Journal of Thermal Biology 36 (2011) 492–497

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Journal of Thermal Biology

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Thermal adaptability and disease association in common carp

(Cyprinus carpio communis) acclimated to different (four) temperatures
S.M. Ahmad a,n, F.A. Shah a, F.A. Bhat a, J.I.A. Bhat b, M.H. Balkhi a
a
Faculty of Fisheries, Sher-e-Kashmir University of Agricultural Sciences and Technology—Kashmir, Shuhama, Srinagar 19006, India
b
Division of Environmental Sciences, Sher-e-Kashmir University of Agricultural Sciences and Technology—Kashmir, Shalimar, Srinagar 19006, India

a r t i c l e i n f o a b s t r a c t

Article history: The present study was designed to investigate the effect of temperature (20 1C, 24 1C, 28 1C and 32 1C)
Received 6 June 2011 on the heamato–biochemical and histological alterations of Cyprinus carpio communis. Increase in the
Accepted 30 August 2011 temperature showed significant decrease in the serum protein, while a reduced level of blood glucose at
Available online 8 September 2011
high temperature of 32 1C was observed leading to hypoglycemic conditions in the experimental fishes.
Keywords: A significant correlation (P o 0.01) was observed between cholesterol (Cho) and triglycerides (TG) for
Temperature different temperature treatments. Elevated blood urea nitrogen (BUN) at high temperatures was a good
Acclimation indicator of gill osmoregulatory failure. A variation of 86.40% and 38.33%, respectively, was noticed in
Biochemistry alanine aminotransferase (ALT) and aspartate aminotransferase (AST) at 32 1C over minimum experi-
Haemotology
mental temperature of 20 1C. The increase in red blood cell (RBC) and Heamoglobin (Hb) concentration
Poikilo-anisocytic anemia
is associated with the decrease of mean corpuscular hemoglobin (MCH) and mean corpuscular
Histology
Parasitic infection hemoglobin concentration (MCHC), could be the reason for observed poikilo-anisocytosis. Histological
Cyprinus carpio communis studies of different organs of experimental fishes showed accumulation of MMC’s (melanomacrophagic
centers) and atrophy of the interrenal tissue on exposure to various levels of temperature. These
changes were related to severity of thermal stress, being most marked when high temperature was
prolonged during acclimatization. Some fishes were found infested by protozoan parasite at elevated
temperature of 32 1C. Increased levels of certain biochemical and haemotological parameters studied
were strongly correlated with disease in the Cyprinus carpio communis species.
& 2011 Elsevier Ltd. All rights reserved.

1. Introduction viewed as an environmental factor, which evokes multiple effects

Physiological processes in fish are often carried out in a harsh
aquatic environment, and fish face conditions and challenges that
do not exist for terrestrial animals. The maintenance of internal
homeostatic equilibrium is essential for the normal functioning of
the animal and in case of any disturbance the fish will try to
establish a new equilibrium. Any environmental disturbance can
be considered a potential source of stress, as it prompts a number
of responses in the animal to deal with the physiological changes
triggered by exterior changes. However, the harmful effects of
many of these stress-induced factors on the physiological condi-
tion significantly increases with even a moderate change in stress
factors, such as a shift of a few degrees in water temperature or a
reduction by a few mg per liter of dissolved oxygen (Pickering,
1993; Wedemeyer, 1997). Temperature affects virtually all bio-
chemical and physiological activities of an animal. It should be
n
Corresponding author. Tel.: þ91 9906402583; fax: þ 91194 2262214.
E-mail address: mudasirbio@gmail.com (S.M. Ahmad).
on animals (Magnuson et al., 1997).
A long-term change in temperature causes fishes to display
acclimatory responses, which may include enzymatic changes to
mitigate the effect of temperature on metabolism (Hazel and
Prosser, 1974). Temperature is one of the important abiotic
factors, which influences biochemical reactions and therefore
has significant impact on the physiology and biochemistry of
exothermic organisms (Woiwode and Adelman, 1991; Jobling,
1996; Person-Le Ruyet et al., 2004). Temperature beyond opti-
mum limits of a particular species adversely affects the health of
aquatic animals due to metabolic stress (Smith, 1989) hence
increases susceptibility to diseases that have suppressive effects
on growth, reproduction and immune capacity. Temperature of
aquatic environment is important for ensuring survival, distribu-
tion and normal metabolism of fish, failure to adapt to tempera-
ture fluctuations is generally ascribed to the inability of fish to
respond physiologically with resultant mortality, which is related
to changes in the metabolic pathways (Forghally et al., 1973).
According to Donaldson (1981) the physiological responses
can be detected in fish and in other vertebrates in the form of
changes in hormonal or substrate concentrations in the plasma or

0306-4565/$ - see front matter & 2011 Elsevier Ltd. All rights reserved.
doi:10.1016/j.jtherbio.2011.08.007
 S.M. Ahmad et al. / Journal of Thermal Biology 36 (2011) 492–497
alterations in erythrocyte parameters, such as cell volume or
enzyme activities. Measurement of serum biochemical para-
meters can be especially useful to help identify general health
status of animals, and is advocated to provide early warning of
potentially damaging changes in stressed organism (Folmar,
1993; Jacobson-Kram and Keller, 2001). Hematological values
of fishes can be affected by environmental and biological
factors such as age, weight, sex, food, bacteria, parasites and
water quality parameters including water temperature, salinity,
oxygen, availability and pH (Steinhagen et al., 1990 and Haider,
1973).
Common carp (Cyprinus carpio communis) has been one of the
oldest domesticated species of fish for food, dwells well in middle
to lower reaches of rivers and shallow confined waters. Best
growth is obtained at water temperature of 23–30 1C. The fish can
survive cold winter periods, salinity up to about 5%, pH (6.5–9.0),
low oxygen concentration (0.3–0.5 mg/1) as well as super satura-
tion (Flajšhans and Hulata, 2006). However, due to ongoing global
warming conditions about one-fifth of all freshwater fishes are
considered to be endangered (Heywood, 1995).
Kashmir is the north most part of India and lies in north
western part of greater Himalayas and is well known coldwater
fisheries sector of the country. It is the temperate region where
the air temperature fluctuates from 8 to 351C. Common carp
(C. c. communis) is one of the major pond-culture fish of this region
with high commercial value in domestic market. It is highly preferred
fish of this region owing to its high nutritive value. Being commer-
cially important and amenable to aquaculture, it is important to have
knowledge of the acclimatory response of C. c. communis to thermal
stress and this experiment is expected to provide better know-how
for its aquacultural practices.
2. Materials and methods
Fish samples were purchased from commercial fish ponds and
were acclimatized in the laboratory for a week at 20 71 1C.
Healthy fishes (6/aquarium) with a mean weight and length of
20375 g and 1973 cm, respectively, were subjected to experi-
mental conditions. Fishes were stocked at 20 1C (control), 24 1C,
28 1C and 32 1C for a period of one month in 24 tanks (200 L
capacity), six replicates to each treatment. Acclimation procedure
followed in this experiment was based on Manush et al. (2004).
Temperature in each treatment was maintained using tempera-
ture controller fitted with sensors (Selectron process controls Pvt.
Ltd., Mumbai, India). Dissolved oxygen concentration was main-
tained at 670.5 mgl  1 by continuous aeration. Rearing experi-
ments tanks were placed in an indoor experimental area and the
light source was naturally enhanced with fluorescent light. Fishes
were fed once in a day with practical diet (35% crude protein).
Each day, dead fish (if occurred) was removed and in each tank
half of the water volume was renewed every day with filtered
water to assure water quality. The daily removal of water helped
in maintaining the alkaline side of neutrality ranging from 7.5 to
8.00. Similarly the nitrate—nitrogen of water tanks remained in
normal range between 480 mg and 720 mg (APHA, 1998). The
supplemental aeration was also provided to maintain dissolved
oxygen levels near saturation levels. The dissolved oxygen was
measured by Winkler’s Modified method (APHA, 1998) and was
maintained above 6 mgl  1.
2.1. Sample collection and analysis
2.1.1. Sampling
Heamato–biochemical parameters were assessed after a
period of one month rearing experimental fishes at these
493
temperatures. Individual animals were anaesthetized before
sampling with clove oil (150 ml/L) in water. Once the fish
lost equilibrium, the blood samples were drawn by cardiac
puncture, the 25-gage needle of disposable syringes were passed
through the posterior wall of the branchial chamber directly into
the lumen of the bulbus arteriosus. The ammonium salt of
heparin, used in combination with ethylenediaminetetraacetic
acid (di-sodium EDTA) was found adequate to prevent
coagulation.
2.1.2. Blood analysis
For the serum biochemical analysis, blood samples were
prepared using the method described by Bernet et al. (2001) with
slight modification. Blood was allowed to coagulate at room
temperature for 2 h. Serum was obtained by centrifugation of
blood samples at 1500  g (for 10 min at 4 1C) and then stored at
 80 1C for several weeks until analysis. The concentration of
glucose, total protein, cholesterol, triglyceride, blood urea nitro-
gen (BUN), alanine aminotransferase (ALT) and aspartate amino-
transferase (AST) were measured by using a Semi-automatic
blood chemistry analyzer (model—ERBA CHEM—PRO.) and test
kits from Accurex Biomedical Ltd. Immediately after sampling,
the red blood cell (RBC), hemoglobin (Hb), hematocrit (Hct), mean
corpuscular volume (MCV), mean corpuscular hemoglobin (MCH),
mean corpuscular hemoglobin concentration (MCHC), white
blood cells (WBC), lymphocytes and granulocytes were deter-
mined with the help of Automated Hematology Analyzer Hema
Screen—18 (Hopitex Diagnostics, Italy).
2.2. Histology
After 30 days of exposure, six fishes from the experimental and
control groups were euthanized with overdose of MS222 and then
the second gill arch, liver, spleen, trunk kidney and head kidney
were carefully removed and preserved in 10% neutral-buffered
formalin (NBF) for 48 h. Organs were rinsed in 4 changes of 70%
ethanol (ETOH), and stored in 70% ETOH until further processing.
They were dehydrated in isopropanol, cleared in xylene, infil-
trated in paraffin and sectioned at a thickness of 5 mm. Sections
were stained with hematoxylin and eosin and examined with a
light microscope. Sections were stained with hematoxylin and
eosin (Luna, 1968), and examined with a light microscope.
2.3. Data analysis
Data was analyzed using one way analysis of variance
(ANOVA) as described by Gomez and Gomez (1984) and Tukeys’s
multiple range test was carried out for post hoc comparison of
means (Po0.05), if they were significantly different.
3. Results
3.1. Haemato–biochemical profile
Data pertaining to biochemical and hematological parameters
of C. c. communis at 20 1C (control), 24 1C (T24), 28 1C (T28) and
32 1C (T32) are presented in Tables 1 and 2. The result based on
biochemical parameters showed that with the increase in tem-
perature the blood glucose of the experimental fishes increased
from 84.8375.4 mg% (20 1C) to 97.14 73.1 mg% (28 1C) and
decreased to 59 76.6 mg% at 32 1C. The decline in the concentra-
tion of total serum protein at elevated acclimation temperatures
was observed. The mean total serum protein concentration
(1.1670.26 g/dl) at the highest temperature employed (33 1C),
is 66.47% lower than that of the lowest temperature group (20 1C,
494 S.M. Ahmad et al. / Journal of Thermal Biology 36 (2011) 492–497

Table 1
Effect of acclimation temperatures on biochemical parameters in the Cyprinus carpio communis (mean 7S.E).

Temperature 20 1C (Control) 24 1C 28 1C 32 1C

Glucose (mg%) 84.83 7 2.24c 88.76 7 1.81c 97.14 7 1.28b 59.66 7 2.57a
Protein (gram/dl) 3.46 7 0.16c 3.137 0.17c 2.48 7 0.16b 1.167 0.49a
Cholesterol (mg/dl) 142.877 5.88c 144.537 2.66c 167.287 3.40b 194.50 7 5.09a
Triglycerides (mg/dl) 108.50 7 1.52c 112.227 2.34c 125.927 3.15b 149.677 2.74a
Blood urea nitrogen (mg/dl) 1.11 7 0.01d 1.387 0c 2.14 7 0.05b 2.987 0.15a
Alanine amino transferase (ALT, mkat l  1) 0.147 0.05c 0.157 0.09c 0.357 0.18b 1.037 0.25a
Aspartate amino transferase (AST, mkat l  1) 1.93 7 0.09b 1.987 0.17b 2.34 7 0.44b 3.137 0.67a

*Different superscripts (a, b, c, d) within the row differ significantly (P o 0.05).

Means were compared using ANOVA and Tukeys’s multiple range test for post hoc comparison.

Table 2
Effect of acclimation temperatures on heamtological parameters in the Cyprinus carpio communis (mean7 S.E).

Temperature 20 1C (Control) 24 1C 28 1C 32 1C

WBC (  103/mm3) 26.31 7 0.99b c

26.98 7 1.02c 23.63 7 0.58b 16.71 7 1.15a
Lymphocytes (  103/mm3) 41.12 7 0.90d 38.56 7 0.75c 33.56 7 1.02b 25.12 7 0.70a
Granulocytes (  103/mm3) 0.54 7 0.02c 0.51 7 0.16c 0.367 0.14b 0.237 0.08a
RBC (  106/mm3) 1.32 7 0.03b 1.35 7 0.05b 1.61 7 0.05a 1.777 0.04a
HCT (%) 26.65 7 0.15c 27.04 7 0.39c 30.67 7 0.60b 34.71 7 0.17a
Hb (g/dl) 4.37 7 0.14b 5.83 7 0.20a 6.057 0.16a 6.087 0.10a
MCV (fl) 198.767 1.54a 175.437 2.13c 188.547 2.50b 201.77 7 1.43a
MCH (Pg) 43.67 7 1.21a 42.88 7 1.82a 42.16 7 0.90a 39.62 7 1.60a
MCHC (gdl  1) 24.16 7 0.60c 22.73 7 0.76bc 21.44 7 0.79b 19.32 7 0.53a

*Different superscripts (a, b, c, d) within the row differ significantly (P o 0.05).

Means were compared using ANOVA and Tukeys’s multiple range test for post hoc comparison.

Fig. 1. Peripheral blood film from C. c. communis at high temperature (32 oC) for a period of 1 month revealing poiklo-anisocytic, hypochromic RBC’s, (a)  40 and
(b)  100 (Giemsa stain).

3.4670.11 g/dl). Except 20 1C and 24 1C a significant decrease
(Po0.05) existed among different temperature treatments for
protein levels. Unlike serum protein the concentrations of both
these enzymes (ALT and AST) used in this study increased with
the increase in temperature. The concentration of ALT at tem-
perature 32 1C increased 8–9 times (0.14 70.01 mkat l  1 to
1.0370.03 mkat l  1), while as the concentration of AST increased
2–3 times (1.9370.09 mkat l  1 to 3.13 70.67 mkat l  1) over the
control. A significant increase at the 0.05 level of probability was
found for both the enzymes at 28 1C and 32 1C temperature. As
might be anticipated, cholesterol varies in the same fashion as
triglycerides. The highest values for both these parameters were
observed at 32 1C. A significant correlation (Po0.01) was
observed between cholesterol and triglycerides for different
temperature treatments. Blood urea nitrogen (BUN) concentra-
tions at higher temperature exposure exhibited higher values
than those of the control group. The significant increase (P o0.05)
of BUN was found throughout all the temperature treatments. The
difference between the lowest (BUN, 20 1C; 1.11 70.41 mg/d) and
highest blood urea nitrogen (BUN, 32 1C; 2.98 71.43 mg/d) was
62.75%.
The hematological data of C. c. communis (Table 2), showed
decrease in leukocytes (WBC) with the rise in acclimation
temperature. A decrease of 36.48% was noticed at 32 1C over an
ambient temperature of 20 1C. The lymphocytes and granulocyte
number of the experimental animals decreased significantly
(Po 0.05) with the rise in temperature. The peripheral blood film
of thermally stressed fishes revealed poikilo-anisocytosis and
hypochromia (Fig. 1). This type of anemia was only found at
protracted exposure of fishes at 32 1C. The results of the study
upon Hb, RBC and Hct are evident in Table 2, showed a common
trend of increase with respect to the increase in temperature.
Although the highest mean Hb level was that encountered at
32 1C (6.08 70.46 g/dl), this was not greatly different from that of
 S.M. Ahmad et al. / Journal of Thermal Biology 36 (2011) 492–497
28 1C (6.05 70.88 g/dl) group. The RBCs at 32 1C (1.7770.05 
106/mm3) were 25.42% higher than lowest value recorded (RBC
20 1C¼1.32 70.05  106/mm3). Effect of increasing temperature
on Hct showed significant difference at 0.05 level of probability
between different groups. Values derived for MCV showed wide
variation with the rise in temperature. Compared to 24 1C and
28 1C, the variation of only 1.51% was observed between 20 1C
(198.7675.99) and 32 1C (201.77 719.95). The values for both
MCH and MCHC tend to decline at higher acclimation tempera-
tures. The value for MCH at 20 1C ¼43.67 72.26 showed 99.21%
and 98.49% similarity with 24 1C¼42.88 72.47 and 28 1C¼
42.16 71.72, while as 9.2% variation was observed with 32 1C¼
39.62 76.68. The level of variation between different values of
MCHC however, ranged from 5.91% to 20%.
3.2. Histological examination of fish tissues
No specific histological alterations were found in the organs of
the fishes exposed to control (20), 24, and 28 1C. Most of the
histopathological manifestations were marked after the pro-
longed exposure of 32 1C. In the present study, exposure of carp
to thermal stress resulted interrenal cell hypertrophy and mela-
nomacrophage centers were scattered throughout kidney and
spleen. The kidney of the fish often showed vacuolar degeneration
of tubular epithelium associated with lysis and loss of tubular
structure. In some cases organization of necrosed tubules was
observed (Fig. 2). Trunk kidney of fish showed enlarged sinusoids
and decreased amount of hematopoietic tissue. Vacuolar change
was the only lesion observed in liver. Gills revealed separation of
epithelium from lamellae, lamellar fusion and swelling of the
epithelial cells. Further histopathological examination of their
gills revealed that the fishes were seen infected with protozoan
parasite, which was identified as Trichodina (Fig. 3). The highest
infestation of the parasite was seen in the branchial cavity.
The parasitic infection was associated with excessive mucus
production, which resulted in respiratory problems. Due to this
problem the fish piped most of the time on the surface of the
water tank followed by rhythmic flashing and bouts of fringed
activity.
4. Discussion
Temperature beyond optimum limits for a particular species,
adversely affects the health of aquatic animals due to metabolic
stress hence increases susceptibility to diseases that have sup-
pressive effects on growth, reproduction and immune capacity
(Cnaani, 2006). Our results clearly showed that with increase in
Fig. 2. Histology section of trunk and portion of head kidney from thermally stressed C. c. communis at 32 oC for a period of 1 month revealing (a) aggregation of
melanomacrophagic centers (MMC’s, H and E, bar ¼200 mm) (b) hypertrophy of renal tubules following thermal stress (bar¼ 100 mm).
495
temperature C. c. communis got stress, which affected all bio-
chemical and physiological activities. High stress conditions were
observed at a temperature of 32 1C. At this temperature the fish
showed less appetite for feed, irregular behavior of swimming
and always gulped for air. However, the adaptive capability of
C. c. communis enabled it to survive through stressful temperature
conditions.
Measurement of serum biochemical parameters gave general
health status, and provided early warnings of damaged changes in
stressed fishes. The results depicted in Table 1 showed increase in
the blood glucose up to 28 1C, a sharp decrease was noticed at
32 1C. A prolonged exposure of high temperature of 32 1C showed
negative effect on blood glucose of C. c. communis. Blood glucose
does not show significant correlation with other biochemical
parameters studied. The highest blood glucose level at low
temperatures is indicative of retarded carbohydrate metabolism,
and is also an index of sub-lethal stress (Smith et al., 1976;
Connors et al., 1978; Best et al., 2001). Further decreased serum
glucose concentrations could be the result of undernourishment
as well as impaired hormonal control. A continuous decrease in
total serum protein with increase in temperature was observed in
our studies. As shown in results 66.47% decrease of total serum
protein was observed at 32 1C over the control temperature of
20 1C. Decreased concentrations of serum protein are common in
many disease status and may result from impaired synthesis
(liver disease), reduced absorption or protein loss (Bernet et al.,
2001). As reported by Walsh et al. (2003) the liver is the primary
organ for urea production and gills appearing to be the main
organ of excretion. A significant effect of temperature on BUN was
Fig. 3. Gill photomicrography of C. c. communis revealing presence of Trichodina and
degenerative changes in lamellae at 321C for a period of one month (bar¼ 100 mm).
496 S.M. Ahmad et al. / Journal of Thermal Biology 36 (2011) 492–497
found throughout all the treatments. Except for blood glucose,
BUN showed significant correlation with all other biochemical
parameters studied. Mensinger et al., 2005 reported that diseased
fish showed significantly elevated BUN levels prior to dying, likely
due to osmoregulatory failure. In our studies a variation of 28% in
BUN was observed between 28 1C and 32 1C is an indicator of gill
osmoregulatory problems. The elevated BUN is more likely
associated with gill or liver disease and probably not indicative
of renal disease (Mensinger et al., 2005). Osmoregulatory failure
has often been demonstrated to be an important contributor to
death in fish (Wood et al., 2003).
Increased concentration of cholesterol and triglycerides with
the rise in water temperature was observed in our study. This
indicates disorders of lipid and lipoprotein metabolism and
especially liver disease. The results suggest that the cells are able
to adjust membrane fluidity by varying the concentration of
cholesterol. These results are compatible with the concept of
homeoviscous adaption: that fish strive to maintain an optimal
level of membrane fluidity and when grown at different tem-
perature will alter lipid composition in order to maintain this
level. These results are consistent with that of Robertson and
Hazel, 1995. Further, increased serum cholesterol concentrations
can result from damage of liver or nephritic syndrome (Yamawaki
et al., 1986; Seyit et al., 2000). Bernet et al., 2001, however
reported that high concentrations of triglycerides may occur with
nephritic syndrome or glycogen storage disease. Among the most
sensitive and widely used liver enzymes are the aminotrans-
ferases. They include alanine aminotransferase (ALT or SGPT) and
aspartate aminotransferase (AST or SGOT). These enzymes are
normally contained within liver cells. The increased level of these
enzymes in blood at high temperature in our study may be due to
liver injury or hepatic metabolism hyperactivity of the experi-
mental animals. The increased SGOT and SGPT values in fishes
reveal enzymes exporting from liver into bloodstream (Yang and
Chen, 2003; Perez-Rostro et al., 2004). According to Chen et al.,
2004 increased level of SGOT and SGPT in Tilapia is associated
with hepatic injury, acute injuries in trunk kidney, bacterial
infection and myocardial infarction. Barcellos et al., 2003 sug-
gested that acute stress condition of Rhamdia quelen Quoy and
Gaimard Pimelodidae resulted in increased SGOT and SGPT due to
hyper liver metabolism.
Hematological profiles have often been used as stress indica-
tors (Dobšikova et al., 2009). Major shifts in the haemogram are
found in fish exposed to acute or chronic stress. The statistical
analysis showed non-significant increase of RBC and Hb with the
increase in acclimation temperature. These results are in agree-
ment with those found by Zarejabad et al. (2009) who studied the
effects of rearing temperature on hematological and biochemical
parameters of great sturgeon and Carine Luı́sa et al. (2004) who
studied the effect of different temperature regimes on metabolic
and blood parameters of silver catfish. Naidu et al. (1989)
reported that due to stress, fish tried to cope with adverse
conditions by enhancing their respiratory capabilities through
evaluated RBC and Hb synthesis. Our results also showed sig-
nificant increase in Hct concentration over the control. In stressed
fish an increase in RBC, hemoglobin and hematocrit concentration
is often observed (Svobodova et al., 1994). Elevated hemoglobin
and hematocrit increase the oxygen carrying capacity of blood,
and thus the oxygen supply to major organs, in response to higher
metabolic demand is the manifestation of stress (Ruane et al.,
1999). However, prolonged exposure of stress may pose a great
threat to their survival. The other hematological parameters
affected by temperature fluctuations include MCH and MCHC.
The results showed drop in MCH and MCHC is associated with the
decrease in RBC and Hb. While examining the peripheral blood
film after period of one month at 32 1C revealed poikilo-anisocytic
and hypochromic RBCs. These results clearly indicate stress and
undernourishment (due to stopping of feeding in elevated tem-
peratures) leading to anisocytic hypochromic anemia of experi-
mental animals at highest temperature. This further suggests that
C. c. communis has an adaptive ability upto certain limit of
temperature. Alteration of hemoglobin leads to loss of hemoglo-
bin solubility, resulting in structural damage to erythrocytes and
it can subsequently cause rapid lyses of the erythrocytes (Bloom
and Brandt, 2001; Everse and Hsia, 1997). As shown in Table 2,
the results obtained for MCV are temperature-independent as it
tends to stabilize at high temperatures. This also shows thermal
adaptabity of experimental fishes to varied temperature condi-
tions. The values of MCV, MCH and MCHC of the present study
correspond to the results of Adeyemo et al. (2003) and Zarejabad
et al. (2009).
In present study the organs examined showed marked
changes, which correlate well with the haemato–biochemical
observation. Kidney (Bucher and Hofer, 1993), liver (ICES, 1997)
and Gills (Poleksic and Mitrovic-Tutundzic, 1994) have been
considered as suitable organs for histological examination in
order to determine the effect of stress. The histopathological
results of kidney showed vacuolar degeneration of tubular
epithelium, loss of tubular structure and necrosis of the renal
tubules. The present results are in agreement with those observed
in Labeo rohita (Hamilton) acclimated to various temperatures
(Dash et al., 2011). The liver plays a key role in the metabolism
and biochemical transformations, which inevitably reflects on its
integrity by creating lesions and other histopathological altera-
tions of the liver parenchyma or the bile duct (Roberts, 1978). We
found vacuolar change was the only lesion observed in liver. As
discussed above the increase in aspartate aminotransferase (AST)
and alanine aminotransferase (ALT) may be due to hepatic
pathological alterations of thermally stress fishes. Increased AST
and ALT activities have been associated with hepatic pathology in
fishes (Casillas et al., 1983). High temperatures can injure gills,
thus reducing the oxygen consumption and disrupting the
osmoregulatory function of fish (Ghate and Mulherkar, 1979;
Saravana Bhavan and Geraldine, 2000). The present results
showed degenerative changes of gills. These results are in con-
gruence with the findings of earlier workers (Sunitha and Sahai,
1983; Roy and Munshi, 1991; Saravana Bhavan and Geraldine,
2000; Dash et al., 2011).
The observation of parasitic infestation of gills in present study
indicated the increased susceptibility of the thermally stressed
fishes to such infections. Stress has been recognized as one of the
important factors favouring parasitic infestation in fishes
(Mcintyre, 1996). The pathological alterations observed in the
gills of affected fishes may be ascribed partly to high parasitic
load. The heavy load of parasite in branchial cavity associated
with excess mucus production may be one of the factors causing
osmoregulatory disturbance. Fish with severe gill infections of
trichodina will have respiratory and osmoregulatory difficulty
and may ‘‘pipe’’ as well as ‘‘flash’’ if there is cutaneous involve-
ment (http://ag.ansc.purdue.edu/courses/aq448/diseases/para
sites.htm).
In summary, knowledge of the hematological and biochemical
values is important tool that can be used as an effective and
sensitive index to monitor physiological and pathological changes
in a particular fish. Further, this study also supports the fact
that fishes have a remarkable resilience to altered temperature
especially with a prolonged time course and physiological
changes during acclimation, which may help common carp of
cold waters to accommodate climatic change. However, the study
indicated the upper thermal limit as 32 1C for common carp as
they couldn’t survive hotter conditions (above 32 1C) for more
than a few hours.
 S.M. Ahmad et al. / Journal of Thermal Biology 36 (2011) 492–497
Acknowledgments
We greatly appreciate all the help and support provided by
Dr. M.H Balkhi, Dean Faculty of Fisheries SKUAST-K, Shalimar. The
help of Dr. Masood Saleem Mir, for some valuable suggestions
regarding histopathology is greatly acknowledged. We also thank
an anonymous reviewer for very helpful comments. This work is
the part of RCM, supported by Sher-e-Kashmir University of
Agricultural sciences and Technology of Kashmir, India.
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