Evaluation of metallothionein and Ki-67 expression in chronic cholangiohepatitis in cats.

Chronic cholangiohepatitis (CCH) is a common pathological condition in cats with a guarded prognosis and unknown etiology. Recently, in human medicine, there has been increased interest in enhancing liver defense mechanisms as an effective treatment strategy to control liver diseases that have a poor prognosis. Metallothionein (MT) is a ubiquitous protein, which has been widely researched for its role in liver defense through heavy metal detoxification, neutralization of reactive oxygen species, and liver regeneration. In this study, immunohistochemistry was used to evaluate the role of MT in CCH and hepatocellular regeneration in 34 cats histologically diagnosed with this condition by assessing the correlation between hepatocellular MT and Ki-67 (marker for cellular proliferation) expression with histological parameters of CCH, such as inflammation, fibrosis, and bile duct proliferation. Statistical analysis was performed using the Spearman-rank correlation test. A significant positive correlation was observed between inflammation and the number of MT-positive hepatocytes (r = 0.36, P = 0.03) and MT labelling intensity (r = 0.37, P = 0.03). In 16 of 34 cases (47%) MT labelling intensity was noted to be pronounced towards the centrilobular zone and very weak or absent towards the portal zone. The results suggest that MT is induced in the liver during chronic inflammatory conditions, which could be speculated as a host defensive mechanism to protect the liver from inflammation-mediated liver injury. Therapeutic interventions utilizing MT, therefore, may have a positive effect on cats with chronic cholangiohepatitis.

La cholangiohépatite chronique (CCH) est une affection pathologique courante chez les chats avec un pronostic réservé et une étiologie inconnue. Récemment, en médecine humaine, il y a eu un intérêt accru pour l'amélioration des mécanismes de défense hépatique en tant que stratégie de traitement efficace pour contrôler les maladies du foie qui ont un mauvais pronostic. La métallothionéine (MT) est une protéine omniprésente, qui a été largement étudiée pour son rôle dans la défense du foie par la détoxification des métaux lourds, la neutralisation des espèces réactives de l'oxygène et la régénération du foie. Dans cette étude, l'immunohistochimie a été utilisée pour évaluer le rôle de la MT dans la CCH et la régénération hépatocellulaire chez 34 chats diagnostiqués histologiquement avec cette condition en évaluant la corrélation entre l'expression hépatocellulaire de la MT et du Ki-67 (marqueur de la prolifération cellulaire) avec les paramètres histologiques de la CCH, comme l'inflammation, la fibrose et la prolifération des voies biliaires. L'analyse statistique a été réalisée à l'aide du test de corrélation de rang de Spearman. Une corrélation positive significative a été observée entre l'inflammation et le nombre d'hépatocytes MT-positifs (r = 0,36, P = 0,03) et l'intensité de marquage MT (r = 0,37, P = 0,03). Dans 16 des 34 cas (47 %), l'intensité du marquage MT était prononcée vers la zone centrolobulaire et très faible ou absente vers la zone porte. Les résultats suggèrent que la MT est induite dans le foie pendant les états inflammatoires chroniques, ce qui pourrait être supposé comme un mécanisme de défense de l'hôte pour protéger le foie contre les lésions hépatiques induites par l'inflammation. Les interventions thérapeutiques utilisant la MT peuvent donc avoir un effet positif sur les chats atteints de cholangiohépatite chronique.(Traduit par Docteur Serge Messier).

Metabolic derangements and reduced survival of bile-extracted Asiatic black bears (Ursus thibetanus).

BACKGROUND: Across China and Southeast Asia, an estimated 17,000 bears are currently farmed for bile, primarily for traditional medicines. Depending on country, bile is extracted daily via transabdominal gallbladder fistulas, indwelling catheters, or needle aspiration. Despite claims that bears do not develop adverse effects from bile extraction, health issues identified in bears removed from bile farms include bile-extraction site infections, abdominal hernias, peritonitis, cholecystitis, hepatic neoplasia, cardiac disease, skeletal abnormalities, and abnormal behaviors. We present a comprehensive assessment of the effects of bile farming by comparing serum biochemical and hematological values of bears from farms that were bile-extracted (BE) and bears from farms not bile-extracted (FNE) with bears from non-farm captive (ZOO) and free-range (FR) environments. We hypothesized BE bears would have significant laboratory abnormalities compared to all non-extracted bear groups. We also hypothesized BE bears would have reduced long-term survival compared to FNE bears despite removal from farms. RESULTS: BE bears exhibited the highest values and greatest variation (on a population level) in laboratory parameters compared to all non-extracted bear groups particularly for alanine transaminase, gamma glutamyltransferase (GGT), total bilirubin (TBIL), alkaline phosphatase (ALKP), blood urea nitrogen, creatinine (CREA), and total white blood cell count. Significant differences were detected between bear groups when accounting for season, sex, and/or age. BE bears exhibited greater mean serum GGT compared to all non-extracted bear groups, and the odds of having elevated TBIL were 7.3 times greater for BE bears, consistent with hepatobiliary disease. Biochemical parameter elevations in BE bears persisted up to 14 years post-rescue, consistent with long-term effects of bile-extraction. BE bears that arrived with elevated CREA and ALKP had median survival times of 1 and 4 years respectively, and regardless of laboratory abnormalities, BE bears had significantly shorter survival times compared to FNE bears. CONCLUSIONS: Our results provide strong evidence that bile extraction practices not only represent a temporary constraint for bears' welfare, but confer distinct long-term adverse health consequences. Routine laboratory panels may be insensitive to detect the extent of underlying illness in BE bears as these bears have significantly reduced survival regardless of biochemical assessment compared to FNE bears.

Spontaneous Cholangiofibrosis in a Wistar Rat.

In a 2-year carcinogenicity study, we identified a spontaneous cholangiofibrosis in a control male Wistar rat. This lesion has long been considered as a compound-related change, with no spontaneous cases reported in the Wistar rat. In addition to routine hematoxylin and eosin stains evaluation, we applied Masson's trichrome staining, Alcian blue-periodic acid-Schiff staining, and OV-6 immunohistochemistry staining. The special staining demonstrated the fibrous component in the interstitium and intestinal metaplasia of the epithelium (presence of goblet cells), while the positive anti-OV-6 reaction indicated the bile duct origin of the epithelium. These results help to confirm the diagnosis of cholangiofibrosis in this case. We report this rare case to alert pathologists that spontaneous cholangiofibrosis does occur in Wistar rats.

Biliary Tract-Infecting Myxosporeans from Estuarine and Reef Stonefish (Scorpaeniformes: Synanceiidae) Off Eastern Australia, with Descriptions of Sphaeromyxa horrida n. sp. and Myxidium lapipiscis n. sp. (Myxosporea: Bivalvulida).

Two new species of myxosporeans are described from the gallbladders of estuarine stonefish, Synanceia horrida, and reef stonefish, Synanceia verrucosa, from localities off Cairns, in tropical north Queensland and in Moreton Bay in southern Queensland, Australia. Sphaeromyxa horrida n. sp. can be distinguished from congeners in the morphologically distinct "balbianii" species group within Sphaeromyxa on the basis of morphometric differences in length and width of mature spores, length and width of polar capsules, and unique small-subunit (SSU) ribosomal (rDNA) sequence composition relative to other taxa. Replicate SSU rDNA sequences generated from Sph. horrida n. sp. collected from Sy. horrida and Sy. verrucosa in tropical north Queensland and from Sy. horrida in Moreton Bay were identical, suggesting that this species is widely distributed along the east coast of Australia. Myxidium lapipiscis n. sp. can be distinguished from the majority of described Myxidium species on the basis of its relatively small mature spore size (6.1-7.9 µm long × 3.1-3.9 µm wide), and its unique SSU rDNA sequence. Specimens putatively identified as M. lapipiscis n. sp. were found in Sy. horrida from both tropical north Queensland and Moreton Bay, suggesting that this taxon is also widely distributed along the east coast of Australia. However, no molecular data were available for the specimens from tropical north Queensland for comparative genetic analyses. Bayesian inference and maximum likelihood analysis of the SSU rDNA sequences for these 2 new species revealed that Sph. horrida n. sp. formed a strongly supported clade with Sphaeromyxa zaharoni Diamant, Whipps, and Kent, 2004, which was described from the scorpaeniform, Pterois miles, from the Red Sea. This is the first report of myxozoans infecting stonefish (Synanceiidae).

Histopathological frequency of feline hepatobiliary disease in the UK.

OBJECTIVES: To determine the histopathological frequency of feline hepatobiliary diseases in the UK and to identify breed, age and gender predispositions to developing individual diseases. METHODS: Histopathology results from 1452 feline liver biopsies were assessed. A control population of microchipped cats was used for breed comparison. Data were retrospectively categorised into hepatobiliary diseases according to World Small Animal Veterinary Association standards. Odds ratios and 95% confidence intervals were calculated to determine breed predispositions to the 10 most frequent diseases. Gender and age distributions were also evaluated. RESULTS: The most frequent diseases based on histopathology were neutrophilic cholangitis (20·5%), reactive hepatitis (20·4%), reversible hepatocellular injury (8·4%), lymphocytic cholangitis (6·8%), biliary cysts (5·7%), acute hepatitis (5·6%), haematopoietic neoplasia (5·6%), hepatocellular neoplasia (4·9%), congenital portosystemic shunt (3·8%) and cholangiocellular neoplasia (3·1%). Some previously unreported breed and age predispositions were identified. CLINICAL SIGNIFICANCE: This is the first study to document the histopathological frequency of hepatobiliary diseases in a large cohort of cats in the UK, as well as novel breed and age predispositions. These data may help increase the index of suspicion of a particular disease in the absence of a biopsy-confirmed diagnosis.

Clinical relevance of radiographic linear branching mineral opacities in the canine liver.

OBJECTIVES: To assess the prevalence, clinical significance and breed distribution of linear branching mineralisation superimposed on the hepatic radiographic silhouette in dogs. MATERIALS AND METHODS: Retrospective review of radiographs or ultrasound images of dogs showing branching mineralisation in the liver. RESULTS: Over the 30-year review period, 17 cases were identified and the mineralisation had a predominantly ventral distribution. Seven of the 17 were cavalier King Charles spaniels, and four of the total 17 dogs were diagnosed with hepatobiliary system disease. Five dogs had repeat radiographs, of which four showed no change in the pattern and one developed the pattern 6 years after being diagnosed with cholangiohepatitis. Serum calcium concentrations were normal in all patients. Liver enzymes were markedly elevated only in the dog diagnosed with cholangiohepatitis. Histology performed on three patients showed no convincing evidence of primary liver disease. CLINICAL SIGNIFICANCE: Branching mineralisation in the liver parenchyma is a rare finding in dogs with little or no clinical significance and cavalier King Charles spaniels may be predisposed. Biopsy of the liver of affected dogs with no clinical or clinicopathological evidence of liver disease is unlikely to be helpful in these cases.

Feline biliary tree and gallbladder disease: Aetiology, diagnosis and treatment

Practical relevance Diseases of the biliary tree and gallbladder are more common in cats than diseases of the liver parenchyma. The parenchyma is usually affected secondarily to systemic illnesses, while the biliary system is the prime target for infectious agents (eg, bacteria and flukes) and non-infectious conditions (eg, neoplasia and cysts). Clinical approaches Cats with biliary disease are evaluated because of common feline clinical signs such as anorexia, nausea, vomiting and lethargy. Icterus may or may not be obvious. Biopsies for histological evaluation, and bile aspirates for culture and cytological evaluation are helpful diagnostically. Antibiotics and immunosuppressive drugs have been used successfully. Hepatosupportive drugs may help in liquefying thick bile and protecting hepatic tissue from damage. Ultrasound is a noninvasive diagnostic tool that may help in identifying dilated bile ducts, liver cysts and choleliths. It is also used to guide percutaneous bile aspiration. Audience This review, written for all veterinarians who treat cats, describes the various conditions that can affect the feline biliary tree and gallbladder. Treatment options are discussed, and brief summaries provided of surgical techniques and diagnostic approaches. Evidence base The veterinary literature pertaining to feline biliary disease is comprehensively reviewed. When appropriate, data on dogs and humans has been included to provide background information. Based on the available literature, more research into feline biliary diseases is needed.

Laboratory Evaluation of the Liver.

Laboratory evaluation of the hepatobiliary system has an important role in the diagnosis, monitoring, and assessment of patients with hepatobiliary diseases. Serum liver enzyme activities can be divided into markers of hepatocellular injury and cholestasis. Liver function can be assessed in several ways, including assessment of synthetic capacity, measurement of ammonia, and measurement of bile acids. It is essential to have an understanding of the performance characteristics and limitations of these tests in order to use them appropriately. This article reviews the laboratory parameters commonly used to aid diagnosing hepatobiliary disorders in dogs and cats.

Diagnostic Imaging of the Hepatobiliary System: An Update.

Recent advances in diagnostic imaging of the hepatobiliary system include MRI, computed tomography (CT), contrast-enhanced ultrasound, and ultrasound elastography. With the advent of multislice CT scanners, sedated examinations in veterinary patients are feasible, increasing the utility of this imaging modality. CT and MRI provide additional information for dogs and cats with hepatobiliary diseases due to lack of superimposition of structures, operator dependence, and through intravenous contrast administration. Advanced ultrasound methods can offer complementary information to standard ultrasound imaging. These newer imaging modalities assist clinicians by aiding diagnosis, prognostication, and surgical planning.

Hemostatic Disorders Associated with Hepatobiliary Disease.

The liver plays a crucial role in all aspects of coagulation because most factors that regulate procoagulation, anticoagulation, and fibrinolysis are produced, cleared, and/or activated in the liver. Establishing the coagulation status of an individual patient with hepatobiliary disease can therefore be challenging. Although, classically, patients with hepatobiliary disease were thought of as potentially hypocoagulable, hypercoagulability also occurs. The article summarizes the breadth of coagulation abnormalities that have been reported in dogs and cats with hepatobiliary disease and provides strategies to respond to bleeding and thrombotic risk.

Emergence of the Zoonotic Biliary Trematode Pseudamphistomum truncatum in Grey Seals (Halichoerus grypus) in the Baltic Sea.

The biliary trematode Pseudamphistomum truncatum parasitizes a wide range of fish-eating mammals, including humans. Here we report the emergence of this parasite in grey seals (Halichoerus grypus) in the Baltic Sea. One hundred eighty-three of 1 554 grey seals (11.9%) examined from 2002-2013 had detectable hepatobiliary trematode infection. Parasite identification was confirmed as P. truncatum by sequencing the ITS2 region of a pool of five to 10 trematodes from each of ten seals collected off the coast of seven different Swedish counties. The proportion of seals parasitized by P. truncatum increased significantly over time and with increasing age of seals. Males were 3.1 times more likely to be parasitized than females and animals killed in fishery interactions were less likely to be parasitized than animals found dead or hunted. There was no significant difference in parasitism of seals examined from the Gulf of Bothnia versus those examined from the Baltic Proper. Although the majority of infections were mild, P. truncatum can cause severe hepatobiliary disease and resulted in liver failure in at least one seal. Because cyprinid fish are the second intermediate host for opisthorchiid trematodes, diets of grey seals from the Baltic Sea were analysed regarding presence of cyprinids. The proportion of gastrointestinal tracts containing cyprinid remains was ten times higher in seals examined from 2008 to 2013 (12.2%) than those examined from 2002 to 2007 (1.2%) and coincided with a general increase of trematode parasitism in the host population. The emergence and relatively common occurrence of P. truncatum in grey seals signals the presence of this parasite in the Baltic Sea ecosystem and demonstrates how aquatic mammals can serve as excellent sentinels of marine ecosystem change. Investigation of drivers behind P. truncatum emergence and infection risk for other mammals, including humans, is highly warranted.

Ultrasound Imaging of the Hepatobiliary System and Pancreas.

Ultrasound is an extremely valuable diagnostic modality for the diagnosis of hepatobiliary and pancreatic disease. Normal appearance and normal variations are important to understand to avoid misinterpretation. Although ultrasound can identify a lesion, cytology and histopathology are usually needed for a final diagnosis.

Computed Tomography and MRI of the Hepatobiliary System and Pancreas.

MRI and computed tomographic (CT) imaging are becoming more common in the diagnosis of hepatobiliary and pancreatic disorders in small animals. With the advent of multislice CT scanners, sedated examinations in veterinary patients are feasible increasing the use of this imaging modality. CT and MRI provide additional information for dogs and cats with hepatobiliary and pancreatic diseases because of lack of superimposition of structures, operator dependence, and through intravenous contrast administration. This added value provides more information for diagnosis, prognosis, and surgical planning.

Canine hyperlipidaemia.

Hyperlipidaemia refers to an increased concentration of lipids in the blood. Hyperlipidaemia is common in dogs and has recently emerged as an important clinical condition that requires a systematic diagnostic approach and appropriate treatment. Hyperlipidaemia can be either primary or secondary to other diseases. Secondary hyperlipidaemia is the most common form in dogs, and it can be a result of endocrine disorders, pancreatitis, cholestasis, protein-losing nephropathy, obesity, as well as other conditions and the use of certain drugs. Primary hyperlipidaemia is less common in the general canine population but it can be very common within certain breeds. Hypertriglyceridaemia of Miniature Schnauzers is the most common form of primary hyperlipidaemia in dogs but other breeds are also affected. Possible complications of hyperlipidaemia in dogs include pancreatitis, liver disease, atherosclerosis, ocular disease and seizures. Management of primary hyperlipidaemia in dogs is achieved by administration of ultra low-fat diets with or without the administration of lipid lowering drugs such as omega-3 fatty acids, fibrates, niacin and statins.

A Laboratory Diagnostic Approach to Hepatobiliary Disease in Small Animals.

Routine biochemical tests generally include serum enzymes, proteins, and other markers useful for identifying hepatobiliary disease in dogs and cats. Obtaining results outside the reference intervals can occur with direct hepatocellular injury, enzyme induction by hepatocytes or biliary epithelium, or decreased hepatic function. However, detection of biochemical abnormalities does not necessarily indicate clinically significant disease. For a comprehensive approach to detection and treatment of hepatobiliary disease, the laboratory results must be correlated with the history and physical examination findings, diagnostic imaging results, and other assays.

Botulinum toxin A as a treatment for delayed gastric emptying in a dog.

A toy Australian shepherd dog was referred for bile peritonitis following excision of a biliary mucocele. Subsequent delayed gastric emptying was refractory to prokinetic therapy but responded to injection of botulinum toxin A into the muscularis layer of the pylorus; a novel therapy for delayed gastric emptying in dogs.

Toxine botulinique : Traitement de la vidange gastrique retardée chez un chien. Un chien Berger australien miniature a été référé pour une péritonite biliaire après l'excision d'une mucocèle biliaire. La vidange gastrique retardée subséquente a été réfractaire au traitement stimulant la motilité gastrique mais a répondu à l'injection de la toxine botulinique A dans la couche musculeuse du pylore; un traitement innovateur pour la vidange gastrique retardée chez les chiens.(Traduit par Isabelle Vallières).

Presumed primary and secondary hepatic copper accumulation in cats.

OBJECTIVE: To determine signalments, clinical features, clinicopathologic variables, imaging findings, treatments, and survival time of cats with presumed primary copper-associated hepatopathy (PCH) and to determine quantitative measures and histologic characteristics of the accumulation and distribution of copper in liver samples of cats with presumed PCH, extrahepatic bile duct obstruction, chronic nonsuppurative cholangitis-cholangiohepatitis, and miscellaneous other hepatobiliary disorders and liver samples of cats without hepatobiliary disease. DESIGN: Retrospective cross-sectional study. ANIMALS: 100 cats with hepatobiliary disease (PCH [n = 11], extrahepatic bile duct obstruction [14], cholangitis-cholangiohepatitis [37], and miscellaneous hepatobiliary disorders [38]) and 14 cats without hepatobiliary disease. PROCEDURES: From 1980 to 2013, cats with and without hepatobiliary disease confirmed by liver biopsy and measurement of hepatic copper concentrations were identified. Clinical, clinicopathologic, and imaging data were compared between cats with and without PCH. RESULTS: Cats with PCH were typically young (median age, 2.0 years); clinicopathologic and imaging characteristics were similar to those of cats with other liver disorders. Copper-specific staining patterns and quantification of copper in liver samples confirmed PCH (on the basis of detection of > 700 µg/g of liver sample dry weight). Six cats with PCH underwent successful treatment with chelation (penicillamine; n = 5), antioxidants (5), low doses of elemental zinc (2), and feeding of hepatic support or high-protein, low-carbohydrate diets, and other hepatic support treatments. One cat that received penicillamine developed hemolytic anemia, which resolved after discontinuation of administration. Three cats with high hepatic copper concentrations developed hepatocellular neoplasia. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that copper accumulates in livers of cats as primary and secondary processes. Long-term management of cats with PCH was possible.

Climatic variables are associated with the prevalence of biliary trematodes in otters.

Parasites with complex life cycles are expected to be disproportionately affected by climate change. Knowledge of current associations with weather and host-parasite interactions is therefore essential for the inference of future distributions. The Eurasian otter, Lutra lutra, is exposed to a range of parasites due to its large home range and use of terrestrial, freshwater and marine habitats. As such, it can act as a sentinel species for generalist parasites. Here we consider two biliary parasites recently reported in the United Kingdom, Pseudamphistomum truncatum and Metorchis albidus (Trematoda, Opisthorchiidae), and ask whether there are associations between abiotic factors (season, temperature, rainfall and the North Atlantic Oscillation) and the prevalence and intensities of these parasites in otters (n = 586). To control for biotic interactions we first examined whether particular sub-groups of the otter population (grouped by sex, age-class and condition) are more prone to infection and whether any damage is associated with the presence of these parasites. Even though mean intensities of the smaller trematode, P. truncatum (28.3 worms/host), were much higher than M. albidus (4.1), both parasite species had similar impacts on the otter. The distributions of parasites on host sexes were similar, but males suffered greater damage and regardless of sex, parasite intensity increased in older hosts. The probability of infection with either parasite was negatively associated with ground frost, minimum temperatures and rainfall, but was positively associated with warm long-term average temperatures. Although it is widely accepted that multiple variables influence parasite distributions, to our knowledge this is one of only a few studies to examine the combined impact of biotic and abiotic variables on parasites with complex life cycles within their wild definitive host. Identifying such associations can give greater accuracy to predictions concerning the distribution and spread of trematodes with future climate change.

Prevalence, risk factors, and biochemical markers in dogs with ultrasound-diagnosed biliary sludge.

Regarded as an incidental finding, biliary sludge is often diagnosed in dogs on abdominal ultrasound. The aims of the present study were to assess the risk factors, biochemical markers and ultrasonographic findings and to estimate the prevalence and influence of different breeds, sexes, and ages on biliary sludge in dogs. Results demonstrate that the prevalence of biliary sludge is high, especially in senior dogs. The biochemical markers did not have a significant correlation with biliary sludge, and the type of diet was not considered to be the major risk factor. Hepatomegaly was frequently observed on the ultrasound scan of affected animals and of dogs on different systemic drugs and with cardiopathies, which have been referred to as risk groups for the development of inspissated bile.

Interlaboratory agreement and handling variability for consistency of results in paired serum bile acid assays.

Paired serum bile acid (SBA) samples from 30 dogs with presumptive hepatobiliary disease and 30 apparently healthy dogs were submitted to three diagnostic laboratories to validate agreement between these facilities. Sensitivities and specificities ranged from 50% to 70% and from 74% to 85%, respectively, for all laboratories. The κ index confirmed almost perfect agreement between laboratories. Variation in sample handling and processing by different laboratories should not be considered a primary factor when aberrant SBA values are encountered. Documenting agreement between laboratories is essential as inconsistencies could direct unnecessary medical or surgical intervention when patients are evaluated at different hospitals during the course of their disease.