Canine serum thermostable alkaline phosphatase isoenzyme from a dog with hepatocellular carcinoma

A dog histopathologically diagnosed with hepatocellular carcinoma (HCC) showed very high serum alkaline phosphatase (ALP) activity. A supernatant of ascitic fluid and tumor tissue extracted from the dog also showed much higher ALP activity than normal. ALP isoenzyme analysis of samples was performed using polyacrylamide gel disk electrophoresis, and a wide, broad abnormal band was observed. By various treatments, the abnormal band showed thermostability, which is a characteristic of tumor-associated ALP that has only been reported in humans. The thermostable ALP isoenzyme was not found in sera from 39 dogs with several types of tumor that originated from the liver, except for HCC, nor was it found in 10 dogs with hepatic diseases that did not include hepatic tumors. The thermostable ALP isoenzyme seemed to be associated with canine HCC.     

Ectopic hepatocellular carcinoma in a dog

CASE HISTORY: A 14-year-old neutered male Bearded Collie was presented with a history of recurrent, intermittent urinary incontinence of 7 years duration.

CLINICAL FINDINGS: A large, firm, non-painful mass was found in the mid-abdominal region on palpation. Ultrasonography of the mass revealed a compartmentalised structure with mixed echogenicity, and which did not appear to be associated with any of the abdominal organs. Ultrasound-guided fine needle aspirates contained several clusters of epithelial cells with cytological features of hepatocytes. At exploratory laparotomy, the mass was found in the gastrosplenic ligament within the greater omentum.

PATHOLOGICAL FINDINGS AND DIAGNOSIS: Histopathologically, the mass consisted of sheets of hepatocytes, but without the characteristic hepatic architecture. The cells showed moderate variation in nuclear size and were sometimes binucle- ate. A diagnosis of hepatocellular carcinoma (HCC) in the mesentery was made.

CLINICAL RELEVANCE: The presence of ectopic hepatic tissue has been reported rarely in man and cats, but not in the dog. Neoplastic transformation of ectopic hepatic tissue is seen in man. This is the first report of the presentation, clinical findings and treatment of a dog with ectopic HCC.     

Diabetes mellitus after resection of hepatocellular carcinoma with hypoglycemia in a dog

A 10-year-old male beagle was referred to us with seizure related to hypoglycemia and a large intraabdominal mass. Based on various types of imaging and a laparoscopic biopsy, the intraabdominal mass was diagnosed as a hepatocellular carcinoma (HCC) of the quadrate lobe. The hypoglycemia was suspected to be associated with the HCC. After lobectomy of the quadrate lobe was performed, blood glucose levels continued to increase to higher than normal values and sugar was detected in the urine. The dog was diagnosed as diabetes mellitus (DM) and was treated with insulin for over two years after the surgery.     

Ectopic hepatocellular carcinoma in a dog

CASE HISTORY: A 14-year-old neutered male Bearded Collie was presented with a history of recurrent, intermittent urinary incontinence of 7 years duration. CLINICAL FINDINGS: A large, firm, non-painful mass was found in the mid-abdominal region on palpation. Ultrasonography of the mass revealed a compartmentalised structure with mixed echogenicity, and which did not appear to be associated with any of the abdominal organs. Ultrasound-guided fine needle aspirates contained several clusters of epithelial cells with cytological features of hepatocytes. At exploratory laparotomy, the mass was found in the gastrosplenic ligament within the greater omentum. PATHOLOGICAL FINDINGS AND DIAGNOSIS: Histopathologically, the mass consisted of sheets of hepatocytes, but without the characteristic hepatic architecture. The cells showed moderate variation in nuclear size and were sometimes binucleate. A diagnosis of hepatocellular carcinoma (HCC) in the mesentery was made. CLINICAL RELEVANCE: The presence of ectopic hepatic tissue has been reported rarely in man and cats, but not in the dog. Neoplastic transformation of ectopic hepatic tissue is seen in man. This is the first report of the presentation, clinical findings and treatment of a dog with ectopic HCC.     

En block resection of a large hepatocellular carcinoma involving the caudal vena cava in a dog

A 13-year-old neutered female Shih Tzu was referred for investigation of a cranial abdominal mass. Investigations including conventional radiography, abdominal ultrasonography and computed tomography confirmed the mass in the caudate lobe of the liver. As a collateral vein originating from the caudal vena cava (CVC) communicated with the azygos vein, the CVC was ligated and transected cranial to the right renal vein and cranial to the mass under temporary occlusion of the thoracic descending aorta and posthepatic CVC. The mass combined with the CVC was excised. The mass was confirmed as hepatocellular carcinoma (HCC). This report describes the first case with successful en bloc resection of a large HCC involving the CVC in a dog.     

Establishment and characterization of primary canine hepatocellular carcinoma cell lines producing alpha-fetoprotein

A primary cultured cell line named CHKS was established from a hepatocellular carcinoma (HCC) of a dog showing a high level of serum alpha-fetoprotein (AFP). CHKS secreted a 66 KDD AFP into the growth medium regardless of the presence or absence of fetal bovine serum (FBS). Cloning CHKS with limiting dilution produced 4 clones, CHKS-1, -2, -3, and -4, which secreted 826, 471, 70, and less than 10 ng/ml, respectively, of AFP into the culture medium. In culture, these cell lines were similar in morphology and proliferation pattern to epithelial cells and positive to periodic acid-Schiff (PAS) staining. The presence of mRNA for canine albumin was demonstrated by nested PCR. The doubling times of the clone cell lines were 21, 45, 36, and 35 h, saturation densities 34, 18, 22, and 24 x 10(4)/cm(2), and plating efficiencies 18, 45, 46, and 45%, respectively. Chromosome analysis of these cell lines showed near triploidy. These results show that CHKS and its clones have hepatic cell functions and are useful for carcinogenetic and clinical studies of canine HCC.     

Isolation and identification of hepatitis contagiosa canis virus (HCC virus) from an enzootic outbreak in a dog kennel

Using a continuous cell line of canine kidney (MDCK-cell line) cells, HCC virus was isolated from 4 of 6 puppies affected in an outbreak of the disease in a dog kennel. The virus was most easily, and rapidly, isolated in primary cultures from the liver samples, but also from other organs such as the spleen and lung, although isolation from these organs could require 1 or 2 passages in addition to the primary culture. Attention is therefore drawn to the necessity of using a sufficient number of passages when organs other than the liver are available. Among the bacteria identified in addition to the virus infection, were Staphylococcus aureus, Bordetella bronchiseptica and β-hemolytic streptococci. Emphasis is made on the significance of isolation and identification of the virus in pure culture, thus enabling its various properties to be studied in detail, and possible new serotypes to be recognized.Keyword: canine hepatitis virus, canine kidney cell line     

Risk factors for nuisance barking in dogs

Objective   To determine the key variables related to the environment, animal and owner that influence nuisance barking by dogs in a city with a subtropical climate.
Design and Population   A case–control survey of dog owners in Brisbane, Queensland, was conducted using a questionnaire investigating key variables connected to nuisance barking. Owners of dogs exhibiting nuisance barking were obtained from a list of dogs being treated in a Brisbane behaviour clinic, and those of control dogs were selected from a telephone directory.
Results   Univariate analysis showed that animal, owner and environmental factors all potentially influence the occurrence of nuisance barking. Multivariate analysis identified the following factors, with the relevant odds ratios (OR) as significant: age of the dog (young dog vs old dog, OR 11.2); multiple dogs in the household vs single (OR 5.6); origin of the dog (home bred vs obtained from breeder or friend, OR 4.0); type of dog, (herding vs other types, OR 3.2) and dog with access to the home vs dog without access (OR 2.5).
Conclusion   The greatest risk for nuisance barking occurs with a young dog of the herding type that is home bred and with access to the house in a multiple dog household.     

Reduced Klotho expression and its prognostic significance in canine hepatocellular carcinoma

Klotho is an anti-ageing gene and is known to act as a tumour suppressor in human hepatocellular carcinoma (HCC). According to a previous study, Klotho is present in normal canine mammary glands, and down-expression in tumours is positively associated with negative prognosis. However, the presence and significance of Klotho in canine HCC has not yet been reported. This study aimed to confirm Klotho expression in normal canine liver tissues using western blotting and immunohistochemistry, and whether the expression differed in non-neoplastic liver disease and HCC. Furthermore, correlation between clinicopathologic features and expression of Klotho was evaluated. All of the normal liver tissues showed the presence of Klotho, and Klotho expression was significantly decreased in the HCC tissue as compared to the non-neoplastic hepatic tissue. Additionally, Klotho expression was significantly associated with tumour size (P = .045), liver enzyme (alanine aminotransferase (ALT)) (P = .018), and metastasis (P = .024). Analysis of the survival curve revealed that reduced Klotho expression was significantly associated with poor disease-free survival (P = .041) in HCC. These results show that Klotho expression is present in normal canine liver tissue and that reduced Klotho expression is associated with poor prognosis in canine HCC. Thus, Klotho was presumed to be a potential clinical prognostic marker for canine HCC.     

Preliminary study of the impact of different dog features on humans in public

Revealing why pedestrians are afraid of some dogs while passing them in public is essential to assess the dog–owner–stranger relationship. Therefore, the effect of different dog features on passersby was gathered. Four dog features differing in size and color were investigated. The dog features were classified as small-dark, small-pale, large-dark, and large-pale. Dog's effect on passersby was recorded in 2 ways: (1) observing the behavioral responses of passersby while passing nearby a dog, and (2) revealing the passersby's perception of that dog by a short interview. Direct observations of 427 passersby's behavioral responses while passing a dog on a sideway were acquired, and 126 persons were interviewed. It was suggested that large-dark dogs seemed to be more threatening than pale or small dogs. Passersby confronted with pale dogs of equal size changed their way far less than when confronted with dark dogs. Significant differences were noted for the type of effect assessed and the reason for this behavioral response. The results showed that only the small-pale dog appeared to be friendly because of its size (P = 0.000). All other dogs were stated as friendly because of individual factors including breed and own dog experiences (P = 0.000). None of the dogs were mentioned as threatening. Our results show that some dog features have a significant effect on behavioral responses and the subjective perception of dogs by people in public.     

Two cases of acute polyradiculoneuritis in dogs consuming a raw poultry diet

A 9-year-old female mixed-breed dog presented with ascending flaccid tetraparesis, and a 5-year-old castrated male Poodle dog presented with ventroflexion of neck, dysphonia, and hindlimb weakness, which progressed to acute ascending tetraparesis. Both dogs were fed raw poultry for over 9 and 5 years, respectively. Blood examination and other test results were normal or unrelated to the present case. Fecal polymerase chain reaction analysis in the Poodle dog was positive for Clostridium perfringens and Campylobacter jejuni. Tetraparesis improved with supportive care in both dogs. Human IV immunoglobulin was only administered to the Poodle dog, which showed a shorter recovery (12 days compared to 34 days in the mixed-breed dog). Both dogs returned to normal conditions eventually.     

Cloning of the short-tailed Gyeongju Donggyeong dog via SCNT: conserving phenotypic inheritance

Somatic cell nuclear transfer is a useful tool to maintain genetic information of animals. The Gyeongju Donggyeong dog is a breed registered as natural monument in Korea. The unique feature of the Donggyeong dog is its tail, as the Donggyeong dog can be classified as either short tailed or tailless. The aim of this study was to preserve the Donggyeong dog’s unique feature by cloning. Fibroblasts were obtained from a short-tailed Donggyeong dog. In vivo matured oocytes were enucleated, microinjected with a donor cell and fused electrically. Reconstructed embryos were transferred to six recipient dogs. One surrogate became pregnant, and one short-tailed Donggyeong dog was delivered. This study demonstrated that the phenotype of the Donggyeong dog could be conserved by somatic cell nuclear transfer.     

A case of juvenile form of dilated cardiomyopathy in a 6-month-old Shiba Inu dog

A 6-month-old Shiba Inu dog was brought to the Veterinary Medical Teaching Hospital because of a cough, exercise intolerance, and pulmonary edema. The dog had a Levine 2/6 systolic murmur. Transthoracic echocardiography revealed left atrial and ventricular dilatation (left atrium to aortic ratio: 2.8), mitral and tricuspid valve regurgitation, and severe left ventricular myocardial hypokinesia (fractional shortening was 11.8%). Bubble contrast echocardiography did not reveal a congenital shunt; therefore, the dog was clinically diagnosed with early onset dilated cardiomyopathy. From the first visit, the dog was treated with pimobendan, taurine, torasemide, and isosorbide dinitrate. After 435 days, echocardiography revealed that systolic function had not improved. On Day 465, atrial fibrillation was confirmed via electrocardiogram, and treatment with diltiazem hydrochloride was initiated. The dog continued to appear clinically stable thereafter, until it died suddenly 1087 days after the initial visit. A postmortem histopathological examination identified severe enlargement of the left atrial and ventricular chambers as well as attenuated wavy fibers in the ventricular myocardium, which confirmed dilated cardiomyopathy in a juvenile. This is the first report of a juvenile form of dilated cardiomyopathy in a Shiba Inu dog. This case report provides evidence that the extended prognosis of this dog differed from that in previously reported cases of dilated cardiomyopathy in young dogs.Key clinical message:This is the first reported case of a juvenile form of dilated cardiomyopathy in a Shiba Inu dog. This report provides evidence that the prognosis of this dog differed from that in previously reported cases of dilated cardiomyopathy in young dogs.     

Cellular characterization of multidrug resistance P-glycoprotein, alpha fetoprotein, and neovascular endothelium-associated antigens in canine hepatocellular carcinoma and cirrhotic liver

P-glycoprotein (P-gp), which is encoded by the multidrug resistance gene (MDR-1); alpha fetoprotein (AFP); and vascular endothelium-associated antigens are well-known markers for human and canine hepatic diseases. We obtained liver tissues from 5 dogs with hepatocellular carcinoma (HCC) and 12 dogs with cirrhosis, and we performed histopathologic and immunohistochemical evaluations using anti-P-gp, anti-AFP, anti-CD31, and anti-CD34 antibodies. P-gp was expressed at higher levels in HCC than in cirrhotic livers ( P < .01), and was most commonly localized in biliary canaliculi and small ductuli. AFP was localized mainly in the cytoplasm in HCC ( P < .01) and in a few cases of cirrhosis. In both HCC and cirrhosis, the AFP-positive cells were morphologically similar to normal hepatocytes and showed an even cytoplasmic distribution of AFP. The endothelial markers CD31 and CD34 were used to investigate vascular distribution. CD31 was expressed strongly in the portal area and parenchyma in HCC, but it was rarely observed in the parenchyma in cirrhosis. CD34 expression could not be detected in both HCC and cirrhosis. This study constitutes the first comprehensive study of P-gp, AFP, and endothelial markers in canine HCC and cirrhosis. The importance of these markers in HCC and cirrhosis in dogs was demonstrated and provides a more accurate basis for a definitive diagnosis of HCC and cirrhosis in dogs.     

Evaluating the contact rate between companion dogs during dog walking and the practices towards potential cases of rabies among dog owners in Japan

This study aimed to examine the habits of dog walking in Japan using an internet survey of insured dog owners. About 96.4% of the respondents (n = 1,151) reported that they would take their dogs for a walk and they most frequently walk their dogs once or twice a day (75.9%) for 30 min to 1 hr (83.1%). The probability of a companion dog having contact with other dogs during dog walking was estimated to be 0.83 (95% confidence interval [CI]: 0.81–0.85), and the associated daily contact rate was estimated using log‐normal distribution with a mean of 2.73 (95% CI: 2.42–3.11) and a standard deviation (SD) of 6.39 (95% CI: 5.18–7.84). Multiple linear regression revealed that the contact rate is mainly influenced by the social behaviour of the owner and to a lesser degree by his/her demographic characteristics including the area of residence, the breed size of dog and the age of the owner. In addition, ten Likert items measured on a 5‐point scale were designed to assess the practices towards potential cases of rabies among dog owners. The respondents (n = 972) achieved a mean score of 2.99 (out of a full score of 4) with a SD of 0.90 in responding to situations related to dog bite incidents and injury from stray cat scratches during dog walking. They achieved a higher score in responding to situations related to sighting a stray or wild animal during dog walking and situations related to non‐specific clinical signs of rabies and bite injuries from stray dogs or wild animals during dog walking with a mean of 3.70 (SD = 0.58) and 3.84 (SD = 0.34), respectively. The level of best practice was also proved to be significantly associated with the demographic characteristics of the dog owner.     

Ehrlichia canis infection in a dog with no history of travel outside the United Kingdom

A two‐year‐old female neutered Tibetan terrier was referred following a one‐month history of lethargy, inappetence and pancytopenia, which had been poorly responsive to immunosuppressive and fluoroquinolone treatment. The dog was diagnosed with pure red cell aplasia and was found to be positive for Ehrlichia canis by both antibody titre measurement and polymerase chain reaction. The dog lived in London and had not travelled outside the UK. The dog was treated with doxycycline, prednisolone and ciclosporin, but died as a result of gastrointestinal tract haemorrhage. To the authors’ knowledge, this represents the first reported case of Ehrlichia canis in a dog in the UK with no previous travel history.     

Completeness of the dog registry and estimation of the dog population size in a densely populated area of Rome

In most European countries, registration and identification of dogs is compulsory. In Italy, the national dog registry is composed of regional dog registries. Although dog registries have been established for many years, the issue related to completeness of data has not been addressed so far. The objective of this study was twofold: first to assess the completeness of data of the dog registry through telephone interview of a sample of dog owners drawn from the dog registry, then to estimate the total owned dog population in 4 boroughs of Rome. For the second objective, a capture–recapture method was applied using data from the dog registry and data from a face-to-face questionnaire submitted to people waiting in the sitting room of 5 points of access for booking and payment of primary and specialist care. Different scenarios are proposed to verify the assumptions of the estimation procedure and potential biases are discussed. The completeness of data of the dog registry was 88.9% (95% CI: 85.8–91.9%) and the owned-dog population was estimated at 26,244 dogs (95% CI: 24,110–28,383). The dog registry is an important source of information especially when it is properly updated and completeness of data is known.     

北极狐及乌苏里貉抑制素α亚基基因克隆及生物信息学分析

试验旨在克隆北极狐及乌苏里貉抑制素α（inhibin α,INHα）亚基基因并对其进行生物信息学分析。根据GenBank中犬科INHα预测mRNA序列（登录号：XM_545660.5）设计1对引物,用RT-PCR技术从北极狐及乌苏里貉的卵巢组织中扩增出INHα亚基基因,同时将其插入到克隆载体中,进行测序及生物信息学分析。测序结果表明,北极狐及乌苏里貉的INHα亚基基因CDS序列全长为1 107 bp,编码369个氨基酸。北极狐及乌苏里貉的INHα亚基基因与犬的同源性最高,分别为97.9%与97.6%。系统进化树分析表明,北极狐及乌苏里貉与犬亲缘关系较近,同时也说明INHα亚基基因在不同物种及进化过程中具有高度保守性。对INHα亚基蛋白的高级结构预测发现,由于半胱氨酸间形成的二硫键导致其采用"蝴蝶形"或"开放手"构型,其中α-螺旋形成分子的"手腕"结构,β-折叠形成分子的"手指"结构。本研究成功克隆了北极狐及乌苏里貉的INHα亚基基因,同时进行了系统的生物信息学分析,为今后研究抑制素在卵母细胞-颗粒细胞同步发育过程中的生物学功能奠定了基础。     

Baclofen intoxication in a dog successfully treated with hemodialysis and hemoperfusion coupled with intensive supportive care

Objective: To describe a case of confirmed baclofen intoxication in a dog that was successfully treated with hemodialysis and hemoperfusion (HD/HP) and to report the serum baclofen kinetics. Case summary: A 2.5‐year‐old, 23 kg, spayed female Brittany Spaniel‐mix was treated after ingesting 21‐52 mg/kg of baclofen. The dog was comatose and was receiving manual ventilation at the time of presentation. Extracorporeal HD/HP was started 10 hours after admission. Within 3 hours of starting HD/HP the dog began initiating breaths and was extubated 18 hours after admission. Serial serum samples that were obtained during the first 24 hours of hospitalization were later analyzed for baclofen concentrations. The dog had elevated creatine phosphokinase and liver enzymes that correlated with an agitated recovery period. The dog had thrombocytopenia that resolved by 10 days after presentation. New or unique information provided: HD/HP shortened the baclofen serum elimination half‐life from 5 to 1.5 hours in the initial 2 hours of treatment. The intrinsic elimination rate constant (K_intr) for this dog was 0.138/hour and the total elimination rate constant (K_tot) during the first 2 hours of HD/HP treatment was 0.458/hour. In this dog, HD/HP was an effective method for rapidly decreasing serum baclofen concentration after an acute overdose.