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Increased Caspase-3 Immunoexpression and Morphology Alterations in Oenocytes and Trophocytes of Apis mellifera Larvae Induced by Toxic Secretion of Epormenis cestri

María H. Viotti
María H. Viotti
, 
Pablo Juri
Pablo Juri
, 
Paula Lombide
Paula Lombide
, 
María L. Presentado
María L. Presentado
, 
José M. Verdes
José M. Verdes
, 
Enrique Nogueira
Enrique Nogueira
, 
Ciro Invernizzi
Ciro Invernizzi
 y 
Graciela Pedrana
Graciela Pedrana
   | 26 abr 2021
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Article Category: Original Article
Publicado en línea: 26 abr 2021
Páginas: 101 - 108
Recibido: 30 ene 2020
Aceptado: 15 nov 2020
DOI: https://doi.org/10.2478/jas-2021-0006
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© 2021 María H. Viotti et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Fig. 1

Morphological effect of toxic honeydew from bee colonies affected by “River Disease” in oenocytes. a) fat body cells from healthy group. Healthy oenocyte (white arrow). b) Fat body cells from toxic group. Oenocyte showing a size decrease and heterochromatic nucleus (black arrow). Scale bar=10μm. Magnification: 400x. Morphometry results of oenocytes: c) area (μm2), d) diameter (μm). Larvae healthy group (white bars) or with toxic group (black bars). Error bars represent values of mean±SEM. Different literals in columns indicate significant differences (P<0.05). Histological images stained with Hematoxylin-Eosin.
Morphological effect of toxic honeydew from bee colonies affected by “River Disease” in oenocytes. a) fat body cells from healthy group. Healthy oenocyte (white arrow). b) Fat body cells from toxic group. Oenocyte showing a size decrease and heterochromatic nucleus (black arrow). Scale bar=10μm. Magnification: 400x. Morphometry results of oenocytes: c) area (μm2), d) diameter (μm). Larvae healthy group (white bars) or with toxic group (black bars). Error bars represent values of mean±SEM. Different literals in columns indicate significant differences (P<0.05). Histological images stained with Hematoxylin-Eosin.

Fig. 2

Effects of toxic honeydew in trophocytes morphology in honeybee larvae at day 5 of development. Histological images with Hematoxylin-Eosin a) trophocytes cytoplasm in healthy larvae fed with honey (white arrow), b) trophocytes in larvae fed with toxic honeydew (black arrow), notice the decrease in cell size and its heterochromatic nucleus compared to healthy larvae. Morphometry results of trophocytes c) trophocyte area (μm2), d) diameter (μm). Larvae fed with honey (white bars) or with toxic honeydew (black bars). Error bars represent values of mean ±SEM. Different literals in columns indicate significant differences (P<0.05).
Effects of toxic honeydew in trophocytes morphology in honeybee larvae at day 5 of development. Histological images with Hematoxylin-Eosin a) trophocytes cytoplasm in healthy larvae fed with honey (white arrow), b) trophocytes in larvae fed with toxic honeydew (black arrow), notice the decrease in cell size and its heterochromatic nucleus compared to healthy larvae. Morphometry results of trophocytes c) trophocyte area (μm2), d) diameter (μm). Larvae fed with honey (white bars) or with toxic honeydew (black bars). Error bars represent values of mean ±SEM. Different literals in columns indicate significant differences (P<0.05).

Fig. 3

Effect of toxic honeydew from colonies affected by “River Disease” on honeybee 5 days old larvae. Immunoexpression of caspase-3 in a) oenocytes in healthy larvae showing light intensity of immunostaining in cytoplasm (white arrow), b) oenocytes in larvae with toxic honeydew (black arrow) showing intense brown immunostaining in cytoplasmic region. Scale bar=10μm. c) Results of caspase-3 immunostaining area (%) and d) immunostaining intensity (mean gray value of pixels) in oenocytes in larvae fed with honey (white bars) or with toxic honeydew (black bars). Error bars represent mean±SEM. Different literals in columns indicate significant differences (P<0.05).
Effect of toxic honeydew from colonies affected by “River Disease” on honeybee 5 days old larvae. Immunoexpression of caspase-3 in a) oenocytes in healthy larvae showing light intensity of immunostaining in cytoplasm (white arrow), b) oenocytes in larvae with toxic honeydew (black arrow) showing intense brown immunostaining in cytoplasmic region. Scale bar=10μm. c) Results of caspase-3 immunostaining area (%) and d) immunostaining intensity (mean gray value of pixels) in oenocytes in larvae fed with honey (white bars) or with toxic honeydew (black bars). Error bars represent mean±SEM. Different literals in columns indicate significant differences (P<0.05).

Fig. 4

Effects of toxic honeydew in trophocytes of honeybees at day 5 of development. Immunoexpression of active caspase-3: a) trophocytes showing perinuclear slight immunostaining in healthy larvae fed with honey (white arrow) and negative basophilic nucleus without immunostaining, b) trophocytes from larvae fed with toxic honeydew showing intense brown immunostaining at nuclear and cytoplasmic region (black arrow). Scale bar=10μm. Immunostaining area of caspase-3 larvae fed with honey (white bars) or with toxic honeydew (black bars) in trophocytes c) immunostaining area (%), d) immunostaining intensity (mean gray values of pixels). Error bars represent values of mean±SEM. Different literals in columns indicate significant differences (P<0.05).
Effects of toxic honeydew in trophocytes of honeybees at day 5 of development. Immunoexpression of active caspase-3: a) trophocytes showing perinuclear slight immunostaining in healthy larvae fed with honey (white arrow) and negative basophilic nucleus without immunostaining, b) trophocytes from larvae fed with toxic honeydew showing intense brown immunostaining at nuclear and cytoplasmic region (black arrow). Scale bar=10μm. Immunostaining area of caspase-3 larvae fed with honey (white bars) or with toxic honeydew (black bars) in trophocytes c) immunostaining area (%), d) immunostaining intensity (mean gray values of pixels). Error bars represent values of mean±SEM. Different literals in columns indicate significant differences (P<0.05).
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