HISTOCHEMICAL CHARACTERIZATION OF CELL DEATH IN HONEYBEE LARVAE MIDGUT AFTER TREATMENT WITH <i>PAENIBACILLUS LARVAE</i>, AMITRAZ AND OXYTETRACYCLINE

Gregorc, Aleš; Bowen, I. D.

doi:10.1006/cbir.1999.0490

Cited by 53 publications

(49 citation statements)

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“…This suggests that there may have been only a background level of early apoptosis-positive product in these cells as demonstrated by the Annexin-V assay. In IHNV infected EPC cells ISCDDK appears to show apoptotic and necrotically induced changes similar to those found in bee larvae (Gregorc and Bowen 2000;Silva de Moraes and Bowen 2000). IHNV infection does appear to cause increased apoptosis as was found in fish cell line infected with infectious pancreatic necrosis virus (IPNV) (Hong et al 1998) and precedes necrosis as was assayed by the ISCDDK and propidium iodide.…”

Section: Discussionsupporting

confidence: 65%

“…Cell death was previously characterised in our laboratory using terminal deoxynucleotidyl transferase (TdT)-mediated biotin-dUTP nick end labelling (TUNEL) in honey bee midgut (Gregorc and Bowen 2000) and Annexin V in salivary gland of honeybee larvae (Silva de Moraes and Bowen 2000). A series of approaches to detect cell death were conducted in different cultured cells using TUNEL (Wutzler et al 2001) and Annexin V or both techniques (Bell et al 2001;Buemi et al 2001).…”

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confidence: 99%

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Infectious Haematopoietic Necrosis Virus-Induced Cell Death in Fish Cell Culture

Gregorc¹,

Bowen²,

JENâIâ³

et al. 2005

Acta Vet. Brno

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Cell death development was investigated in the epithelioma papulosum cyprini (EPC) cell line after infection with infectious haematopoietic necrosis virus (IHNV). Cytopathic effect (CPE) was evaluated, immunohistochemistry for virus localisation and the development of cell death were studied. Modes of cell death were characterised by staining with TdT-mediated dUTP nick end labelling (TUNEL), and with Annexin V and propidium iodide.CPE was evaluated as 25%, 50%, 75% and 100% at the virus culture infective concentration doses (TCID50/0.1 ml) of 1. 25, 6.25, 31.25 and 156.25, respectively, and no CPE was found at the concentration lower than 0.25 TCID50/0.1ml. At the PBS:virus concentration 0.25 TCID50/0.1 ml only a few cells were found anti IHNV antibody positive, and no CPE was observed. In the range of virus concentrations from 6.25 to 156.25 TCID50/0.1 ml all cells were positive. The level of positive cells using the TUNEL technique increased 30% and 55% after the increasing the CPE up to 50% and 75%, respectively. At the higher levels of CPE, all the remaining cells were positive.Infected cells exhibited features of early stage of apoptosis as demonstrated reactivity with annexin V. Apoptotic cells expose external phosphatidylserine residues while preserving membrane integrity. Red fluorescence with propidium iodide is associated with increased levels of necrosis and appeared in cells simultaneously with FITC marker and also alone in vacuolated and enlarged necrotic cells. Apoptosis appears before the pathological changes of necrosis and it is correlated to the spread of infection. The relevance of the current view that IHNV infection includes at least two modes of cell death: apoptosis and necrosis, is discussed. Epithelioma papulosum cyprini, cell death, necrosis, immunohistochemistry, annexin V, TUNELCellular death can occur either by accident, referred to as necrosis, or by design variously described as physiological cell death, programmed cell death or apoptosis (Bowen et al. 1996). Apoptosis presents a range of morphological symptoms including cell shrinkage, chromatin margination followed by DNA fragmentation and a florid break-up of the cell into spherical apoptotic bodies (Wyllie 1981). It has been shown that apoptosis can be induced by genetic means (Ellis et al. 1991), but can also be induced by non-genetic means (Arends and Wyllie 1991). Necrotic cell death appears to be induced under extreme conditions such as ischaemia, hypoxia, toxin, hyperthermia. The subsequent morphological changes encompass oedema following calcium overload, dilatation of the endoplasmic reticulum, polysome disaggregation, and mitochondrial swelling. The chromatin condenses, and clumps are formed at the nuclear periphery (Bowen et al. 1996). Necrosis refers to the post mortem changes that occur following the death of the cell (Trump and Berezesky 1998).A range of approaches are available to demonstrate the apoptosis of cells in living tissue or cell suspensions. Immunocytochemical methods assaying DNA fragmentation (Trauth ...

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Section: Discussionsupporting

confidence: 65%

mentioning

confidence: 99%

Infectious Haematopoietic Necrosis Virus-Induced Cell Death in Fish Cell Culture

Gregorc¹,

Bowen²,

JENâIâ³

et al. 2005

Acta Vet. Brno

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“…In our previous experiments (Gregorc and Bowen, 2000), higher cell levels were found when an "In situ cell death detection kit, AP" (Roche) was used as compared to an ApopTag kit (Oncor). The "In situ cell death detection kit, AP" was unable to differentiate between apoptosis 454 A.…”

Section: Introductionmentioning

confidence: 96%

Cell death in honeybee (Apis mellifera) larvae treated with oxalic or formic acid

2004

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-The effects of oxalic (OA) and formic acids (FA) on honeybee larvae in colonies were assessed and evaluated. Cell death was detected by the TUNEL technique for DNA labelling. In 3-and 5-day-old larvae exposed to OA, cell death was found in 25% of midgut epithelial cells 5 h after the treatment, using an "In situ cell death detection kit, AP" (Roche). The level of cell death increased to 70% by the 21st hour and the morphology of the epithelium remained unchanged. Fifty hours after the application, cell death was established in 18% of the epithelial cells of the 3-day-old larvae and had increased to 82% in the 5-day-old larvae. A "DeadEnd" apoptosis detection kit (Promega) showed sporadic cell death mainly in the larval fat body 5 h after treatment. Twenty-one hours after the OA application cell death was found in 4% of the larval midgut epithelial cells. Evaporated formic acid induced extensive apoptotic cell death in the peripheral, cuticular and subcuticular tissues that preceded the cell death of the entire larval body.Apis mellifera / cell death / oxalic acid / formic acid / immunochemical method

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“…The 7-µm sections were then deparaffinized and processed as per the instructions of the two kits used: the 'In situ cell death detection kit, AP' (ISCDDK) (Roche) and the 'DeadEnd colorimetric apoptosis detection system' (Promega). Cell death is a general term describing apoptosis and necrosis and Gregorc and Bowen (2000) showed that the two kits had different levels of sensitivity in detecting these two forms of cell deletion. The highly sensitive ISCDDK assay detects both in different tissues (Matylevitch et al, 1998), whereas the DeadEnd kit is more specific to apoptotic cell death.…”

Section: Immunohistochemical Analysesmentioning

confidence: 99%

“…Using an ISCDDK assay Gregorc and Bowen (2000) found that amitraz -another acaricide -increased the level of cell death in bee larvae and that an OA application triggered necrotic cell death in the midgut (Pulkkanen et al, 2000). Using this assay, we established the level of epithelial cell death in the midgut caused by a 3% OA application to an individual adult bee was 69% after 24 h which was 40% more than the control.…”

Section: Cell Deathmentioning

confidence: 99%

Toxicological and immunohistochemical testing of honeybees after oxalic acid and rotenone treatments

Gregorc

Škerl

2007

Apidologie

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-Bees removed capped brood and young larvae from combs at a greater rate after a rotenone treatment than after an oxalic acid (OA)/sucrose treatment. Rotenone (1%) caused 75.2% of capped brood to be removed, OA (3%) 18.7% and a control treatment, 13.3%. Caged worker bees treated with a 1% rotenone powder, a 3% OA or with a control solution had mortality rates of 10.9%, 5.1% and 1.9% respectively. Rotenone (1%) significantly affected the mortality of brood and adult bees whereas OA (3%), did not. Solutions of 3% OA/32% sucrose, 3.4% OA/47.6% sucrose, 3.7% OA/27.1% sucrose (w/w) and a 32% sugar solution applied to adult bees resulted in death rates of 11%, 14%, 11.2% and 6.5% respectively. Individually treated bees consumed more of a 3% OA solution than solutions with higher OA concentrations. A TUNEL assay detected necrotic cell death in 69% of bee midgut cells 24 h after an OA treatment. Normal cell turnover is approximately 8%.Treatment / Apis mellifera / acaricide / TUNEL / cell death / mortality

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HISTOCHEMICAL CHARACTERIZATION OF CELL DEATH IN HONEYBEE LARVAE MIDGUT AFTER TREATMENT WITH PAENIBACILLUS LARVAE, AMITRAZ AND OXYTETRACYCLINE

Cited by 53 publications

References 13 publications

Infectious Haematopoietic Necrosis Virus-Induced Cell Death in Fish Cell Culture

Infectious Haematopoietic Necrosis Virus-Induced Cell Death in Fish Cell Culture

Cell death in honeybee (Apis mellifera) larvae treated with oxalic or formic acid

Toxicological and immunohistochemical testing of honeybees after oxalic acid and rotenone treatments

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