ABSTRACT Fluoride has been considered to cause hepatic and renal tissue damages in animals and humans. The present study investigated the effect of varying concentrations of fluoride on hepatic and renal enzyme profile in domestic chicken (n=80). Chicken were distributed into 4 groups. Group A was kept unexposed while group B, C and D were exposed to 10, 20 and 30 µg/g body weight of NaF respectively on weekly basis for four weeks. Alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine amino-transferase (ALT) and bilirubin were determined as indicators of liver function test (LFT), while uric acid was as a parameter for renal function test (RFT). All LFT and RFT parameters showed high values (P&lt; 0.05) after one, two three and four weeks in all groups. 579.4 ± 1.55, 355.0 ± 2.13, 246.2 ± 2.45 and 0.83 ± 1.46 were the ALP, AST, ALAT and bilirubin values for LFT and uric acid was 6.74 ± 2.92 in D group at the end of four weeks. All these results indicate the probability of severe effect on the physiology of the liver and kidneys in the exposed birds

A Khalique

A Yaqub

M A Rashid

M Z Yousaf

M.S K M Anjum

Mughal

N Mumtaz

U Sayyed

CiteSeerX

Anjum et al., J. Anim. Plant Sci. 24(1):2014
77
INFLUENCE OF INCREASING FLUORIDE DOSE RATES ON SELECTED LIVER AND
KIDNEY ENZYMES PROFILE IN DOMESTIC CHICKEN (Gallus domesticus)
K. M. Anjum, M.S. Mughal*, U. Sayyed**, A. Yaqub**, A. Khalique***, M.A. Rashid***, M. Z. Yousaf*†, and N. Mumtaz*††
Department of Wildlife and Ecology, University of Veterinary and Animal Sciences, Lahore, Pakistan
*Department of Fisheries and Aquaculture, University of Veterinary and Animal Sciences, Lahore, Pakistan.
**Department of Zoology, GC University, Lahore, Pakistan
***Department of Animal Nutrition, University of Veterinary and Animal Sciences, Lahore, Pakistan
†Nano-Biotechnology Lab, Department of Advanced Materials, College of Engineering, Peking University, Beijing, China
*††Department of Zoology, University of Education, Bank Road Campus, Lahore, Pakistan
Corresponding author email: raviankhalid@gmail.com
ABSTRACT
Fluoride has been considered to cause hepatic and renal tissue damages in animals and humans. The present study
investigated the effect of varying concentrations of fluoride on hepatic and renal enzyme profile in domestic chicken
(n=80). Chicken were distributed into 4 groups. Group A was kept unexposed while group B, C and D were exposed to
10, 20 and 30 µg/g body weight of NaF respectively on weekly basis for four weeks. Alkaline phosphatase (ALP),
aspartate aminotransferase (AST), alanine amino-transferase (ALT) and bilirubin were determined as indicators of liver
function test (LFT), while uric acid was as a parameter for renal function test (RFT). All LFT and RFT parameters
showed high values (P< 0.05) after one, two three and four weeks in all groups. 579.4 ± 1.55, 355.0 ± 2.13, 246.2 ± 2.45
and 0.83 ± 1.46 were the ALP, AST, ALAT and bilirubin values for LFT and uric acid was 6.74 ± 2.92 in D group at the
end of four weeks. All these results indicate the probability of severe effect on the physiology of the liver and kidneys in
the exposed birds.
Key words: Fluoride intoxication, domestic chicken, LFT, RFT, bilirubin.
INTRODUCTION
Although fluoride intake is necessary for the
development of teeth and body skeleton but requirements
are in traces. However, a few recent studies indicated that
more fluoride intake might cause toxic effects in animals
and human being (Bhatnagar et al., 2011). Fluoride
intake (fluorosis) usually occurs in two forms; endemic
fluorosis, related to intake of drinking water with high
fluoride contents (Li and Cao, 1994) and industrial
fluorosis, related to exposure to air rich in fluoride
contents (Czerwinski et al., 1988; Grandjean et al.,
1985). Fluorosis has been found to cause severe side
effects, not only to skeletal parts of the body (Finkelman
et al., 1999; Wei, 1979) but also to the soft tissues like
brain, liver, kidney and spinal cord (Guan et al., 1998;
Wang et al., 2004).
Earlier work by Guan et al. (2000) reported that
50-80% of the absorbed fluorides are eliminated by the
kidneys indicating the chances of kidney damages due to
fluorosis. Liver is the main organ for fluoride
detoxification and, therefore, is highly susceptible to the
fluoride intoxication (Wang and Li, 2002). Various
studies demonstrated that elevated levels of serum
hepatic and renal enzymes have been found following
fluoride intoxication indicating degenerative and
inflammatory damages to the liver and kidney
(Shivashankara et al., 2000; Wang and Li, 2002; Shashi
and Thapar, 2001). Other histopathological findings
include hyperplasia, dilatations of hepatic sinusoids
(Kour et al., 1981), and accumulation of amorphous and
crystalline bodies in the hepatocytes around the hepatic
vein (Phillips et al., 1934), and in some rare cases the
hepatocholan-giocarcinomain (Maurer et al., 1990). It has
been found that fluoride intoxication induced hepatic
hemangiosarcoma, hepatocellular adenoma and
carcinoma, metastatic lung tumors, and Zymbal’s gland
tumors in rats (Bogdanffy et al., 1995).
Commercial poultry production is being done
extensively throughout the Pakistan. Further,
management and production varies with the region which
might be due to changes in the water and the presence of
industrial wastes. Fluoride intoxication could be one of
the intoxicants. Therefore, considering the growth of
poultry sector and intensification of poultry farms around
the urban areas, we hypothesized that higher levels of
fluoride can affect the profile of liver and kidney
enzymes. Selected serum liver enzymes were; alkaline
phosphatase, aspartate aminotransferase, alanine
aminotransferase, bilirubin and uric acid and for kidneys.
Therefore, objective of current experiment was to
evaluate the effect of fluoride intoxication on changes in
liver and kidney enzymes of domestic chicken (Gallus
domesticus).
The Journal of Animal & Plant Sciences, 24(1): 2014, Page: 77-80
ISSN: 1018-7081
Anjum et al., J. Anim. Plant Sci. 24(1):2014
78
MATERIALS AND METHODS
Eighty, a day old, domestic chickens (Gallus
domesticus) were purchased from a local market. Chicks
were reared till the age of four weeks before the start of
experiment. The experimental protocols were approved
by the ethical committee on animal research, GC
University, Lahore (GCU).
Experimental Design: The birds were reared at the
Animal House, Government College University, Lahore.
After adaptability period, chicks were assigned one of the
four dietary treatments namely A, B, C and D with 20
chicks per treatment. Group A served as a control group
that was not treated with any fluoride intoxication;
whereas, in group B, C and D, chicks were injected a
weekly dose of NaF 10, 20, and 30 µg per gram of body
weight, respectively. Chicks were sacrificed at the end of
experiment and blood samples were collected.
Determination of Activities of Enzymes: Blood was
collected from wing vein of chicken and then centrifuged
at 5,000 rpm for five minutes to harvest the serum that
were stored at - 20°C till further analysis. At the time of
analyses, the serum samples were thawed and
concentrations of alkaline phosphatase, aspartate
aminotransferase, alanine aminotransferase, bilirubin and
uric acid were determined using (MEDPHARM)
commercial kits.
Data Analysis: Data, presented as mean ± S.D, were
analyzed using one-way analysis of variance (ANOVA)
and Tukey’s posthoc multiple comparison test. Data
analysis was performed using statistical package SPSS
(Version 13.0 SPSS Inc., Chicago, IL, USA) and results
were declared significant at P< 0.05.
RESULTS AND DISCUSSION
When chicks were exposed to acute fluoride
intoxication, higher (P<0.05) values of ALP were
observed in all treatment groups. The ALP in control
group was reported as 339.2 ± 1.44. The present study
shows that ALP increased in an increased dose of
fluoride manner, in group B ALP was 444.8 ± 2.32;
whereas 507.1 ± 1.54 and 579.4 ± 1.55 was observed
ALP in group C and D. These results clearly indicate that
the ALP was higher in those groups receiving the higher
dose of fluoride intoxicant as shown in Table 1.
The serum AST also increased (P<0.05) with the
increasing dose of fluoride. In control group, AST was
198.4 ± 2.44; whereas, in group B concentration was
268.2 ± 1.90. Highest values 310.9 ± 2.12 and 355.0 ±
2.13 of AST were observed in C and D groups
respectively. Furthermore, serum ALT was 168.7 ± 1.55,
204.7 ± 4.18, 220.0 ± 3.56 and 246.2 ± 2.45 in the control
B, C and D groups respectively shown (Table 1 & Fig1).
The bilirubin level in control was 0.52 ± 0.04 U/L which
increased in all treatment groups. The highest level of
bilirubin (0.83 ± 1.46 U/L) was reported in the group D at
the end of one month exposure to NaF as shown in Fig 2.
Increased serum bilirubin level indicates the
impairment of liver functions as it is generally considered
as a true test for proper liver functioning. It reflects the
liver s’ ability to take up and secrete bilirubin into the
bile. The increase in ALP, AST, ALT and bilirubin was
due to the degeneration and necrosis of liver cells. Shashi
and Thapar (2001) reported that necrosis depends on the
time of fluoride exposure. They showed that hepatic cell
necrosis and other structural changes were more
pronounced in the rabbits exposed to fluoride intoxication
for three months than those exposed for one month.
Similar study was conducted in human by Michael et al.
(1996) to investigate the effect of high dose of fluoride
intake from drinking water on some soft tissues and
found that high levels of fluoride disturbed the normal
ALT and AST values. The elevated ALP, AST, ALT and
bilirubin is an indication of the impairment of liver
functions. Most of the liver function enzymes were found
abnormal in the children having skeletal fluorosis
(Shivashankara et al., 2000). All these studies support our
present findings of higher AST, ALP, ALT and bilirubin
levels in the blood serum of chicken exposed to high
doses of fluoride.
RFT parameter, uric acid also increased (P<
0.05) in treatment groups. The uric acid level in the
control was 3.63 ± 2.45; whereas 4.54 ± 2.86, 5.55 ± 1.88
and 6.74 ± 2.92 (Table 1) were the reported values of uric
acid after one month in group A, B, C and D respectively
as shown in Fig 2. High uric acid indicates the
impairment in the kidney functions as there are several
evidences that hyperuricemia is the causative agent of
kidney diseases (Sowers, 2004). High uric acid also aids
in the development of other diseases like the albuminuria,
microvascular disease, glomerulosclerosis, and
tubulointerstitial fibrosis (Nakagawa et al., 2002). There
are evidences that uric acid is a potent risk factor for
kidneys in normal population as well as the population
having kidney disease due to immunoglobulin A (IgA)
nephropathy. A comparative study also indicate that
hyperuricemia have more severe effects to the kidneys
than proteinuria and make the kidneys less efficient and
develop the kidney insufficiency (Sanchez-Lozada et al.,
2005). High uric acid is a prediction of hypertension,
metabolic syndrome (Masuo et al., 2003), diabetes
(Nakanishi et al., 2003) and stroke (Bos et al., 2006). The
present study provides us informations about the serum
enzymes which are basic indicators of liver and kidney
functioning at different fluoride exposure. It will be
helpful to devise some strategies to protect the liver,
kidneys and skeleton before they are fully damaged and
get irreversible loss.
Anjum et al., J. Anim. Plant Sci. 24(1):2014
79
Table 1. Effect of increasing levels of fluoride on liver and kidney serum enzymes of domestic chicken.
Treatment Groups1
A B C D
ALP 339.2a ± 1.14 444.8b ± 2.32 507.1c ± 1.54 579.4d ± 1.55
AST 198.4a ± 2.44 268.2b ± 1.90 310.9c ± 2.12 355.0d ± 2.13
ALT 168.7a ± 1.55 204.7b ± 4.18 220.0c ± 3.56 246.2d ± 2.45
Bilirubin 0.52a ± 3.20 0.61b ± 3.21 0.65c ± 2.58 0.83d ± 1.46
Uric Acid 3.63a ± 2.45 4.54b ± 2.86 5.55c ± 1.88 6.74d ± 2.92
a-d within a row means for each variable without a common superscript differs significantly at P < 0.05.
1 Treatments: Group A (control) without fluoride; whereas, Group B, C and D, chicks were injected a weekly dose of
NaF 10, 20, and 30 µg per gram of body weight, respectively.
Figure 1. Means ALP, AST and ALT of liver function
enzymes injected increasing dose rate of
fluoride in domestic chicken: Group A (control)
without fluoride, and a dose of NaF 10, 20, and
30 µg/g of BW injected to chickens on a weekly
basis in treatment Group B, C and D,
respectively. Means for each treatment are
presented as: ALP; solid bars, AST; dots, and
ALT; upward cross bars.
Figure 2. Means of bilirubin and uric acid injected
increasing dose rate of fluoride in domestic
chicken: Group A (control) without fluoride, and a
dose of NaF 10, 20, and 30 µg/g of BW injected to
chickens on a weekly basis in treatment Group B,
C and D, respectively.  Means for each treatment
are presented as: Bilirubin; solid bars, and Uric
Acid; dots.
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INFLUENCE OF INCREASING FLUORIDE DOSE RATES ON SELECTED LIVER AND KIDNEY ENZYMES PROFILE IN DOMESTIC CHICKEN (Gallus domesticus)

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INFLUENCE OF INCREASING FLUORIDE DOSE RATES ON SELECTED LIVER AND KIDNEY ENZYMES PROFILE IN DOMESTIC CHICKEN (Gallus domesticus)

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