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187
N
S
ave Nature to
S
urvive
ISSN: 0974 - 0376
: Special issue, Vol. 1;
Paper presented in 3rd International Conference on
Climate Change, Forest Resource and Environment
(ICCFRE, 2011)
December 09 - 11, 2011, Thiruvananthapuram,
organized by
Department of Environmental Sciences,
University of Kerala
in association with
National Environmentalists Association, India
www.neaindia.org
QUARTERLY
www.theecoscan.in
KEYWORDS
Saptadeepa Roy et al.
EDC
Bisphenol A
Histopathology
187 - 190
HISTOPATHOLOGICAL EFFECTS OF BISPHENOL A ON LIVER OF
HETEROPNEUSTES FOSSILIS (BLOCH)
188
SAPTADEEPA ROY*, JOGEN CHANDRA KALITA AND MEHNAZ MAZUMDAR1
Physiology and Biochemistry Laboratory, Department of Zoology, Gauhati University, Guwahati - 781 014
1Department of Zoology, M.C.D College, Sonaimukh, Cachar - 788 785
E-mail: saptadeepa_roy@yahoo.co.in
INTRODUCTION
Considerable scientific evidence indicates that a multitude of environmental
contaminants can modulate or mimic the actions of steroid hormones and, in
some cases, produce biological responses qualitatively similar to those produced
by endogenous hormones. Hormonal activity has become a widely recognized
mechanism of toxicity, and a number of laboratory studies have indicated that
exposure to endocrine-modulating substances can impair reproductive function
in adults of either sex, lead to irreversible abnormalities when administered during
development ,or cause cancer. It is entirely possible that exposures to mixtures of
endocrine modulating substances, at the concentrations present in the
environment, may cause additive (Jobling et al., 1995, Soto et al., 1995) or even
synergistic effects.
Indeed, fishes are one of the most thoroughly studied groups of wildlife in terms
of the effects of chemicals on developmental and reproductive processes (Kime,
1998).
Bisphenol-A (BPA) or 2, 2-bis (hydroxy - phenyl)propane is the building block of
polycarbonate plastics, a hard plastic used to make numerous consumer products,
including most baby bottles and water bottles. Its final product includes adhesives,
coatings, paints, building materials. BPA waste may enter the environment during
handling, loading and unloading, heating or releases. BPA leaches out in trace
amount from resins and polycarbonates plastics of food packages (Knaak and
Sullivan, 1966; Krishnan et al., 1993). BPA can enter the human body through
reusable baby bottles (Biles et al., 1997), food packing materials (Krishnan et al.,
1993), liquid of canned vegetables (Brotons et al., 1995), and dental sealants
(Olea et al., 1996).
BPA is one of the major flame retardants and is also a known endocrine-disrupting
chemical (EDC) (Meerts et al., 2001). It has been known that BPA has the efficacy
of the hormone estradiol in some tissues (Dodds and Lawson, 1936), and it has
recently been shown to also antagonize thyroid hormones action (Moriyama et
al., 2002) and antagonize androgen action (Lee et al., 2003). The objective of the
present investigation was to examine the endocrine disruptor Bisphenol A on the
liver of Heteropneustes fossilis (Bloch).
MATERIALS AND METHODS
Experimental fish specimens
Healthy specimens of freshwater fish H. fossilis (Bloch) were procured from local
outlets. The specimens had an average weight of 14 ±2 g and an average length
of 12 ±3 cm. They were then treated with 0.05% KMnO4 solution for 2 min to
avoid any dermal infection. Test water used in the experiment was analyzed for
physico-chemical properties. The fishes were then acclimatized in the laboratory
conditions for 15 days and were fed with boiled egg white, goat liver and processed
*Corresponding author
N
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QUARTERLY
Exposure to endocrine-disrupting chemicals
(EDCs) is currently regarded as one of the most
serious anthropogenic threats to biodiversity
and ecosystems. EDCs are defined as an exog-
enous substance that causes adverse health
effects in an intact organism, or its progeny,
consequent to changes in endocrine function.
Bisphenol A is a well known endocrine
disruptor and is widely used in the produc-
tion of polycarbonates, epoxy resins and flame
retardants. It has also been detected in marine
and freshwater habitats around the world at
considerable concentrations. Low levels of
BPA have been found to cause harmful bio-
logical effects. The present work dealt with
the in vivo study on the Histopathologic ef-
fects of Bisphenol A on liver of Heteropneustes
fossilis. Treatment for 7 days resulted in hepa-
tocellular degeneration. Swollen hepatocytes
with pale vacuolated cytoplasm and loss of
nuclei in some were also observed.
ABSTRACT
189
Figure 1: Representative light photomicrographs of H&E-stained liver sections(40 X) of Heteropneustes fossilis showing normal structural
appearance [A, B] and 7 days treated with 17â-estradiol [C] 0.8 ppm BPA [D], 1ppmBPA [E] and 5 ppm BPA [F]. Unlike normal control, the
estradiol treated group had bile depositions (bd). Hepatocellular degeneration [D], pycnotic nuclei[E] and Fibrosis[F] . Abbreviations: bd-bile
deposition ; pn-pycnotic nuclei
food. The fecal matter and other waste materials were
siphoned off daily to reduce ammonia content in water.
Chemical
All chemicals used in the present study were obtained from
Sigma (Sigma-Aldrich Corporation, Missouri, USA), Merck
(Merck Limited, Mumbai, India), Qualigens (Qualigens Fine
Chemicals, Mumbai, India), HiMedia (HiMedia Laboratories)
Determination of Lethal concentration (LC50)
The acute toxicity bioassay to determine the LC50-96 h value
of Bisphenol A was conducted in a semi static system with the
change of test water on every alternate day. The acute toxicity
bioassay was based on standard methods (APHA, 1998). The
stock solution of Bisphenol A was prepared in analytical grade
ethanol.
Experimental design
After acclimatization in laboratory conditions, fishes were
transferred into six aquaria containing 15L of dechlorinated
tap water. They were then divided into six groups (n=4).
Group I and Group II were Control (untreated) and Solvent
control (ethanol) respectively. Group III was Positive control
receiving 0.1 ppm estradiol Group IV, Group V and Group VI
received 0.8 ppm, 1 ppm and 5ppm BPA. Exposure was given
for 7 days. Animals were constantly observed throughout the
experimental period. Sacrifice was done on the 8th day.
RESULTS
Effect of BPA on liver of H.fossilis
Effect of BPA on liver of the fish was quite evident after 7 days
of treatment. There were stark differences which were marked.
Fishes exposed with Bisphenol-A showed degenerative
changes in hepatic cells. The control and solvent control groups
showed normal compact structure of hepatocytes, while the
group treated with estradiol (positive control) showed bile
deposition. In the groups treated with BPA hepatocellular
degeneration was observed. At the lowest dose of 0.8 ppm
BPA caused the loosening of cells which led to the
disintegration of the compact structure of liver cells. Pycnotic
nuclei were evident in the group treated with 1 ppm BPA. The
highest dose of 5 ppm BPA caused fibrosis in the liver.
DISCUSSION
BPA, an environmental estrogenic chemical BPA has been
reported to be hydroxylated in vivo (Knaak and Sullivan, 1966)
and to bind to DNA (Atkinson and Roy, 1995).
The liver of fish can be considered a target organ to pollutants,
alterations in its structure can be significant in the evaluation
of fish health (Myers et al., 1998), and exhibit the effects of a
variety of environmental pollutants (Hinton et al., 1992).
HISTOPATHOLOGICAL EFFECTS OF BISPHENOL
A
DEF
BC
190
SAPTADEEPA ROY et al.,
Moreover, the liver has play a major role in complex enzymatic
processes of tetraiodothyronine (thyroxine)-tri-iodothyronine
(T4-T3) conversion.
The fibrosis, steatosis, hyperemia and necrosis; which were
formed a rectangle of hepatic tissue changes, are similar to
those reported for fish caught in contaminated water or
exposed ones to various chemicals in laboratory conditions
(Wahbi and El-Greisy, 2007; Aniladevi et al., 2008). Fibrosis
and local blood congestions in the liver sinusoids of the
flounder, Platichthys flesus, the ruffe, Gymnocephalus cernua,
and the smelt, Osmerus eperlanus, were reported as a
consequence of pollution by Peters et al. (1987). Radhaiah
and Rao (1992) reported moderate cytoplasmic degeneration
in hepatocytes, formation of vacuoles, rupture in blood vessels,
and pyknotic nuclei in the liver of Tilapia mossambica exposed
to fenvalerate. Tilak et al. (2005) observed the same changes
in liver of Catla catla ex-posed to chlorpyrifos.
In present study; it has been observed that BPA showed
degenerative changes in hepatic cells after 7 days exposure
compared to control. Thus BPA has toxic properties and further
research is required in this field.
ACKNOWLEDGEMENT
The authors are very thankful to Head, Department of Zoology,
Gauhati University for permitting to carry out the experiments
and University Grants Commission (UGC) for providing SRF
to the fellow.
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