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Summary

  1. Top of page
  2. Summary
  3. Introduction
  4. Materials and methods
  5. Results
  6. Discussion
  7. Acknowledgement
  8. References

Hematological and plasma chemistry indices are simple and essential diagnostic tools for monitoring the physiological and health status of fish. Aim of the present study was to obtain reference values for the hematological and plasma chemistry of wild populations of Labeo rohita captured in a freshwater pond between July 2008 and June 2010. These reference values and the mean were evaluated according to sex and season. In summer, the red blood cells (1.84 × 106 38 per cubic mm), haemoglobin (8.52 gm dl−1) and haematocrit (31.49%) were highest in males, whereas the maximum values for white blood cells (5.635 × 103 40 per cubic mm) were found in females, however, no significant variation of mean corpuscular volume (MCV) or mean corpuscular haemoglobin (MCH) was observed between sexes. Various blood parameter levels between the sexes in summer were notably different from those measured in other seasons except for MCH and MCHC values (p < 0.05). Compared to most teleosts, the L. rohita has similar mean values for PCV and Hb. Throughout summer the glucose (76.0 mg dl−1), lipid (3.41 gm dl−1) and cholesterol (145.0 mg dl−1) levels were highest. In spring the plasma protein levels were higher in males, but higher in winter for females. Consequently, the seasons are key factors when using blood parameters as biomarkers for environmental alterations.


Introduction

  1. Top of page
  2. Summary
  3. Introduction
  4. Materials and methods
  5. Results
  6. Discussion
  7. Acknowledgement
  8. References

Knowledge of the hematologic and biochemical parameters of aquatic species is important for assessing and managing their health status in natural habitats. Evaluation of the clinical status is compromised by the limited diagnostic techniques whereby hematologic and plasma chemistry parameters can provide predictive, although highly variable, information, depending on the fish species, age, cycle of sexual maturity, and health condition (Hrubec et al., 2001; Rey Vázquez and Guerrero, 2007). Gabriel et al. (2004) demonstrated that hematological parameters are closely related to the response of an animal to the environment, an indication that the environment of the fishes could exert an influence on the hematological characteristics. Many publications report on the hematological aspects of vertebrate species in order to correlate their physiology and evolution; a thorough knowledge of the fish physiology is thus imperative.

Periodic hematological analyses provide a simple means of evaluating stress, metabolic disorders, reproductive dysfunctions, and disease caused by environmental husbandry conditions in wild and cultured fish (Filiciotto et al., 2012; Pradhan et al., 2012b). Hematological indices have been employed in effectively monitoring fish responses to stressors, and thus their health status, for the quality of fish under adverse conditions (Rey Vázquez and Guerrero, 2007). A difficulty in assessing the propagation of a natural fish population has been the paucity of reliable reference values in a natural habitat. Pursuant to this objective, many fish physiologists have concentrated on hematological studies as proven valuable diagnostic tools in evaluating fish quality (Kori-Siakpere et al., 2005; Oluyemi et al., 2008). Determination of hematological parameters can provide substantial diagnostic information, once standardized reference values are established (Jamalzadeh et al., 2008). Moreover, recent attention has been given to the biochemical characterization of fish blood as an internal index (Edsall, 1999). Many studies have demonstrated the usefulness of hematology and blood biochemistry in the assessment of fish health and as biomarkers of exposure to pollution (Handy and Depledge, 1999).

Evaluation of the hemogram involves determining the total erythrocyte count (RBC), total white blood cell count (WBC), hematocrit (PCV), hemoglobin concentration (Hb), erythrocyte indices (MCV, MCH, MCHC), white blood cell differential count, and the evaluation of stained peripheral blood film (Campbell, 2004). Hematocrit, erythrocytes counts and haemoglobin concentrations are the most readily determined haematological parameters in field and hatchery conditions (Bhaskar and Rao, 1990). Wedemeyer et al. (1983) demonstrated that the hematocrit value is not as easily altered as other parameters, and should be used in conjunction with erythrocyte and leucocyte counts, hemoglobin contents, osmotic fragility and differential leucocyte counts. Variations in hematological parameters of fishes are caused by malnutrition (Ighwela et al., 2012), gender (Collazos et al., 1998), seasonal differences and breeding efficiency (Pradhan et al., 2012b), environmental stress (Singh and Tandon, 2009), and fish size (Garcia et al., 1992). The influence of environmental factors on blood parameters of fishes (Orun et al., 2003) has generally been used as an effective and sensitive index to monitor the physiological and pathological changes in fish health (Chakrabarthy and Banerjee, 1988). Interestingly, the hematology of fish continues to offer a valuable diagnostic tool as well as progress in establishing normal range values for blood parameters in different fish species, but information regarding Cyprinidae species is incomplete. L. rohita belongs to the Cyprinidae family and is common in rivers and freshwater lakes of South Asia and South-East Asia. Among all Indian carps, L. rohita is the most popular and delicious food fish in Asia including India, and rich in protein content. To better understand the health status of L. rohita in a natural habitat, hematology and plasma biochemistry parameters were investigated for two consecutive years with the objective that L. rohita might provide some useful information that could be used as a biomarker associated with stressor agents or as an available tool to diagnose and monitor disease in this species, collect baseline data on hematology and blood biochemistry of the freshwater L. rohita and calculate normal reference values, as well as determine if there are seasonal or annual physiologic differences in male and female fish living in natural environments.

Materials and methods

  1. Top of page
  2. Summary
  3. Introduction
  4. Materials and methods
  5. Results
  6. Discussion
  7. Acknowledgement
  8. References

Study site and fish collection

Fish and water samples were collected seasonally in the natural habitat of a freshwater pond in Khurda District, Bhubaneswar (19º40″−20º25″N; 24º55″−36º05″E; area, 2889 km2), India, from July 2008 to June 2010. The area and pond were chosen considering the tropical climate with an average ambient temperature range between 20.5°C (winter) to 30.5°C (summer), an annual rainfall of 1449.1 mm, as well as for the agricultural, domestic, fish farming and human activities. Water samples were collected in triplicate in different areas of the pond in 1-L glass containers prewashed with diluted hydrochloric acid and rinsed thoroughly with the sample water before filling to capacity and labelling accordingly. Meanwhile, pond water temperature and pH were measured in the field. For water quality and dissolved oxygen concentration analyses, fish samples were transported on the same day in 300-L tanks filled with properly oxygenated pond water to the laboratory at Utkal University, where the fish were maintained for 15-day acclimatization to laboratory conditions (APHA, 1998). Ten adult specimens (34.5 ± 1.5 cm standard length, 1050 ± 50 g weight) of each sex were sacrificed for the experiment.

Blood sample collection

Blood samples were collected as described by Lavanya et al. (2011). Briefly, blood was taken from the caudal peduncle and sometimes from the heart by cardiac puncture using a disposable plastic syringe fitted with a 26-gauge needle moistened with heparin; each sample was immediately deposited in a separate heparinised vial on ice (Lavanya et al., 2011). Whole blood was used for the estimation of hemoglobin, and RBC and WBC counts.

Hematological analysis

For hemoglobin concentration estimation the cynmethaemoblobin method was used (Lavanya et al., 2011). To count the number of RBCs and WBCs we followed standard procedures as published in Gabriel et al. (2004). The results were expressed as the number of RBCs or WBCs per 1 mm3 of blood. Total red blood corpuscles or erythrocytes were counted in an improved Neubauer hemocytometer using Hayem's diluting fluid. The number of counted cells was determined following the method of Oluyemi et al. (2008). Total RBC count per mm³ = 200 × 50 × N = 10 000 N (N = Number of RBC counted, Dilution factor = 200). Total WBC or leucocytes were counted in an improved Neubauer hemocytometer using Tukey's diluting fluid. Total count per mm3 was 20 × 1 × L/0.4 cells = 50 × L (L = number of WBCs counted, dilution factor = 50). Determination of the packed cell volume (PCV) was performed according to Adebayo et al. (2007). The PCV was estimated using a microhaemotocrit reader and expressed as a percentage. Erythrocyte indices MCV, MCH, and MCHC were calculated as per formulae (MCV = PCV × 100/erythrocyte count, MCH = hemoglobin × 10/erythrocyte count, MCHC = hemoglobin × 100/PCV).

Biochemical analysis

Plasma protein estimation was according to the method of Lowry et al. (1951) and the amount of protein expressed as gm dl−1. The blood glucose level was determined by GOD/POD method using Kit (Ceral clinical system, India) and the amount of glucose expressed as mg dl−1 (Triander, 1969). Lipid concentration was estimated as per Frings and Dunn (1970) and the amount of lipid expressed as gm dl−1. The cholesterol level was determined by the CHOD/PAP method using Kit (Crest Biosystem, India) and the amount of cholesterol expressed as mg dl−1 (Triander, 1969).

Statistical analysis

Hematological data from the experiment were subjected to one-way analysis of variance (anova) using SPSS 17 for Windows. Differences between means were determined by Duncan's multiple range test (DMRT) (p < 0.05). The correlation between hematological variables was analysed by the Pearson coefficient for linear correlation at p < 0.05.

Results

  1. Top of page
  2. Summary
  3. Introduction
  4. Materials and methods
  5. Results
  6. Discussion
  7. Acknowledgement
  8. References

Differences in hematological parameters of the freshwater L. rohita were statistically analysed by one-way anova. As shown in Table 1, there were significant seasonal differences between sexes in hematological parameters. In males, the Hb concentration varied from 5.5 to 8.5 gm dl−1, with an annual mean of 7.03 ± 0.2 gm dl−1. In females, the values ranged from 4.8 to 7.3 gm dl−1, with an annual mean of 6.1 ± 0.3 gm dl−1. The higher concentration of Hb in both sexes was in summer; there were otherwise significant differences between males and females (Fig. 1), whereby the total number of erythrocytes ranged from 1.4 × 106 to 1.8 × 106 mm−3 blood in males and 1.2–1.7 × 106 mm−3 blood in females during the annual study period. Male fish always had higher RBC numbers than the females, and with highest overall values in summer (Fig. 2). The seasonal total leukocyte count (TLC) value was higher in summer and lower in winter for both sexes of L. rohita (Fig. 3). TLC varied between 2.8 × 103 and 5.1 × 103 mm−3 and 3.0 × 103 and 5.6 × 103 mm−3 blood in males and females, respectively. Annual mean values for males and females were 3.7 × 103 and 4.1 × 103 per cubic mm of blood, respectively. As shown in Fig. 4, the seasonal PCV value was higher in the summer and lower in the winter, irrespective of the fish sex. The PCV value ranged from 24.1 to 33.2% in males, and varied between 22.5 and 27.6% in females; the mean value for males and females was 29.6 and 24.5%, respectively, indicating that this was higher for males throughout the study period. MCV value was higher in summer and lower in winter for both sexes (Fig. 5). The MCV value for males ranged between 173.0 and 180.7 μ3 and 173.0–180.7 μ3 for females. There were no major differences in male and female Labeo rohita. The value of MCH throughout the experimental period varied between 51.8 and 54.8 pg in males and 55.5–57.7 pg in female L. rohita. MCH was higher in females (annual mean 55.2 pg) than in males (54.4 pg). In both sexes, a higher value was observed in spring, with lower values in summer and autumn (Fig. 6). The MCHC value ranged from 30.7 to 32.2% in males, and between 31.7 and 32.5% in females. For male L. rohita, higher values were observed in the rainy season and lower values in winter; for females, lower values were in spring (Fig. 7).

Table 1. Analysis of one-way Anova (anova) on mean blood parameters, e.g. hemoglobin (Hb), erythrocytes (RBCs), leucocyte (WBCs), hematocrit or packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) and mean corpuscular hemoglobin (MCH) by sex (N = 10 per sex per month) and seasons in Labeo rohita, July 2008 to June 2010
VariablesSourceType I sum of squaresd.f.Mean squareFSignificance
HbSeason27.546.883.30.0
Sex7.817.895.20.0
Season * sex0.440.11.40.2
Error1.6200.0  
Corrected total37.529   
RBCsSeason0.740.12.10.1
Sex0.410.45.10.0
Season * sex0.040.00.20.9
Error1.7200.0  
Corrected total3.029   
WBCsSeason23.945.938.10.0
Sex1.111.17.10.0
Season * sex0.140.00.20.9
Error3.1200.1  
Corrected total28.329   
PCVSeason236.2459.0231.30.0
Sex195.11195.1764.50.0
Season * sex7.441.87.30.0
Error5.1200.2  
Corrected total443.929   
MCVSeason301.6475.40.90.4
Sex7.017.00.00.7
Season * sex11.242.80.00.9
Error1580.62079.0  
Corrected total1900.629   
MCHCSeason4.741.14.10.0
Sex0.710.72.60.1
Season * sex2.440.62.10.1
Error5.6200.2  
Corrected total13.529   
MCHSeason26.846.718.10.0
Sex14.9114.940.50.0
Season * sex24.746.116.70.0
Error7.3200.3  
Corrected total73.929   
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Figure 1. Average concentration of blood Hb of Labeo rohita between sexes and seasons. Bars = means, vertical lines SE (N = 10 per sex per month)

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Figure 2. Average number of erythrocyte counts of Labeo rohita between sexes and seasons. Bars = means, vertical lines SE (N = 10 per sex per month)

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Figure 3. Average number of leukocyte counts of Labeo rohita between sexes and seasons. Bars = means, vertical lines SE (N = 10 per sex per month)

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Figure 4. Average hematocrit value of Labeo rohita between sexes and seasons. Bars = means, vertical lines SE (N = 10 per sex per month)

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Figure 5. Average number, mean cell volume (MCV) of Labeo rohita between sexes and seasons. Bars = means, vertical lines SE (N = 10 per sex per month)

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Figure 6. Average mean cell hemoglobin concentration (MCHC) of Labeo rohita between sexes and seasons. Bars = means, vertical lines SE (N = 10 per sex per month)

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Figure 7. Average mean cell hemoglobin (MCH) of Labeo rohita between sexes and seasons. Bars = means, vertical lines SE (N = 10 per sex per month)

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For selective plasma biochemistry analysis of each adult fish, sex and seasonal variations were considered during our annual study period (Table 2). Annual mean biochemical parameters were indicated as slightly higher in females (31.9%) in comparison to males (31.6%) such as protein, lipid, cholesterol, and glucose for both sexes in the different seasons. Seasonal variation in amounts of protein among the test fishes ranged from 4.0 (autumn) to 4.8 (spring) gm dl−1. Higher seasonal values were observed in the spring for males and winter in for females. Similarly, higher amounts of protein were found in male groups. Lipid content of fish showed seasonal variations, ranging from 2.7 to 3.4 gm dl−1. The highest lipid value was in summer for females; the lowest value was in the rainy season for males. Cholesterol ranged from 143.3 (winter) to 159.0 (summer) mg dl−1. Seasonally, a higher amount of cholesterol was observed in spring. The amount of glucose varied from 49.9 (winter) to 59.0 (summer) mg dl−1.

Table 2. Seasonal variations in plasma biochemical parameter parameters including protein, lipid, cholesterol and glucose of male (N = 10 per month) and female (N = 10 per month) Labeo rohita, July 2008 to June 2010
SeasonSex ProteinLipidCholesterolGlucose
  1. Mean values followed by different letters in rows are significantly different, p = 0.05.

SpringMaleMean4.43.2129.058.0
SEM0.2b0.1b0.5c0.1a
FemaleMean4.23.2136.356.3
SEM0.2b0.2b2.3c0.1a
SummerMaleMean4.13.3133.374.3
SEM0.4a0.2c1.6b0.9c
FemaleMean3.63.5140.071.0
SEM0.4a0.2b2.6a0.2c
RainyMaleMean3.82.8130.362.1
SEM0.1ab0.1a1.2b0.9ab
FemaleMean3.22.7133.061.3
SEM0.3ab0.1ab2.1b1.0ab
AutumnMaleMean3.62.8130.053.7
SEM0.3a0.1c0.5b0.1d
FemaleMean3.52.8133.259.9
SEM0.3a0.1c0.5b0.1d
WinterMaleMean4.02.9122.450.2
SEM0.2a0.1a0.5c0.4c
FemaleMean4.22.9127.353.0
SEM0.3c0.1a2.4b0.2c

Hematological variables are valuable tools in diagnosis procedures. Correlations among hematological parameters are shown in Tables 3-5. The highest positive correlation was for Hb-Hct (r = 0.9, p < 0.01), RBC-MCV (r = 0.8, p < 0.01), and MCHC-RBC (r = 0.8, p < 0.01). A negative correlation found between MCH-WBC (r = −0.4). The highest positive correlations for females were Hb-Hct (r = 0.9, p < 0.01), WBC-Hb (r = 0.9, p < 0.01), and Hct-Hb (r = 0.9, p < 0.01). A negative correlation was found between MCH-MCHC (r = −0.2). The analysis among plasma biochemical parameters showed a strong and positive correlation of protein-glucose (r = 0.4 at p ≤ 0.01) and also lipid-cholesterol (r = 0.4 at p ≤ 0.01) (Table 6).

Table 3. Correlation analyses of blood parameters including hemoglobin (Hb), erythrocytes (RBCs), leucocyte (WBCs), hematocrit or packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) and mean corpuscular hemoglobin (MCH) by sex (N = 10 per sex per month) and seasons in Labeo rohita, July 2008 to June 2010
SeasonHbRBCsWBCsPCVMCVMCHCMCH
  1. a

    Correlation is significant at 0.05 level (2-tailed).

  2. b

    Correlation is significant at 0.01 level (2-tailed).

Spring
Hb10.7−0.50.9b0.40.9b−0.6
RBC0.710.0710.50.9b0.9b−0.0
WBC−0.50.01−0.70.4−0.20.9b
PCV0.9b0.5−0.710.10.7−0.8a
MCV0.40.9b0.40.110.70.3
MCHC0.9b0.9b−0.20.70.71−0.4
MCH−0.6−0.00.9b−0.8a0.3−0.41
Summer
Hb10.5−0.00.9b0.40.1−0.8a
RBC0.510.70.30.9b0.8a0.0
WBC−0.00.71−0.30.8a0.9b0.5
PCV0.9b0.3−0.310.1−0.0−0.9b
MCV0.40.9b0.8a0.110.9b0.1
MCHC0.10.8a0.9b−0.00.9b10.3
MCH−0.8a0.00.5−0.9b0.10.31
Rainy
Hb10.7−0.10.9b0.20.10.0
RBC0.710.40.60.70.60.5
WBC−0.10.41−0.30.8a0.9b0.9b
PCV0.9b0.6−0.310.0−0.1−0.2
MCV0.20.70.8a0.010.9b0.9b
MCHC0.10.60.9b−0.10.9b10.9b
MCH0.00.50.9b−0.20.9b0.9b1
Autumn
Hb10.9b0.00.9b0.5−0.10.5
RBC0.9b10.20.70.8a0.10.7
WBC0.00.21−0.30.70.9b0.8a
PCV0.9b0.7−0.310.2−0.40.2
MCV0.50.8a0.70.210.70.9b
MCHC−0.10.10.9b−0.40.710.7
MCH0.50.70.8a0.20.9b0.71
Winter
Hb10.8a−0.10.9b0.6−0.60.6
RBC0.8a10.20.8a0.8a−0.20.9b
WBC−0.10.21−0.20.70.8a0.5
PCV0.9b0.8a−0.210.4−0.70.6
MCV0.60.8a0.70.410.20.9b
MCHC−0.6−0.20.8a−0.70.210.0
MCH0.60.9b0.50.60.9b0.01
Table 4. Correlation analysis of blood parameters, e.g. hemoglobin (Hb), erythrocytes (RBCs), leucocyte (WBCs), hematocrit or packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) and mean corpuscular hemoglobin (MCH) for male Labeo rohita (N = 10 per month), July 2008 to June 2010
ParametersHbRBCWBCPCVMCVMCHCMCH
  1. a

    Correlation significant at 0.05 level (2-tailed).

  2. b

    Correlation significant at 0.01 level (2-tailed).

Hb10.5a0.9b0.9b0.40.7b−0.4
RBC0.5a10.5a0.5a0.8b0.8b0.1
WBC0.9b0.5a10.9b0.5a0.6a−0.4
PCV0.9b0.5a0.9b10.30.7b−0.3
MCV0.40.8b0.5a0.310.6a0.0
MCHC0.7b0.8b0.6a0.7b0.6a10.1
MCH−0.40.1−0.4−0.30.00.11
Table 5. Correlation analysis of blood parameters including hemoglobin (Hb), erythrocytes (RBCs), leucocyte (WBCs), hematocrit or packed cell volume (PCV), mean corpuscular volume (MCV), mean corpuscular hemoglobin concentration (MCHC) and mean corpuscular hemoglobin (MCH) for female Labeo rohita (N = 10 per month), July 2008 to June 2010
ParametersHbRBCWBCPCVMCVMCHCMCH
  1. a

    Correlation significant at 0.05 level (2-tailed).

  2. b

    Correlation significant at 0.01 level (2-tailed).

Hb10.8b0.9b0.9b0.5a0.10.7b
RBC0.8b10.8b0.7b0.8b0.40.6b
WBC0.9b0.8b10.9b0.6b0.10.6b
PCV0.9b0.7b0.9b10.4−0.00.7b
MCV0.5a0.8b0.6b0.410.7b0.3
MCHC0.10.40.1−0.00.7b1−0.2
MCH0.7b0.6b0.6b0.7b0.3−0.21
Table 6. Correlation matrix between plasma biochemical parameters including protein, lipid, cholesterol and glucose by sex (N = 10 from each sex in each month) and seasons in Labeo rohita from July 2008 to June 2010
ParameterProteinLipidCholesterolGlucose
  1. a

    p ≤ 0.05 (0.205 table value).

  2. b

    p ≤ 0.01 (0.267 table value).

Protein1.0   
Lipid0.2a1.0  
Cholesterol0.3b0.4b1.0 
Glucose0.4b−0.00.4b1.0

Discussion

  1. Top of page
  2. Summary
  3. Introduction
  4. Materials and methods
  5. Results
  6. Discussion
  7. Acknowledgement
  8. References

In recent years there has been a growing interest in the study of hematological and biochemical parameters of fish blood, regarded as important for aquaculture purposes. The study of blood characters can corroborate the diagnoses and prognoses of morbid conditions in fish populations (Tavares-Dias and Moraes, 2004) and thereby contribute to better comprehension of the comparative physiology, phylogenetic relations, feeding conditions and other ecological parameters. Baseline data regarding hematological and plasma biochemical reference values facilitates the fish health condition assessment of diagnostics in freshwater fish (Pradhan et al., 2012b). There are abundant and excellent references for guidance regarding abnormal fish health, and the incisive expert needs to use all available diagnostic skills to manage these cases whereby the use of clinical pathology is a great enhancement for this challenge. Once reference values are determined in L. rohita, they can facilitate monitoring of the fish stress response, their nutritional condition, reproductive state, tissue damage due to frequent handling procedures, detection, and diagnosis of metabolic disturbances or disease processes in the fish population (Congleton and Wagner, 2006). Many hematological reports are available for small and young Cyprinidae fishes by different researchers using various approaches; the present work basically focused on reference ranges for hematological and plasma biochemical parameters in adult species of L. rohita. Our hematological data showed marginal differences in comparison with Clarias isheriensis (Kori-Siakpere, 1985) and Clarias gariepinus (Oluyemi et al., 2008), and some Cyprinidae species such as Catla catla (Pradhan et al., 2012a) and Cirrhinus mrigala (Pradhan et al., 2012b). Many publications demonstrated that variations in blood index values are attributable to factors such as age, fish size, nutritional state, season, spawning, sex, and genetic variation in different fish species. The present study findings on hematological parameters in L. rohita show statistical differences in seasonal and sex variations. Moreover, this study further attributes that because of a high body metabolic rate during summer, the high ambient temperature and reproductive activities, most of the hematological parameters showed higher values than in other seasons. The lowest value, on the other hand, was during winter, which might be because of the low ambient temperature and low metabolic rate. These findings are supported by our own research and by other researchers in other species (Orun et al., 2003; Adebayo et al., 2007). Numerous publications demonstrated that variation in hematocrit value and other hematological parameters between sexes and their diversity might be due to the higher metabolic rates of male fish. Findings in the present work also support this idea, which is related to an increase in fish activity with an increase in size. Considering the sex variations, many studies demonstrated that the male fish has higher values in almost all hematological parameters except in TLC (Orun et al., 2003). These higher values favouring males may be attributed to their physiological activeness. During our 2-year study, MCHC and MCH values did not show any marked differences between seasons and sexes. In freshwater fish, the blood glucose level may vary due to different external as well as internal factors, including diet. Declines in glucose levels during winter probably reflect the reduced feed intake in comparison to spring and summer as well as the increase in tissue uptake that is mediated by pancreatic hormones (Sheridan and Mommsen, 1991). Previous studies by various researchers demonstrated that basal levels of glucose varied in ecologically-distinct species, in part influenced by environmental and non-environmental factors such as feeding habits and life mode of the fish, particularly related to locomotion (Tavares-Dias and Moraes, 2004). In the present L. rohita study, the plasma glucose values were lower in autumn and winter, but higher in spring and summer. Similarly, total protein in these fishes varied slightly throughout the year in males compared to females, with higher values in winter and spring. Our results corroborate the reports of Hrubec et al. (2001) and our earlier reports in two other carp species (Pradhan et al., 2012a,b). The decrease in protein content in the summer may be due to augmented proteolysis and possible utilization of the product for metabolic purposes and/or utilization in the development of gonads. Many publications demonstrated that the seasonal fluctuations of lipids might be controlled by an endogenous biological clock, which exerts an independent effect on fish blood; they reported that the comparative increase in lipid during summer might be due to the demands of energy and environmental alterations, which enhance metabolic stress that reduces the metabolic reserves. In female fish, the cholesterol level is highest in the summer and lowest in the winter, which may be due to reproductive activity.

In conclusion, this study has established a reference value regarding the selected hematological and plasma biochemical parameters of L. rohita, similar to the reference values for the other tropical freshwater species, C. catla and C. mrigala. Establishment of hematological and plasma biochemical parameters is highly dependent on endo- and exogenous factors, including living conditions, season, age, gender, origin, breeding system, physiological and nutritional status, and genetics of each individual that governs reference values for fish, which can be useful in many fields. Although most wild fishes in a freshwater pond were sampled, the small numbers of any specific fish population limits the interpretation of the results, thus further validation is required. In summation, there is an urgent need for a reliable database for this economically important species.

Acknowledgement

  1. Top of page
  2. Summary
  3. Introduction
  4. Materials and methods
  5. Results
  6. Discussion
  7. Acknowledgement
  8. References

We are extremely grateful to Head, Utkal University, Bhubaneswar, India for providing the necessary facilities to carry out this work.

References

  1. Top of page
  2. Summary
  3. Introduction
  4. Materials and methods
  5. Results
  6. Discussion
  7. Acknowledgement
  8. References
  • Adebayo, O. T.; Fagbenro, O. A.; Ajayi, C. B.; Popoola, O. M., 2007: Normal haematological profile of Parachanna obscura as a diagnostic tool in aquaculture. Int. J. Zool. Res. 3, 193199.
  • APHA (American Public Health Association), 1998: Standard methods for the examination of water and wastewater, 20th edn. APHA, Washington, DC.
  • Bhaskar, B. R.; Rao, K. S., 1990: Use of haematological parameters as diagnostic tools in determining the health of milk fish, Chanos chanos (Forskal), in brackish water culture. Aquacult. Res. 21, 125130.
  • Campbell, R. A., 2004: CPUE standardisation and the construction of indices of stock abundance in a spatially varying fishery using general linear models. Fish. Res. 70, 209227.
  • Chakrabarthy, P.; Banerjee, V., 1988: Effect of sublethal toxicity of three organophosphere pesticides on the peripheral haemogram of the fish (Channa punctatus). Environ. Ecol. 6, 151158.
  • Collazos, M. E.; Ortega, E.; Barriga, C.; Rodriguez, A. B., 1998: Seasonal variation in hematological parameters of male and female Tinca tinca. Mol. Cell. Biochem. 183, 165168.
  • Congleton, J. L.; Wagner, T., 2006: Blood-chemistry indicators of nutritional status and food intake in juvenile salmonids. J. Fish Biol. 69, 473490.
  • Edsall, C. C., 1999: A blood chemistry profile of lake trout. J. Aquatic Anim. Health 11, 8186.
  • Filiciotto, F.; Fazio, F.; Marafioti, S.; Buscaino, G.; Maccarrone, V.; Faggio, C., 2012: Assessment of hematological parameter range values using an automatic method in European sea bass (Dicentrarcbus labrax L.). Natura Rerum 1, 2936.
  • Frings, C. S.; Dunn, R. T., 1970: A colorimetric method for determination of total serum lipids based on the sulfo-phospho-vanillin reaction. Am. J. Clin. Pathol. 53, 8991.
  • Gabriel, U. U.; Ezeri, G. N. O.; Opabunmi, O. O., 2004: Influence of sex, source, health status and acclimation on the haematology of Clarias gariepinus (Burch, 1822). Afr. J. Biotechnol. 3, 463467.
  • Garcia, M. P.; Echevarrin, G.; Martinez, F. J.; Zamora, Z., 1992: Influence of blood sample collections on the haematocrit value of two teleosts, rainbow trout (Oncohrhynchus mykiss) and European sea bass (Dicentrachus labrax L.). Comp. Biochem. Physiol. 101, 733736.
  • Handy, R. D.; Depledge, M. H., 1999: Physiological responses: their measurement and use as biomarkers in ecotoxicology. Ecotoxicology 8, 329349.
  • Hrubec, T. C.; Smith, S. A.; Robertson, J. L., 2001: Age-related changes in hematology and plasma chemistry values of hybrid striped bass (Morone chrysops x Morone saxatatilis). Vet. Clin. Pathol. 30, 815.
  • Ighwela, K. A.; Ahmad, A. B.; Abol-Munafi, A. B., 2012: Haematological changes in Nile tilapia (Oreochromis niloticus) fed with varying dietary maltose levels. World J. Fish Mar. Sci. 4, 376381.
  • Jamalzadeh, H. R.; Oryan, S.; Ghomi, M. R., 2008: Hematological characteristics and correlations of diploid and triploid Caspian salmon Salmo trutta caspius in juvenile stage. J. Cell Anim. Biol. 2, 195198.
  • Kori-Siakpere, O., 1985: Haematological characteristics of Clarias isheriensis Sydenham. J. Fish Biol. 27, 259263.
  • Kori-Siakpere, O.; Ake, J. E. G.; Idoge, E., 2005: Haematological characteristics of the African snakehead, Parachanna obscure. Afr. J. Biotechnol. 4, 527530.
  • Lavanya, S.; Ramesh, M.; Kavitha, C.; Malarvizhi, A., 2011: Hematological, biochemical and ion regulatory responses of Indian major carp Catla catla during chronic sublethal exposure to inorganic arsenic. Chemosphere 7, 977985.
  • Lowry, O. H.; Rosebrough, N. J.; Farr, A. L.; Randall, R. J., 1951: Protein measurement with the folin phenol reagent. J. Biol. Chem. 193, 265275.
  • Oluyemi, K. G.; Adecparusi, E. A.; Olanrewage, J., 2008: Basic hematological parameters in African catfish, Clarias gariepinus (Burchell, 1822) fed ascorbic acid supplemented diets. Res. J. Anim. Sci. 2, 1721.
  • Orun, I.; Dorucu, M.; Yazlak, H., 2003: Hematological parameters of three Cyprinid fish species. Onl. J. Biol. Sci. 3, 320328.
  • Pradhan, S. C.; Patra, A. K.; Sarkar, B.; Pal, A., 2012a: Seasonal changes in hematological parameters of Catla catla (Hamilton 1822). Comp. Clin. Pathol. 21, 14731481.
  • Pradhan, S. C.; Patra, A. K.; Mohanty, K. C.; Pal, A., 2012b: Hematological and plasma biochemistry in Cirrhinus mrigala (Hamilton 1822). Comp. Clin. Pathol. doi:10.1007/s00580-012-1642-z.
  • Rey Vázquez, G. R.; Guerrero, G. A., 2007: Characterization of blood cells and hematological parameters in Cichlasoma dimerus (Teleostei, Perciformes). Tissue Cell 39, 151160.
  • Sheridan, M. A.; Mommsen, T. P., 1991: Effects of nutritional state on lipid and carbohydrate metabolism of Oncorhynchus kisutch. Gen. Comp. Endocrinol. 81, 473483.
  • Singh, B. P.; Tandon, P. K., 2009: Effect of river water pollution on hematological parameters of fish, Wallago attu. Res. Environ. Life Sci. 2, 211214.
  • Tavares-Dias, M.; Moraes, F. R., 2004: Hematologia de peixes teleósteos. Ribeirão Preto, São Paulo.
  • Triander, P., 1969: Determination of glucose in blood using glucose oxidase with an alternate oxygen receptor. Ann. Clin. Biochem. 6, 2427.
  • Wedemeyer, G. A.; Gould, R. W.; Yasutake, W. T., 1983: Some potentials and limits of the leucocrit test as a fish health assessment method. J. Fish Biol. 23, 711716.