布鲁菌外膜蛋白OMP10表达及其抗原性的研究

设计1对特异性引物对羊布鲁菌16M总DNA进行外膜蛋白omp10的PCR扩增,得到了一个大小为330 bp的目的基因片段(去掉17个氨基酸编码的信号肽),测序证实它与国外报道的羊布鲁菌omp10基因完全一致.将其克隆到表达载体PET-30a中,经酶切、PCR扩增和测序分析,表明重组表达载体构建成功.将此重组质粒转化入大肠埃希菌BL21(DE3)中,IPTG诱导表达,该基因以包涵体的形式在大肠埃希菌中表达,经过包涵体的变性、复性和亲和层析纯化,成功获得大小为14.2 ku的融合蛋白,与理论推测的蛋白分子质量一致;Western blot和间接ELISA试验证明,纯化之后的OMP10重组蛋白可以被布鲁菌阳性血清识别.     

A canine case of gliosis with cyst formation in the posterior fossa

A 5-month-old male Great Pyrenees with symptoms of convulsions, circling, and a head tilt was referred to the Animal Medical Center of Nihon University. On a magnetic resonance image (MRI), a cyst in the posterior fossa was noted and a part of the cyst enhanced by gadoteridol. Based on MRI and clinical findings, the patient was tentatively diagnosed with a cyst formation tumor, and an operation to open the cyst and remove the part enhanced by contrast was performed. Postoperatively, the clinical course was good. Pathologically, the removed tissue was diagnosed as a gliosis with cyst formation.     

Endoscopic balloon dilation of benign esophageal strictures in dogs and cats

Endoscopic balloon dilation of benign esophageal strictures was performed in 18 dogs and 10 cats with a median age of 4 years. Stricture formation was associated with a recent anesthetic episode in 18 patients. Regurgitation was the most common clinical sign and was present a median of 4 weeks before dilation. Most animals had a single stricture; median diameter was 5 mm, and median length was 1 cm. Esophagitis and mucosal fibrosis were detected in 9 patients each. Dilation was performed with progressively increasing diameter balloons, from 6 to 20 mm. After dilation, mucosal hemorrhage was mild to moderate in most patients. Esophageal perforation was the only serious complication and occurred in 1 patient. Postdilation therapy consisted of administration of cimetidine, metoclopramide, sucralfate, and prednisone in most animals. The median number of dilation procedures performed in each animal was 2, with a range of 1-5. The median interval between dilations was 13 days. Stricture diameter markedly increased with subsequent dilations. Median duration of follow-up was 131 weeks. A successful outcome occurred in 88% of patients, with most animals able to eat canned, mashed, or dry food without regurgitation. Mucosal fibrosis was associated with a better clinical response score, while increasing age was weakly associated with fewer dilations. The dilation protocol used in this group of animals was safe and efficacious.     

Prepubic urethrostomy and vaginoplasty in a female cat

A 1-year-old, intact female, domestic shorthaired cat was presented for dysuria resulting from perineal scarring subsequent to injuries incurred during a cat fight. A prepubic urethrostomy was performed to manage the dysuria. Eleven months later, the cat was re-presented with a 3-day history of pyrexia and inappetence. A pinpoint opening extending 0.5 cm ventral to the anus on midline, exuding a clear discharge, was noted in the perineal region. A contrast fistulogram was performed, and a vaginoperineal fistula was diagnosed. The fistulous tract was a result of vulvar stricture from the trauma of the cat fight. A vaginoplasty was performed to create an opening for vaginal secretions. This is the first published report of a prepubic urethrostomy performed in a female cat.     

A Combined Tension Band and Lag Screw Technique for Fixation of Olecranon Osteotomies

A combined tension band and lag screw technique for fixation of olecranon osteotomies was used in six canine clinical patients weighing 4.5 to 19 kg. After the proximal part of the ulnar shaft was exposed, a screw hole was drilled and tapped just cranial to the caudal cortex of the olecranon. An osteotomy was performed and the hole in the olecranon fragment was overdrilled to form a gliding hole. For reconstruction, the olecranon was reduced anatomically and compressed with a screw placed in lag fashion. With the trochlear notch exposed, a Kirschner wire was inserted cranial to the screw, using care not to enter the elbow joint. A double-twist figure-eight tension band wire was placed around the Kirschner wire in five dogs and around the screw in one dog. In this dog, a spiked washer was used with the screw because a small olecranon fragment had been produced by incorrect osteotomy position. A painful soft tissue swelling over the prominent washer, which resolved after implant removal, was the only complication attributed to the technique. The combined tension band wire and lag screw technique was a rapid and reliable method for fixation of olecranon osteotomies.     

LbCas12a蛋白的原核表达及活性检测

【目的】获得具有体外切割活性的来自毛螺旋菌属(Lachnospiraceae) ND2006的LbCas12a蛋白,以期为LbCas12a的应用研究提供重要的生物学工具。【方法】根据pMBP-LbCas12a质粒(Addgene, 113431)的基因序列合成LbCas12a基因,并将其与pET-28a(+)线性化载体进行同源重组,构建重组质粒pET28a-LbCas12a,经测序和NheⅠ、SalⅠ双酶切鉴定后将阳性重组质粒转入大肠杆菌BL21(DE3)感受态细胞中进行IPTG诱导表达。通过12%SDS-PAGE凝胶电泳检测确定IPTG诱导的最佳浓度及温度,鉴定重组蛋白的表达形式,通过Ni-NTA树脂亲和层析的方法进行纯化、超滤法浓缩,BCA法检测蛋白浓度。将浓缩后的LbCas12a与猪细小病毒(PPV)靶标DNA、CRISPR RNA (crRNA)及偶联荧光基团的非特异性单链DNA(ssDNA)探针FQ共孵育,设置1个无LbCas12a蛋白的对照组、4个不同蛋白浓度(125、250、500、1 000 nmol/L)的试验组,检测不同组别ssDNA探针的荧光强度,检测测试位点PP...     

Gastric emptying of liquids in the dog: serial test meal and modified emptying-time techniques

A serial test meal technique was used in the dog to evaluate the gastric emptying of a liquid test meal. On separate days, a phenol red test meal was placed in the stomach and completely aspirated at different times. A composite representation of the gastric emptying pattern was obtained for each dog, and a set of normative data was generated. Three emptying determinants (emptying time, half-life, and starting index) were calculated. The basic pattern of canine gastric emptying was linear and declined exponentially or as a function of the square root of the gastric volume. A modified emptying-time technique (METT) correlated well with the serial test meal and a set of normative values was determined. The METT was performed by aspirating test meal from the stomach 30 minutes after instillation. The METT was a simple and inexpensive technique that could be performed in 1 short testing period. With the METT, the half-life of gastric emptying was calculated, using the log of the percentage of retention of test meal; gastric emptying time was calculated, using the square root of the percentage of retention of test meal. Acetylpromazine was shown not to alter gastric emptying of a liquid test meal.     

IMAGING DIAGNOSIS—INTRAPERICARDIAL RIGHT AURICLE ANEURYSM IN A DOG

An 11‐year‐old, male, Pekinese was examined because of a sporadic, nonresponsive, nonproductive cough. Radiographically there was cardiomegaly and a soft tissue mass in the cranial mediastinum. Echocardiographically, chronic degenerative valvular disease was diagnosed. Furthermore, a large anechoic structure located cranial to the right atrium was detected. Computed tomography was performed to define the mediastinal mass and a right auricle aneurysm was diagnosed. The dog died suddenly 1 day after the diagnostic imaging procedures. At necropsy, a right auricular aneurysm without discontinuity of the pericardial sac was confirmed. Intrapericardial right auricular aneurysm should be included in the differential diagnoses of cranial mediastinal masses.     

IMAGING DIAGNOSIS—EXTRAHEPATIC BILIARY TRACT OBSTRUCTION SECONDARY TO A DUODENAL FOREIGN BODY IN A CAT

A 13-month-old, neutered female domestic shorthaired cat was evaluated for vomiting, anorexia, and lethargy. The cat was icteric and hyperbilirubinemic. Radiographically a partially radiolucent proximal duodenal foreign body was suspected. Ultrasonographically, there was a foreign body at the level of the duodenal papilla and dilation of the common bile duct and cystic duct; a diagnosis of extrahepatic biliary tract obstruction secondary to a duodenal foreign body was made. Sonographic findings were confirmed at surgery and a duodenal foreign body was removed. This information defines duodenal foreign body as a cause of extrahepatic biliary obstruction in cats.     

Traumatic elbow luxation repair in a common squirrel monkey (Saimiri Sciureus) and a bonnet macaque (Macaca radiata).

Traumatic elbow luxation was diagnosed in a common squirrel monkey (Saimiri sciureus) and a bonnet macaque (Macaca radiata). In the common squirrel monkey, the luxation was complicated by an ulnar fracture and would not stay reduced when attempts at closed reduction were made. The luxation was surgically reduced and a transarticular type I external skeletal fixator was placed. In the bonnet macaque, closed reduction was performed and the elbow was stabilized using a cast. Both monkeys regained good function of the injured elbows.     

Pyloric outflow obstruction secondary to sclerosing encapsulating peritonitis in a dog

A 6-year-old, male neutered mixed breed dog was presented emergently with a three-week history of hyporexia, vomiting, diarrhoea and weight loss. Upon examination, the patient was dull, had generalised muscle atrophy, moderate abdominal pain and a mild amount of peritoneal effusion. A fluid-filled, distended, corrugated small bowel with marked gastroparesis and moderate peritoneal effusion was noted on abdominal ultrasonography. Endoscopy revealed hyperaemic and friable mucosa and a subjectively narrowed pylorus. Emergency exploratory celiotomy was performed due to worsening patient condition and revealed thick, diffuse, fibrous adhesions of the abdominal cavity. Based on these findings, sclerosing encapsulating peritonitis (SEP) was suspected. A large mass of omentum adjacent to the greater curvature of the stomach had caused a pyloric outflow obstruction. Adhesiolysis was attempted but was unsuccessful due to the friability of the small intestines. The dog was humanely euthanased under anaesthesia. A diagnosis of SEP was confirmed via necropsy. No underlying cause was identified. This is the first known case of a pyloric outflow obstruction secondary to SEP in a dog. Although rare, this condition should be considered as a differential for dogs with signs of a pyloric outflow obstruction with concurrent ascites and abdominal pain, hyporexia, vomiting and diarrhoea.     

Removal of an intracoelomic hook via laparotomy in a sandbar shark (Carcharhinus plumbeus)

A twenty-one-year-old male sandbar shark (Carcharhinus plumbeus) demonstrated signs of chronic weight loss even though its appetite remained unchanged. In addition, the gradual development of a red, circular lesion on its ventral body was noted. After months without any significant changes, a ventral fistula appeared revealing the tip of a wire exposed within its center. The shark was immobilized via intramuscular remote injection through dartgun with etomidate; removed from its exhibit tank; and clinically examined with radiographic imaging, a cell blood count/serum chemistry evaluation, and the use of a metal detector along the body wall. A metallic hook was identified in the coelom about 10 cm cranial to the external fistula. The shark was transferred to an isolation pool for 1 mo. A second immobilization via immersion in eugenol was conducted in order to perform a celiotomy. The hook was located in a liver lobe and was surgically removed. After a prolonged recovery from anesthesia, the shark was released into its primary tank and recovered uneventfully, although some of the skin sutures sloughed prematurely.     

SPONTANEOUS PANCREATIC ISLET CELL ADENOMA WITH PERIPHERAL NEUROPATHY IN A PET RAT (RATTUS NORVEGICUS)

A pancreatic islet cell adenoma was suspected in a 2.5-year-old pet rat that was presented for lethargy and progressive paraparesis. Hypoglycemia was confirmed using a handheld glucometer. Mild improvement was noted after initial corticosteroid treatment. No evidence of a pituitary mass was identified on magnetic resonance imaging. Although a slight treatment response was observed following drug administration the patient's clinical condition deteriorated to the point that the owners elected to have the rat euthanized. Histopathologically, an islet cell adenoma was identified within the pancreas, along with peripheral neuropathy and muscle atrophy, which were consistent with clinical findings. Immunohistochemical staining of neoplastic cells was positive for insulin. Pancreatic islet cell adenomas are not commonly reported, and their clinical prevalence in pet rats is undetermined. The current report describes a case of pancreatic islet cell adenoma in a rat with concurrent peripheral neuropathy secondary to hypoglycemia.     

A giant parovarian cyst in a dog with a granulosa cell tumor

A 13-year old intact female poodle showed persistent signs of estrus. Ultrasonographically, a large cystic mass containing echogenic fluid with a thin wall was observed, and it was 8 cm in diameter. The cystic mass was located at the mesovarium between the left ovary and uterine horn, and it was identified as a giant parovarian cyst. A right ovarian mass was histologically diagnosed as a granulosa cell tumor. Persistent estrogen hormone secretion by the granulosa cell tumor was thought to cause a rapid increase in the size of the parovarian cyst. This rare cystic mass originated from the genital system and was differentiated from the uterus.     

Chest wall reconstruction with latissimus dorsi and an autologous thoracolumbar fascia graft in a dog

A new technique for autogenous chest wall reconstruction using a latissimus dorsi muscle flap and a free graft of thoracolumbar fascia was utilised in a two‐year‐old Dobermann after resection of a high‐grade osteosarcoma from the left thoracic wall. En bloc excision of the chest wall mass, including six ribs, was performed. The resulting chest wall defect was too large to be reconstructed with only a pedicled muscle flap and was reconstructed with a latissimus dorsi muscle flap cranially and a free graft of thoracolumbar fascia caudally. The graft was harvested easily, and there was no donor site morbidity or postoperative complications. A free graft of thoracolumbar fascia can be considered as an option to supplement autogenous reconstruction of the chest wall.     

Phacofragmentation for morgagnian cataract in a horse

A 3‐year‐old Paint gelding was presented for sudden onset of cataract affecting the right eye. Ophthalmic examination revealed subtle to mild microphthalmia and a mature cataract affecting the right globe. The right eye was treated with an antibiotic‐corticosteroid ophthalmic ointment and phacofragmentation was recommended. On re‐examination 5 months later, resorption of the cortex of the right lens and a morgagnian cataract were evident. The nucleus was positioned ventronasally in the lens capsule. The right eye had light perception, and a menace response was present. Examination of the right fundus via binocular indirect ophthalmoscopy was not possible. Ophthalmic ultrasonography revealed that the right globe was 4 mm shorter in axial length when compared with the left globe. There was a marked reduction in the size of the right lens vs. the left lens, and the presence of a moderate amount of hyperechoic debris was identified within the vitreal compartment of the right globe. Phacofragmentation, irrigation and aspiration of the morgagnian cataract and an anterior vitrectomy were performed. On postoperative examination, the eye was comfortable and a menace response was present. The horse regained useful vision and continues to have a comfortable, visual eye 7 months postoperatively.     

Transvenous retrograde portography for identification and characterization of portosystemic shunts in dogs

Transvenous retrograde portography for identification and characterization of portosystemic shunts in dogs A method for transvenous retrograde portography (TRP) in dogs suspected to have a portosystemic shunt (PSS) and results in 20 dogs are described. For TRP, dogs were anesthetized and positioned in left lateral recumbency A dual-lumen balloon-tipped catheter was inserted into the right jugular vein and advanced into the azygos vein. The balloon was inflated to occlude the azygos vein, and contrast material was injected during fluoroscopic evaluation. The catheter was then positioned in the caudal vena cava just cranial to the diaphragm. The balloon was again inflated to occlude the vena cava, and contrast material was again injected. Once a shunt was identified, selective catheterization was attempted with a guide wire and angled catheter. A PSS was identified in 18 of the 20 dogs. In 10 of the 18, the shunt vessel could be selectively catheterized, allowing measurement of portal pressures while the shunt was occluded with the balloon. In 1 dog, results of TRP were normal, but subsequent exploratory celiotomy revealed a single extrahepatic PSS, which was surgically attenuated. The other dog in which results of TRP were normal did not have a macroscopic PSS. In dogs suspected to have a PSS, TRP may be a useful adjunctive diagnostic test that is less invasive than operative mesenteric vein portography and allows measurement of portal pressures before and after temporary shunt occlusion.     

Megaesophagus in a cat

Megaesophagus in an eight month old Siamese cat is described. Initially, a cause for the vomiting was not discovered and the cat was treated for pyloric spasm. Several months later the same cat, in poor physical condition, was presented with a palpable bulge along its ventral neck. At this time a very dilated and flaccid esophagus was found. An exploratory thoracotomy was done but a cause for the megaesophagus was not discovered.     

Testicular neoplasia in the retained testicles of an intersex male dog

This case describes the presentation and management of an 8 yr old phenotypically female intersex male dog presented for evaluation of a mass in the right inguinal region. The right inguinal space was surgically explored, and a large irregular mass resembling a fully developed testicle was identified in the right vaginal tunic. A second mass resembling an atrophied, but anatomically mature testicle, was identified in the left tunic. The larger mass was identified as a Sertoli cell tumor that had replaced all normal testicular tissue. The smaller mass was identified as a testicle that contained a small intratubular seminoma. The patient was diagnosed as having a phenotypic female sex, chromosomal male sex, and a gonadal male sex. Hormone assays completed before and after the gonadectomy and mass removal document an elevation of circulating progesterone presurgically that returned to baseline by 1 mo postsurgically. The source of the progesterone was identified to be the Leydig cells of the atrophied testicle.