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A PhD was conducted at the University Marine Biological Station Millport (UMBSM). Tigriopus brevicornis inhabits a high shore environment and therefore must endure extreme physiochemical changes in salinity, temperature and oxygen. The study was conducted to investigate the previously unknown physiological mechanisms for tolerance of the physiochemical extremes which is experienced by the harpscticoid copepod Tigriopus brevicornis. The osmotic physiology, respiratory physiology, functional morphology and aspects of the cryobiology of Tigriopus brevicornis were investigated.
University Marine Biological Station (UMBS), Millport
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Other details | ||
Internal code | Internally assigned metadata identifier | 4523 |
Title | The title is used to provide a brief and precise description of the dataset such as 'Date', 'Originating organisation/programme', 'Location' and 'Type of survey'. All acronyms and abbreviations should be reproduced in full. | 1998 UMBSM Clyde Sea Life at it's Limits: The Ecophysiology of the High Shore Rockpool Inhabitant Tigriopus Brevicornis |
File Identifier | The File Identifier is a code, preferably a GUID, that is globally unique and remains with the same metadata record even if the record is edited or transferred between portals or tools. | a3536c49ff6cf8a3471b793ee9f5c3fb |
Resource Identifier | This is the code assigned by the data owner. | UMBSM9454 |
Resource type | The resource type will likely be a dataset but could also be a series (collection of datasets with a common specification) or a service. | dataset |
Start date | This describes the date the resource starts. This may only be the year if month and day are not known | 1995-12-01 |
End date | This describes the date the resource ends. This may only be the year if month and day are not known | 1996-09-01 |
Spatial resolution | This describes the spatial resolution of the dataset or the spatial limitations of the service. | inapplicable |
Frequency of updates | This describes the frequency with which the resource is modified or updated i.e. a monitoring programme that samples once per year has a frequency that is described as 'annually'. | notPlanned |
Abstract | The abstract provides a clear and brief statement of the content of the resource. | A PhD was conducted at the University Marine Biological Station Millport (UMBSM). Tigriopus brevicornis inhabits a high shore environment and therefore must endure extreme physiochemical changes in salinity, temperature and oxygen. The study was conducted to investigate the previously unknown physiological mechanisms for tolerance of the physiochemical extremes which is experienced by the harpscticoid copepod Tigriopus brevicornis. The osmotic physiology, respiratory physiology, functional morphology and aspects of the cryobiology of Tigriopus brevicornis were investigated. |
Lineage | Lineage includes the background information, history of the sources of data, data quality statements and methods. | The study investigated the ecology of the high shore rockpools which are inhabited by Tigriopus brevicornis. The environmental conditions of five high shore rockpools were measured during a 2 year environmental monitoring programme which was conducted in December 1995. Five rockpools were selected on the Isle of Cumbrae. Maximum pool diameter and depth was recorded using a 30cm ruler for the smaller rockpools whereas a metre rule was utilized to measure larger pools. A simple surveying technique was used to determine rockpool position above Chart Datum (CD). The salinity, temperature and oxygen concentrations were measured every 4h for a 24h period on a selected day once a month. Additionally, measurements were collected on days of extreme weather. Salinty was recorded using a pre-calibrated Atago hand refractometer. Temperature and oxygen levels were measured using a pre-calibrated Ciba Corning Checkmate 90 combined probe. No rockpool readings were taken during the months of April and May 1997. Associated rockpool fauna were recorded. The changes in the population dynamics of T. brevicornis were recorded in 2 high shore rockpools. The first rockpool survey was conducted in April 1996. The comparative second survey was conducted in September 1996. Monitoring was conducted around the 20th of each month. Sample areas within the rockpool were stirred up using a glass rod. Five sweeps were conducted over the sample area at varying heights in the water column using a 250ml plastic beaker. Contents on the beaker were emptied into a 100 micron sieve between each sweep. If less than 200 individuals were obtained further sweeps were performed. The sieve was immersed in seawater to keep animals alive and returned to the relevant pool after the survey. Two hundred individuals were selected from the sieve classified into groups (male, egg/ovary-bearing females, ovary-bearing females, 'clean' females, immature individuals). Percentage populations were calculated and animals were returned to their rockpool. The role which Enteromorpha has on the ecology of T. brevicornis was investigated. Nine replicate artificial rookpools were set up using 10cm diameter plastic boats which were coated with ground sandstone (epoxy adhesive). The bowls were washed in running water for 24h. One bowl was filled with Enteromorpha, a second was filled with ground sandstone (1cm deep). A third (with original sandstone coating) was used as a control. Twenty five ml of seawater (33 psu) and approximately 50 T. brevicornis were added to each bowl. Bowls were placed infront of a Glen electric fan heater 2000 series blowing on a cold setting (to evaporate seawater at a comparable natural rate). Sixteen replicates were performed for each treatment and left to dry for 0.5h, then hourly for 1 to 6h, then every two hours from 6 to 24h. Six replicates were performed for the control treatment. and left to dry for 10, 20, 30, 60, 120, 180 minutes. All pools were then reimmersed in seawater and left for 24h. Mortality was assessed and analyzed using Probit analysis (Finney 1971) to assess median lethal time. Fouling microbiota was assessed for a role influencing T. brevicornis ecology. Biofouled individuals were assessed using a Olympus Phase Contrast Inverted IMT-2 and prepared for observation under scanning electron microscope (SEM). Samples were fixed in 3 percent glutaraldhyde for 30 minutes and rinsed in distilled water twice for 5 minutes. Samples were dried with filter paper and placed into a Edwards Pirani 10 freeze drier at -40 degrees Celsius for 48h. Samples were coated in pure gold using a BIo-Rad SEM sputter coating system. Observations were performed using a Jeol 5200 SEM. The presence of bacteria in high shore rock pools was assessed. Seawater (400ml) was collected in 600ml beakers from the three previous high shore rockpools. Twelve fine mesh bags (5x5cm) with 1.0g of practical grade chitin (Sigma Co.). Three mesh bags were placed into each beaker and kept for 18 degrees Celsius. A control beaker with 2 micron filtered seawater was set up. After 7days a bag was removed from each beaker and dried at 50 degrees Celsius and reweighed. This was repeated after 2 and 3 weeks. Bacteria of T. brevicornis was assessed to determine whether it was a generalist or chitin-specific bacteria. Bacteria were plated oto chiti-agar plates and normal seawater plates ad observed. Marine Agar 2216 (Difco Laboratories) (1.8g) was mixed into each 100ml of filtered seawater. One gram of chitin was added to the solution for the agar plates. Solutions were steamed for 15 minutes followed by 20 minutes autoclaving in a portable autoclave (Dixons Ltd) and left to set. A sterile scalpel was used to cut around the agar edge and turned over, exposing the chitin. A heavily colonised individual was wiped over the agar surface and the plates were incubated at 37 degrees Celcius for a week. Bacteria grown were subcultered onto five plates chitin and non-chitin agar and wrapped in cling film and again incubated for three weeks. The osmotic physiology of T. brevicornis was assessed. Animals were collected by handnet (100 micron mesh) in rockpools at or above HW mark, close to Keppel Pier, Cumbrae. Batch samples were placed into beakers with 100ml of seawater (33psu) at 10 degrees Celsius for 2 days and fed on Enteromorpha spp. and Isochrysis galbana. Palemon elegans (4-5cm) were collected and held 10 degrees Celsius for 2 days. Artemia nauplii were hatched from cysts (Interpet Ltd. Brine shrimp eggs) and left at 20 degrees Celsius for 2 days then held for 2 days at 10 degrees Celsius. Approximately 150 eggless adult copepods were transferred to culture media (50-100 psu) and left for three days at 10 degrees Celsius. Cultures were left at salinity steps for 12h. Animals were filtered through 100 micron mesh and rinsed in distilled water. Animals were transferred to filter paper and squashed. Paper was placed into a calibrated vapour pressure osmometer (Wescor 5100B) and 'haemolymph' concentration was determined. Osmolalities of 5 micron samples of each of the culture media were recorded. Samples utilizing Artemia and P.elegans followed the same procedure. With P.elegans, haemolymph was extracted from the pericardium with 10 micron microcapillary pipettes (Sigma Co.) Twelve P.elegans were used with the squash technique (by pestle and mortar). The concentration of sodium ions in body fluids of T. brevicornis ans A. napulii was determined by using culture media (5-100 psu). Samples were processed at previously stated however animals were not squashed. Once samples were dried at 50 degrees celsius, each sample was digested in 2ml of a 1:1:1 mixture of 70 percent nitric acid, 3 percent hydrogen peroxide and distilled water (McDonagh and Stiffler, 1981) and then placed into boiling water for 5 minutes. Concentrations were determined by dilution with a calibrated flame photometer (Corning model 410). Density of T. brevicornis was assessed based on the methodology of Moore et al., 1997. Cryobiology of T. brevicornis is presented in McAllen and Block (1997). Cryobiology of A. napulli is presented in McAllen (1997). Samples were collected using a handnet (100 micron mesh) in rockpools at or above HW mark, close to Keppel Pier, Cumbrae, during late September 1996. The oxygen uptake of T. brevicornis was assessed at different temperatures. Oxygen consumption was measured using an oxygen electrode technique described by Davenport (1976). A 12ml perpex vessel with plankton mesh mounted 1cm from the bottom was used. The oxygen electrode (Strathkelvin 1302) was connected to a chart recorder (Rikadeki R22) via a radiometer PHM71 acid-base analyzer. The electrode was zeroed in seawater bubbled with oxygen-free nitrogen for 30 minutes. The electrode was calibrated in air-saturated filtered seawater. The electrode was inserted into a chamber with a sample of animals from 33psu and 10 degrees Celsius treatment. The assembly was placed onto the magnetic stirrer which was immersed in a Grant KD waterbath fitted with a Haake EK 51-2 dip chiller. Ten replicates were performed at 0,5,10,15,20,25,30,35 degrees Celsius. Temperatures above 20 degrees Celsius the dip chiller was replaced by a controlled heating element. Experiments were continued until a 40 percent decrease in oxygen tension was recorded. After experimentation, T brevicornis were removed and blot dried and dried overnight at 50 degrees Celsius. The dry weight of individuals were recorded. Control experiments were conducted using empty respirometer chambers. After each replicate, chambers were thoroughly cleaned. The rate of oxygen uptake was calculated from the traces produced using the nomogram of Green and Caritt (1967). Spectrophotometry was used to investigate the absence or the presence a respiratory pigment in T. brevicornis. Approximately 1000 individuals were placed into a 1.5ml epindorf tube and sonicated using a Microson Cell Disrupter for 2 minutes. The sample was then centrifuges on a Heraeus Biofuge for 10 minutes at 10,000g. Expressed body fluid was diluted with saline of the same concentration as normal seawater at a ration of 10 microlitres fluid to 1000 microlitres of saline. The diluted fluid sample was scanned using a Philips PU8700 spectrophotometer at wavelengths between 250-400nm. Haemocyanin results were checked by deoxygenising the sample with nitrogen and sodium sulphite. The tolerance of T. brevicornis of anoxic conditions at different temperatures was investigated. Samples were placed into conical flasks in which oxygen-free nitrogen was bubbled through for 30 minutes and oxygen concentration was measured using a Ciba Corning Checkmate 90 oxygen meter. When anoxic the flask was held at 0,5,10,18,25,30 degrees Celsius. Controls were set up with bubbled air instead of nitrogen. Anerobic pathways in T. brevicornis was investigated. Samples were placed into jars containing 200ml of 33 psu seawater and left to acclimate for 24h. Nitrogen was bubbled continuously through one jar for 24h. Air was bubbled through the other. Animals were filtered through 100 micron sieve and transferred to a crucible of liquid nitrogen. Comparative studies were performed on P.elegans. Once liquid nitrogen had evaporated, samples were transferred to 1.5ml epindorf tubes and weighed. 0.3M of chilled perchloric acid (500 microlitres) were added. Samples were sonicated on a Microson Ultrasonic Cell Disrupter for 2 minutes and lactate was retained. Epindorf tubes were centrifuged at 10,000g for 10 minutes on a Heraeus Biofuge. Supernatant was removed and stored on ice. A further addition of 0.3M of chilled perchloric acid (500 microlitres) was added to the pellet and mixed and centrifuged for a further 20 minutes. Supernatant was added to the previous collections ad samples were neutralised by the addition of 2M potassium biocarbonate. The pH was checked using indicator paper. The method of lactate determination is based on that of Gutmann and Wahlefeld (1974). The effect of low temperature dormancy on T. brevicornis was investigated. Samples were placed in 2ml of filtered seawater in a capped glass vial (10ml) Thirty replicate trials were set up and 15 were placed in a Neslab RTE-211 refrigerated bath filled with ethylene glycol at 0 degrees Celsius. The other 15 samples were placed into a Grant KD waterbath with a Haake EK51-2 dip chiller at 5 degrees Celsius. All animals were starved during the experiment. A replicate vial was removed from each temperature bath each week and animals were placed in fresh aerated seawater (33 psu) for 24h before mortality was assessed. Finally the morphology of T. brevicornis was investigated. Samples were collected as previously stated. All animals were placed in seawater (33 psu) at 10 degrees Celsius and held until required. Harpacticus flexus were collected during ELWS on Kames Bay, Millport. Samples were collected by scraping the top layer (2cm) of sand at extreme low water mark and placed into a bucket and left for an hour with a few centimetres of seawater. H. flexus were gathered when the came to the surface of the sand by pipette and stored similarly to T. brevicornis. Dr P.R.O. Barnett (UMBSM) confirmed the identity of H.flexus. Both species were observed by SEM using the procedure previously stated. Video image analysis was achieved using two narrow viewing chambers (6mm long by 1mm wide) which were constructed from two 22 x 50 mm glass cover slips, held apart by two 22 x 22mm glass cover slips glued at each end. The viewing chamber was one third filled with ground sandstone. The viewing chamber was filled with seawater (33 psu) by a hypodermic needle. Animals were then placed into the chamber using a fine tungsten needle. General behaviour was recorded (movement and position) on video tape. The unit was help in position by two clamps attached to the mounting block under the chamber. A Toshiba CCD 1K-M36PK compact colour camera with x20 lens was mounted on a micropostioner attached to an Ealing optical bench. The camera was connected to a Panasonic NV-SD450 video cassette recorder and Sony KX14109M colour video monitor. All footage was analysed visually and significant fields were saved as video prints obtained from a Hitachi VV-150E video printer. |
Related keywords | ||
Keyword | General subject area(s) associated with the resource, uses multiple controlled vocabularies | Marine Environmental Data and Information Network |
General subject area(s) associated with the resource, uses multiple controlled vocabularies | Species distribution | |
General subject area(s) associated with the resource, uses multiple controlled vocabularies | Zoobenthos taxonomy-related abundance per unit area of the littoral zone | |
Geographical coverage | ||
North | The northern-most limit of the data resource in decimal degrees | 55.7502 |
East | The eastern-most limit of the data resource in decimal degrees | -4.9054 |
South | The southern-most limit of the data resource in decimal degrees | 55.7491 |
West | The western-most limit of the data resource in decimal degrees | -4.9068 |
Regional sea | Irish Sea | |
Responsible organisations | ||
Role | The point of contact is person or organisation with responsibility for the creation and maintenance of the metadata for the resource. | pointOfContact |
Organisation name | University Marine Biological Station (UMBS), Millport | |
Individual name | Carly Cassidy | |
Postal code | KA28 0EG | |
City | Millport | |
Role | The distributor is the person or organisation that distributes the resource. | distributor |
Organisation name | Data Archive for Seabed Species and Habitats (DASSH) | |
Position name | Data Manager | |
Phone | 01752 633102 | |
Fax | 01752 633291 | |
Delivery point | Marine Biological Association of the UK, The Laboratory, Citadel Hill | |
Postal code | PL1 2PB | |
City | Plymouth | |
Role | The originator is the person or organisation who created, collected or produced the resource. | originator |
Organisation name | University Marine Biological Station (UMBS), Millport | |
Individual name | Philip Smith | |
Postal code | KA28 0EG | |
City | Millport | |
Role | The custodian is the person or organisation that accepts responsibility for the resource and ensures appropriate care and maintenance. If a dataset has been lodged with a Data Archive Centre for maintenance then this organisation is be entered here. | custodian |
Organisation name | University Marine Biological Station (UMBS), Millport | |
Individual name | Kathryn Stevenson | |
Postal code | KA28 0EG | |
City | Millport | |
Dataset constraints | ||
20 Limitations on Public Access - Access constraints | ISO restriction code chosen from ISO 19115-1 Codelist | otherRestrictions |
20 Limitations on Public Access – Other constraints | Any restriction on the use of the resource such as the need to agree to certain licence conditions. | DASSH terms and conditions apply |
21 Conditions for Access and Use - Use limitation | Any restrictions imposed on accessing the resource such as the need to agree to certain licence conditions. | Permission to copy, or reproduce the contents of this report is granted subject to citation of the source of this material. |
Available data formats | ||
Data format | Format in which digital data can be provided for transfer | Documents |
Version info | ||
Date of publication | The publication date of the resource or if previously unpublished the date that the resource was made publicly available via the MEDIN network. | 1998-08-01 |
Metadata date | The date when the content of this metadata record was last updated. | 2019-11-04 |
Metadata standard name | The name of the metadata standard used to create this metadata | MEDIN Discovery metadata standard |
Metadata standard version | The version of the MEDIN Discovery Metadata Standard used to create the metadata record | 2.3.8 |
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