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Short Communication · Volume 54, Issue 5, 19 July 2022 · Open Access

Toxicity of Iron-Oxide Nano-Particles on the Hematology of Cyprinus carpio

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Affiliations & Notes ⌄
  1. 1Department of Zoology, The Islamia University of Bahawalpur, Bahawalpur, Pakistan 2Department of Zoology, Cholistan University of Veterinary and Animal Sciences, Bahawalpur, Pakistan 3Department of Wildlife and Ecology, University of Veterinary and Animal Sciences, Lahore 4Department of Fisheries and Aquaculture, University of Veterinary and Animal Sciences, Lahore, Pakistan 5Department of Food Science and Technology, Cholistan University of Veterinary and Animal Sciences Bahawalpur, Pakistan 6D
Article Info ⌄
Journal:
Pakistan Journal of Zoology
Published:
19 July 2022
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From the article

  • The maximum immunity response with increased number of WBCs was revealed in T1 (282.32±9.32), followed by T0 (275.55±13.64) and T2 (258.62±2.92) groups.
  • The counts of RBCs, Hct, MCV and PLT were found to be increased in both experimental groups as compared to control one.
  • The changes in RBCs indicate variation in all red cell indices; Hb, MCV, Hct, MCH, MCHC.
  • There is recorded a significant decrease in the values of MCV, MCH and MCHC in experimental fishes as compared to control one (Table I).

Selected from the article's own text.

Abstract

Owing to growing exploration in nanotechnology, nano-particles application in various fields of industries and medicine is growing. However, limited study is available on their toxicological impacts which can adversely affect the organism’s health and their environment. Current research was designed to study the possible toxicity of iron oxide nano-particles on common carp, C. carpio after exposure period of 96 hours by the analysis of their haematological parameters. Fish were segregated into three groups (10fish/treatment). The iron oxide nano-particles at 0.25g/l doze were administered orally to treatment T1 and intravenously from caudal vein of fish to treatment T2. After 96h of exposure period, the blood was collected from fishes by the caudal vein puncture method into tubes containing anticoagulants EDTA and transferred immediately to the laboratory for the assessment of their blood profile. There was recorded no mortality after 96h of exposure period. IONPs significantly affected the counts of WBCs, Hb, MCH and MCHC of experimental and control groups. The analysis of blood profile demonstrated that haematological parameters; the number of WBCs and Hb were increased in T1 and decreased in T2 in relation to T0. The counts of RBCs, HCT, MCV and PLT were found to be increased in both experimental groups as compared to control one. It is inferred from current results that IONPs have toxic effect on the haematology of C. carpio. Moreover, oral route of administering IONPs was found to be more toxic than intravenous one. Present study can be beneficial for the safety of aquatic environment and management of aquatic toxicity.

Materials and methods

For preparation of nanoparticles, the seeds of Withania coagulans were washed, dried, pulverized into coarse powder, 10g mixed with 100ml of distilled water and boiled on hot plate stirrer for 45 min. This extract was filtered and then stored at 4°C until further processing. The seed extract (20ml) was mixed with 100ml of FeCl2.4H2O and FeCl3.6H2O (2:1 molar ratios) solution heated at 90◦C and stirred for 2 min. After 15 min, 20ml of aqueous solution of sodium hydroxide with ratios of 3 ml per min was added in the mixture and change in color was observed.

Experimental fish (C. carpio) of average weight of 115±7g and length of 19±4cm were purchased from Punjab Government Fish Hatchery Bahawalpur, were stocked in well aerated water glass tanks of 92×46×46 cm and dissolved oxygen at 7.0 ± 0.5 mg/L. Fish were fed twice a day with commercial fish food at the rate of 3% of their body weight and acclimatized for 15 days. Fish were divided into 3 groups (T0, T1, T2) each of 10 fish. Fishes were exposed to sonicated iron oxide nanoparticles for 30 min at room temperature orally (T1) at 0.25g/l doze and intravenously from the caudal vein (T2) for 96 h. The blood was collected from caudal vein of fish and transferred immediately into tubes containing anticoagulants ethylene diaminetetra acetic acid (EDTA) to prevent blood coagulation. Fish blood hematological analysis was carried out by using digital analyzer from a local laboratory.

The collected data achieved from experimental and control groups were analyzed using SPSS. The difference of p˂0.05 was exhibited as statistically significant between treatment groups and data was presented as mean ± standard error of mean (SEM).

Results and discussion

Table I shows the effect of nanoparticles administrated orally and intravenously for 96 h on hematological parameters of fish. The maximum immunity response with increased number of WBCs was revealed in T1 (282.32±9.32), followed by T0 (275.55±13.64) and T2 (258.62±2.92) groups. The mean number of RBCs (0.66±0.36) was found to be highest in T1 and lowest in T0 (0.38±0.05). T1 demonstrated peak Hb level of 9.64±0.99 as compared to 7.95±0.41 in T2. There was observed extreme Hct and MCV value of 11.42±6.94 and 168.98±7.74, respectively in T1 of treatment groups followed by T2 and T0. The level of MCH and MCHC was recorded highest in control group and lowest in T1. Unlike other heamotological parameters, there was found uppermost increase in platelets of 17.25±13.91 in T2, 15.20±8.29 in T1 and 13.00±10.03 in T0.

In relation to control group, the number of WBCs and Hb were increased in T1 and decreased in T2. The counts of RBCs, Hct, MCV and PLT were found to be increased in both experimental groups as compared to control one. However, reversed was recorded in case of MCH and MCHC. The oral route of administering IONPs was found to be more toxic than intravenous one.

Current study exhibited a substantial increase in the value of total RBCs, Hct, MCV and PLT in both experimental groups; while WBCs and Hb counts were high in T1 and low in T2 in relation to control one. In previous toxicological studies, significant rise of RBCs, Hb and Hct was explored in C. carpio on exposure to copper NPs by Sevcikova et al. (2016) and in Labeo rohita treated with IONPs , While authors found reduction in value of total WBCs, MCHC, MCV and MCH. Channappa et al. (2018) recorded a considerable rise of WBCs and fall in number of RBCs and Hb in C. carpio on exposure to calcium zincate nanoparticles at the dose of 3.53 mg/L and 7.05mg/L in 14 and 21 days. Similarly, WBCs increase and RBCs, Hb, Hct and PLT decline in fish Labeo rohita was studied by Keerthika et al. (2017). The value of RBCs, WBCs, Hb and Hct was increased and MCH, MCV, MCHC counts were declined in Oreochromis mossambicus when exposed to IONPs .

Table I. The effect of biologically synthesized IONPs administered orally at 0.25g/l (T1) and intravenously in caudal vein (T2) for 96 h on hematological parameters of C. carpio.

Parameters

T0

T1

T2

WBCs (103/ul)

275.55±13.64a

282.32±9.32a

258.62±2.92b

RBCs (103/ul)

0.38±0.05a

0.66±0.36a

0.43±0.05a

Hb (g/dl)

9.43±1.18a

9.64±0.99a

7.95±0.4b

HCT (%)

5.78±1.20a

11.42±6.94a

7.05±1.11a

MCV (fl)

157.17±10.35a

168.98±7.74a

163.50±6.92a

MCH (pg)

249.97±50.60a

167.16±52.89b

186.53±18.90ab

MCHC (g/dl)

171.15±55.37a

99.94±33.21b

114.55±15.99ab

PLT (103/ul)

13.00±10.03a

15.20±8.29a

17.25±13.91a

WBSs, white blood cells; RBCs, red blood cells; Hb, hemoglobin; HCT, hematocrit, MCV, mean corpuscular volume; MCH, mean corpuscular hemoglobin; MCHC, mean corpuscular hemoglobin concentration; PLT, platelets.

The changes in RBCs indicate variation in all red cell indices; Hb, MCV, Hct, MCH, MCHC. The variation in RBCs counts in metal exposed gills is an adaptive response of a fish towards the impairment of gaseous exchange prompting an increased energetic demand . In response to chronic metal exposure in the blood of fish facing stress condition, an increase in number of total RBCs, Hb concentrations and Hct values is frequently observed due to enhanced erythropoiesis , as has been studied in present research. The increased concentration of Hb reflects the increased oxygen supply to the tissues leading to enhanced physical activities . The decline in RBCs counts is an indication of severe anemia and decreased Hb and Hct level demonstrates the shrinkage of RBCs due to microcytic anemia or hypoxia or some toxicant stress on the erythropoietic tissues .

WBCs counts in all of the vertebrates including fish may rise or fall in response to many stressors i.e. chemical pollutants and infections . There is recorded rise of WBCs in T1 and fall of these in T2 in comparison to T0 (Table I). The high level of WBCs reflects immune system response against nanoparticles as a foreign-agent and their decline might be a response against various infectious diseases or can be attributed to unhealthy functioning of blood forming tissues (kidney and spleen) . WBCs reduction (leucopenia) can also be a response of toxic metals accumulation in different biological tissues and its adverse effect on cell manufacturing from spleen due to the rise in the level of corticosteroid hormones which play a great role for preventing and healing inflammation .

The values of erythrocyte indices i.e. MCH, MCV and MCHC exhibit a wide range of physiological changes as a result of exposure of fish to different inorganic form of supplemented diet. There is recorded a significant decrease in the values of MCV, MCH and MCHC in experimental fishes as compared to control one (Table I). Kori-Siakpere et al. (2008) demonstrated that change in the level of RBCs, Hb and HCT affect other hematological indices; MCV, MCH and MCHC. The decline in the level of Hb and MCH cause hypocromia as revealed in current study. The increase level of MCH and MCHC results from spherocytosis .

Present research confirmed that IONPs have toxic impacts on the hematology of fish, C. carpio which can adversely disturb the survival of fish. Therefore, there is a need of proper and safe handling of IONPs in research laboratories and their proper disposal should be carried out while promoting their application in various fields.

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