Evaluation of hemorrhage, sample size, and collateral damage for five hepatic biopsy methods in dogs

OBJECTIVES: To compare the volume of hemorrhage, number of lobules, and portal triads available for histologic evaluation, and resultant collateral damage between 5 hepatic biopsy methods: biopsy punch, biopsy needle, ligature method, laparoscopic biopsy forceps, and ultrasonically activated scalpel (UAS). STUDY DESIGN: Experimental, repeated measures, block. ANIMALS: Twelve adult dogs. METHODS: Biopsies were obtained from the periphery and center of the left lateral liver lobe of each dog using each of 5 biopsy techniques. Hemorrhage was quantified and compared between methods and sites. Biopsy samples were evaluated histologically to characterize collateral damage and determine the number of lobules and portal triads sampled. RESULTS: Regardless of technique, liver biopsy resulted in minimal hemorrhage (<2 mL). For peripheral biopsies, UAS was comparable with the ligature method, but caused significantly less hemorrhage than all other methods, whereas for central biopsies, UAS caused significantly less hemorrhage than other methods. Except for the laparoscopic biopsy forceps, UAS caused significantly more collateral damage than other methods. UAS and ligature biopsy methods yielded specimens that had more portal triads per sample than other methods. Eight of 48 biopsy needle samples were inadequate for histologic evaluation, whereas other methods yielded adequate specimens. CONCLUSIONS: All biopsy methods produced minimal hemorrhage and except for needle biopsy yielded adequate tissue samples for histologic evaluation. CLINICAL RELEVANCE: Use of UAS is a reliable, safe alternative technique for liver biopsy and can be used laparoscopically to obtain large tissue samples.     

Oral hirudiniasis in a stray dog,first report in Italy

In June 2014, a male stray dog was recovered at Ente Nazionale di Protezione Animali (ENPA) kennel of Manfredonia, Apulia region, showing oral bleeding and physical prostration. The dog fell in a water canal and was trapped. During the clinical examination, a specimen of leech was revealed into its oral cavity. The parasite, probably entered by drinking unfiltered and contaminated water, has been identified as an adult of aquatic leech Limnatis nilotica. Leeches could overrun wide variety of animals, and few reports about blood sucking leech infestations in mammals are available in literature. This paper describes here the first oral hirudiniasis in a dog in Italy and highlights the possibility of human nasopharyngeal leech-related infection in Apulia region.     

Colchicine treatment in a dog with hepatoportal fibrosis

A two-year-old female spayed English setter with hepatic encephalopathy was found to have hepatoportal fibrosis and acquired portosystemic shunts on laparotomy and liver biopsy. The dog was treated symptomatically with a low-protein diet, lactulose and metronidazole, while colchicine was given as antifibrotic therapy. There was a gradual improvement in clinical signs and liver function until 30 months after first presentation, when the dog developed severe haematuria caused by renal infarction of unknown aetiology. Post mortem examination revealed hepatic fibrosis to be unchanged. This case illustrates that long term survival may be possible in canine hepatoportal fibrosis and suggests that colchicine may be effective in improving liver function and slowing progression of hepatic fibrosis.     

Clinical and histologic outcome in a dog surviving massive hepatic necrosis

This report describes the clinical and histologic recovery of a 2‐year‐old mixed‐breed dog presented with hypovolemic shock, markedly increased serum alanine amino transferase activity, and hemoabdomen. Emergency exploratory surgery revealed a friable liver with multiple capsule hemorrhages necessitating removal of the left lateral lobe. Histologic evaluation showed acute massive hepatic necrosis with centrilobular and midzonal distribution. The dog survived, and all monitored laboratory values normalized within 7 weeks. A liver biopsy taken 8 weeks after presentation revealed normal hepatic architecture with a few, randomly distributed neutrophilic foci. Follow‐up included intermittent determination of liver variables including liver function tests for a period of 7 years. The dog's health status, and all test results remained normal during this time. Complete recovery and good long‐term quality of life after life‐threatening acute liver failure secondary to massive hepatic necrosis is possible in dogs.     

Use of recombinant human factor VIIa in 2 patients with postoperative noncompressible,abdominal hemorrhage

Objective

To describe the clinical presentation, clinical course, and successful management of noncompressible, abdominal hemorrhage with recombinant human factor VIIa (rFVIIa) in 2 postoperative patients.

Case Summary

A 14-year-old neutered female Border Terrier and a 9-year-old neutered male domestic shorthair were treated with rFVIIa to treat noncompressible abdominal hemorrhage in the postoperative period. The dog presented for a septic abdomen following endoscopic intestinal biopsies 10 days prior and was found to have a jejunal perforation along with a fractured liver lobe and hepatic lymphoma at the time of exploratory laparotomy. The cat presented for a spontaneous hemoabdomen associated with hepatic amyloidosis. Clinically significant hemorrhage occurred in the perioperative and postoperative period and both patients received massive transfusions and antifibrinolytic therapy. Despite these interventions, the patients continued to have ongoing abdominal hemorrhage and surgical attempts at hemostasis were not attempted due to the friable nature of the liver at the time of surgery. Both patients received rFVIIa intravenously every 3 hours at a dose between 70 and 90 μg/kg as indicated by the clinical picture, which subsequently decreased transfusion requirements.

New or Unique Information Provided

This case report describes the use of rFVIIa in a cat and a dog with severe, noncompressible abdominal hemorrhage in combination with standard hemostatic interventions.     

In ovo leptin administration affects hepatic lipid metabolism and microRNA expression in newly hatched broiler chickens

Background

A leptin-like immunoreactive substance has been found in chicken eggs and has been implicated in serving as a maternal signal to program offspring growth and metabolism. In the present study, we investigated the effects of in ovo leptin administration on hatch weight, serum and hepatic concentrations of metabolites and hormones, as well as on the expression of genes involved in hepatic lipid metabolism and the predicted microRNAs (miRNAs) targeting the affected genes. To this end we injected fertile eggs with either 0.5 μg of recombinant murine leptin or vehicle (PBS) before incubation.

Results

Prenatally leptin-exposed chicks showed lower hatch weight, but higher liver weight relative to the body weight, compared to the control group. In ovo leptin treatment increased the hepatic content and serum concentration of leptin in newly hatched chickens. The hepatic contents of triglycerides (TG) and total cholesterol (Tch) were decreased, whereas the serum levels of TG, Tch and apolipoprotein B (ApoB) were increased. The hepatic mRNA expression of sterol regulator element binding protein 1 (SREBP-1c), SREBP-2, hydroxy-3-methylglutaryl coenzyme A reductase (HMGCR) and cholesterol 7α-hydroxylase 1 (CYP7A1) was significantly up-regulated, as was the protein content of both SREBP-1c and SREBP-2 in hepatic nuclear extracts of leptin-treated chickens. Moreover, out of 12 miRNAs targeting SREBP-1c and/or HMGCR, five were significantly up-regulated in liver of leptin-treated chicks, including gga-miR-200b and gga-miR-429, which target both SREBP-1c and HMGCR.

Conclusions

These results suggest that leptin in ovo decreases hatch weight, and modifies hepatic leptin secretion and lipid metabolism in newly hatched broiler chickens, possibly via microRNA-mediated gene regulation.     

Plasma concentration of transforming growth factor-β1 and hepatic fibrosis in dogs

Liver fibrosis is a morphologic alteration that accompanies chronic liver diseases. Apart from analysis of liver biopsy specimens, there has been no means of diagnosing and evaluating the course of liver fibrosis in the dog. Several plasma markers, including transforming growth factor beta-1 (TGF-β1), are used to indicate liver fibrosis in humans, but none has been validated for use in dogs. There is a significant correlation between the presence and severity of hepatic fibrosis and the plasma concentration of TGF-β1 in humans with hepatic fibrosis and cirrhosis. The feasibility of using TGF-β1 as a marker for hepatic fibrosis in dogs was evaluated by comparing plasma concentrations in 29 healthy dogs and 18 dogs with liver disease. The plasma concentrations of TGF-β1, were 193 to 598 pg/mL in the healthy dogs, 143 to 475 pg/mL in the 7 dogs with mild hepatic fibrosis or none at all, and 427 to 1289 pg/mL in 11 dogs with moderate to severe hepatic fibrosis. The plasma concentrations of TGF-β1 in the dogs with moderate to severe fibrosis differed significantly (P < 0.001) from those in the other 2 groups, whereas the concentrations in the dogs with mild or no fibrosis did not differ significantly from those in the healthy dogs (P > 0.05). It was concluded that TGF-β1 is a potential plasma marker for hepatic fibrosis in dogs.     

Ultrasonographic differentiation between portal venous and parenchymal gas may be important for the prognosis of canine and feline hepatic emphysema: 37 cases

The aim of this retrospective, cross‐sectional, study was to evaluate clinical findings and outcomes for different ultrasonographic patterns of hepatic emphysema in dogs and cats. Dogs and cats with an ultrasonographic diagnosis of hepatic emphysema and a known outcome, from January 2010 to January 2018, were enrolled. The following data were recorded from medical and ultrasonographic records: ultrasonographic patterns of hepatic emphysema (parenchymal, portal venous, biliary), clinical signs, laboratory findings, and outcomes (favorable, poor). A total of 33 dogs and four cats met the inclusion criteria. Among these, 23 cases were classified as hepatic portal venous gas, 10 as parenchymal emphysema, and four as biliary emphysema. Clinical diagnosis categories were as follows: infection/sepsis (9), gastro‐intestinal disease (9), iatrogenic (9), trauma (5), and liver neoplasia (5). An increase in serum liver enzymes was significantly associated with parenchymal emphysema (P = .03). Other clinical and laboratory findings were not associated with the type of hepatic emphysema. Hepatic portal venous gas was mostly transient in patients with ultrasonographic follow‐up. The overall mortality was 40.5%. A significant difference was found between mortality by portal venous gas (21.7%) and mortality by parenchymal emphysema (90%) (P = .003). In conclusion, the ultrasonographic differentiation of hepatic emphysema between hepatic portal venous gas and parenchymal emphysema may be important for the prognosis of hepatic emphysema. The presence of parenchymal emphysema may be a poor prognostic indicator, while hepatic portal venous gas may be more benign. However, ultrasound findings should be carefully evaluated in the context of clinical findings.     

Chronic hepatitis in the dog

Chronic hepatitis in the dog encompasses a diverse group of liver diseases characterised by a mixed inflammatory infiltrate within the hepatic parenchyma but with multiple causes. The aetiology is often unknown. Potential causes include hepatic copper accumulation, drugs, infections, and, speculatively, immune-mediated disease. Diagnosis is made based on liver biopsy. Treatment is predominantly supportive, while corticosteroids may be indicated in selected cases with idiopathic chronic hepatitis.     

Mycotic aneurysm caused by Graphium species in a dog

A 10-year-old castrated male mixed-breed dog exhibited vomiting, lethargy and anorexia. An abdominal ultrasound examination revealed a focal dilation of the abdominal aorta. The dog died 2 days after examination, and necropsy revealed a ruptured aneurysm at the abdominal aorta between the kidneys. Histological examination revealed severe granulomatous and necrotizing pan-arteritis with fungal organisms. Graphium species was identified through DNA sequence analysis of the PCR product from formalin-fixed paraffin-embedded samples. To our knowledge, this is the first report of aortic aneurysm caused by Graphium species in a dog.     

Accelerated neutrophil apoptosis in 2 canine cases of hepatic disorder

Accelerated neutrophil apoptosis was confirmed by TUNEL assay in two canine cases of hepatic disorder. One dog was diagnosed as having lymphocytic hepatitis and the other lymphocytic cholangitis by histopathology of liver biopsy specimen.     

Peliosis hepatis and hemoperitoneum in a dog with diphacinone intoxication

Objective: To describe the clinical course of a dog presented with peliosis hepatis and hemoperitoneum in concert with anticoagulant rodenticide intoxication.
Case summary: A 7.75-year-old spayed female Shetland Sheepdog presented with clinical signs consistent with hypovolemia, hemoperitoneum, and a history of bright green stool 3 days before the onset of clinical signs. Initial packed cell volume/total solids were consistent with acute hemorrhage. A coagulation profile showed prolongation in activated clotting time and prolongation of both prothrombin and activated partial thromboplastin time, suggesting abnormal coagulation. Abdominal hemorrhage persisted in the face of normalization of the hemodynamic status and coagulation profile, and treatment with Vitamin K₁. Abdominal ultrasound revealed multiple patchy hypoechoic areas throughout the caudate liver lobe. An exploratory laparotomy was performed 24 hours after presentation and revealed the caudate liver lobe as the source of the hemorrhage. Histopathologic examination of a specimen of the liver was consistent with peliosis hepatis. Toxicologic testing identified diphacinone levels in the blood consistent with anticoagulant rodenticide intoxication. Postoperative recovery was uneventful, and within 48 hours the dog was discharged. The dog returned to full function and a hepatic ultrasound performed 15 months postoperatively showed no significant abnormalities.
New or unique information provided: Exposure to anticoagulant rodenticides may be associated with the development of peliosis hepatis in dogs.     

Fatal hemoptysis after bronchoscopic biopsy in a dog

An 8‐year‐old 24.6 kg mixed breed dog underwent bronchoscopy for evaluation of a persistent progressive cough. Bronchoscopy documented a markedly thick and irregular, cobblestone appearance of the mucosa. A bronchoscopic biopsy was obtained; immediately after the biopsy, a large amount of hemorrhage poured from the endotracheal tube. Multiple efforts to control the hemorrhage were unsuccessful and the dog suffered a cardiopulmonary arrest and could not be revived. A necropsy was performed, which was significant for pallor, evidence of prior heartworm disease, prominent bronchial arteries, and erosion of the submucosal vessels at the site of the biopsy. The cause of death was hemorrhage associated with transbronchial biopsy of an enlarged bronchial artery associated with heartworm disease. This report describes a rare complication of a routine diagnostic procedure.     

IMPROVED DETECTION OF METASTATIC HEPATIC HEMANGIOSARCOMA NODULES WITH CONTRAST ULTRASOUND IN THREE DOGS

Three dogs with a splenic hemangiosarcoma were imaged with conventional gray-scale ultrasound and no lesions were identified in the liver. After administration of intravenous ultrasound contrast medium (Definity) small, poorly enhanced, hypoechoic nodules were identified in the liver in each dog. The spleen and liver lesions were identified at surgery and the dogs underwent splenectomy and nodule biopsy. All lesions were identified histologically as hemangiosarcoma. These preliminary results suggest that contrast ultrasound may result in improved detectability of metastatic hepatic hemangiosarcoma.     

Protective Effects of Platycodon grandiflorum Aqueous Extract on Thioacetamide-induced Fulminant Hepatic Failure in Mice

The aim of the present study was to evaluate the protective activity of aqueous extract from Platycodon grandiflorum (BC703) on thioacetamide (TA)-induced hepatotoxicity in mice. We found that BC703 significantly decreased mortality and the change in serum transaminase following TA administration. The group treated with BC703 at doses of 1, 5, and 10 mg/kg produced significant hepatoprotective effects against TA-induced liver damage by decreasing the activities of serum enzymes, nitric oxide and lipid peroxidation in dose-dependent manners. Histopathological studies further substantiated the protective effect of BC703. These results show the hepatoprotective activity of aqueous extract from Platycodon grandiflorum on thioacetamide-induced fulminant hepatic failure.     

Evaluation of Canine-Derived Fibrin Sealant as a Hemostatic Agent

The purpose of this study was to determine whether canine-derived fibrinogen concentrate applied with bovine thrombin was a safe and effective topical hemostatic agent. A canine liver biopsy model was selected to test this product. Cryoprecipitate was prepared from frozen canine plasma using two freeze/thaw/centrifugation cycles. Six healthy adult dogs (weighing more than 18 kg) were used in the fibrin sealant study, and an additional three dogs were used as controls for the liver biopsy. A 1 × 3 cm liver biopsy specimen was obtained, digital pressure was applied to reduce bleeding, and the fibrinogen concentrate was immediately sprayed on the bleeding surface simultaneously with bovine thrombin (1,000 IU/mL). The mean ± standard error of the mean (SEM) blood pressure at time of biopsy was 98 ± 9 mm Hg, and the rate of hemorrhage from the cut liver edge was 8.0 ± 1.1 mL/min. The total blood loss during fibrin sealant application was 37 ± 9 mL and total time for hemostasis was 5.5± 1.3 minutes. There was no additional hemorrhage after application of the fibrin sealant. In the three control dogs, fibrin sealant was not applied and only digital compression was used to decrease hemorrhage. Before digital compression, the rate of hemorrhage from the cut liver edge was 13.1 ±3.1 mL/min. Bleeding had not stopped after 10 minutes of compression and the mean postcompression rate of hemorrhage was 4.0 ± 2.6 mL/min. Signs of secondary bleeding after fibrin sealant was applied were not evident during the immediate postoperative period or over the next 14 days. The coagulation profile, alanine amino transferase (ALT), alkaline phosphatase (AP), and bile acids were not significantly different on day 14 from preoperative values, suggesting that the liver was only mildly affected. The dogs were euthanatized and gross and histological examinations of the biopsy site were performed on day 14. There were minimal or no adhesions at the biopsy site. In all dogs, the hepatic capsule was less than 1 mm thick and histological signs of secondary bleeding, hepatocellular damage, thrombosis, infection, or inflammation were not observed. In this study, canine-derived fibrin sealant was a safe and effective topical hemostatic agent.     

Intracaval Repair of a Persistent Ductus Venosus in a Dog

A persistent ductus venosus was diagnosed in a 4-month-old Old English sheepdog. Total occlusion of blood flow to the liver was achieved through heparinization, hepatic artery occlusion, and placement of bypass cannulas from the portal vein and caudal vena cava to the right atrium. Intracaval closure of the ductus venosus then was accomplished. Despite postoperative complications of continued hemorrhage and acute renal uremia, the animal recovered. The neurologic abnormalities exhibited before surgery resolved, and the dog became more active and gained weight. The dog died due to undetermined causes three months after surgery. The ductus venosus was found to be closed, and the liver was markedly increased in size at necropsy.     

IATROGENIC BILOMA (BILIARY PSEUDOCYST) IN A CAT WITH HEPATIC LIPIDOSIS

An eight-year-old neutered female domestic longhair cat was presented with icterus and a palpable cranial abdominal mass. The cat had undergone an open biopsy of the liver two years previously. Abdominal radiographic findings included a solitary soft tissue mass originating from the right cranial hepatic region. An anechoic cystic structure was identified on ultrasound examination. Surgical exploration revealed the cyst to be filled with bile. The fibrous capsule of the cyst originated in the liver. Severe hepatic lipidosis was also present. It is believed that the bilorna was a complication of the previous hepatic biopsy and not a sequela of hepatic lipidosis.     

Liver disease in the geriatric patient.

Normal functioning of the liver does not seem to change significantly in dogs and cats as a result of age. Despite this, older dogs and cats are at greater risk for the development of liver disease. The diagnosis of liver disease is initiated by the veterinarian's suspicion that liver disease might be present, followed by the case history and a physical examination. The initial workup for the older dog or cat with suspected liver disease should begin with a CBC, serum chemistry profile, and urinalysis. This may be followed by a liver function test, radiographic or ultrasonographic imaging studies, hepatic fine-needle aspiration, and, ultimately, liver biopsy.     

EVALUATION OF HEPATOBILIARY SCINTIGRAPHY AS AN INDICATOR OF HEPATIC FUNCTION IN DOMESTIC PIGEONS (COLUMBA LIVIA) BEFORE AND AFTER EXPOSURE TO ETHYLENE GLYCOL

This study investigated the use of quantitative hepatobiliary scintigraphy to assess liver function in 14 white Carneaux pigeons (Columba livia). Liver scintigraphy using 99mTc-mebrofenin was performed and liver function was quantified using deconvolutional analysis and the area under the normalized heart time-activity curve as previously described in the dog and horse. Liver biopsies were performed in all birds before and after toxin-induced liver damage with ethylene glycol. Before the induction of liver disease, all biopsy specimens showed varying degrees of granulomatous inflammation. After ethylene glycol administration, hepatic lesions were scored and compared with scintigraphic findings. Scintigraphic results showed a significant decrease (P = 0.04) in hepatic function using the area under the normalized time-activity curve. There was good correlation between the overall histologic score posttoxin exposure and scintigraphic measures of liver function (P < 0.03). Based upon these preliminary results, the area under the heart time-activity curve can determine hepatic extraction as a measure of hepatic parenchymal cell function. The results also showed that worsening hepatic cellular function correlated with increased histologic damage to the liver. The use of hepatobiliary scintigraphy using 99mTc-mebrofenin to determine liver function in pigeons has not been previously reported. Additional studies are warranted to evaluate the application of this technique in clinical patients and to establish the sensitivity of this technique.