy dogs. ANIMALS: 20 conditioned purpose-bred research Beagles. PROCEDURE: Seven days prior to treatment, dogs were anesthetized for endoscopic evaluation of the upper portion of the gastrointestinal tract (ie, the gastric and duodenal mucosa). Five regions of the gastroduodenal area were scored by 2 investigators. Dogs were randomly assigned to 1 of 4 treatment groups as follows: saline-saline, dexamethasone-saline, saline-meloxicam, and dexamethasone-meloxicam groups. On days 1, 2, and 3, dogs received either dexamethasone or saline (0.9% NaCl) solution injections SC twice daily. On days 2, 3, and 4, dogs received either meloxicam or saline solution injections SC once daily. On day 2, dogs were anesthetized for a sham surgery (ie, electrostimulation). On day 5, the gastroduodenal area of each dog was reevaluated by use of endoscopic evaluation and histologic examination of biopsy specimens. RESULTS: The total endoscopic score of the dexamethasone-meloxicam group was significantly greater than the scores of the other groups. The dexamethasone-saline group had a mean cumulative score that was significantly greater than the saline-meloxicam or saline-saline groups. Endoscopic scores of the saline-meloxicam group were not significantly different from scores of the saline-saline group. No significant differences in histologic findings were found between groups. CONCLUSIONS AND CLINICAL RELEVANCE: In healthy dogs, meloxicam appears to be safe with regard to adverse effects on the gastrointestinal tract. Concurrent administration of dexamethasone and meloxicam is more likely to cause gastric erosions than meloxicam administration alone.

Delivery of recombinant gene product to canine brain with the use of microencapsulation.

An alternative approach to somatic gene therapy is to deliver a therapeutic protein by implanting "universal" recombinant cells that are immunologically protected from graft rejection with alginate microcapsules. This strategy has proved successful in reversing pathologic conditions in several rodent models of human disease (dwarfism, lysosomal storage disease, hemophilia, cancer). In particular, neurologic disease and behavioral deficit in the mouse model of a neurodegenerative disease (mucopolysaccharidosis [MPS] VII) were significantly improved through the intraventricular implantation of the recombinant encapsulated cells. Here we report the feasibility of delivering recombinant gene products to the central nervous systems (CNSs) of dogs, first using human growth hormone as a marker for delivery in normal dogs and then using alpha-iduronidase as a therapeutic product for delivery in the MPS I dog that is genetically deficient in this lysosomal enzyme. Madin-Darby canine kidney cells were genetically modified to express either human growth hormone or canine alpha-iduronidase, then enclosed in alginate-poly-l-lysine-alginate microcapsules of about 500 microm in diameter. The encapsulated cells were implanted into the brain under steoreotaxic guidance. The brains were monitored with computed tomographic scans before and after surgery and examined biochemically and histologically. Delivery of gene products, as measured in the plasma and cerebrospinal fluid sampled periodically through 21 days or in various regions of the brain after death showed that the delivery of both gene products was extremely low but detectable. However, we noted extensive inflammatory reactions, both at the sites of implantation and in the immediate vicinity of the implanted microcapsules. Hence for this technology to be applicable to the CNSs of larger animals and human beings, a more accurate and less invasive neurosurgical procedure, more biocompatible microcapsule-recombinant cell combinations, and higher output of recombinant products must be developed.