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Ascites is a pathological accumulation of free fluid
within the peritoneal cavity (Hall and German 2005) and it
is mostly accompanied with sever chronic liver disease due
to portal hypertension, hypoalbuminenia and increased renal
sodium and water retention (Rutgers and Biourge 2007).
The diagnosis of liver disorders is a challenge to clinicians
because clinical signs are often very vague and non-specific,
especially in the early stage of hepatobiliary diseases.
Because of varied functions of liver, no single test can
accurately identify the hepatic disease or its underlying
cause hence, a battery of test is required to assess ascites
due to hepatobiliary disorder. So, comprehensive study of
ascitic fluid, serum biochemical and serum marker is the
need of the present scenario for proper diagnosis of ascites
due to hepatic damage. Therefore the present study was
taken up to detect the haemato-biochemical, ascitic fluid
analysis and ultrasonography changes of ascites due to
hepatobiliary disorders in dog.
MATERIALS AND METHODS
Dogs having signs of distended abdomen presented at
Referral Veterinary Polyclinic (RVP), Indian Veterinary
Research Institute, Izatnagar, was taken under study. History
of each dog was noted in relation to breed, age, sex, body
weight etc. for epidemiological study. The samples of ascitic
fluid and venous blood samples were obtained on the same
day. Collected serum samples were stored in deep freezer
at –20°C for biochemical estimation.
Haematology: Haemoglobin, PCV, total erythrocyte and
leukocyte counts and red blood cell indices (MCV, MCH,
MCHC).
Serum biochemistry: BUN, creatinine, ALT, AST, ALP,
total bilirubin, total protein (TP), albumin, sodium and
potassium and prothrombin time (PT).
Ultrasonography: Ultrasonographic examination was
carried out with 2.5–5.0 MHz transducers.
Abdominal paracentesis: Abdominal paracentesis was
performed on standing and or left lateral recumbency and
the surrounding area was clipped and full surgical
preparation was done. Site was chosen 2 to 3 cm caudal to
the umbilicus and 2 to 3 cm left of the midline. Urinary
bladder was emptied before centesis. A 22 or 20 G needle
or catheter was inserted at an angle of 45o to the body wall.
Fluid was collected directly into the sterile vials with or
without EDTA for further analysis (Alleman 2003, Rudolff
2005).
Ascitic fluid analysis: (a) Physical examination: Colour/
turbidity and specific gravity. (b) Microscopic examination:
1. Cell count was done by haemocytometer counting
chamber. 2. Cytological examination: Ascitic fluid cytology
smear was stained by Giemsa. (c) Biochemical test: Total
protein, albumin and serum ascitic albumin gradient
(Alleman 2003).
Ph.D. work of first author, approved by Deemed University,
Indian Veterinary Research Institute, India.
Present address: 1,6Assistant Professor (sara82vet
@yahoo.com, sasivetext@gmail.com), Teaching Veterinary
Clinical Complex, VCRI Orathanadu, Thanjavur. 2Principal
Scientist (dbmondal@gmail.com). 3Assistant Professor (SS;
kalyan_srm@rediffmail.com); 4Scientist (mahivet2002
@yahoo.co.in). 5Assistant Professor (vijayvet1985@gmail.com),
MVC, Chennai.
Indian Journal of Animal Sciences 84 (5): 503–506, May 2014/Article
Comprehensive study of haemato-biochemical, ascitic fluid analysis and
ultrasonography in the diagnosis of ascites due to hepatobiliary disorders in dog
M SARAVANAN1, D B MONDAL2, K SARMA3, K MAHENDRAN4, H VIJAYAKUMAR5 and V SASIKALA6
Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh 243 122 India
Received: 1 December 2012; Accepted: 18 January 2014
ABSTRACT
Two percent of the dogs with ascites of hepatic insufficiency were recorded. Decline in the values of Hb, PCV,
RBCs and increased TLC and prothrombin time were recorded. Hypoproteinemia, hypoalbuminemia with elevated
bilirubin and liver enzymes were observed. Abdominal ultrasound revealed anechoic structure with focal and diffused
hyper echoic, cirrhosis of liver and gall bladder disorders were recorded. Ascitic fluid was clear/transudate with a
few mesothelial cells, lymphocytes, monocytes and neutrophils in most of the ascitic dogs. Decreased serum total
protein, albumin and elevated serum ALT, AST, ALP, total bilirubin, prolonged prothrombin time and SAAG was
good indicator for diagnosis of ascites due to HBD. Ascitic fluid analysis and serum ascites albumin gradient
(SAAG) may be a key indicator for diagnose etiopathogenesis of ascites in dog.
Key words: Abdominal paracentesis, Ascites, Liver, Portal hypertension
504 SARAVANAN ET AL. [Indian Journal of Animal Sciences 84 (5)
32
Serum ascitic albumin gradient (SAAG): It was
calculated by subtracting the albumin concentration of the
ascitic fluid from the albumin concentration of a serum
obtained on the same day (Beg et al. 2001).
Statistical analysis: All data were expressed as
means±SE. The analyses were performed with SPSS 16.0.
The Student’s t-test was used to analyze the study
parameters.
RESULTS AND DISCUSSION
Incidence: Total number of dogs brought to RVP, IVRI,
Izatnagar irrespective of the nature of the diseases were
3,600 during the course of the study (March 2011– 2012).
Out of these, 72 dogs (2.0 %) were observed with ascites
due to hepatic insufficiency. Spitz dogs (54.2%) had higher
incidence followed by Mongrel (16.4%), Labrador Retriever
(12.5%), German Shepherd (6.9%), Doberman pinscher
(6.9%), Great Dane (1.4%) and Golden Retriever (1.4%).
Higher incidence was recorded in male mongrel dogs
(75.0%) whereas in female Doberman pinscher (80.0%)
showed higher incidence. Most of the ascities were noticed
more than 5 years old (33.3%) followed by 4–5 years
(15.3%), 1–2 years (13.9%), 2–3 years (12.5%), 3–4 years
(12.5%) and less than one year (12.5%) of age. 2–3 years
old male (88.9%) had higher incidence whereas in female
more than 5 years (66.7%) old dogs. In the present study
the overall sex-wise distribution revealed that male dogs
(54.2%) had higher incidences of ascites. Most of the ascites
dogs were fed with homemade vegetarian diet. Cases of
ascites were recorded from time to time by various
researchers in different breeds of dog namely Labrador
Retriever (James et al. 2008), German Shepherd (Nottidge
et al. 2003), Doberman pinscher (Fuentealba et al. 1997)
and Great Dane (Raffan et al. 2009). Various authors have
reported ascites in different age groups, viz. from 16 weeks
to 9 years (mean 3.12 years) and from 1 to 13 (mean 5)
years (James et al. 2008). Ascites is being considered to be
a common complication of chronic hepatitis in dogs (Raffan
et al. 2009) and it could be aggravated by the activation of
hepatic stellate cells with subsequent presinusoidal collagen
accumulation and fibrosis, that results in sinusoidal
occlusion followed by portal hypertension and promote the
formation of fluid accumulation (Hall and German 2005).
Clinical signs: All the dogs showed abdominal distension
followed by inappetance (69.4%), lethargy (63.9%), pale
mucous membrane (45.8%), respiratory dyspnoea (15.3%),
pedal oedema (12.5%), diarrhoea (8.3%), vomiting (6.9%),
melena (6.9%) and polyuria/polydipsia (4.2%). The clinical
signs of hepatobiliary dysfunctions (HBD) are relatively
vague and highly variable in nature. The findings of present
study were similar to the observations of various authors
(Watson and Bunch 2009, Saravanan et al. 2012).
Haematology: Highly significant (P<0.001) decrease in
Hb, PCV and RBCs count were noticed in ascitic dogs.
Significant (P<0.05) increase in TLC, neutrophil and
basophil count and decreased lymphocyte count were
observed as compared to healthy dogs (Table 1). Liver being
the prime organ, involved in the production of erythropoietin
and other factors required for erythropoiesis and so the
dysfunction of liver may lead to anaemia (Sharma et al.
2001). Washabau (2010) observed severe leukocytosis and
neutrophilia in dogs with granulomatous hepatitis, hepatic
cirrhosis, hepatic abscess and hepatic neoplasia and
emphasized the probable causes for significant increase in
total leukocytes, neutrophil and basophil counts in the study.
Serum biochemical profile: Significant increase (P<0.05)
in ALT, AST, ALP, total bilirubin and prolonged
prothrombin time activities were observed in ascitic dogs.
Serum AST and ALT measurements were highly useful in
detecting hepatocellular injury and monitoring clinical
progress (Tennant and Center 2008). Solter et al. (1994)
stated that the high serum ALP as a sensitive marker for
cholestasis in most of the mammalian species including
dogs. Highly significant (P<0.001) decrease in total protein
and albumin level were noticed in ascitic dogs (Table 2),
which could be due to the primary role of liver in the
synthesis of major plasma protein as well as site of
degradation and synthesis of many other proteins that is
influenced by liver diseases in many ways (Webster 2005).
Ascites leads to increased albumin distribution and lowers
the blood albumin concentration, which decreases the
plasma oncotic pressure and aggravates the ascites (Richter
Table 1. Mean±SE of haematological profile in ascitic dogs
Parameters Control (n=6) Ascites dog (n=72)
Hb (g/dl) 12.28±0.23 8.90±0.25**
PCV % 43.0±0.83 27.37±0.68**
RBC (×106/ /μl) 4.31±0.07 2.73±0.10**
MCV (fl) 100.29±2.95 104.83±2.44NS
MCH (pg) 30.03±0.74 33.73±0.68NS
MCHC (%) 29.97±0.18 32.46±0.36NS
WBC (×103/ /μl) 10.39±0.42 18.57±0.87*
Lymphocyte (%) 27.67±1.05 18.50±1.01*
Monocyte (%) 1.67±0.33 1.93±0.13NS
Neutrophil (%) 69.50±0.92 78.19±1.07*
Basophil (%) 0.17±0.17 0.87±0.09*
Eosinophil (%) 1.0±0.26 0.61±0.11NS
Table 2. Mean±SE of serum biochemical profile in ascitic dogs
Parameters Control (n=6) Ascites dog (n=72)
BUN (mg/dl) 26.26±1.89 31.29±3.26NS
Creatinine (mg/dl) 0.62±0.15 1.17±0.15NS
Prothrombin time (sec) 5.0±0.25 8.50±0.31*
ALT (IU/L) 41.55±3.37 173.47±12.29*
AST (IU/L) 29.72±2.0 161.18±11.15*
ALP (IU/L) 48.51±2.28 120.74±7.89*
Total protein (g/dl) 6.63±0.09 5.15±0.10**
Albumin (g/dl) 3.31±0.09 2.02±0.03**
Globulin (g/dl) 3.32±0.04 3.13±0.10NS
A: G ratio 1.0±0.03 0.83±0.14NS
Total bilirubin (mg/dl) 0.36±0.03 0.99±0.07*
Na+ (mEq/L) 136.3±0.8 139.2±1.7NS
K+ (mEq/L) 4.9±0.1 4.3±0.1NS
May 2014] DIAGNOSIS OF ASCITES DUE TO HEPATOBILIARY DISORDERS IN DOG 505
33
2003, Tennant and Center 2008). Estimation of serum
albumin levels and prothrombin time are often considered
tests of liver function. These findings are agreeing with the
present study.
Ultrasonography: All the 72 dogs were observed with
anechoic structure (ascitic fluid) in abdominal cavity
followed by 18 dogs (25%) with focal hyper echoic and 33
dogs (45.8%) showed diffused hyper echoic liver
parenchyma (Fig. 1) with normal size of liver. Liver
cirrhosis (Fig. 2) were found in 19 dogs (26.38%) as
evidenced by hyper echoic bright small size liver with
irregular margin. Gall bladder disorders (cholecystitis/
cholelithiasis/ distension) (Fig. 3) were noticed in 10 dogs
(13.90%). Also multiple hepatic cysts in 1 case (1.4%) and
multiple hepatic nodules in 1 case (1.4%) were recorded.
In the present study, liver, kidney (right and left), spleen,
intestine and urinary bladder were suspended in ascitic fluid
during USG examination.The presence of ascitic fluid in
the abdominal cavity greatly enhanced the image of various
abdominal organs, viz. liver, spleen, intestine, urinary
bladder and right and left kidney. Mc Grothy and Doust
(2004) stated that the abdominal ultrasound is being
considered as more sensitive than survey radiography to
detect peritoneal fluid. Cholecystitis as generalized gall
bladder wall thickening is associated with acute
pyelonephritis, portal hypertension, chronic renal diseases,
and hepatitis (Mattoon and Nyland 2002).
Ascitic fluid analysis: Ascitic fluid was aspirated in
severely distended/ tensed abdomen to relive respiratory
dysponea. Colour/turbidity of ascitic fluid was clear/
transudate in 68 dogs followed by clear/straw colour in 2
dogs and clear reddish yellow in 2 dogs. Cytologically,
ascitic fluid revealed a few mesothelial cells, lymphocytes,
monocytes and neutrophils (Fig. 4) in most of the ascitic
dogs, whereas two dogs showed tumor cells in the ascitic
fluid. Specific gravity (1.014±0.001), total nucleated cell
count (388.93±66.32/cmm), total protein (1.96±0.08 g/dl),
albumin (0.83±0.05 g/dl) and serum ascites albumin
gradient (1.20±0.05 g/dl) of ascitic fluid were showed
transudative ascites. Paracentesis could assist in timely
identifying the pathological causes which are responsible
for the fluid accumulation or it can help further investigation
to diagnose the diseases condition (Papasouliotis and
Dewhurst 2005). The mean specific gravity, total nucleated
cell count, total protein, albumin and serum ascites albumin
gradient found to be a transudative type of ascites of the
present study. These findings are in line with the findings
of Burgees (2004).
The SAAG reflects the oncotic pressure gradient between
the vascular bed and elevated gradient (greater than or equal
to 1.1 g/dl) usually being associated with increased portal
pressure, whereas a low gradient (<1.1 g/dl) is associated
with conditions where the ascites is not related to portal
hypertension (Tarn and Lapworth 2010). Burgees (2004)
and Saravanan et al. (2012) opined that the serum-ascites
albumin gradient (SAAG) provides better discrimination
between ascites of chronic liver diseases from other origin.
Hence, SAAG > 1.1 g/dl is suggestive of the presence of
portal hypertension due to chronic liver disease.
Refractory form of ascites were noticed in 16 dogs
(22.22%), of which 3 dogs showed recurrence within a
month and the remaining 13 dogs after 1 month from the
day of clinical presentation and standard therapeutic
intervention. Among these 13 recurrent ascitic dogs, one
dog exhibited recurrence of ascites after 6th, 8th, 11th, 13th
and 15th month during standard therapeutic intervention.
However, 3 dogs exhibited ascites recurrence at 4th, 6th, 8th
and 10th month of standard therapeutic intervention.
Reoccurrence of ascites in dogs is mostly because of the
pet owners discontinuing therapy once after their pets
getting clinical aid, response within few day of therapy.
Hence, this also could be one of the reasons for recurrence
and diuretic resistance ascites which were observed in the
present study. Cirrhotic patients with ascites were
characterized by marked alterations in their splanchnic and
systemic haemodynamics leading to spanchnic congestion,
central hypovolaemia and arterial hypertension with
activation of rennin-angiotensin and vasoconstrictor
systems with renal increase of sodium reabsorption. One
of the most adverse consequences of such hemodynamic
dysfunctions were the development of refractory ascites in
which standard medical treatment with low sodium diet and
Figs. 1–3. 3. USG: Diffused hyper echoic liver parenchyma with massive ascitic fluid (sagittal view). 2. USG: Cirrhotic liver
parenchyma with massive ascitic fluid (sagittal view). 3. USG: Cirrhotic liver parenchyma, gall stone (cholelithiosis) with ascitic fluid
(sagittal view).
506 SARAVANAN ET AL. [Indian Journal of Animal Sciences 84 (5)
34
diuretics are unable to resolve the ascites. Further massive
ascites and hepatorenal syndrome (HRS) are frequent
complication of liver cirrhosis (Gerbes and Gulberg 2006).
Refractory ascites generally affects with advanced cirrhosis
that may also develop HRS. Both the refractory ascites and
HRS are the independent predictors of short survival
(Salerno et al. 2010). Decreased serum total protein,
albumin level and increased ALT, AST, ALP, total bilirubin,
prothrombin time and serum ascites albumin gradient
(SAAG) were good indicator for diagnosis of ascites due
to HBD.
ACKNOWLEDGMENTS
Authors are extremely grateful to the Director, Indian
Veterinary Research Institute, Izatnagar, for providing all
facilities for the study.
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