See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/277970624

Scanning electron microscopy study of the gills, scales and erythrocytes of

Anabas testudineus upon exposure to chlorpyrifos

Article in Toxicological and Environmental Chemistry · May 2015

DOI: 10.1080/02772248.2015.1041527

CITATIONS READS

8 635

4 authors:

Vel Murugan M. Selvanayagam

Sir Theagaraya College Loyola ICAM College of Engineering and Technology (LICET), SENESCYT,Ecuador, …
27 PUBLICATIONS 999 CITATIONS 106 PUBLICATIONS 1,534 CITATIONS

SEE PROFILE SEE PROFILE

Elif Ipek Cengiz Pelin Ugurlu

Dicle University, Faculty of Pharmacy Dicle University
34 PUBLICATIONS 1,829 CITATIONS 13 PUBLICATIONS 302 CITATIONS

SEE PROFILE SEE PROFILE

All content following this page was uploaded by Pelin Ugurlu on 16 December 2015.

The user has requested enhancement of the downloaded file.

 Toxicological & Environmental Chemistry

ISSN: 0277-2248 (Print) 1029-0486 (Online) Journal homepage: http://www.tandfonline.com/loi/gtec20

Scanning electron microscopy study of the gills,

scales and erythrocytes of Anabas testudineus
upon exposure to chlorpyrifos

Babu Velmurugan, Mariadoss Selvanayagam, Elif Ipek Cengiz & Pelin Ugurlu

To cite this article: Babu Velmurugan, Mariadoss Selvanayagam, Elif Ipek Cengiz & Pelin Ugurlu
(2015) Scanning electron microscopy study of the gills, scales and erythrocytes of Anabas
testudineus upon exposure to chlorpyrifos, Toxicological & Environmental Chemistry, 97:2,
208-220, DOI: 10.1080/02772248.2015.1041527

To link to this article: http://dx.doi.org/10.1080/02772248.2015.1041527

Published online: 26 May 2015.

Submit your article to this journal

Article views: 23

View related articles

View Crossmark data

Full Terms & Conditions of access and use can be found at

http://www.tandfonline.com/action/journalInformation?journalCode=gtec20

Download by: [Dicle University] Date: 30 October 2015, At: 05:01

 Toxicological & Environmental Chemistry, 2015
Vol. 97, No. 2, 208 220, http://dx.doi.org/10.1080/02772248.2015.1041527

Scanning electron microscopy study of the gills, scales and

erythrocytes of Anabas testudineus upon exposure to chlorpyrifos
Babu Velmurugana, Mariadoss Selvanayagama, Elif Ipek Cengizb* and Pelin Ugurluc
a
Department of Zoology, Environmental Science and Biotechnology Research Unit, Loyola College,
Chennai, India; bFaculty of Pharmacy, Department of Pharmaceutical Toxicology, Dicle
University, Diyarbakır, Turkey; cScience and Technology Application and Research Center, Dicle
University, Diyarbakır, Turkey
(Received 20 November 2014; accepted 13 April 2015)
Downloaded by [Dicle University] at 05:01 30 October 2015

The present study was undertaken to assess the toxicity of sublethal concentrations
(125, 250, and 375 mg L¡1) of chlorpyrifos on Anabas testudineus for 21 days. The
morphological changes on the gills, scales, and erythrocytes of A. testudineus were
observed using scanning electron microscopy. Gill alterations included highly active
mucous cells, epithelial hyperplasia, and fusion of secondary lamellae. The scales
showed damaged lepidonts. Oozed out cytoplasmic content and lobopodial projections
were observed in the erythrocytes after exposure to chlorpyrifos.
Keywords: chlorpyrifos; Anabas testudineus; scanning electron microscopy (SEM);
gill; scale; erythrocytes

Introduction
Chlorpyrifos (O,O-diethyl-O-(3,5,6-trichloro-2-pyridyl)phosphorothioate) is an organo-
phosphorus insecticide for a wide range of pests (O’Brien 1967). The chemical enters
water bodies through intentional application, runoff from farms, aerial drift, and acciden-
tal and illegal release (Zelikoff 1994). Due to its lipophilic nature, fish are able to absorb
and bioconcentrate chlorpyrifos from moderate to high levels.
Anabas testudineus has been selected as test animal because of its euryhaline and
eurythermal nature, and its position in the food chain. They are quite sturdy and ideally
suited for experimentation in the laboratory for longer periods. These fish are cultured in
ponds and they have a good commercial value because of their nutritive value and taste
(Afsar 2012).
Gills are generally considered as good indicators of water quality (Rankin, Stagg, and
Bolis 1982), being models for studies of environmental impact (Mallatt 1985). Because
the gills perform several vital functions and have a large surface area in contact with the
external environment, they are particularly sensitive to physical and chemical changes in
the aquatic environment and are the main target organ in fish for toxic waterborne pollu-
tants (Mallatt 1985).
Organic xenobiotics such as pesticides pass quickly through the gills and contaminate
other organs through the blood stream. A few studies are available on the effects of pesti-
cides on gill surface morphology (Dutta et al. 1997; Machado and Fanta 2003; Rao,
Shilpanjal, and Kavitha 2003; Johal, Sharma, and Ravnet 2007).

*Corresponding author. Email: ecengiz@dicle.edu.tr

Ó 2015 Taylor & Francis

 Toxicological & Environmental Chemistry 209

Since blood cells play a central role in the physiology of fish respiration, they
represent an outstanding model to study xenobiotic-induced damage. The erythrocytes
of lower vertebrates preserve a nucleus and mitochondria, unlike the anucleated
erythrocytes of mammals (Tiano et al. 2003). Blood cell profiles has been
considered as an important indicator of diseases and other toxicants (Tripathi and
Srivastav 2010).
Scales have been used for classification and identification of growth in studies on dif-
ferent fishes. Although scales come into immediate contact with the environment and pol-
lutants, they have rarely been used as bioindicators of pollution (Tandon and Johal 1993).
However, any sudden change in the fish environment causes alterations in circuli shape
and pattern in elemental deposition of scales (Johal and Dua 1994). The scanning electron
microscopy (SEM) study on scales in relation to pollution and environmental conditions
is a recent approach (Khanna et al. 2007).
SEM is a useful tool for assessing the effects of environmental stressors on fish struc-
Downloaded by [Dicle University] at 05:01 30 October 2015

tures critical to their fitness and/or survival (Palaniappan et al. 2008), for instance, by
revealing the surface ultrastructure of various organs, including the gills (Dutta et al.
1997). This is of great importance with regard to respiratory organs since gas exchange
takes place across the surface, and surface morphology may well determine the efficiency
of the system. Damage to surface ultrastructure cannot be identified by light or transmis-
sion electron microscopy; therefore, it is necessary to use SEM to reveal changes and
damage that may occur in tissues (Dutta et al. 1997).
There are few SEM studies about adverse effects of pesticides on surface structure on
fish scales, gills, and blood tissues (Johal and Dua 1994; Dutta et al. 1997; Rao, Shilpan-
jal, and Kavitha 2003). In the present investigation, morphological changes induced by
chlorpyrifos at sublethal concentrations on gills, scales, and erythrocytes are studied using
SEM.

Materials and methods

Experimental design
Renewal toxicity test methods (APHA 1995) were done to find out the 96-h LC50 concen-
tration. The 96-h LC50 for chlorpyrifos in A. testudineus was found to be 2500 mg L¡1.
The fish were divided into four groups and placed in separate glass aquaria. The present
study was performed in three replicates in order to ensure the reproducibility of the
results. Ten fish were used for each group per replicate. Groups I, II, and III were exposed
to sublethal concentrations of chlorpyrifos. Group IV was maintained in pesticide-free
water to serve as control. The nominal concentrations of chlorpyrifos tested were 125,
250, and 375 mg L¡1.

Gill and scale sample collection and processing

The experimental and control fish were sacrificed at the end of 21 days. The scales were
removed from the second row, just above the lateral line directly under the anterior ray of
the dorsal fin with the help of tweezers.
Gill tissues and scales were preserved in 2.5% glutaraldehyde buffered with 0.1 mol L¡1
phosphate buffer (pH 7.4) at 4  C for 24 h, then washed in phosphate buffer (pH 7.4) for
three times, and dehydrated in a graded series of ethanol (30% 100%) for 1 h each. The
 210 B. Velmurugan et al.

dehydrated tissues were dried in a desiccator. Dried specimens were mounted on a stub with
double-sided tape and coated with a thin layer of platinum in a sputter coating unit.

Blood sample collection and processing

After 21 days of treatment, blood samples were collected by severing the caudal peduncle
of the fish and mixed immediately with 15 mL of K2EDTA (di-potassium ethylene
diamine tetraacetic acid), prepared as 10% solution in distilled water, for 1 mL of blood
as anticoagulant for further SEM studies.
Blood samples were fixed with 2.5% glutaraldehyde in 0.1 mol L¡1 phosphate buffer,
pH 7.2, at 4  C for 2 h. Fixed blood was centrifuged at 1500 g for 5 10 min. Extra fixa-
tive was removed and the erythrocyte pellet was washed thrice with 0.1 mol L¡1 phos-
phate buffer till the odor of glutaraldehyde disappeared completely. The pellet was then
gently suspended in a small volume of triple distilled water, and a drop of erythrocyte sus-
Downloaded by [Dicle University] at 05:01 30 October 2015

pension was deposited on glass coverslips and air-dried. Air-dried samples in the cover
slips were mounted on a stub with double-sided tape and then coated with a thin layer of
platinum in a sputter coating unit. The coated specimens (gill, scale, and erythrocyte)
were observed by SEM (JSM 6360, JEOL, USA) and photographed with computer inte-
grated with software.

Semiquantitative scoring
The structural, morphological changes and deformities in the scales, gill, and blood were
examined in randomly selected three fish from each group per replicate. The mean preva-
lence of each ultrastructural parameters was categorized as none (¡), mild (C, <25% of
sections), moderate (CC, 25% 50% of sections), and severe (CCC, >50% of sections)
(Table 1) (Korkmaz et al. 2009).

Table 1. Semiquantitative evaluation of alterations on the gills, scales, and erythrocytes of A.

testudineus exposed to sublethal concentrations of chlorpyrifos.

Chlorpyrifos

Alterations Control 125 mg L¡1 250 mg L¡1 375 mg L¡1

Gill
Epithelial hyperplasia ¡ C CC CCC
Fusion of secondary lamellae ¡ C CC CCC
Highly active mucous cells ¡ ¡ C CC

Scale
Damaged lepidonts ¡ C CC CCC
Extensive damage of circuli ¡ C CC CCC

Erythrocytes
Lobopodial projections ¡ C C C
Oozed out cytoplasmic content ¡ C CC CC

Note: none (¡), mild (C), moderate (CC), severe (CCC).

 Toxicological & Environmental Chemistry 211

Results
Gills
In the control fish, the gills had four arches on each side of the body and each one sup-
ported many filaments (primary lamellae). On the upper and lower surface of each fila-
ment, there was a row of secondary lamellae. Inter-lamellar water spaces were
prominent. The surfaces of primary and secondary lamellae were covered by microridged
epithelial cells. Mucous cells were seen in between these cells. Epithelial cells were
polygonal in shape. Ridges were in a maze-like pattern. The primary and secondary
lamellae were normal (Figures 1 and 2).
Gills of fish exposed to 125 mg L¡1 chlorpyrifos for 21 days showed epithelial
hyperplasia and fusion of secondary lamellae (Figure 3). After exposure to 250 mg L¡1
(Figures 4 and 5) and 375 mg L¡1 chlorpyrifos (Figure 6), fusion of secondary lamellae,
epithelial hyperplasia, and highly active mucous cells were observed.
Downloaded by [Dicle University] at 05:01 30 October 2015

Scales
Normal scale structure had circuli present all over, with signs of calcium deposition or
growth rings. In the anterior region, the number of circuli was maximum. The fully
formed circuli had rows of lepidonts, teeth-like structures present on their ridge. The lepi-
donts assist in the attachment of scales to the skin and prevent their detachment. Scales of
the control group had normal morphology (Figures 7 and 8).
Scales of fish exposed to 125 mg L¡1 chlorpyrifos showed damaged lepidonts
(Figure 9), and after exposure to 250 mg L¡1 (Figure 10) and 375 mg L¡1 chlorpyrifos
(Figure 11), extensive damage of circuli was observed.

Figure 1. SEM micrograph of gill of control fish Anabas testudineus: (a) interlamellar water
spaces; (b) primary lamella; (c) secondary lamella.
 212 B. Velmurugan et al.
Downloaded by [Dicle University] at 05:01 30 October 2015

Figure 2. SEM micrograph of gill of control fish Anabas testudineus: (a) primary lamella;
(b) secondary lamella.

Erythrocytes
Each erythrocyte appeared elliptical under SEM. The micrograph of control samples
showed regularly shaped erythrocytes with the usual smooth surface (Figures 12 and 13).
Fish exposed to 125, 250, and 375 mg L¡1 chlorpyrifos for 21 days exhibited lobopo-
dial projection and oozed out cytoplasmic contents (Figures 14 16).

Figure 3. SEM micrograph of gill of fish Anabas testudineus exposed to 125 mg L¡1 chlorpyrifos
for 21 days: (a) epithelial hyperplasia; (b) fusion of secondary lamellae.
 Toxicological & Environmental Chemistry 213
Downloaded by [Dicle University] at 05:01 30 October 2015

Figure 4. SEM micrograph of gill of fish Anabas testudineus exposed to 250 mg L¡1 chlorpyrifos
for 21 days: (a) epithelial hyperplasia; (b) fusion of secondary lamellae.

Discussion
In the present study, highly active mucous cells, epithelial hyperplasia, and fusion of sec-
ondary lamellae may be related to a defense mechanism which reduces the water surface
around the gill and increases the barrier distance for diffusion of toxicants from outside to
the blood capillaries. Although this mechanism protects the fish from toxicants, it also
reduces the oxygen supply, which leads to suffocation of the fish. Kossakowski and

Figure 5. SEM micrograph of gill of fish Anabas testudineus exposed to 250 mg L¡1 chlorpyrifos
for 21 days: (a) active mucous cells.
 214 B. Velmurugan et al.
Downloaded by [Dicle University] at 05:01 30 October 2015

Figure 6. SEM micrograph of gill of fish Anabas testudineus exposed to 375 mg L¡1 chlorpyrifos
for 21 days: (a) epithelial hyperplasia; (b) fusion of secondary lamellae.

Ostaszewska (2003) concluded that the increase of mucous secretion is the first reaction
of the external tissues of fish to the toxic compounds in water. This defense mechanism
helps in the removal of any bound pathogens, toxicants, or foreign materials (Parashar
and Banerjee 2002).
Gills of bluegill sunfish, Lepomis macrochirus, exhibited structural damage following
exposure to sublethal concentrations of diazinon. The thickness of the microridge on the
gill arch, mucus extrusion, lamellar swelling, and reduced microridges were reported
(Dutta et al. 1997). The gills of fish, Metynnis roosevelti, exposed to methyl parathion

Figure 7. SEM micrograph of scale of control fish Anabas testudineus: (a) focus; (b) interior cir-
culi towards focus; (c) marginal ciruli; (d) radii.
 Toxicological & Environmental Chemistry 215
Downloaded by [Dicle University] at 05:01 30 October 2015

Figure 8. SEM micrograph of scale of control fish Anabas testudineus: (a) circuli; (b) radii.

showed cellular degeneration, collapse of the blood spaces, and decrease in the amount of
microridges at the epithelial cells surface of the primary lamellae (Machado and Fanta
2003). The changes including thick coat of mucus covering the entire gill filaments,
decreases in diameter of the primary gill lamellae, distorted shapes of gill rakers, mild
degenerative changes in the interlamellar region, tissue erosion at gill arch, epithelial
necrosis, and rupture of the gill epithelium were responses induced by the action of profe-
nofos in fish Oreochromis mossambicus (Rao, Shilpanjal, and Kavitha 2003). The degen-
eration of microridges, detachment of the epithelial layer, inflammation in the centre of
the pavement cells, thickening of the boundary walls, and the reduction in the number of
mucous and chloride cells were reported in the gills of Cyprinus carpio communis
exposed to monocrotophos (Johal, Sharma, and Ravnet 2007). In a previous study carried

Figure 9. SEM micrograph of scale of fish Anabas testudineus exposed to 125 mg L¡1 chlorpyrifos
for 21 days: (a) damaged lepidonts.
 216 B. Velmurugan et al.
Downloaded by [Dicle University] at 05:01 30 October 2015

Figure 10. SEM micrograph of scale of fish Anabas testudineus exposed to 250 mg L¡1 chlorpyri-
fos for 21 days: (a) damaged lepidonts and extensive damage of circuli.

out by Ba-Omar, Al-Jardani, and Victor (2011), an increase in mucous secretion at the gill
filament’s surface after exposure of Aphanius dispar to temephos was shown.
The results of the present study indicate that chlorpyrifos induced a significant alter-
ation in the shape of the erythrocytes. Kalair, Mishra, and Singh (1993) observed coarsely
crenated erythrocytes after exposure of Amphipnous cuchia to sumithion. SEM micro-
graphs of erythrocytes following fenvalerate exposure revealed various degrees of distor-
tion in shape, ruptured membranes, and echinocyte formation (Prasanthi, Muralidhara,
and Rajini 2005).
More importantly, it is speculated that oxidative stress in erythrocytes may lead to
alterations in their structural conformation which may compromise effective blood flow,
oxygen uptake, and release (Youdim et al. 2000).

Figure 11. SEM micrograph of scale of fish Anabas testudineus exposed to 375 mg L¡1 chlorpyri-
fos for 21 days: (a) damaged lepidonts and extensive damage of circuli.
 Toxicological & Environmental Chemistry 217

Figures 12 and 13. SEM micrograph of erythrocyte of control fish Anabas testudineus: (a)
erythrocyte.
Downloaded by [Dicle University] at 05:01 30 October 2015

In the present study, it was confirmed that the lepidonts on the scales of A. testudineus
were damaged. Detailed study of marginal circuli and mineral composition of various
parts of the scale of Channa punctatus (Bloch) had been used as an indicator of pollution
on exposure to different concentrations of endosulfan (Johal and Dua 1994). It was obvi-
ous that this disorganization was due to the altered chemical composition in response to
the toxic effect of endosulfan. Johal and Sawhney (1997) observed that lepidonts on the
scale of C. punctatus showed various degrees of damage upon exposures to malathion.

Figure 14. SEM micrograph of erythrocyte of fish Anabas testudineus exposed to 125 mg L¡1
chlorpyrifos for 21 days: (a) lobopodial projections; (b) oozed out cytoplasmic content.
 218 B. Velmurugan et al.
Downloaded by [Dicle University] at 05:01 30 October 2015

Figure 15. SEM micrograph of erythrocyte of fish Anabas testudineus exposed to 250 mg L¡1
chlorpyrifos for 21 days: (a) lobopodial projections; (b) oozed out cytoplasmic content.

Figure 16. SEM micrograph of erythrocyte of fish Anabas testudineus exposed to 375 mg L¡1
chlorpyrifos for 21 days: (a) lobopodial projections; (b) oozed out cytoplasmic content.
 Toxicological & Environmental Chemistry 219

Conclusion
The findings of the present SEM investigations demonstrate a direct correlation between
pesticide exposure and morphological alterations observed in several tissues. These
results have showed that SEM is a sensitive tool for determining early effects of toxicants
in the organisms. And, also, the study showed up a simple and inexpensive procedure uti-
lizing scales without sacrificing the fish in an effort to assess the environment. From the
present observations, it is suggested that the scales can be successfully employed as a reli-
able biological marker for pesticide pollution. The scale, thus, can be used as a biomarker
of pollution. An added advantage is that these can be used without sacrificing the animal.
As a conclusion, morphological alterations in scale, gill, and blood demonstrate a
direct correlation between pesticide exposure and morphological alterations.

Disclosure statement
Downloaded by [Dicle University] at 05:01 30 October 2015

No potential conflict of interest was reported by the authors.

Funding
The work was supported by Jawaharlal Nehru Memorial Fund, New Delhi [grant number SU/1/192/
2006-2007/646].

References
Afsar, S. 2012. “Glucose Post Exposure Recovery from Lead Intoxicated Fresh Water Fish Anabas
testudineus.” International Journal of Biomedical and Advance Research 3: 59 63.
APHA (American Public Health Association). 1995. Standard Methods for the Examination of
Water and Wastewater. 19th ed. Washington, DC: APHA.
Ba-Omar, T.A., S. Al-Jardani, and R. Victor. 2011. “Effects of Pesticide Temephos on the Gills of
Aphanius dispar (Pisces: Cyprinodontidae).” Tissue and Cell 43: 29 38.
Dutta, H.M., J.S.D. Munshi, P.K. Roy, N.K. Singh, L. Motz, and S. Adhikari. 1997. “Effects of
Diazinon on Bluegill Sunfish, Lepomis macrochirus, Gills: Scanning Electron Microscopy
Observations.” Experimental Biology Online 2: 1 11.
Johal, M.S., and A. Dua. 1994. “SEM Study of the Scales of Freshwater Snakehead Channa puncta-
tus (Bloch) upon Exposure to Endosulfan.” Bulletin of Environmental Contamination and Toxi-
cology 52: 718 721.
Johal, M.S., and A.K. Sawhney. 1997. “Lepidontal Alterations of the Circuli on the Scales of Fresh
Water Snake Head, Channa punctatus (Bloch) upon Exposure to Malathion.” Current Science
72: 367 368.
Johal, M.S., M.L. Sharma, and K.U. Ravnet. 2007. “Impact of Low Dose of Organophosphate,
Monocrotophos on the Epithelial Cells of Gills of Cyprinus carpio communis Linn. SEM
Study.” Journal of Environmental Biology 28: 663 667.
Kalair, J.S., V.S. Mishra, and R.K. Singh. 1993. “Pesticide Sumithion Induced Haematological
Changes in Mud Eel Amphipnous cuchia.” Biological Memoirs 19: 41 48.
Khanna, D.R., P. Sarkar, A. Gautam, and R. Bhutani. 2007. “Fish Scales as Bio-Indicator of Water
Quality of River Ganga.” Environmental Monitoring and Assessment 134: 153 160.
Korkmaz, N., E.I. Cengiz, E. Unlu, E. Uysal, and M. Yanar. 2009. “Cypermerthrin-Induced Histo-
pathological and Biochemical Changes in Nile Tilapia (Oreochromis niloticus), and the Protec-
tive and Recuperative Effect of Ascorbic Acid.” Environmental Toxicology and Pharmacology
28: 198 205.
Kossakowski, M.K., and T. Ostaszewska. 2003. “Histopathological Changes in the Juvenile Carp
Cyprinus carpio.” Archives of Polish Fisheries 1: 57 67.
Machado, M.R., and E. Fanta. 2003. “Effects of the Organophosphorous Methyl Parathion on the
Branchial Epithelium of a Freshwater Fish Metynnis roosevelti.” Brazilian Archives of Biology
and Technology 46: 361 372.
 220 B. Velmurugan et al.

Mallatt, J. 1985. “Fish Gill Structural Changes Induced by Toxicants and Other Irritants: A Statisti-
cal Review.” Canadian Journal of Fisheries and Aquatic Sciences 42: 630 648.
O’Brien, R.D. 1967. Insecticides, Action and Metabolism. New York: Academic Press.
Palaniappan, P.L.R.M., S. Sabhanayakam, N. Krishnakumar, and M. Vadivelu. 2008.
“Morphological Changes due to Lead Exposure and the Influence of DMSA on the Gill Tissues
of the Freshwater Fish Catla catla.” Food and Chemical Toxicology 46: 2440 2444.
Parashar, R.S., and T.K. Banerjee. 2002. “Toxic Impact of Lethal Concentration of Lead Nitrate on
the Gills of Air-Breathing Catfish Heteropneustes fossilis (Bloch).” Veterinary Archives 72:
167 183.
Prasanthi, K., Muralidhara, and P.S. Rajini. 2005. “Morphological and Biochemical Perturbations in
Rat Erythrocytes Following In Vitro Exposure to Fenvalerate and Its Metabolite.” Toxicology In
Vitro 19: 449 456.
Rankin, J.C., R.M. Stagg, and L. Bolis. 1982. “Effects of Pollutants on Gills.” In Gill, edited by D.F.
Houlihan, J.C. Rankin, and T.J. Shuttleworth. New York: Cambridge University Press.
Rao, J.V., D.I. Shilpanjal, and P. Kavitha. 2003. “Toxic Effects of Profenofos on Tissue Acetylcho-
linesterase and Gill Morphology in a Euryhaline Fish, Oreochromis mossambicus.” Archives of
Toxicology 77: 227 232.
Downloaded by [Dicle University] at 05:01 30 October 2015

Tandon, K.K., and M.S. Johal. 1993. “Mineral Composition of Different Regions of the Scale of an
Endangered Fish Tor putitora (Hamilton) Using Energy-Dispersive X-Ray Microanalysis
Technique.” Current Science 65: 572 573.
Tiano, L., D. Fedeli, G. Santoni, I. Davies, and G. Falcioni. 2003. “Effect of Tributyltin on Trout
Blood Cells: Changes in Mitochondrial Morphology and Functionality.” Biochimica et Biophy-
sica Acta 1640: 105 112.
Tripathi, S., and A.K. Srivastav. 2010. “Alterations in the Profile of Blood Cells of Wistar Rats
Induced by Long-Term Ingestion of Chlorpyrifos.” International Journal of Pharma and Bio
Science 1: 315 322.
Youdim, K.A, B. Shukitt-Hale, S. MacKinnon, W. Kalt, and J.A. Joseph. 2000. “Polyphenolics
Enhance Red Blood Cell Resistance to Oxidative Stress: In Vitro and In Vivo.” Biochimica et
Biophysica Acta 1523: 117 122.
Zelikoff, J.T. 1994. “Fish Immunopharmacology.” In Immunotoxicology and Immunopharmacol-
ogy, edited by J.H. Dean, M.I. Luster, and A.E. Munson, 71 79. New York: Raven Press.

View publication stats