Hematological Parameters in Red tilapia (Oreochromis sp.) exposed to different concentrations of Lead (Pb)

 

Van-Thanh Vo*, Thai-Minh-Long Le, Thi-Quynh-Anh Duong, Nhat-Anh-Thu Mai

Department of Animal and Human Physiology, Biology Faculty, Ho Chi Minh University of Education, 280 an Duong Vuong str., Ward 4, Dist. 5, Ho Chi Minh city, Vietnam

*Corresponding Author E-mail: thanhvo@hcmup.edu.vn, thanhvv@hcmue.edu.vn

 

ABSTRACT:

The study investigated the effect of lead at concentrations of 0.5mgL-1, 1.0mgL-1, 1.5mgL-1 on the hematocrit index, hemoglobin content, number of red blood cells, white blood cells in Oreochromis sp. Experimental results showed that the concentration of lead is inversely proportional to hematocrit, hemoglobin content, the number of red blood cells of Oreochromis sp. and is directly proportional to the number of white blood cells. When lead exposure in another groups at the investigated concentrations, the hematocrit index was reduced to 13.67%, 28.66% and 33.67%, respectively compared to the control; hemoglobin content decreased by 24.69%, 29.46%, 30.68% compared to control; Red blood cells decreased by 17.25%, 21.18% and 26.67%, respectively, compared to controls; increased white blood cell count by 9.81%, 24.61%, 34.48% compared to control.

 

KEYWORDS: Hematological parameters, Oreochromis sp., lead (Pb), red blood cells, white blood cells.

 

 

 

INTRODUCTION:

Heavy metals are popular such as: Copper (Cu), Lead (Pb), Mercury (Hg), Arsen (As), Zinc (Zn) have a significant influence on physiology [5] the development process of fish and cause many diseases in fish and poisoning fish death [4]. The blood system directly or indirectly reacts to changes in the environment, objectively reflects the physiological state and allows predicting the direction of adaptive response in the body [21,22]. Changes in hemoglobin, hematocrit, fluctuations in the number of red blood cells and white blood cells clearly show the physiological reactions of the blood under the toxic effects from the environment [6]. The blood system is one of the major organ systems in which heavy metals impact [1]. In 2005, Al-Attar AM et al. showed that the average hemoglobin content decreased, the number of red blood cells and hematocrit decreased when the fish Oreochromis niloticus exposed to Cadmium (Al-Attar AM et al., 2005) [2], similar results (Kori-Siakpere O. et al., 2008) when Heteroclarias sp. exposed to Zn [14]. In 2014, Sharma J. and Langer S.

 

 

exposed Garra gotyla to the Manganese resulting in the hematocrit index, hemoglobin content and the number of red blood cells decreased, and the number of white blood cells increased [29]. Lead (Pb) causes abnormalities in hemoglobin synthesis by affecting hematopoietic organs systems [12]. Hrubec T. C. et al. (2000) [8], Mauel M. J. và et al. (2007) [19] have published a reference interval table of blood systems in red tilapia, as a criterion for evaluating the normal state of the indicators hematology in Oreochromis sp. However, the study of the effects of Lead at different infectious concentrations on hematological indicators in fish is limited.

 

The objective of the research is to evaluate hematological parameters in red tilapia (Oreochromis sp.) exposed to different concentrations of lead (Pb)

 

MATERIAL AND METHODS:

This research was performed at the Department of Animal and Human Physiology, Biology Faculty, Ho Chi Minh University of Education, Ho Chi Minh City, Vietnam. Red tilapia with an average weight 158.25±9.31g and average length 20.68±2.21cm were purchased from Research Institute for Aquaculture no.2, Ministry of Agriculture & Rural Development Vietnam. During the research, fishes were fed 3 times/day, using the formula food commercial Tilatech, Cargill Vietnam company, Vietnam.

Fish is raised in tanks with size of each tank 90 cm × 70 cm × 40 cm. Fish (n=60) were randomly divided into 4 groups with numbers of 15 fish/group, causing lead infection with concentrations of 0.5mgL-1; 1.0mgL-1 and 1.5mgL-1, noninfectious control treatments. Before infection, fish are kept stable under the same conditions within 14 days. Experimental blood samples were collected 96 hours [3, 17, 22, 25] after exposure to lead in culture tanks with 5 fish/collection.

 

The blood of red tilapia is collected by 3mL syringe in the tail vein and placed in a dedicated blood vessel with anticoagulant Heparin. Hematocrit index was determined by blood centrifugation and measurement of the ratio of red blood cells/ plasma sedimentation. Hemoglobin index is determined using Sahli hemoglobin. The number of red blood cells is counted on Neubauer counting chamber. The number of white blood cells of red tilapia is counted indirectly through the consumption of Giemsa staining. Data were statistically processed by Minitab 18, the difference was statistically significant with p≤0.05.

 

RESULTS AND DISCUSSION:

The experiment investigated the fluctuation of hematological indices in Oreochromis sp. (hematocrit index, hemoglobin content, number of red blood cells, white blood cells) under the action of lead at infectious concentrations: 0.5mgL-1, 1.0mgL-1 and 1.5mgL-1 after 4 days of infection. Survey results are shown in the following table.

 

 

Table. Heamatology parameters of Oreochromis sp. exposed to different concentrations of lead

 

Control

Concentration of Lead (mgL-1)

0.5

1.0

1.5

Hematocrit (%)

39.29±6.04a

33.92±4.33b

28.07±5.26c

26.06±1.71c

Hemoglobin (g%)

8.180±0.535a

6.160±0.972b

5.773±0.997b

5.673±0.573b

Red blood cells (×106/mm3)

2.554±0.267a

2.107±0.425b

2.011±0.369b, c

1.872±0.310c

White blood cells (×103/mm3)

60.27±3.59a

66.18±4.08b

75.10±8.25c

81.05±8.23d

Notes: a, b, c, d – The difference is statistically significant (p<0.05).

 

 

The results from the Table show that the concentration of lead-contaminated metal has a significant effect on the hematological indices of Oreochromis sp. The higher the concentration of Lead (Pb), the lower the hematocrit index, decrease the hemoglobin content, the number of red blood cells and the number of white blood cells.

 

After 96 hours causing infection, in different concentration of lead 0.5mgL-1; 1.0mgL-1; 1.5mgL-1, hematocrit index of red tilapia decreased from 39.29±6.04(%) to 26.06±1.71(%). At the higher concentrations of lead, the hematocrit index is more effected: with the concentration 0.5mgL-1 decreases by 13.67%, with the concentration 1.0mgL-1 decreases by 28.56% and with concentration 1.5mgL-1 reduce by 33.67% compared to survey index. All differences are statistically significant p<0.05. The hematocrit index at concentration 1.5mgL-1 has decreased compared with concentration 1.0mgL-1 but the difference is not statistically significant with p>0.05.

 

The higher concentration of lead, the lower hematocrit index, the results of this research are in line with other results of the study that mention the effect of metal on hematocrit index such as: Sharma J. and Langer S. (2014) research Manganese influence on Garra gotyla hematology index [29] and research by Ololade I. A. and Oginni O. (2010) on the influence of Nickel on fish hematological index Clarias gariepinus [23]. All of these studies show that hematocrit index changes are inversely proportional to the concentration of heavy metal surveyed.

Hemoglobin content decreases from 8.180±0.535 (g%) to 5.673±0.573 (g%) after 96 hours of infection at different lead concentrations. With higher concentrations of lead, the hemoglobin content is more affected, example: in the concentrations 0.5mgL-1 decreases by 24.7%, in the concentrations 1.0mgL-1 decreases by 29.4% and in the concentrations 1.5mgL-1 reduce by 30.6% compared to the control. All differences are statistically significant (p<0.001). Hemoglobin content is reduced through survey concentrations of lead although differences don’t have statistically significance with p>0.05. Results show that after 96 days at the survey concentrations, there are not many significant effects on hemoglobin content.

 

Higher concentration of lead effect more decreasing hemoglobin content, the results of this research is suitable other results of the study about effect of heavy metals on hemoglobin content such as: Al-Attar AM (2005) in the study of hematological indices of Oreochromis nioticus fish under different Cadmium concentrations [2], results of the research Singh D. et al (2007) after water fish sweet Channa punctatus exposed heavy metal Cu [30] and with the research results of Ololade I. A. and Oginni O. (2010) in the study of toxicity and the influence of Nickel metal on hematological indices in African catfish (Clarias gariepinus) [23],  all their results reduce hemoglobin content. The hemoglobin index at concentrations of 0.5mgL-1, 1.0mg L-1 and 1.5mgL-1 was prolonged after 96 hours of infection, lower than the reference interval of Hrubec T. C. et al (2000) [8] and Mauel M. J. et al (2007) [19] on hemoglobin content of normal fish (7-9.8g%). Decreased hemoglobin levels indicate that abnormal red blood cells or red blood cells produce inadequate, consistent in this study when the hematocrit index is also reduced, indicating blood loss caused by lead. Stress-induced anemia caused by heavy metals is due to damage blood cells, dysfunctional hemoglobin [20], Pamila D. et al. (1991) explained that reduced hemoglobin content is caused by heavy metals direct impact on the enzyme system with the function of synthesizing and regulating hemoglobin content [24], Joshi P. K. et al. (2002) explained the impact of heavy metal to reduce the ability of iron absorption to heme nuclei formed and reduced hemoglobin value [10].

 

At different concentrations of lead, the quantity of red blood cells after 96 hours decreased from 2.554±0.267×106/mm3 to 1.872±0.310×106/mm3. The number of red blood cells of Tilapia in concentration 0.5mgL-1 decreased by 17.5%; in concentration 1.0mgL-1 decreased by 21.3% and in concentration 1.5mgL-1 decreased by 26.7% compared with survey, all differences are statistically significant with p<0.001. The quantity of red blood cells in concentration 0.5mgL-1 compared with concentration 1.0mgL-1 don’t have statistically significant (p>0.05), similar with the quantity of red blood cells in concentration 1.0mgL-1 compared with concentration 1.5mgL-1. The difference doesn’t have statistically significant (p>0.05).

 

The results of research are consistent with other results such as: Dick P. T. and Dixon D. G. (1985) [7] Fish Sulmo gairdneri live in the conditional of copper metal contamination (Cu), Shakoori A. R. and partners (1994) [28] in grass carp research, Ctenopharyngodon idella adapt with HgCl2, results of Kim J. H. and Kang J. C. (2014) [13] about red carp  Pagrus major live in the Selen contamination; Sharma J. and partners (2014) [29] researched about the effects of Manganum on hematological parameters in fish Garra gotyla, all the red blood cells decrease with increasing concentration and prolongs lead exposure. The number of red blood cells in concentration 1.5mgL-1 after 96 hours lower than the reference range (1.91-2.83×106/mm3) of Hrubec T. C. with partners (2000) [8] and Mauel M. J. with partners (2007) [19]. Sharma J. (2014) researched that the heavy metal toxicity effects on blood transport organs, blood thinning, releases normal red blood cells in blood circulation[29]. This is also the reason about the appearance of many kinds red blood cells, such as: adult red blood cells, malformed red blood cells. Hematopoietic organs may be toxic to heavy metals, reduce red blood cells reproduction, this is cause why the number of red blood cells decrease sharply when concentrations higher. Similar with others objects in the research of Santhakumar M. with partners (2000) [26], Kadambri G. with partners (2009) [11]. 

 

The quantity of white blood cells in different concentrations after 96 hours increased from 60.27±3.59×103/mm3 to 81.05±8,23×103/mm3. With high concentration of lead, the number of white blood cells are more increased, example: in concentration 0.5mgL-1 the number of white blood cells increased by 9.8%; in concentration 1.0mgL-1 increased by 24.6%; in concentration 1.5mgL-1 increased by 34.5% compared with survey, all of the differences are statistically significant with p<0.05.

 

Concludes that high concentrations of lead cause significant effects on the increase in quantity of white blood cells in Red Tilapia.

 

The quantity of white blood cells increased significantly compared to the control after 96 hours with different concentrations. The results of research are consistent with the research of Nussey G. and partners (1995) about the effects of heavy metals on white blood cells differentiation in tilapia (Oreochromis mossambicus) [21], the research of Singh D. and partners (2007) suggested that after Channa punctatus exposed heavy metals, the number of neutrophils  would increase [30], Kossai P. and partners (2011) studied about the lead resistance of Calcium in tilapia’s blood (Oreochromis niloticus) [15], Results of Kotsanis N. with partners (2000) about salmon Oncorhynchus mykiss is exposed to Arsenic and Cadmium, also showed the increase of white blood cells compared with the control [16]. Javed M. and Usmani N. (2014), the research about the snakehead is exposed to water contaminated with heavy metals mixture (Cu, Ni, Fe, Co, Mn, Cr, Zn), the number of white blood cells is higher than the control [9]. The quantity of white blood cells in all treatments are consistent with the reference range (18.75-154.69×103/mm3) of Hrubec T. C. with partners (2000) [8] and Mauel M. J. with partners (2007) [19]. In reality, when the fish is affected by harmful agents, the number of white blood cells is increased. Functions of white blood cells are against foreign agents entering, creating immunity and protecting. When the foreign agents entered the body, a sufficient amount of white blood cells will be created to protect the body. These changes can be explained by Sayed A. E. D. H. with Moneeb R. H. (2015) [27] and Martins M. L. with partners (2008) [18].

 

CONCLUSION:

Thus, from above results it can be inferred that lead toxicity has a marked effect on the hematological parameters of red tilapia (Oreochromis sp.). At the lead concentrations of 0.5mgL-1, 1.0mgL-1, 1.5mgL-1, lead toxicity reduced the number of erythrocytes leading to a decrease in the hematocrit index and hemoglobin content. This is because lead has a direct effect on hematopoietic organs, reducing their function. Besides, under the effect of lead at the survey concentrations, the immune system is activated by increasing the white blood cells to fight lead toxins. This is an adaptive response to the body's harmful agents at the cellular level.

 

ACKNOWLEDGEMENT:

The authors are special grateful to the Department of Animal and Human Physiology, Biology Faculty, Ho Chi Minh University of Education, Ho Chi Minh City, Vietnam for making facilities available for this research.

 

CONFLICT OF INTEREST:

The authors declare no conflict of interest.

 

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Received on 02.04.2019           Modified on 21.05.2019

Accepted on 28.06.2019         © RJPT All right reserved

Research J. Pharm. and Tech. 2019; 12(11): 5281-5284.

DOI: 10.5958/0974-360X.2019.00914.4