Morphologic, phenotypic, and genotypic similarities between primary tumors and corresponding 3D cell cultures grown in a repeatable system-preliminary results.

BACKGROUND: Three-dimensional (3D) cell cultures are the new frontier for reproducing the tumor micro-environment in vitro. The aims of the study were (1) to establish primary 3D cell cultures from canine spontaneous neoplasms and (2) to demonstrate the morphological, phenotypic and genotypic similarities between the primary canine neoplasms and the corresponding 3D cultures, through the expression of tumor differentiation markers. RESULTS: Seven primary tumors were collected, including 4 carcinomas and 3 soft tissue sarcomas. 3D cell cultures reproduced the morphological features of the primary tumors and showed an overlapping immunophenotype of the primary epithelial tumors. Immunohistochemistry demonstrated the growth of stromal cells and macrophages admixed with the neoplastic epithelial component, reproducing the tumor microenvironment. Mesenchymal 3D cultures reproduced the immunophenotype of the primary tumor completely in 2 out of 3 examined cases while a discordant expression was documented for a single marker in one case. No single nucleotide variants or small indel were detected in TP53 or MDM2 genes, both in primary tumors and in 3D cell cultures specimens. In one sample, MDM2 amplicons were preferentially increased in number compared to TP53 ones, indicating amplification of MDM2, detectable both in the primary tumor and in the corresponding cell culture specimen. CONCLUSION: Here we demonstrate a good cell morphology, phenotype and genetic profile overlap between primary tumors and the corresponding 3D cultures grown in a repeatable system.

Retrospective evaluation of a hand-sewn side-to-side intestinal anastomosis technique in dogs and cats.

Background: Hand-sewn intestinal resection and anastomosis are commonly performed in veterinary medicine. The outcome of the hand-sewn side-to-side anastomosis (SSA) technique has never been described and compared to other techniques in dogs and cats. Aim: The study aims to describe the side-to-side hand-sewn anastomosis technique in small animals and to compare it with the end-to-end technique. Methods: A retrospective evaluation of the clinical records of dogs and cats that underwent enterectomy between 2000 and 2020 and were treated with side-to-side or end-to-end anastomosis (EEA) was performed. Results: Of the 52 dogs and 16 cats included in the study, 19 dogs and 6 cats received an SSA, and the remaining received an EEA. No intraoperative complication was reported. However, short-term complication rates were comparable, and mortality rates in the EEA group were higher. At the same time, stenosis was a frequent complication of SSA and was never reported following EEA. Conclusion: End-to-end technique remains the gold standard for hand-sewn intestinal anastomosis in small animals. However, SSA can be considered for selected cases with acceptable morbidity and mortality rates.

Perforated duodenal ulcer in a dog with gallbladder agenesis.

Background: Duodenal ulceration (DU) in dogs derives from different causes but has never previously been related to gallbladder agenesis (GA). GA is a rare congenital disorder in dogs and is considered a predisposing factor for DU in humans. Case Description: A 5-month-old intact female Maltese was presented for acute vomiting and diarrhea. Abdominal ultrasound suggested duodenal perforation and absence of the gallbladder. Exploratory laparotomy was performed to treat the perforation and confirmed GA. Hepatic ductal plate malformation (DPM) was histologically diagnosed in liver biopsy, but no signs of liver dysfunction were detected by blood work at first admission. Two months later, the dog developed signs of portal hypertension and medical treatment was started. However, the clinical condition gradually worsened until liver failure and the dog was euthanized 8 months after surgery. Necropsy confirmed hepatic abnormalities. Conclusion: This report describes a case of DU associated with GA and DPM in a dog. As in humans, GA may represent a hepatobiliary disease predisposing to gastroduodenal ulcerations.

Automated Wound Image Segmentation: Transfer Learning from Human to Pet via Active Semi-Supervised Learning.

Wound management is a fundamental task in standard clinical practice. Automated solutions already exist for humans, but there is a lack of applications regarding wound management for pets. Precise and efficient wound assessment is helpful to improve diagnosis and to increase the effectiveness of treatment plans for chronic wounds. In this work, we introduced a novel pipeline for the segmentation of pet wound images. Starting from a model pre-trained on human-based wound images, we applied a combination of transfer learning (TL) and active semi-supervised learning (ASSL) to automatically label a large dataset. Additionally, we provided a guideline for future applications of TL+ASSL training strategy on image datasets. We compared the effectiveness of the proposed training strategy, monitoring the performance of an EfficientNet-b3 U-Net model against the lighter solution provided by a MobileNet-v2 U-Net model. We obtained 80% of correctly segmented images after five rounds of ASSL training. The EfficientNet-b3 U-Net model significantly outperformed the MobileNet-v2 one. We proved that the number of available samples is a key factor for the correct usage of ASSL training. The proposed approach is a viable solution to reduce the time required for the generation of a segmentation dataset.

Surgical findings and outcomes after unilateral adrenalectomy for primary hyperaldosteronism in cats: a multi-institutional retrospective study.

CASE SERIES SUMMARY: Twenty-nine cats from different institutions with confirmed or highly suspected primary hyperaldosteronism treated by unilateral adrenalectomy were retrospectively included in this study. The most frequent clinical signs were lethargy (n = 20; 69%) and neck ventroflexion (n = 17; 59%). Hypokalaemia was present in all cats, creatinine kinase was elevated in 15 and hyperaldosteronism was documented in 24. Hypertension was frequently encountered (n = 24; 89%). Preoperative treatment included potassium supplementation (n = 19; 66%), spironolactone (n = 16; 55%) and amlodipine (n = 11; 38%). There were 13 adrenal masses on the right side, 15 on the left and, in one cat, no side was reported. The median adrenal mass size was 2 × 1.5 cm (range 1-4.6 × 0.4-3.8); vascular invasion was present in five cats, involving the caudal vena cava in four cats and the renal vein in one. Median duration of surgery was 57 mins. One major intraoperative complication (3%) was reported and consisted of haemorrhage during the removal of a neoplastic thrombus from the caudal vena cava. In 4/29 cats (14%), minor postoperative complications occurred and were treated medically. One fatal complication (3%) was observed, likely due to disseminated intravascular coagulation. The median duration of hospitalisation was 4 days; 97% of cats survived to discharge. The potassium level normalised in 24 cats within 3 months of surgery; hypertension resolved in 21/23 cats. Follow-up was available for 25 cats with a median survival of 1082 days. Death in the long-term follow-up was mainly related to worsening of comorbidities. RELEVANCE AND NOVEL INFORMATION: Adrenalectomy appears to be a safe and effective treatment with a high rate of survival and a low rate of major complications. Long-term medical treatment was not required.

Congenital Lobar Emphysema in a Kitten with Concomitant Hiatal Hernia and Nutritional Secondary Hyperparathyroidism.

A 2 mo old domestic shorthair kitten was presented for acute respiratory distress and severe ambulatory difficulties. Thoracic radiography revealed hyperinflation of the left cranial lung lobe and a mass with soft-tissue/gas opacity in the caudal mediastinum, leading to the suspicion of congenital lung lobe emphysema and hiatal hernia. Decreased bone radiopacity and suspected pathological fractures were also present. Complete clinicopathological analyses showed significant ionized hypocalcemia and suspicion of secondary hyperparathyroidism related to an inadequate diet. Lung lobectomy and reduction of the hiatal hernia following a median sternotomy and a cranial laparotomy were performed. IV and oral supplementation of calcium led to a full recovery and improvement in the kitten's walking. A histopathological analysis revealed pulmonary emphysema associated with hypoplastic and irregular bronchial cartilage. Congenital lobar emphysema is a rare disease in both humans and animals. This is the first veterinary report describing a kitten affected by congenital lobar emphysema combined with a hiatal hernia and additionally complicated by secondary nutritional hyperparathyroidism with a good long-term outcome.

A Target Animal Effectiveness Study on Adjuvant Peptide-Based Vaccination in Dogs with Non-Metastatic Appendicular Osteosarcoma Undergoing Amputation and Chemotherapy.

Despite efforts to develop novel treatment strategies, human and canine osteosarcomas continue to have poor prognosis and limited overall survival. The aim of this clinical trial was to test the antitumor effect and safety of multiple dermal administrations of a peptide-based anticancer vaccine in dogs with non-metastatic appendicular osteosarcoma undergoing standard of care (SOC), consisting of limb amputation and adjuvant chemotherapy. Salmonella-infected canine osteosarcoma cells were induced to release immunogenic peptides in the extracellular space via Cx43 hemichannels opening; the secretome was collected and constituted the vaccine. Dogs with non-metastatic appendicular osteosarcoma were eligible for recruitment. Following limb amputation and adjuvant carboplatin, dogs were vaccinated on a monthly basis for six times and followed up with serial thoracic radiographs. A population of dogs undergoing SOC treatment (amputation and adjuvant carboplatin) before the vaccine was available served as controls. Primary endpoints were time to metastasis (TTM) and tumor-specific survival (TSS). Secondary endpoints were feasibility, toxicity, T-cell and humoral immune responses. A total of 20 dogs were vaccinated along with SOC and 34 received SOC only. Vaccine-specific humoral and T-cell responses were observed; their amplitude correlated with TSS. Vaccine-associated toxicity was not recorded. TTM and TSS were significantly longer in vaccinated versus unvaccinated dogs (TTM: 308 vs. 240 days, respectively; p = 0.010; TSS: 621 vs. 278 days, respectively; p = 0.002). In dogs with non-metastatic osteosarcoma undergoing SOC, the addition of a bacteria-based vaccination strategy increased TTM, thereby prolonging survival, while maintaining a safe profile. Additionally, vaccinated dogs developed a long-term tumor-specific response, as documented by the immunomonitoring of these patients over time. These results hold promise for future management of canine osteosarcoma.

Prediction of vascular invasion using a 7-point scale computed tomography grading system in adrenal tumors in dogs.

BACKGROUND: Previous studies evaluating the accuracy of computed tomography (CT) in detecting caudal vena cava (CVC) invasion by adrenal tumors (AT) used a binary system and did not evaluate for other vessels. OBJECTIVE: Test a 7-point scale CT grading system for accuracy in predicting vascular invasion and for repeatability among radiologists. Build a decision tree based on CT criteria to predict tumor type. METHODS: Retrospective observational cross-sectional case study. Abdominal CT studies were analyzed by 3 radiologists using a 7-point CT grading scale for vascular invasion and by 1 radiologist for CT features of AT. ANIMALS: Dogs with AT that underwent adrenalectomy and had pre- and postcontrast CT. RESULTS: Ninety-one dogs; 45 adrenocortical carcinomas (50%), 36 pheochromocytomas (40%), 9 adrenocortical adenomas (10%) and 1 unknown tumor. Carcinoma and pheochromocytoma differed in pre- and postcontrast attenuation, contralateral adrenal size, tumor thrombus short- and long-axis, and tumor and thrombus mineralization. A decision tree was built based on these differences. Adenoma and malignant tumors differed in contour irregularity. Probability of vascular invasion was dependent on CT grading scale, and a large equivocal zone existed between 3 and 6 scores, lowering CT accuracy to detect vascular invasion. Radiologists' agreement for detecting abnormalities (evaluated by chance-corrected weighted kappa statistics) was excellent for CVC and good to moderate for other vessels. The quality of postcontrast CT study had a negative impact on radiologists' performance and agreement. CONCLUSIONS AND CLINICAL IMPORTANCE: Features of CT may help radiologists predict AT type and provide probabilistic information on vascular invasion.

Disseminated aspergillosis in a German shepherd mixed breed dog with unusual initial localization to the iliac wing.

A female, 1.5 years old, mixed­breed dog, was presented for left hind limb lameness. Radiographs revealed an irregular periosteal proliferation on the left iliac wing. The clinical condition worsened with generalised enlargement of the lymph nodes, azotaemia, and pyelonephritis. The magnetic resonance imaging of the pelvis and a surgical biopsy diagnosed a mycotic myositis and osteomyelitis of the iliac wing and gluteal muscles. Aspergillus terreus was isolated from culture of urine and lymph nodes aspirates. The antifungal susceptibility test showed moderate sensitivity to Itraconazole. After one month of therapy with itraconazole, the dog presented discospondylitis of L1­L2 and partial ureteral obstruction due to mycotic bezoar that was resolved with medical treatment and itraconazole dose elevation. After twelve months, itraconazole was suspended; a severe osteomyelitis of the left femur developed, and the dog was euthanised. The necropsy confirmed the presence of mycotic osteomyelitis of the iliac wing and femur, discospondylitis, lymphadenitis and severe granulomatous pyelonephritis. Systemic aspergillosis has rarely been reported in the literature, especially in Italy. The pelvic bone involvement is rare both in dogs and humans. Although itraconazole treatment allowed remission of the clinical signs for one year, it was not able to cure the dog.

Clinical Features and Outcome of 79 Dogs With Digital Squamous Cell Carcinoma Undergoing Treatment: A SIONCOV Observational Study.

In dogs, digit squamous cell carcinoma (SCC) is uncommon. Clinical signs are frequently underestimated, leading to a diagnostic delay. The purpose of this retrospective study was to report our experience regarding the clinical presentation, diagnostic work-up, treatment and outcome of 79 client-owned dogs with SCC of the digit. The greatest majority (84.8%) of dogs was dark-coated. Schnauzers represented approximately one third of the study population, and had a poorer outcome compared with other breeds. The majority of SCCs occurred in the front limbs (61%), and bone lysis was frequently observed (92.4%). Approximately 9% of dogs had involvement of multiple digits, and this was associated with a shorter time to progression (TTP; P = 0.047). Similarly, a duration of clinical signs >90 days was associated with a shorter TTP (P = 0.02). Regional lymph node metastases were documented in 17.7% of dogs at admission and were significantly associated with tumor-related death (P < 0.001). At presentation, none of the dogs had evidence of distant metastasis. Digit amputation achieved adequate local tumor control in the majority of cases. Adjuvant chemotherapy and radiation therapy were carried out in 21.5% of cases, with uncertain benefit. Due to the relatively non-aggressive clinical behavior of digit SCC, chemotherapy should only be offered in the case of metastatic disease. Approximately one fourth of dogs developed de novo SCCs during the follow-up. Careful examination of the digits should be encouraged in breeds considered at high risk and in dogs with a previous history of digital SCC.

The use of small-bore wire-guided chest drains for the management of feline pyothorax: A retrospective case series.

Background: Pyothorax in cats is routinely managed, at least initially, with thoracic tube placement associated with systemic antimicrobial administration. Traditionally, large-bore trocar-type thoracostomy tubes have preferentially been used for the drainage of thick material from the pleural space. In recent years, the use of small-bore wire-guided thoracic drains has increased in both small animals and in humans. Few studies have highlighted the efficacy of small-bore wire-guided thoracostomy tubes. Aim: The purpose of this study was to describe the use of small-bore wire-guided thoracostomy tubes in feline pyothorax in terms of efficacy, safety, and outcome. Methods: Cats with pyothorax managed with small-bore thoracostomy tubes (SBTTs) (2015-2018) were retrospectively studied. The number of drains inserted, the need for anesthesia and analgesia for chest tube placement and maintenance, and related major and minor complications were reviewed. Clinical data, diagnostic results, treatment, and outcome were recorded. Results: Ten cats were enrolled. Thoracostomy tube placement was unilateral in 7/10 cats, despite the presence of bilateral effusion in 9/10 cats, and required sedation (8/10) or anesthesia (2/10). Three cats experienced minor complications during the chest tube insertion, including self-limiting pneumothorax (1/3) and malpositioning (2/3). One cat had a major complication (non-functional malposition) requiring reposition of the drain. Pain management was adequately achieved using opioids (8/10) or opioids plus nonsteroidal anti-inflammatory drugs (2/10). Partial chest tube occlusion occurred in three cases and it was resolved with lavage. In one case, the occlusion was complete, requiring drain removal. Three out of 10 cats were treated medically, combining thoracostomy tubes and antibiotics, while 7/10 cats underwent surgery. All the cats survived. Conclusion: SBTTs represent a safe and effective option for the initial management of feline pyothorax. In fact, mainly minor complications were reported during insertion and usage. The SBTTs were well tolerated by the cats with a satisfactory performance in terms of exudate drainage in most cases. The combined use of a small-bore thoracostomy drain together with the common practice of surgical treatment might have resulted in the successful management of the cases presented.

Incidence of Gastroesophageal Reflux in Dogs Undergoing Orthopaedic Surgery or Endoscopic Evaluation of the Upper Gastrointestinal Tract.

Gastroesophageal reflux (GER) is a common event during general anaesthesia but is often underdiagnosed in veterinary medicine. The oesophageal pH in anaesthetised dogs undergoing endoscopic evaluation of the upper gastrointestinal tract (END group; n = 12) or orthopaedic surgery (ORT group; n = 12) was measured using an oesophageal probe. The dogs were sedated with acepromazine or with methadone or butorphanol, and anaesthesia was induced with propofol and maintained with isoflurane. Of the 24 dogs in this study, 21 (87.5%) had an episode of GER during anaesthesia. The incidence of GER, as well as the first, the minimum, and the maximum pH values, did not differ significantly between the groups. The mean maximum difference versus the first pH value was higher for dogs in the END group (-2.6 ± 3.5) as compared with those in the ORT group (-0.7 ± 2.5), although they were not statistically significant (p = 0.25). The administration of methadone or butorphanol had no significant effect on the development of acidic reflux or biliary reflux. In the acepromazine-sedated dogs, the incidence of GER did not differ significantly between patients undergoing an endoscopic procedure and those undergoing orthopaedic surgery; however, during endoscopy, fluctuations in the oesophageal pH can be expected, even without any clinical signs of GER.

Use of Contrast-Enhanced Ultrasonography for the Characterization of Tumor Thrombi in Seven Dogs.

Tumors of adrenal and thyroid glands have been associated with vascular invasions-so-called tumor thrombi, both in humans and dogs. The detection and characterization of venous thrombi is an important diagnostic step in patients with primary tumors for both surgical planning and prognosis. The aim of this study was to describe the use of contrast-enhanced ultrasonography (CEUS) for the characterization of tumor thrombi. Dogs with tumor thrombus who underwent bi-dimensional ultrasound (B-mode US) and CEUS were included. Seven dogs were enrolled in this retrospective case series. On B-mode US, all thrombi were visualized, and vascular distension and thrombus-tumor continuity were seen in three and two cases, respectively. On color Doppler examination, all thrombi were identified, seemed non-occlusive and only two presented vascularity. On CEUS, arterial-phase enhancement and washout in the venous phase were observed in all cases. Non-enhancing areas were identified in the tumor thrombi most likely representing non-vascularized tissue that could potentially be embolized in the lungs after fragmentation of the tumor thrombi. On the basis of these preliminary study, CEUS appeared to be useful for the characterization of malignant intravascular invasion.

Validation of an optical, computer-assisted technique for intraoperative tracking of 3-dimensional canine stifle joint motion.

Background: Cranial cruciate ligament (CCL) rupture is the most common orthopedic pathology in dog and in men. In human, optical computer-assisted technique is considered as a repeatable and reliable method for the biomechanical assessment of joint kinematics and laxity in case of CCL surgery. Aim: To evaluate the repeatability and reliability afforded by clinical tests in terms of laxity measured by means of a computer-assisted tracking system in two canine CCL conditions: CCL-Intact, CCL-Deficient. Methods: Fourteen fresh frozen canine stifles were passively subjected to Internal/External (IE) rotation at 120° of flexion and Cranial drawer test (CC). To quantify the repeatability and the reliability, intra-class correlation coefficient (ICC) and the mean percent error were evaluated (Δ r %). Results: The study showed a very good intra-class correlation, before and after CCL resection for kinematics tests. It was found a minimum ICC = 0.73 during the IE rotation in CCL-Intact and a maximum value of ICC = 0.97 for the CC displacement in CC-Deficient. IE rotation with CCL-Intact is the condition with the greatest Δ r % = 14%, while the lowest Δ r % = 6% was obtained for CC displacement in CCL-Deficient. Conclusion: The presented work underlined the possibility of using a computer-assisted method also for biomechanical studies concerning stifle kinematics and laxity.

Kidney-Sparing Surgery for Renal Subcapsular Abscess Caused by Staphylococcus pseudintermedius in a Dog.

An 11 yr old male Drahthaar dog was presented for dysuria, pollakiuria, and history of uroliths. Abdominal ultrasound revealed a subcapsular fluid-filled lesion of the left kidney, suspected cholecystitis, and a splenic infarct. The renal lesion was fully drained and cytology of the renal subcapsular and perirenal fluids revealed septic exudate. Bacterial culture of the urine, bile, and perirenal and subcapsular fluids were all positive for Staphylococcus pseudintermedius. Antimicrobial therapy was instituted based on culture sensitivity. After 7 days the dog re-presented for vomiting and abdominal pain, and a focal intestinal injury was suspected based on abdominal ultrasound. Enterectomy of an ischemic jejunal loop, a partial splenectomy, and excision of the left renal subcapsular abscess were performed. The renal parenchyma was left intact. Histopathology confirmed the diagnosis of a renal subcapsular abscess, intestinal infarction, and focal pyogranulomatous splenitis. Cholecystitis was confirmed by bile cytology and culture. No major complications and no recurrences were encountered at 1 yr follow-up. This is the first report of a renal subcapsular abscess in the dog, with septic complications, and treated with a kidney-sparing surgery.

Surgical outcomes of six bulldogs with spinal lumbosacral meningomyelocele or meningocele.

OBJECTIVE: To report the surgical treatment and outcome of six bulldogs with spina bifida (SB) and meningocele (MC) or meningomyelocele (MMC). STUDY DESIGN: Case series. ANIMALS: Five French bulldogs and one English bulldog with MC or MMC. METHODS: Medical records of dogs with spinal MC or MMC diagnosed by MRI at two institutions between 2013 and 2016 were reviewed for surgical treatment and outcomes. RESULTS: Meningocele was diagnosed in two dogs, and MMC was diagnosed in four dogs. A lumbosacral dimple was noted in all dogs along with neurological deficits most commonly consisting of urinary and fecal incontinence (n = 6) and mild/moderate paraparesis (n = 3). Dorsal laminectomy was performed in all dogs to allow dissection of the meningeal sac to the vertebral column defect. In dogs with MMC, nerves were repositioned and protruded meninges were removed prior to suturing remaining meninges. Adhesions and filum terminale were resected in two dogs with suspected tethered cord syndrome. Urinary and fecal incontinence improved in two dogs and remained unchanged in four. Paraparesis improved in two dogs. CONCLUSION: Surgical treatment resulted in partial improvement of the urinary and fecal incontinence (2/6 dogs) and paraparesis (2/3 dogs) or stable neurological condition (3/6 dogs), with only minor temporary complications. CLINICAL SIGNIFICANCE: In the absence of published data comparing surgical and conservative treatment of puppies affected by SB and MC or MMC, early surgical treatment can be considered to prevent deterioration of neurological signs and, eventually, facilitate improvement of neurological signs.

Deep Vegetal Foreign Bodies in Cats: A Retrospective Study of 10 Cases.

The aim of this study was to describe the anatomic locations and treatments available for deeply located vegetal foreign bodies in cats. Ten cases of migrating vegetal foreign bodies (vFBs) requiring surgical removal are reported. The diagnoses of vFBs in the thoracic cavity (4/10), retroperitoneal space (2/10), and paratracheal region (1/10) were obtained by ultrasound examination; however, in the perineum or penile urethra (2/10) and peritoneum (1/10), the vFBs were found during surgery. Intraoperative ultrasound guidance helped remove vFBs from the retroperitoneal space and paratracheal tissues. Clinical signs resolved in 8 out of 10 cases, 1 out of 10 cases had recurrent draining fistula, and 1 out of 10 was euthanized for ethical reasons. The intrathoracic was the most common location in the present study, followed by retroperitoneal space and urethra/perineum. Ultrasound guidance was essential for the diagnosis and/or treatment of vFBs located in the thoracic, retroperitoneal, and cervical regions.

Sequential Treatment of a Large Pituitary Corticotroph Neoplasm and Associated Neurological Signs in a Dog.

No standardized treatment guidelines are reported in veterinary medicine for dogs with large pituitary corticotroph neoplasms causing neurological signs, and such dogs usually have a short overall survival. When these dogs undergo pituitary surgery and the tumor regrows there are few reports of subsequent treatments. A 7 yr old male Maltese diagnosed with pituitary-dependent hypercortisolism developed seizures in conjunction with a large pituitary corticotroph adenoma and underwent transsphenoidal hypophysectomy. After 3 yr of clinical remission, hypercortisolism recurred, and trilostane therapy was initiated. One year later, the dog developed new neurological signs and computed tomography revealed regrowth of a large pituitary mass that was then treated with radiation therapy. The dog lived disease-free for 3 more yr. At postmortem examination, a more aggressive pituitary neoplasm than the one examined at the time of surgery was found, which is suggestive of malignant transformation into a carcinoma despite the absence of convincing metastasis.