Study on the effect of imidacloprid on gill tissue of the fish channa punctatus (perciformes channidae)

© 2018 IJRAR January 2019, Volume 06, Issue 1 www.ijrar.org (E-ISSN 2348-1269, P- ISSN 2349-5138)
IJRAR190N067 International Journal of Research and Analytical Reviews (IJRAR) www.ijrar.org 372
Study on the effect of Imidacloprid on gill tissue of
the fish Channa punctatus (Perciformes:
Channidae) (Bloch.)
Pawara, Ravindra H., Patel Nisar G. and Sarvade Raju C.
P.G. and Research Centre, Department of Zoology,
Pratap College, Amalner, Dist- Jalgaon, (M.S.), India, PIN- 425401.
Email:- rhpawara@gmail.com
ABSTRACT
The present study has been undertaken to have a basic understanding of the acute toxicity of
Imidacloprid insecticide on the experimental fish Channa punctatus. The objective of this study was to
investigate the histomorphological changes and recovery responses for freshwater teleost Channa punctatus
(Bloch.). The fish was exposed to sublethal concentration for 24, 48, 72 and 96 hrs to observe the alteration
in histomorphological structure. These changes were found to be time and dose dependent.
KEYWORDS: Channa punctatus, Imidacloprid, Histomorphology, Gill.
INTRODUCTION
The use of pesticides in the field has the potential to change the aquatic medium, affecting the tolerance
limit of aquatic fauna and flora, as well as creating danger to the ecosystem (Adedeji and Okocha, 2012).
About one billion people worldwide relied on fish as their primary source of animal protein (FAO, 2002).
Imidacloprid (Confidor) is one of the most widely used insecticides in India and other parts of the World.
Imidacloprid enters the target pest via ingestion or direct contact. Imidacloprid is currently the most widely
used insecticide in the world, which acts as an insect neurotoxin with much lower toxicity to mammals.
Imidacloprid is a systemic poison and shows medium soil adsorption. Imidacloprid is manufactured by Bayer
crop science, since its launch in 1991, has gained registration in about 120 countries and is marketed for use
on over 140 agricultural crops with annual sale of more than 600 million Euro (Cox, 2001). It is sold under
many trade names which include Confidor, Admire, Gaucho, Merit, Advantage, Provado, and Winner.
Imidacloprid is a neonicotinoid insecticide in the chloronicotinyl nitroguanidine chemical family (Wismer,
2004; Tomlin, 2006)). Imidacloprid is used for controlling sucking insects, soil insects, termites, and some
chewing insects. It is applied to seeds, soil, crops, and structures, and is used as a topical flea control treatment
on domestic pets. In India, Imidacloprid is registered for use on a wide array of crops including cotton,
cabbage, okra, brinjal, sugarcane, wheat and sunflower. The main crop of the region is cotton, jawar, wheat,
chilli, groundnut, sunflower, paddy, maize, sugarcane, banana and variety of vegetables (tomato, potato,
cabbage, okra, brinjal etc.). Channa punctatus (Bloch.) is a fresh water teleost, which is found in abundance
in the natural water bodies of Maharashtra. The snakehead, Channa punctatus (Bloch.) locally called Dok in
Khandesh and Murrel called in other parts of Maharashtra. Dok is an important and popular fish species. C.

punctatus of Indian capture fishery. Due to presence of accessory respiratory organs, these fish can remain
alive out of water for a considerable time and hence can enter the crop field from adjoining water bodies. It is
one of the air breathing fish and is capable of adapting variety of habitats. Body elongated, mostly rounded,
streamlined body decreases the amount of friction from the water for rapid swimming. Teleosts have five
pairs of gill arches. In the front four pairs, the slender gill filaments form two lines facing towards the back
and these two lines are joined to each other at the base by a gill septum. The gills are the most delicate structure
of the teleost body. The secondary gill lamellae are also known as respiratory lamellae. Gills are the primary
organs for respiration, ion regulation and osmoregulation, acid base regulation and nitrogen excretion. Any
environmental stress influencing the function of gills as gills are the very first site where pesticides and
herbicides induced lesions may occur which may result in an impaired gas and ion exchange. Mucus cells and
pavement cells are also present in the epithelium of the filament and at the base of lamellae, but they are
smaller than chloride cells the gill filaments are covered with squamous pavement cells showing characteristic
concentric patterns of microridges (Devi and Mishra, 2013).
MATERIAL AND METHODS
Finney (1971) Probit Analysis LC50 determination method was used to find out the LC50 concentration.
The fish were grouped into five groups (I, II, III, IV and V) having ten fish each. Different concentrations viz;
0.0 ppm, 0.2 ppm, 0.4 ppm, 0.6 ppm, and 0.8 ppm were labeled as I, II, III, IV and V groups respectively. All
groups were observed for 24, 48, 72 and 96 hrs and mortality was recorded. Experimental groups were
exposed to sublethal concentration of Imidacloprid to an interval of 24, 48, 72 and 96 hrs. Five fish were taken
in each interval for control and each experimental group for microscopic studies. In the present study, after
24, 48, 72 and 96 hrs Imidacloprid exposed fish were taken out, sacrificed and the tissue of gill was excised
out and five fish were kept for recovery study. Bouin’s solution was used as fixative. The attached block was
cut into thin sections of desired thickness (4.0- 6.0 Micron) of the microtome. The clean slides were smeared
with adhesive Mayer’s albumen mounting the material on glass slides. Haematoxylin and Eosin stains were
used for staining and finally mounted in DPX mountant.
RESULTS AND DISCUSSION
In present study authors observed primary gill lamellae consist of centrally placed rod like central axis
with chloride cells and with blood vessels on either side. Numerous semicircular secondary gill lamellae are
lined up along both sides of the primary gill lamellae. The lamellae are lined by a squamous epithelium
composed by pavement and nondifferentiated epithelial cells. Each lamella is made up of two sheets of
epithelium delimited by many pillar cells, which are contractile and separate the blood (capillary) channels in
which one to two erythrocytes are usually recognized within each capillary lumen. Between the lamellae, the
filament is lined by a thick stratified epithelium constituted by several cellular types, such as chloride cell,
mucous or goblet cell and pavement cell (Fig. a.). In present work after treatment of Imidacloprid at 24 hrs
fish gill showed degeneration of epithelium, degeneration of secondary lamellae, inter lamellar region,
separation of epithelium (Fig. b.) as well as curling of secondary lamellae, detached of secondary lamellae,

epithelial lesions and lacuna, collapsed pillar cell (Fig. c.) were also observed with 48 hrs exposure. Swelling
of central axis, swelling of secondary gill lamellae tip and vacuolation (Fig. d.) were observed after 72 hrs.
Bulging tip of gill lamellae, disrupted primary lamellae, erosion of secondary lamellae and swelling of central
axis (Fig. e.) were also noticed in 96 hrs treatment of sublethal concentration. After the recovery period gill
reached the near normal condition central axis, primary lamellae and secondary lamellae, pillar cell were
clearly seen. Epithelial lesions, vacuolation (Fig. f.) were not observed after 10 days recovery period.
Chandra and Banerjee (2004) noticed in the gill of Channa striata the blood capillaries dilated and
showed congestion, causing extensive bulging and protrusion onto the surface, epithelial linings of gill
filaments as well as respiratory lamellae were detached and lifted up, neighbouring secondary gill lamellae
fused, causing decreased surface area, while reducing the efficiency of gills as well as ladder-like
arrangements of the pillar cells and blood capillaries also collapsed after treatment of air exposure fish died
due to anoxia and other physiological disorders. Butchiram, et al. (2009) observed histopathological changes
in the gill include reduce central axis, reduced mucous cells i.e. necrotic condition, which leads into
interlamellar space formation, swelling and pycnotic conditions in epithelial cells, vacuolar degeneration,
fusion and atrophy of primary and secondary gill lamellae bulging of tip of primary gill filaments with
distortion of the shape of secondary filaments, rupture was also observed in some places of secondary gill
lamellae in Channa punctatus with Alachlor treatment. Tilak, et al. (2005) also observed similar findings in
the gill tissue of the fish Channa punctata exposed to sublethal concentration of Butachlor and Machete and
found marked pathological changes such as bulging of tips of primary gill filaments and fusion of disorganized
secondary gill filaments. The mucosubstances secreted by the gill epithelial cells and mucous cells perform
some functions in the life of fish. Some of the studies are concerned with the effects of some toxicants or
change of habitat on mucin secretion in the gill of fish (Patil, et al, 2012). Verma (1997) reported swollen
oedematous as well as atrophic epithelial cells of secondary lamellae and mucosal sloughing along with sub-
mucosal hyperplasia in intestine during acute Aflatoxin toxicity in Channa punctatus. The main clinical
internal sings that can lead to death of fish include neurological disorder and disruption of nerve functions,
respiratory dysfunction and suffocation (Banaee, et al., 2011).
Fig. a) Fig. b)

Fig. c) Fig. d`
Fig. e) Fig. f)
Fig. a) to f) Vertical section of gill of C. punctatus (H-E) (Control, Treated and Recovery)
BT- Bulging tip of gill lamellae CA- Central axis C- Chloride cell CDC- Chondrocytes MC- Mucous cell
CSL- Curling of secondary lamellae DE- Degeneration of epithelium DP- Disrupted primary lamellae L- Lacuna
DSL- Degeneration of secondary lamellae DSLM- Detached secondary lamellae EC- Epithelial cell PC- Pillar cell
EL- Epithelial lesions ESL- Erosion of secondary lamellae EWCL- Erythrocyte within capillary lumen
ILR- Inter lamellar region PGL- Primary gill lamella PLC- Pillar cell collapsed SCA- Swelling of central axis
SGL- Secondary gill lamella SS- Swelling of secondary gill lamellae tip V- Vacuolation
Devi and Mishra (2013) found similar alternations in the gill showed at long term period (7 days) of
Chlorpyrifos exposure were accentuated lifting of the lamellar epithelium, oedema in the filamentary
epithelium and an intense lamellar vasodilatation. Gills treated with 1.46 μl/l Chlorpyrifos exhibited lamellar
fusion in numerous areas of the secondary lamellae with hyperplasia in chloride cell, pillar cell, pavement cell
and mucus secreting globlet cells for short term (3 days) exposure time. Gill showed lamellar aneurism
(lesions of blood vessels), synechiae (adhesion of tip) and congestion and leucocytes infiltration,
interlamellare oedema and congestion in secondary lamellae were noticed at long term treatment of
Chlorpyrifos. Gills of 0.538 μl/l Chlorpyrifos treated group showed extensive hypertrophy and hyperplasia of
epithelial, mucus secreting cell and chloride cells due to complete fusion of secondary lamellae and
haemorrhage in cartilaginous core, at the end of 3 day treatment. At the long term (7 days) 0.538 μl/l
Chlorpyrifos showed hypertrophy and proliferation in the erythrocytes of cartilaginous core, complete
destruction of the secondary lamellae, obliteration of normal lamellae architecture as lifting of epithelial cells

affecting the apical distal ends of the gill and the destructed blood capillaries aggregate and showed lamellar
aneurism.
Lal, et al. (2007) observed the histopathological changes in the gill of snake headed fish, Channa
punctatus exposed to Benzene Hexachloride. The histological changes noticed by Velmurugan, et al. (2009)
in the gill and liver of Cirrhinus mrigala after in both 0.91ppm and 1.82ppm concentrations of Dichlorvos
exposed such as hyperplasia, desquamation as well as necrosis of epithelial, epithelial lifting, oedema,
lamellar fusion, collapsed secondary lamellae, curling of secondary lamellae and aneurism in the secondary
lamellae were observed in gill tissues exposed to Dichlorvos. Senthamilselvan, et al. (2011) observed
histopathological changes showed hypertrophy, necrosis and swollen tips of the primary lamellae after
exposed to nickel as well as in Mercury exposure hyperplasia, lamellae fusion, necrosis also seen and after
above both mix metals showed desquamated epithelium, necrosis and blousing at the tips of the secondary
lamellae, edema and lifting up of the epithelium, swelling, hyperplasia in the gills of fish, Lates calcarifer.
Ezemonye and Ogbomida (2010) showed gill distortion and fusion of adjacent secondary lamella as a result
of hyperplasia and excessive mucus accumulation in Clarias gariepinus fingerlings exposed to Gammalin20.
Several other workers have also noticed lifting of lamellar epithelium, damaged pillar cells, hypoxia and
respiratory failure problems with ionic and acid-base balance in the gills (Alazemi, et al., 1996; and Virtanen,
1986). Mohamed (2009) noticed proliferative, degenerative and necrotic changes in the epithelium of gill
filaments and secondary lamellae, edema in secondary lamellae, dilation and congestion in blood vessels of
gill filaments and mucous cells proliferation of Tilapia zillii and Solea vulgaris by the effect of agricultural
and sewage drainage water. Our results are in concurrence with the observations of above workers.
ACKNOWLEDGEMENTS
Authors are thankful to The Chairman, K.E. Society; Prof. Dr. Rane J.S., Principal, Pratap College,
Amalner for continuous encouragement as well as providing excellent library as well as laboratory facilities
during the tenure of this work. Dr. R. H. Pawara is grateful to UGC, New Delhi for providing Rajiv Gandhi
National Fellowship as SRF under the able guidance of Principal, Dr. Nisar G. Patel for Ph.D.
CONCLUSION
It is observed that the sub lethal concentrations of Imidacloprid are capable of altering significant
histomorphological structure of gill tissue of C. punctatus. It could be concluded that the very lower
concentration of pesticides affect environmental contamination which can induce several histomorphological
changes in the gill tissue of C. punctatus. These chemicals can be a major threat to fish and aquatic
environment by diminishing their fecundity and reproduction potential. Such kinds of studies are useful for
protecting the biota in natural environment. The current study may be primarily used to understand mechanism
of Imidacloprid induced histomorphological changes.

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Study on the effect of imidacloprid on gill tissue of the fish channa punctatus (perciformes channidae)

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