The relationship between histologic lesions in endometrial biopsy specimens and susceptibility to chronic uterine infection (CUI) in mares was investigated. Mares were allotted to 4 groups on the basis of degree of endometrial lesions. Mares in group 1 (n = 6) had no pathologic changes, mares in group 2 (n = 5) had only mild pathologic changes, group-3 mares (n = 7) had moderate changes, and group-4 mares (n = 7) had severe inflammatory and fibrotic endometrial changes. Susceptibility to CUI was determined by the inflammatory response to intrauterine inoculation of 5 X 10(6) Streptococcus zooepidemicus. The inoculum was given on the third day of behavioral estrus and in the presence of a follicle > 30 mm. Mares with > 1 neutrophil/5 high-magnification (400 X) microscopic fields and > 20 colonies of S zooepidemicus at 96 hours after inoculation were considered to be susceptible to CUI. There was a significant association between biopsy grade and susceptibility to CUI among the groups. Histologically normal endometrium was associated with resistance to CUI, and severe histopathologic changes in the endometrium were associated with susceptibility to CUI. Mild to moderate endometrial lesions did not correlate consistently with susceptibility or resistance to CUI.

Inflammatory mammary carcinoma (IMC) is a rare disease with a poor prognosis and one affecting dogs. Inflammatory breast carcinoma (IBC) is a subtype of malignant breast cancer in humans with a high degree of malignancy and a similarly poor prognosis. Since the clinical symptoms and prognoses of both are similar, canine IMC has been considered as a model of human IBC. In this study, we newly established a stable IMC-derived cell line from a patient at the Yamaguchi University Animal Medical Center in Japan. The patient was a female toy poodle presenting with an inflamed mammary gland, which was diagnosed as IMC. The cell line was established from a tissue biopsy. Surface antigen marker (CD24 and CD44) expression was determined. Cystine/glutamate antiporter (xCT) expression was determined by Western blotting, flow cytometry and fluorescence immunostaining, and sulfasalazine was administered to ascertain if it suppressed xCT expression. Stem cell marker (Nanog, Sox2, Myc and Klf4) expression and aldehyde dehydrogenase (ALDH) activity were also investigated. The cultured cells showed xCT, and its suppression showed downregulation of stem cell markers and ALDH activity. Stable cell proliferation was verified. A new canine IMC-derived cell line was established. In the future, we aim to study the effect of xCT on the maintenance of cancer stem cell properties in canine tumours, and propose a new therapeutic method for the treatment of canine IMC by targeting xCT.

Introduction: Chronic gastritis is a common diagnosis in dogs with signs of chronic vomiting. However, there is no data concerning endoscopic and histopathological agreement in dogs with chronic gastritis. Thus, a question should be raised whether taking gastroduodenal biopsies in dogs with chronic gastritis is necessary or not. Consequently, the purpose of the study was to compare the endoscopic and histopathological agreement in dogs with chronic gastritis. Material and Methods: A total of 22 non-pregnant client-owned dogs with the signs of chronic gastritis were enrolled in this prospective study. Procedures including clinical examination, blood analysis, and diagnostic imaging were performed before anaesthesia. Biopsies obtained from gastroduodenal sites were histopathologically evaluated. A total of 110 gastroduodenal samples were examined. Results: Sixtyeight samples had abnormal histopathology and endoscopy while 11 showed normal histopathological and endoscopic evidence. Conclusion: The obtained data demonstrated that it is not necessary to take extra gastroduodenal biopsies in dogs with evidence of endoscopic gastroduodenitis. We also believe that further prospective studies, including cost and time effectiveness and more specific comparison between endoscopic appearance and histopathology, are necessary to make final recommendations regarding the need of using both procedures for definitive diagnosis.

Introduction: Canine transmissible venereal tumour (CTVT) is a sexually transmitted tumour affecting dogs worldwide, imposing a financial burden on dog owners. A stable culture cell line in continuous passages for >18 months has only been achieved once. The present study investigated a stable CTVT cell line isolated from a bitch and its potential as a vaccine. Material and Methods: A biopsy from a 2-year-old mongrel bitch with CTVT was obtained for histopathological confirmation and isolation of tumour cells. The isolated cells were cultured to passage 55 and characterised by flow cytometry, with karyotyping by GTG-banding and by PCR detection of myc S-2 and LINE AS1. The isolated CTVT cell line was also used as a preventive vaccine in a canine model. Results: Histopathological analysis of the isolated tumour cells revealed typical CTVT characteristics. Constant proliferation and stable morphological characteristics were observed during culture. Phenotypic analysis determined the expression of HLA-DR⁺, CD5.1⁺, CD14⁺, CD45⁺, CD83⁺, CD163⁺, and Ly-6G-Ly-6C⁺. GTG-banding revealed a mean of 57 chromosomes in the karyotype with several complex chromosomal rearrangements. LINE-c-myc insertion in the isolated CTVT cell line at 550 bp was not detected. However, a 340-bp band was amplified. Isolated CTVT cell line inoculation at a concentration of 1×10⁸ did not induce tumour growth in bitches, nor did a challenge with primary CTVT cells. Conclusion: The present study successfully identified and isolated a stable CTVT cell line that may be useful in CTVT prevention.

Introduction: The clinical symptoms of portosystemic shunts (PSSs) and hepatic microvascular dysplasia (HMD) – portal vein hypoplasia (PVH) in dogs are similar. PSSs are abnormal vascular connections between the portal vein system and systemic veins. HMD is a very rare developmental vascular anomaly, recognisable during histopathological examination. The study aim was to assess the prevalence of HMD–PVH and hepatocellular and vascular pathologies in the liver. Material and Methods: Liver biopsies from 140 dogs (of different breeds and both sexes) arousing clinical suspicion of PSS were examined histopathologically. Results: An initial PSS diagnosis was confirmed in 125 dogs (89.29%). HMD–PVH was found in 12.32% of dogs, as an isolated disease in 9.29%, especially in Yorkshire terriers, and with extrahepatic PSS in 6.67%. Histopathological analysis of muscles around sublobular veins showed that HMD cases presented hypertrophy or hypertrophy with fibrosis. In 2.17% of all dogs with liver vascular developmental disorders calcification was visible around vessels (without correlation by degenerative changes in those vessels), suggesting prior onset of deep metabolic disorders. Clinical suspicion of PSS was also formed upon quite different pathological processes in young dogs. Conclusion: Histopathological findings diagnosed the type of vascular anomalies (PSS or HMD–PVH) or other pathological changes conclusively, therefore detailed hepatic histopathology is an indispensable component of the clinical diagnostic process.

OBJECTIVE To evaluate histologic changes and gene expression patterns in body and limb wounds in horses in response to bacterial inoculation. SAMPLE Wound biopsy specimens from 6 horses collected on days 7, 14, 21, and 27 after excisional wounds (20 wounds/horse) were created over the metacarpal and metatarsal region and lateral thoracic region (body) and then inoculated or not inoculated on day 4 with Staphylococcus aureus and Pseudomonas aeruginosa. PROCEDURES Specimens were histologically scored for the amount of inflammation, edema, angiogenesis, fibrosis organization, and epithelialization. Quantitative PCR assays were performed to quantify gene expression of 10 inflammatory, proteolytic, fibrotic, and hypoxia-related markers involved in wound healing. RESULTS Except for gene expression of interleukin-6 on day 27 and tumor necrosis factor-α on day 14, bacterial inoculation had no significant effect on histologic scores and gene expression. Gene expression of interleukin-1β and -6, serum amyloid A, and matrix metalloproteinase-9 was higher in limb wounds versus body wounds by day 27. Gene expression of cellular communication network factor 1 was higher in limb wounds versus body wounds throughout the observation period. CONCLUSIONS AND CLINICAL RELEVANCE The lack of clear markers of wound infection in this study reflected well-known difficulties in detecting wound infections in horses. Changes consistent with protracted inflammation were evident in limb wounds, and gene expression patterns of limb wounds shared similarities with those of chronic wounds in humans. Cellular communication network factor warrants further investigation and may be useful in elucidating the mechanisms underlying poor limb wound healing in horses.

OBJECTIVE To assess effects of photobiomodulation, silver sulfadiazine, and a topical antimicrobial product for the treatment of experimentally induced full-thickness skin wounds in green iguanas (Iguana iguana). ANIMALS 16 healthy subadult green iguanas. PROCEDURES Iguanas were anesthetized, and three 5-mm cutaneous biopsy specimens were obtained from each iguana (day 0). Iguanas were randomly assigned to 2 treatment groups, each of which had a control treatment. Wounds in the topical treatment group received silver sulfadiazine, a topical antimicrobial product, or no treatment. Wounds in the laser treatment group received treatment with a class 4 laser at 5 or 10 J/cm2 or no treatment. Wound measurements were obtained daily for 14 days. Iguanas were euthanized, and treatment sites were evaluated microscopically to detect ulceration, bacterial contamination, reepithelialization, necrosis, inflammation, fibrosis, and collagen maturity. RESULTS On day 14, wounds treated with a laser at 10 J/cm2 were significantly smaller than those treated with silver sulfadiazine, but there were no other significant differences among treatments. Histologically, there were no significant differences in ulceration, bacterial infection, reepithelialization, necrosis, inflammation, fibrosis, and collagen maturity among treatments. CONCLUSIONS AND CLINICAL RELEVANCE Photobiomodulation at 10 J/cm2 appeared to be a safe treatment that was tolerated well by green iguanas, but it did not result in substantial improvement in histologic evidence of wound healing, compared with results for other treatments or no treatment.

OBJECTIVE To evaluate effects of pneumoperitoneum created with warmed humidified CO2 (WHCO2) during laparoscopy on core body temperature, cardiorespiratory and thromboelastography variables, systemic inflammation, peritoneal response, and signs of postoperative pain in healthy mature dogs. ANIMALS 6 mature purpose-bred dogs. PROCEDURES In a randomized crossover study, each dog was anesthetized twice, and pneumoperitoneum was created with standard-temperature CO2 (STCO2; 22°C and 0% relative humidity) and WHCO2 (37°C and 98% relative humidity). Data were collected during each procedure, including core body temperature, cardiorespiratory and thromboelastography variables, and inflammatory biomarkers. Peritoneal biopsy specimens were collected and evaluated with scanning electron microscopy. Dogs were assessed for signs of postoperative pain. RESULTS Mean core body temperature was significantly lower (35.2°C; 95% confidence interval, 34.5° to 35.8°C) with WHCO2 than with STCO2 (35.9°C; 95% confidence interval, 35.3° to 36.6°C) across all time points. Cardiac index increased during the procedure for both treatments but was not significantly different between treatments. Thromboelastography variables did not differ significantly between treatments as indicated by the coagulation index. Subjective evaluation of peritoneal biopsy specimens revealed mesothelial cell loss with STCO2. There was no significant difference in circulating C-reactive protein or interleukin-6 concentrations. There was a significant increase in the number of postoperative pain scores > 0 for the WHCO2 treatment versus the STCO2 treatment. CONCLUSIONS AND CLINICAL RELEVANCE Analysis of these data suggested that effects on evaluated variables attributable to the use of WHCO2 for creating pneumoperitoneum in healthy mature dogs undergoing laparoscopy did not differ from effects for the use of STCO2.

Visceral adipose tissue (AT) obtained from surgical waste during routine ovariectomies was used as a source for isolating canine mesenchymal stem cells (MSCs). As determined by cytofluorimetry, passage 2 cells expressed MSC markers CD44 and CD90 and were negative for lineage-specific markers CD34 and CD45. The cells differentiated toward osteogenic, adipogenic, and chondrogenic directions. With therapeutic aims, 30 dogs (39 joints) suffering from elbow dysplasia (ED) and osteoarthritis (OA) were intra-articularly transplanted with allogeneic MSCs suspended in 0.5% hyaluronic acid (HA). A highly significant improvement was achieved without any medication as demonstrated by the degree of lameness during the follow-up period of 1 y. Control arthroscopy of 1 transplanted dog indicated that the cartilage had regenerated. Histological analysis of the cartilage biopsy confirmed that the regenerated cartilage was of hyaline type. These results demonstrate that transplantation of allogeneic adipose tissue-derived mesenchymal stem cells (AT-MSCs) is a novel, noninvasive, and highly effective therapeutic tool in treating canine elbow dysplasia.

OBJECTIVE To evaluate the efficacy and safety of gastroscopy and biopsy of the proventriculus and ventriculus in pigeons (Columba livia). ANIMALS 15 adult pigeons. PROCEDURES Each pigeon was anesthetized, and the upper gastrointestinal tract (from the cervical portion of the esophagus to the ventriculus) was endoscopically evaluated by use of a rigid endoscope inserted orally. Saline (0.9% NaCl) solution was orally infused to achieve lumen dilation and visibility. Two mucosal biopsy specimens were collected from each of the proventriculus and ventriculus, histologically evaluated, and graded for crush artifacts and depth. Pigeons were monitored for adverse effects for 3 to 6 days after the procedure, after which they were euthanized for necropsy. RESULTS Gastroscopy via the oral approach provided excellent visibility of the lumen and mucosal surfaces of the proventriculus and cranial portion of the ventriculus and was safe provided that appropriate precautions were taken. Two intraoperative deaths occurred at the beginning of the study; following procedure refinement, no additional deaths occurred. No major adverse effects of the procedure were detected in the remaining 13 pigeons during the postoperative monitoring period or at necropsy. Diagnostic quality of proventriculus specimens was adequate for 10 of 13 pigeons. Eight of 13 ventriculus specimens were of inadequate quality, and only 3 were of adequate quality. CONCLUSIONS AND CLINICAL RELEVANCE Gastroscopy was useful for evaluating the lumen and mucosal surface of the proventriculus and ventriculus in pigeons, and biopsy of those organs was safely performed with the appropriate technique. Further evaluation of these techniques is needed in birds with clinical disease and birds of other species.