Brief description
This study investigates the expression of hsp70 protein and DNA strand breakage in the black bream (Acanthopagrus butcheri) in Swan-Canning Estuary in 2002. This is undertaken by determing the baseline levels of hsp70 under laboratory conditions and hsp70 levels in black bream from the Swan-Canning Estuary in two different seasons, evaluating seasonal and spatial variability of hsp70 expression in black breamLineage
Maintenance and Update Frequency: notPlanned
Statement: - Fish Collection -
Black bream were collected during late summer (April - May) 2002 and again in the late winter months (August and September) 2002, from four sites in the Swan River (N = 32) and one site in the Canning River (N = 8) (see thumbnail).
Indigenous black bream were captured by a commerical fisherman using a 120 m, 100 mm monofilament haul net and sacrificed within 2 hours of capture. Each fish was killed by the method of Iki Jimi (spike through the brain), the gills and liver taken and immediately immersed in liquid nitrogen then stored at -80 degrees Celsius until analysis.
Juvenile hatchery bred black bream (N = 10) were purchased from the Fremantle Maritime Centre. Five fish were placed into each of two 100 L aquariums and acclimated in seawater (35 ppt) at 17 degrees Celsius (+-0.03) for 10 days. The aquariums were provided with a sponge filter and aeration via an airstone. Ammonia levels were monitored daily to ensure water quality was maintained. At the end of the acclimation period 5 black bream were transferred to a 100 L aquarium with seawater heated to 27 degrees Celsius (positive controls), kept under observation for signs of excessive stress, and then returned to their aquarium (17 degrees Celsius; negative controls); then, one hour later these fish were again netted to ensure both groups of fish experienced the same number of capture events. Twenty-four hours following heat shock treatment all fish were sacrificed, gills removed from all black bream and livers were collected from the 5 negative control fish. All samples were immediately immersed in liquid nitrogen then stored at -80 degrees Celsius until analysis.
Black bream were collected during late summer (April - May) 2002 and again in the late winter months (August and September) 2002, from four sites in the Swan River (N = 32) and one site in the Canning River (N = 8) (see thumbnail).
Indigenous black bream were captured by a commerical fisherman using a 120 m, 100 mm monofilament haul net and sacrificed within 2 hours of capture. Each fish was killed by the method of Iki Jimi (spike through the brain), the gills and liver taken and immediately immersed in liquid nitrogen then stored at -80 degrees Celsius until analysis.
Juvenile hatchery bred black bream (N = 10) were purchased from the Fremantle Maritime Centre. Five fish were placed into each of two 100 L aquariums and acclimated in seawater (35 ppt) at 17 degrees Celsius (+-0.03) for 10 days. The aquariums were provided with a sponge filter and aeration via an airstone. Ammonia levels were monitored daily to ensure water quality was maintained. At the end of the acclimation period 5 black bream were transferred to a 100 L aquarium with seawater heated to 27 degrees Celsius (positive controls), kept under observation for signs of excessive stress, and then returned to their aquarium (17 degrees Celsius; negative controls); then, one hour later these fish were again netted to ensure both groups of fish experienced the same number of capture events. Twenty-four hours following heat shock treatment all fish were sacrificed, gills removed from all black bream and livers were collected from the 5 negative control fish. All samples were immediately immersed in liquid nitrogen then stored at -80 degrees Celsius until analysis.
Statement: - Stress Protein Assay -
Stress protein response was measured using the methods of Martin et al. (1996). For further information see section 7.2.3 of the thesis.
- DNA Alkaline Unwinding Assay -
DNA strand breaks were determined using the methods of Shugart (1996), which were optimised for black bream. All samples, solutions and equipment were maintained at 4 degrees Celsius throughout sample preparation. For further information see section 7.2.4 of thesis.
Stress protein response was measured using the methods of Martin et al. (1996). For further information see section 7.2.3 of the thesis.
- DNA Alkaline Unwinding Assay -
DNA strand breaks were determined using the methods of Shugart (1996), which were optimised for black bream. All samples, solutions and equipment were maintained at 4 degrees Celsius throughout sample preparation. For further information see section 7.2.4 of thesis.
Notes
PurposeTo determine the usefulness of hsp70 expression in black bream as a biomarker of effect in environmental monitoring of contaminants in the Swan-Canning Estuary.
Issued: 27 07 2005
Data time period: 2002-04 to 2002-09
text: westlimit=115.75; southlimit=-32.1; eastlimit=116; northlimit=-31.85
Subjects
37 353003 |
ANIMALS/VERTEBRATES |
Acanthopagrus butcheri |
BIOLOGICAL CLASSIFICATION |
Black bream |
COASTAL PROCESSES |
Contaminants |
DNA strand breakage |
EARTH SCIENCE |
Estuaries |
Fish |
OCEANS |
TERRESTRIAL HYDROSPHERE |
WATER QUALITY/WATER CHEMISTRY |
biomarkers |
oceans |
stress / heat shock proteins |
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Other Information
(PhD Thesis)
uri :
http://adt.curtin.edu.au/theses/available/adt-WCU20061204.135553/
global : d5aac340-5c41-11dc-af0d-00188b4c0af8
Identifiers
- global : 3754a870-5ffa-11dc-ae50-00188b4c0af8
