Pedigree study of the heredity of copper-associated hepatitis in Dalmatians in Japan

The pedigrees of 3 Dalmatian dogs afflicted with copper-associated hepatitis were investigated to discover the mode of inheritance. A composite family pedigree showed that the 3 affected Dalmatians were related. None of the parents of the affected dogs showed clinical symptoms of liver disease, and the disease had no sex predisposition. The estimated segregation ratio was approximately 3:1 based on surviving littermates. These findings suggested that the copper-associated hepatitis in these Dalmatians was an autosomal recessive mode of inheritance. In addition, some male Dalmatians imported from abroad might have been involved in the occurrence of this disease in Japan.     

Copper-associated hepatitis in a young Dalmatian dog in Japan

A male 25-month-old Dalmatian dog attended our veterinary hospital because of anorexia and high circulating liver enzyme activities. Abdominal computed tomography showed a slightly small liver with rounded edges, and laparoscopic examination showed that the liver was yellowish. Histopathological examination revealed multifocal necrosis of hepatocytes and severe chronic hepatitis. Rhodanine staining showed severe copper accumulation in hepatocytes and a quantitative analysis of the copper content of the liver showed substantial accumulation (10.3 mg/g dry mass), suggesting a diagnosis of copper-associated hepatitis. Previously reported canine mutation in the COMMD1, the gene responsible for the copper-associated hepatitis in the Bedlington terrier, was not identified. To our knowledge, this is the first report of copper-associated hepatitis in a Dalmatian in Japan.     

Genotype frequency of ATP7A and ATP7B mutation-related copper-associated hepatitis in a Japanese guide dog Labrador retriever population

The incidence of copper-associated hepatitis in Labrador retriever in Japan has not been examined. This study examined the genotype frequencies of ATP7B:c.4358G>A, a mutation responsible for copper-associated hepatitis, and ATP7A:c.980C>T, a modifier of this disease, in Labrador retrievers of guide dog associations in Japan. Genetic material was collected by buccal swabs from 253 Labrador retrievers and genotyping was performed for the ATP7B and ATP7A mutations. The gene frequency was 0.107 for ATP7B:c.4358A. For ATP7A:c.980C, the gene frequencies were 0.703 in females and 0.368 in males. In this study, we established genotyping methods for the ATP7B:c.4358G>A and ATP7A:c.980C>T mutations. Based on the genotyping results, the risk of copper-associated hepatitis in the study population was 0.80% in males and 1.05% in females.     

Copper-associated chronic hepatitis in Labrador Retrievers

This study summarizes the clinical and pathologic findings in 15 Labrador Retrievers with copper-associated chronic hepatitis (CACH). Our hypothesis was that this form of hepatitis is caused by a defect in hepatic copper metabolism, which most likely originates from a genetic defect. Affected Labradors consisted of 11 female and 4 male Labrador Retrievers. Eight family members of 2 of these patients were examined prospectively, as were 6 unrelated healthy Labrador Retrievers. All dogs were registered at the breed club. The average age at clinical presentation was 7 years (range, 2.5-10.5 years). All dogs were presented for anorexia, which was associated with vomiting in 8 patients. The diagnosis of CACH was based on histologic examination of liver biopsy specimens in all dogs, including semiquantitation of copper. A disproportionate increase in alanine aminotransferase (ALT) activity relative to alkaline phosphatase (ALP) activity, as well as the centrolobular localization of copper and the association of copper accumulation with hepatic lesions, suggested a primary copper storage disease rather than primary cholestatic liver disease causing copper accumulation. Mean hepatic copper concentration measured in related Labradors was 1,317 microg/g dry weight liver (range, 402-2,576 microg/g). Mean hepatic copper concentration of unrelated normal Labradors was 233 microg/g dry weight liver (range, 120-304 microg/g). Our findings support the hypothesis that a hereditary form of hepatitis occurs in Labrador retrievers and is caused by a defect in hepatic copper metabolism.     

Nutritional management of inherited copper-associated hepatitis in the Labrador retriever

Canine hereditary copper-associated hepatitis is characterized by gradual hepatic copper accumulation eventually leading to liver cirrhosis. Therapy is aimed at creating a negative copper balance with metal chelators, of which d-penicillamine is the most commonly used. d-penicillamine often causes gastro-intestinal side effects and life-long continuous therapy may lead to a deficiency of copper and zinc. The aim of the current study was to investigate the effect of a low-copper, high-zinc diet as an alternative to continuous d-penicillamine treatment for the long-term management of canine copper-associated hepatitis.Sixteen affected Labrador retrievers were followed for a median time period of 19.1 months (range, 5.9–39 months) after being effectively treated with d-penicillamine. The dogs were maintained on a diet containing 1.3 ± 0.3 mg copper/1000 kcal and 64.3 ± 5.9 mg zinc/1000 kcal. Liver biopsies were taken every 6 months for histological evaluation and copper determination. Plasma alanine aminotransferase (ALT) and alkaline phosphatase, as well as serum albumin were determined.Dietary treatment alone was sufficient to maintain hepatic copper concentration below 800 mg/kg dry weight liver in 12 dogs during the study period. Four dogs needed re-treatment with d-penicillamine. ALT activity and albumin concentration were not associated with hepatic copper concentration, but showed a significant association with the stage and grade of hepatitis respectively. In conclusion, a low-copper, high-zinc diet can be a valuable alternative to continuous d-penicillamine administration for long-term management of dogs with copper-associated hepatitis. The copper re-accumulation rate of an individual dog should be considered in the design of a long-term management protocol and in determining re-biopsy intervals.     

Copper-associated liver disease in Dalmatians: a review of 10 dogs (1998-2001)

This retrospective study summarizes 10 Dalmatians suspected of having hepatic copper toxicosis. Hepatic copper toxicosis can result from either a primary metabolic defect in hepatic copper metabolism or from altered hepatic biliary excretion of copper. An inherited copper-associated hepatopathy has been documented in Bedlington Terriers, and there is evidence for familial copper-associated liver disease in West Highland White (WHW) Terriers and Skye Terriers. Nine of the 10 Dalmatians in this study presented for gastrointestinal clinical signs, including anorexia and vomiting. All animals had increased alanine aminotransferase (ALT) enzyme activity, and 9 of 10 had increased alkaline phosphatase (ALP) enzyme activity. The relative increase in ALT activity was much greater than the relative increase in ALP activity, suggesting a predominantly hepatocellular rather than cholestatic liver disease. The mean hepatic copper concentration for 9 Dalmatians was 3,197 microg/g dry weight liver (dwl) (normal, <450 microg/g). In 5 of these 9 dogs, hepatic copper concentrations exceeded 2,000 microg/g dwl. Necroinflammatory alterations associated with copper-laden parenchymal cells were the notable histopathologic finding. The inflammatory infiltrate was either primarily lymphocytic or neutrophilic. Morphologic features of cholestasis generally were not prominent except in those dogs with severe pathology. These findings lend support to the hypothesis that a primary metabolic defect in hepatic copper metabolism occurs in the Dalmatian breed. The mechanism and genetic basis of this condition require further study.     

Nutritional management of hepatic disease

The successful management of hepatic diseases of dogs and cats requires an understanding of hepatic metabolism and nutritional processes. General aspects of dietary therapy for hepatic diseases are described, along with specific recommendations for the promotion of tissue regeneration. Special considerations, including the role of diet in encephalopathy, hepatic lipidosis, and copper-associated hepatic toxicosis, are also discussed.     

Survey of copper and zinc concentrations in commercially available dry ferret diets

BackgroundTo survey macronutrients and two micronutrients (copper and zinc) in commercial ferret diets to investigate dietary factors that may influence copper-associated hepatopathy in ferrets.MethodsProximate analysis and copper and zinc concentrations of 12 commercially available dry adult maintenance ferret diets. Medians were calculated on a dry matter and metabolizable energy basis and compared to the Association of American Feed Control Officials (AAFCO) nutrient profiles for cats and the National Research Council (NRC) recommendations for mink.ResultsCalculated metabolizable energy of diets ranged from 3830 to 4305 kcal/kg dry matter (DM). Crude protein ranged from 89.1 to 151.4 g/1000 kcal, nitrogen-free extract ranged from 13.9 to 124.7 g/1000 kcal, and crude fat ranged from 29.6 to 52.4 g/1000 kcal. The median copper concentration was 9.0 mg/1000 kcal and 37.8mg/kg DM, 7 times the AAFCO minimum for cats and 6.3 times the recommendation for mink. Median zinc concentration was 64.2 mg/1000 kcal and 243.1mg/kg DM, 3.4 times the AAFCO minimum for cats and 3.7 times the NRC recommendation for mink. The median zinc to copper ratio was 6.7.Conclusions and Clinical RelevanceThere is considerable variation in micro- and macronutrient composition among tested commercial ferret diets. High copper concentrations in ferret diets may contribute to copper-associated hepatopathy in ferrets.     

Histochemical demonstration of copper and copper-associated protein in the canine liver

Three different histochemical methods for copper detection were compared. Atomic absorption analysis was used to substantiate the tissue stains. There was good correlation between rhodanine staining and rubeanic acid-stained tissue sections. The orcein reaction for copper-associated protein did not consistently correlate with the methods demonstrating copper. Prolonged staining (72 hours) with rubeanic acid more consistently and clearly detected increased copper in canine livers than did staining with rhodanine. Seventy-two hour staining with rubeanic acid is the method of choice for histochemical detection of copper in canine liver.     

鸭肝炎病毒分子生物学研究进展

鸭病毒性肝炎(DVH)是由鸭肝炎病毒(DHV)引起雏鸭的一种以肝脏为主要病理变化的急性、高度致死性、接触性传染病。目前,该病在世界范围内流行,给养鸭业带来了巨大的经济损失,许多地区频频出现DHV免疫鸭群暴发鸭肝炎的报道,严重制约了养鸭业的健康发展。论文就鸭肝炎病毒的生物学特性与病毒基因组结构、分子生物学诊断方法、序列分析与基因分型、结构蛋白VP1、VP3等分子生物学研究进行综述,以期为合理制定鸭肝炎病毒的免疫方法与免疫程序提供参考,为鸭病毒性肝炎的综合防控提供依据。     

鸭病毒性肝炎诊断的研究进展

鸭病毒性肝炎是一种传播迅速的病毒性、高度致死性疾病,是危害养鸭业最为严重的疾病之一。该病主要发生在3周龄以内的雏鸭,致死率高达100%。文章对鸭病毒性肝炎诊断技术研究进展情况进行了综述,并对鸭病毒性肝炎诊断技术的研究前景进行了展望。     

应用RT—PCR方法检测鸭病毒性肝炎新型和I型病毒

鸭病毒性肝炎病毒是危害养鸭业的重要病毒性传染病,主要感染低日龄雏鸭,临床上分离到的病原主要为鸭病毒性肝炎I型病毒和鸭病毒性肝炎新型病毒,为了加快本病的检测速度,尽快确诊病原,根据GeneBank上两种毒株的基因组序列,选择两种毒株的差异基因序列分别设计了一对特异性的检测引物,将两对引物放在一个PCR反应体系中,两种毒株分别进行特异性扩增,根据扩增片段的大小进行鉴别,建立了可以同时检测鸭病毒性肝炎病毒I型和新型毒株的复合RT—PCR方法,该检测方法快速,特异性强,灵敏性高,可以用于鸭病毒性肝炎病毒的分型鉴别检测。     

Campylobacter jejuni is associated with, but not sufficient to cause vibrionic hepatitis in chickens

Vibrionic hepatitis is a disease of poultry which is characterised by the presence of focal lesions in the liver, usually 1-2mm in size and greyish-white in colour. The cause of the disease remains unclear, as do the reasons for its recent re-emergence. We examined the livers of commercial broiler chickens taken during processing and found Campylobacter spp. in both normal livers and those displaying signs indicative of focal hepatitis. Livers with signs of hepatitis had significantly more Campylobacter spp. present than those without and other bacterial genera were infrequently present. We were unable to replicate the disease in a healthy host following experimental infection with a Campylobacter jejuni strain isolated from a liver showing signs of focal hepatitis. However, a significant T cell response to C. jejuni was seen in the liver of Campylobacter infected birds. We conclude that the presence of Campylobacter spp. in the liver alone is not sufficient to cause vibrionic hepatitis, but that a predisposing factor, possibly within the host is required. We also provide evidence that chickens mount an adaptive T cell response to systemic C. jejuni.     

A new transmissible agent causing acute hepatitis, chronic hepatitis and cirrhosis in dogs

There is a hepatitis of dogs which occurs in acute, persistent and chronic forms. Histological studies of spontaneous cases suggested that several apparently diverse hepatic diseases might be stages of one process. This was also implied by follow up studies and case histories: acute non-lethal episodes were followed later by the development of chronic hepatitis, cirrhosis and liver failure. Serum was taken and homogenates of liver were made from three field cases representing different putative temporal stages of the complex. These were injected into experimental dogs and a hepatitis was induced in all. The cytopathological and histological changes were the same in all animals and were identical to field cases. Acute lethal disease and persistent infections were produced. Two second passages were carried out and an identical condition was induced, characterised by recurrent episodes of subclinical hepatitis and persistent infection. It is suggested that the disease might be named canine acidophil cell hepatitis in view of the pathognomonic cytopathology. Specific morphological criteria have been established for this hepatitis.     

Equine parvovirus hepatitis

Equine parvovirus hepatitis (EqPV-H) was first described in 2018 in a fatal case of Theiler's disease which followed the administration of an equine-origin biological product. The virus has since been frequently identified in serum and liver tissue of horses affected by Theiler's disease—an acute, severe hepatitis characterised by fulminant hepatic necrosis with a fatal outcome in most cases. EqPV-H is hepatotropic, appears to be associated with subclinical to severe hepatitis in horses, and is a likely cause of Theiler's disease. Although this disease is most frequently reported following the administration of equine-origin biological products, it can also occur among in-contact horses. Horizontal transmission may be iatrogenic, via contaminated equine-origin biological products such as equine serum, botulism or tetanus antitoxin, and mesenchymal stem cells or by means of the oral route of infection. Other horizontal transmission routes, for example, arthropod vectors, warrant further investigation. A worldwide prevalence of EqPV-H antibodies and DNA has been reported in asymptomatic horses. EqPV-H-positive horses suffering from acute, severe hepatitis have reportedly developed clinical signs including icterus, lethargy, inappetence, and neurological abnormalities and have had increased liver-associated biochemistry parameters recorded. The most common histopathological abnormalities of the liver have been hepatocellular necrosis, collapse of the lobular architecture, and lymphocytic infiltration. Most horses infected experimentally with EqPV-H have developed subclinical hepatitis, and close temporal associations between peak viraemia, seroconversion, and the onset of hepatitis have been observed. Based on strong evidence indicating that EqPV-H causes hepatitis in horses, veterinarians should consider this virus an important differential diagnosis in such cases. Potential risks associated with the administration of equine-origin biological products must be emphasised.     

Granulomatous peritonitis and pleuritis in interferon-gamma gene knockout mice naturally infected with mouse hepatitis virus

OBJECTIVE: To investigate a disease outbreak in a colony of laboratory mice with targeted disruption of the gene for interferon-gamma. FORMAT: A case report based on necropsy, histopathology, serology and immunohistochemistry. RESULTS: Affected mice exhibited depression and variable ascites. Necropsy revealed a granulomatous peritonitis and pleuritis with extensive adhesions although parenchymal lesions were minimal. Serum samples had high concentrations of antibody to mouse hepatitis virus and immunohistochemical examination revealed the presence of mouse hepatitis virus antigen in granuloma macrophages. Sero-logical testing for other infectious agents and bacterial culture were negative and wild type mice kept in the same facility remained healthy. Despite the association between the disease and mouse hepatitis virus infection, the precise role played by mouse hepatitis virus was not determined. While the disease is superficially similar to feline infectious peritonitis (another coronavirus-induced serositis), differences exist between the histopathological findings in these two conditions. CONCLUSION: This unusual disease process illustrates how new diagnostic challenges can arise in novel mouse genotypes created through molecular genetics. Furthermore, the association between the disease and mouse hepatitis virus illustrates the importance of maintaining laboratory animals under specific-pathogen free conditions.     

Obstructed portal venous flow and portal vein thrombus in a dog

A young adult Doberman Pinscher had clinical signs and laboratory data consistent with the copper-associated hepatopathy commonly found in Doberman Pinschers. Hepatic biopsy and hepatic copper analysis did not support this diagnosis. Furthermore, changes seen in the hepatic biopsy specimen did not explain the acquired portosystemic shunting or the portal hypertension that the dog had. A mesentery venoportagram revealed markedly delayed filling of one portal vein, and necropsy revealed a small thrombus partially occluding that hepatic vein. It was not determined whether obstructed portal venous flow caused the portal vein thrombus or vice versa; however, it was theorized that the portal vein thrombus, once formed, contributed to further portal hypertension and/or delayed portal vein filling.     

鸭肝炎病毒的研究进展

鸭肝炎病毒是一种高致死性、高传播性的病毒性传染病病原,临床主要引起急性肝炎,主要侵害3周龄以内的雏鸭,尤以1周龄内的雏鸭为甚,病死率高达100%。20世纪以来,鸭肝炎已成为危害中国养鸭业最为严重的疫病之一,给养鸭业带来了巨大的经济损失。作者对鸭肝炎病毒的发生与分布、病原学、基因组学、诊断与防治等方面进行了综述。