Objective—To determine total glutathione (GSH) and
glutathione disulfide (GSSG) concentrations in liver
tissues from dogs and cats with spontaneous liver
Sample Population—Liver biopsy specimens from
63 dogs and 20 cats with liver disease and 12 healthy
dogs and 15 healthy cats.
Procedure—GSH was measured by use of an enzymatic
method; GSSG was measured after 2-vinylpyridine
extraction of reduced GSH. Concentrations were
expressed by use of wet liver weight and concentration
of tissue protein and DNA.
Results—Disorders included necroinflammatory liver
diseases (24 dogs, 10 cats), extrahepatic bile duct
obstruction (8 dogs, 3 cats), vacuolar hepatopathy (16
dogs), hepatic lipidosis (4 cats), portosystemic vascular
anomalies (15 dogs), and hepatic lymphosarcoma
(3 cats). Significantly higher liver GSH and protein
concentrations and a lower tissue DNA concentration
and ratio of reduced GSH-to-GSSG were found in
healthy cats, compared with healthy dogs. Of 63 dogs
and 20 cats with liver disease, 22 and 14 had low liver
concentrations of GSH (µmol) per gram of tissue; 10
and 10 had low liver concentrations of GSH (nmol) per
milligram of tissue protein; and 26 and 18 had low
liver concentrations of GSH (nmol) per microgram of
tissue DNA, respectively. Low liver tissue concentrations
of GSH were found in cats with necroinflammatory
liver disease and hepatic lipidosis. Low liver concentrations
of GSH per microgram of tissue DNA
were found in dogs with necroinflammatory liver disease
and cats with necroinflammatory liver disease,
extrahepatic bile duct occlusion, and hepatic lipidosis.
Conclusions and Clinical Relevance—Low GSH values
are common in necroinflammatory liver disorders,
extrahepatic bile duct occlusion, and feline
hepatic lipidosis. Cats may have higher risk than dogs
for low liver GSH concentrations. (Am J Vet Res
Objective—To determine whether metal concentrations in canine liver specimens were influenced by specimen size, assay variability, tissue processing (formalin fixation and deparaffinization), or storage in paraffin blocks.
Sample Population—Liver specimens (fresh frozen and deparaffinized) from 2 dogs with chronic hepatitis (high copper but unremarkable iron concentration [liver 1] and unremarkable copper but high iron concentration [liver 2]) as well as fresh and deparaffinized-archived liver specimens from 20 dogs with various hepatopathies.
Procedures—Fresh frozen liver specimens (obtained via simulated needle-core and wedge biopsy), fresh hepatic tissue, and deparaffinized-archived specimens (0.5 to 14 years old) were analyzed for concentrations of copper, iron, and zinc by atomic absorption flame spectrometry. Clinical severity scores were assigned on the basis of tissue metal concentrations.
Results—Interassay variation of metal standards was < 4%. Measurements of liver tissues on 8 consecutive days yielded high coefficients of variation (3.6% to 50%) reflecting heterogenous histologic metal distribution; variation was highest in liver 1 and deparaffinized-archived tissues. Heterogenous metal distribution was confirmed by histologic evaluation. The largest range of metal concentrations was detected in wedge biopsy specimens. In tissues with high metal concentrations, copper and iron concentrations were significantly lower in needle-core versus wedge biopsy specimens. A higher zinc concentration in deparaffinized-archived specimens masked a low zinc concentration in fresh liver tissue of 10 of 20 (50%) dogs.
Conclusions and Clinical Relevance—Retrospective measurement of copper and iron concentrations but not zinc concentrations in deparaffinized-archived liver specimens provided relevant information. The value of needle-core biopsy specimens for measurement of metal concentrations is questionable.
Objective—To assess the influence of meal ingestion and orally administered erythromycin on gallbladder volume in dogs.
Animals—22 healthy dogs.
Procedures—Ultrasonographically determined gallbladder dimensions in unsedated dogs were used to calculate volume. Measurements were recorded after food was withheld for 12 hours (time 0) and 15, 30, 45, 60, 90, and 120 minutes after a 100-g meal without (n = 22) or with erythromycin (1.0 mg/kg , 2.5 mg/kg , and both dosages ). Gallbladder ejection fraction represented the percentage of volume change from time 0. Intraday and interday coefficients of variation determined operator repeatability and physiologic variation.
Results—We did not detect significant differences in gallbladder volume per unit of body weight between treatments at time 0 or in ejection fraction percentage within or between treatments. Median time 0 gallbladder volume was 0.6 mL/kg (range, 0.4 to 1.9) but was > 1.0 mL/kg in 3 of 22 (14%) dogs and ≤ 1.0 mL/kg in 19 of 22 (86%) dogs. Twenty dogs achieved an ejection fraction ≥ 25% with at least 1 treatment, but 2 dogs with a gallbladder volume ≤ 1.0 mL/kg at time 0 did not. Intraday and interday coefficients of variation were 18% and 25%, respectively.
Conclusions and Clinical Relevance—Gallbladder volume ≤ 1.0 mL/kg at time 0 and ejection fraction ≥ 25% were typical. No treatment consistently induced greater gallbladder contraction. Dogs with a gallbladder volume > 1.0 mL/kg and ejection fraction < 25% may require a combined meal and erythromycin protocol.
OBJECTIVE To establish reference limits for hepatic bile duct-to-arteriole ratio (BD:A) and bile duct-to-portal tract ratio (BD:PT) in healthy cats and assess whether these parameters could be used to support a diagnosis of biliary ductopenia in cats.
SAMPLE Hepatic biopsy samples from healthy cats (n = 20) and cats with ductopenia (2).
PROCEDURES Hepatic biopsy samples from healthy cats were used to count the number of bile ducts and hepatic arterioles in 20 portal tracts for each cat. Mean BD:A and mean BD:PT for each cat were calculated, and these values were used to determine reference limits for mean BD:A and mean BD:PT. Results of histologic evaluation, including immunohistochemical staining in some instances, were compared for healthy cats versus cats with ductopenia.
RESULTS Of the 400 portal tracts from healthy cats, 382 (95.5%) and 396 (99.0%) had BD:A and BD:PT, respectively, ≥ 1.0, with less variability in BD:A. Mean BD:A and BD:PT were markedly lower in both cats with ductopenia, compared with values for healthy cats. However, only mean BD:A for cats with ductopenia was below the reference limit of 0.59.
CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that systematic evaluation of BD:A, with a lower reference limit of 0.59 to define biliary ductopenia in cats, may be a discrete and easily applied morphometric tool to enhance detection of ductopenia in cats. However, application of this ratio required evaluation of ≥ 20 portal tracts with cross-sectioned portal elements to determine a mean BD:A value.
To characterize the frequency and type of bacterial infection by culture- and immunohistochemical (IHC)-based methods and determine the impact of infection on clinical features and survival time in cats with suppurative cholangitis-cholangiohepatitis syndrome (S-CCHS).
168 client-owned cats with S-CCHS (cases).
Clinical features, bacterial culture results, culture-inoculate sources, and survival details were recorded. Cases were subcategorized by comorbidity (extrahepatic bile duct obstruction, cholelithiasis, cholecystitis, ductal plate malformation, biopsy-confirmed inflammatory bowel disease, and biopsy-confirmed pancreatitis) or treatment by cholecystectomy or cholecystoenterostomy. Culture results, bacterial isolates, Gram-stain characteristics, and IHC staining were compared among comorbidities. Lipoteichoic acid IHC staining detected gram-positive bacterial cell wall components, and toll-like receptor expression IHC reflected pathologic endotoxin (gram-negative bacteria) exposure.
Clinical features were similar among cases except for more frequent abdominal pain and lethargy in cats with positive culture results and pyrexia, abdominal pain, and hepatomegaly for cats with polymicrobial infections. Bacteria were cultured in 93 of 135 (69%) cats, with common isolates including Enterococcus spp and Escherichia coli. IHC staining was positive in 142 of 151 (94%) cats (lipoteichoic acid, 107/142 [75%]; toll-like receptor 4, 99/142 [70%]). With in-parallel interpretation of culture and IHC-based bacterial detection, 154 of 166 (93%) cats had bacterial infections (gram-positive, 118/154 [77%]; gram-negative, 111/154 [72%]; polymicrobial, 79/154 [51%]). Greater frequency of bacterial isolation occurred with combined tissue, bile, and crushed cholelith inoculates. Infection and gram-positive bacterial isolates were associated with significantly shorter long-term survival times.
S-CCHS was associated with bacterial infection, pathologic endotoxin exposure, and frequent polymicrobial infection in cats. Combined tissue inoculates improved culture detection of associated bacteria.
To characterize clinical features, comorbidities, frequency of bacterial isolation, and survival time in cats with suppurative cholangitis-cholangiohepatitis syndrome (S-CCHS).
168 client-owned cats with S-CCHS.
Data were prospectively (1980 to 2019) collected regarding clinical features, comorbidities, bacterial infection, illness duration, and treatments. Variables were evaluated for associations with survival time.
Median age of cats was 10.0 years, with no breed or sex predilection observed. Common clinical features included hyporexia (82%), hyperbilirubinemia (80%), lethargy (80%), vomiting (80%), jaundice (67%), weight loss (54%), and hypoalbuminemia (50%). Comorbidities included extrahepatic bile duct obstruction (53%), cholelithiasis (42%), cholecystitis (40%), and ductal plate malformation (44%) as well as biopsy-confirmed inflammatory bowel disease (60/68 [88%]) and pancreatitis (41/44 [93%]). Bacterial cultures were commonly positive (69%) despite prebiopsy antimicrobial administration in most cats. Of surgically confirmed choleliths, diagnostic imaging identified only 58%. Among 55 cats with “idiopathic pancreatitis,” 28 (51%) were documented to have transiting choleliths, and 20 had pancreatic biopsies confirming pancreatitis. Cholelithiasis (with or without bile duct obstruction) and cholecystectomy were associated with survival advantages. Survival disadvantages were found for leukocytosis, ≥ 2-fold increased alkaline phosphatase, and hyperbilirubinemia. Cholecystoenterostomy had no survival impact. Cats with ductal plate malformations were significantly younger at diagnosis and death than other cats. Chronic treatments with antimicrobials, S-adenosylmethionine, and ursodeoxycholic acid were common postbiopsy.
S-CCHS in cats was associated with bacterial infection and various comorbidities and may be confused with pancreatitis. Surgically correctable morbidities (ie, cholecystitis, cholecystocholelithiasis) and cholecystectomy provided a significant survival advantage.
Objective—To evaluate the influence of a 1,4-
butanedisulfonate stable salt of S-adenosylmethionine
(SAMe) administered orally on clinicopathologic
and hepatic effects induced by long-term administration
of prednisolone in dogs.
Animals—12 healthy dogs.
Procedure—Following a pilot study (4 dogs), 2 groups
of 4 dogs received prednisolone (2.2 mg/kg) orally
once daily (84-day trial). One group received SAMe
(20 mg/kg/d divided in 2 doses) for 42 days and then
a placebo for 42 days; the other group received treatments
in the reverse order. Before and during the
trial, numerous variables were monitored, including
serum total alkaline phosphatase (ALP) and glucocorticoid-
induced ALP (G-ALP) activities, serum haptoglobin
concentration, and total and oxidized glutathione
(TGSH and GSSG) and thiobarbiturate-reacting
substances (TBARS) concentrations in erythrocytes
and liver tissue (days 0, 42, and 84). Hepatic
specimens also were examined microscopically.
Results—The stable salt of SAMe was biologically
available; plasma concentrations of SAMe or prednisolone
were not affected by coadministration.
Compared with baseline values, serum ALP and GALP
activities and haptoglobin concentrations
increased and erythrocyte GSSG and TBARS concentrations
decreased with both treatments. Erythrocyte
TGSH concentration decreased with the prednisolone-
placebo treatment. Administration of SAMe
appeared to conserve erythrocyte TGSH values and
did not inhibit hepatocyte glycogen vacuolation but
increased hepatic TGSH concentration and improved
the hepatic tissue GSSG:TGSH ratio.
Conclusions and Clinical Relevance—In dogs,
administration of 20 mg of SAMe/kg/d may mitigate
the apparent pro-oxidant influences of prednisolone
but did not block development of classic clinicopathologic
or histologic features of vacuolar hepatopathy.
(Am J Vet Res 2005;66:330–341)
Objective—To investigate the influence of dietary supplementation with l-carnitine on metabolic rate, fatty acid oxidation, weight loss, and lean body mass (LBM) in overweight cats undergoing rapid weight reduction.
Procedures—Cats fattened through unrestricted ingestion of an energy-dense diet for 6 months were randomly assigned to 4 groups and fed a weight reduction diet supplemented with 0 (control), 50, 100, or 150 μg of carnitine/g of diet (unrestricted for 1 month, then restricted). Measurements included resting energy expenditure, respiratory quotient, daily energy expenditure, LBM, and fatty acid oxidation. Following weight loss, cats were allowed unrestricted feeding of the energy-dense diet to investigate weight gain after test diet cessation.
Results—Median weekly weight loss in all groups was ≥ 1.3%, with no difference among groups in overall or cumulative percentage weight loss. During restricted feeding, the resting energy expenditure-to-LBM ratio was significantly higher in cats that received l-carnitine than in those that received the control diet. Respiratory quotient was significantly lower in each cat that received l-carnitine on day 42, compared with the value before the diet began, and in all cats that received l-carnitine, compared with the control group throughout restricted feeding. A significant increase in palmitate flux rate in cats fed the diet with 150 μg of carnitine/g relative to the flux rate in the control group on day 42 corresponded to significantly increased stoichiometric fat oxidation in the l-carnitine diet group (> 62% vs 14% for the control group). Weight gain (as high as 28%) was evident within 35 days after unrestricted feeding was reintroduced.
Conclusions and Clinical Relevance—Dietary l-carnitine supplementation appeared to have a metabolic effect in overweight cats undergoing rapid weight loss that facilitated fatty acid oxidation.
Objective—To evaluate differences in hepatic copper concentrations in Labrador Retrievers with and without chronic hepatitis.
Design—Retrospective case-control study.
Sample—Liver tissue specimens from 36 Labrador Retrievers with chronic hepatitis and 36 age- and sex-matched Labrador Retrievers without chronic hepatitis (control dogs).
Procedures—Liver tissue specimens were obtained during 2 study periods (1980 to 1997 and 1998 to 2010). For each tissue specimen, a histologic score was assigned independently by each of 2 interpreters, and the hepatic copper concentration was qualitatively determined via rhodanine staining and quantitatively determined via atomic absorption spectroscopy.
Results—Mean hepatic copper concentration was significantly higher in dogs with chronic hepatitis (614 μg/g of dry weight [range, 104 to 4,234 μg/g of dry weight]), compared with that in control dogs (299 μg/g of dry weight [range, 93 to 3,810 μg/g of dry weight]), and increased significantly over time. A higher proportion of liver tissue specimens collected during the 1998–2010 study period had hepatic copper concentrations > 400 μg/g of dry weight (the upper limit of the reference range), compared with the proportion of liver tissue specimens collected during the 1980–1997 study period. The qualitative copper score did not accurately predict quantitative hepatic copper concentration in 33% of study dogs.
Conclusions and Clinical Relevance—Results suggested that the increase in hepatic copper concentrations in Labrador Retrievers with and without chronic hepatitis over time may be the result of increased exposure of dogs to environmental copper, most likely via the diet.
Objective—To evaluate prognostic factors, survival,
and treatment protocols for immune-mediated
hemolytic anemia (IMHA) in dogs.
Animals—151 dogs with IMHA not associated with
underlying infectious or neoplastic disease.
Procedure—Information recorded from review of medical
records included signalment at the time of initial
evaluation; vaccination history; 30-, 60-, and 365-day follow-up outcomes; laboratory data; results of imaging
studies; and necropsy findings. Dogs were grouped
according to the presence of spherocytes, autoagglutination,
a regenerative erythrocyte response, and treatments
received (azathioprine, azathioprine plus ultralowdose
aspirin, azathioprine plus mixed–molecular-weight
heparin [mHEP], or azathioprine plus ultralow-dose
aspirin plus mHEP) for comparisons. All dogs received
Results—Cocker Spaniels, Miniature Schnauzers,
neutered dogs, and female dogs were overrepresented.
Alterations in certain clinicopathologic variables were
associated with increased mortality rate. Rates of survival
following treatment with azathioprine, azathioprine
plus ultralow-dose aspirin, azathioprine plus mHEP, and
azathioprine plus ultralow-dose aspirin plus mHEP were
74%, 88%, 23%, and 70%, respectively, at hospital discharge;
57%, 82%, 17%, and 67%, respectively, at 30
days; and 45%, 69%, 17%, and 64%, respectively, at 1
year. In comparison, mean survival rates at discharge
and at 30 days and 1 year after evaluation collated from
7 published reviews of canine IMHA were 57%, 58%,
and 34%, respectively.
Conclusions and Clinical Relevance—Treatment
with a combination of glucocorticoids, azathioprine,
and ultralow-dose aspirin significantly improved short-and
long-term survival in dogs with IMHA. (J Am Vet
Med Assoc 2005;226:1869–1880)