Microscopic Studies on Erythrocytes of Channa punctata Exposed to Commercial Grade Lindane

Bhattacharjee, Debasish; Das, Suchismita

doi:10.1590/1678-4324-2017160341

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Biological and Applied Sciences • Braz. arch. biol. technol. 60 • 2017 • https://doi.org/10.1590/1678-4324-2017160341 linkcopy

Microscopic Studies on Erythrocytes of Channa punctata Exposed to Commercial Grade Lindane

Authorship SCIMAGO INSTITUTIONS RANKINGS

Light and scanning electron microscopic studies of erythrocytes of Channa punctata exposed to two sublethal doses of lindane over a period of 21 days revealed myriads of anomalies. High frequency of micronucleus appeared progressively in the erythrocyte, ranging from 3.8-7.5%, when monitored weekly for 21 d at 0.25 g/L and 1.0-3.0% at 0.025 g/L lindane. Prominent structural anomalies of erythrocytes included abnormal shapes, vacuolation, membrane invagination and disintegration. At higher dose and at later duration, a unique tendency of abnormal cells to stick together was observed. It could be concluded that commercial formulation of lindane could induce genotoxicity and structural anomalies in the erythrocytes of fish.

Key words:
RBC; SEM; light microscopy; micronucleus

Authorship

person Debasish Bhattacharjee

schoolAssam University - LifeScience and Bioinformatics Silchar, Assam, India.Assam UniversityIndiaAssam, IndiaAssam University - LifeScience and Bioinformatics Silchar, Assam, India.

person Suchismita Das *

schoolAssam University - LifeScience and Bioinformatics Silchar, Assam, India.Assam UniversityIndiaAssam, IndiaAssam University - LifeScience and Bioinformatics Silchar, Assam, India.

*Author for correspondence:
drsuchismita9@gmail.com,
das_aus@rediffmail.com

SCIMAGO INSTITUTIONS RANKINGS

Assam University - LifeScience and Bioinformatics Silchar, Assam, India.Assam UniversityIndiaAssam, IndiaAssam University - LifeScience and Bioinformatics Silchar, Assam, India.

Figures | Tables

Figures (4)
Tables (2)

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Fig. 1 Light micrograph of blood of C. punctata (H & E, × 400): (A) control- nucleated erythrocytes (Er), eosinophil (E), neutrophil (N), small lymphocyte (SL) and monocyte (M); (B) abnormal shapes (Ab), vacuolation (v), micronucleus (MN) and cytoplasmic bleb (Cb) after 7 d; (C) abnormal shapes (Ab), erythrocyte membrane invagination (invg), acanthocytes (A), lobopodium (L), micronucleus (MN) and membrane disintegration (arrow) after 14 d; (D) sticking (Sc), abnormal shapes (Ab), membrane invagination (invg), micronucleus (MN) and disintegrated membrane (arrow) after 21 d of exposure to 0.25 g/L lindane (inset-enlarged view).

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Fig. 2 Light micrograph of blood of C. punctata (H & E, × 400): showing (A) abnormal shapes (Ab) and erythrocyte membrane invagination (invg) after 7 d; (B) after 14 d; (C) abnormal shapes (Ab), membrane invagination (invg), echinocytes (Ec) and micronucleus (MN) after 21 days of exposure to 0.025 g/L lindane (inset-enlarged view).

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Fig. 3 SEM of blood of C. punctata (A) under control condition showing oval erythrocytes (× 5000); (B) abnormal shapes (Ab) and lobopodial projections after 7 days (×1,700); (C) membrane invagination (invg) after 14 d (× 2,300); (D) stickiness (Sc), abnormal shapes (Ab), membrane invagination (invg), and disintegrated membrane (arrow) (× 1, 700) after 21 d; (E) Echinocytes (Ec) and slender erythrocyte (Ab) (× 3,700) after 21 days; (F) membrane invagination (invg) and echinocytes (×2, 500) after 21 days of exposure to 0.25 g/L lindane.

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Fig. 4 SEM of erythrocytes of C. punctata showing (A) abnormal shapes (Ab) after 7 days (×1, 900); (B) abnormal shapes (Ab), and clumps after 14 days (× 2, 000); (C) abnormal shapes (Ab) and membrane invagination (invg) (×1,900) after 21d; (D) enlarged abnormal shape (×5,000) after 21 days of exposure to 0.025 g/L lindane.

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Table 1 Semi quantitative analysis of erythrocyte abnormalities from lindane treated C. punctata at various exposure durations and concentrations

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Table 2 Micronucleus frequency (mean ±SD) % in erythrocyte of C. punctata at various exposure durations and concentrations of lindane

imageFig. 1 Light micrograph of blood of C. punctata (H & E, × 400): (A) control- nucleated erythrocytes (Er), eosinophil (E), neutrophil (N), small lymphocyte (SL) and monocyte (M); (B) abnormal shapes (Ab), vacuolation (v), micronucleus (MN) and cytoplasmic bleb (Cb) after 7 d; (C) abnormal shapes (Ab), erythrocyte membrane invagination (invg), acanthocytes (A), lobopodium (L), micronucleus (MN) and membrane disintegration (arrow) after 14 d; (D) sticking (Sc), abnormal shapes (Ab), membrane invagination (invg), micronucleus (MN) and disintegrated membrane (arrow) after 21 d of exposure to 0.25 g/L lindane (inset-enlarged view). open_in_new

imageFig. 2 Light micrograph of blood of C. punctata (H & E, × 400): showing (A) abnormal shapes (Ab) and erythrocyte membrane invagination (invg) after 7 d; (B) after 14 d; (C) abnormal shapes (Ab), membrane invagination (invg), echinocytes (Ec) and micronucleus (MN) after 21 days of exposure to 0.025 g/L lindane (inset-enlarged view). open_in_new

imageFig. 3 SEM of blood of C. punctata (A) under control condition showing oval erythrocytes (× 5000); (B) abnormal shapes (Ab) and lobopodial projections after 7 days (×1,700); (C) membrane invagination (invg) after 14 d (× 2,300); (D) stickiness (Sc), abnormal shapes (Ab), membrane invagination (invg), and disintegrated membrane (arrow) (× 1, 700) after 21 d; (E) Echinocytes (Ec) and slender erythrocyte (Ab) (× 3,700) after 21 days; (F) membrane invagination (invg) and echinocytes (×2, 500) after 21 days of exposure to 0.25 g/L lindane. open_in_new

imageFig. 4 SEM of erythrocytes of C. punctata showing (A) abnormal shapes (Ab) after 7 days (×1, 900); (B) abnormal shapes (Ab), and clumps after 14 days (× 2, 000); (C) abnormal shapes (Ab) and membrane invagination (invg) (×1,900) after 21d; (D) enlarged abnormal shape (×5,000) after 21 days of exposure to 0.025 g/L lindane. open_in_new

table_chartTable 1 Semi quantitative analysis of erythrocyte abnormalities from lindane treated C. punctata at various exposure durations and concentrations

Lesion	Control	7-d	14-d 21-d		7-d 14-d		21-d
		0.255 g/L			0.025 g/L

Abnormal shapes	-	+	++	+++	+	++	++
Membrane invagination	-	++	+++	+++	+	++	+++
Micronucleus	-	+	++	+++	-	+	++
Vacuolation	-	-	+	+	-	-	-
Cytoplasmic blebs	-	-	+	+	-	-	-
Lobopodium	-	-	-	++	-	-	-
Acanthocyte	-	-	-	+	-	-	-
Membrane	-	-	+	+++	-	-	-
disintegration
Sticky cells	-	-	-	++	-	-	-

- none, + mild, ++ moderate, - none, + mild, ++ moderate, +++ severe occurence

table_chartTable 2 Micronucleus frequency (mean ±SD) % in erythrocyte of C. punctata at various exposure durations and concentrations of lindane

Test	7-d	14-d	21-d

Control	0.15±0.23	0.1±0.008	0.12±0.03
0.255 g/L	3.8±0.12^*	4.5±0.11^*	7.5±0.3^*
0.025 g/L	1.0±0.05^*	2.3±0.21^*	3.2±0.03^*

*indicates significance at p<0.05

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