Suggested Citation:
Schlenker, L.. 2019. Electro-olfactogram bicolor damselfish (Stegastes partitus). Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/n7-48jn-7a47
Data Parameters and Units:
Initial and final sum 50 PAH data from 6% 24-hr HEWAF exposures (Damsel EOG PAH DATA.csv); bicolor damselfish lengths, masses, treatment group, and fish ID number (Bicolor Damselfish Data.csv); voltage change across the olfactory epithelium of control and oil exposed bicolor damselfish while various olfactory cues were delivered (Labchart .txt files; e.g., file damsel 8_21_18.txt is EOG recordings from several bicolor damselfish on August 21, 2018). Dataset also includes water chemistry information such as Tank ID, Date, Time, Monitoring Period, Water Temp (Degrees C), pH (S.U.), Dissolved Oxygen (mg/L), Salinity (ppt).
Methods:
Damsel EOG PAH DATA.csv: PAH Data represents the sum PAH concentration (micrograms per liter) of 50 listed PAHs as part of a static 24-hr treatment. Initial and final samples were collected for each 24-hr incubation. All oil treatments were a 6% HEWAF. Sum % refers to the percent of a particular ring class (e.g. 2-ring, 3-ring, 4-ring, and >4-ring). LABCHART TEXT FILES: Labchart text files need to be imported into Labchart (AD Instruments) to be used. File names include the date of recording (e.g. file damsel 8_21_18.txt is EOG recordings from several bicolor damselfish on August 21, 2018) which can be matched to fish information (Bicolor Damselfish Data.csv) and PAH exposure information (Damsel EOG PAH DATA.csv). Two-channels were used for recording, EOG and Filtered EOG. Filtered EOG had a 60hz filter and was used for all measurements and analysis. Electro-olfactogram (EOG) experiments were performed to directly quantify the ability of control (n=14) and oil-exposed (n=11) bicolor damselfish to detect conspecific alarm cue (CAC) as an olfactory cue at the olfactory epithelium (OE). Additional olfactory cues were also tested. These experiments were done blinded, so the researcher did not know whether the fish were oil exposed or control at the time of experiment. All EOG experiments were done in clean seawater. Not all fish included in the Labchart files provided robust data. Information on whether data was included in analysis and notes on rational if not used are included in the Bicolor Damselfish Data.csv file. Bicolor damselfish were anaesthetized with 0.2 g/L MS-222 (tricaine methanesulfonate, Western Chemical, Inc., Ferndale, WA), buffered with NaHCO3 (Sigma Aldrich, St. Louis, MO), and transferred to the submerged experimental chamber where they were ventilated with aerated culture water containing 0.1 g/L MS-222. To access the olfactory epithelium the septum that separates the anterior and posterior nares was removed and a recording electrode, a non-polarizable Ag-AgCl electrode (Warner Instruments, Hamden, CT), fitted with a 1.5 mm glass capillary tube (Warner Instruments) pulled to a fine tip and filled with 3 M potassium chloride (Sigma Aldrich) was placed on the longest lamellae of the anterior dorsal section of the OE. An identical reference electrode was placed directly posterior to the eye. All recordings were performed blind such that the researcher did not know whether the fish was a control or oil exposed individual. The output from the recording and reference electrode was differentially amplified 1000x (DP-311, Warner Instruments), filtered (0.1Hz-0.1kHz, 50/60Hz; DP-311, Warner Instruments), digitized and filtered (60 Hz, Power Lab, AD Instruments, Dunedin, NZ), and recorded (Chart Software v. 8.1.3, AD Instruments, Colorado Springs, CO). All cues were delivered in a randomized order with each cue delivered three times. Each cue delivery was followed by the administration of a blank cue. Cue abbreviations are as follows: food=brine shrimp rinse, food.1=10% dilution of brine shrimp rinse, CAC=conspecific alarm cue, ala-1=10-1 M l-alanine, ala-2=10-2 M l-alanine, ala-3=10-3 M l-alanine, tca-3=10-3 M taurocholic acid, blank=control seawater. Brine shrimp rinse was made by soaking 0.5g of frozen brine shrimp in 200 mL of damselfish culture water for 30 minutes and filtering it through a 100 m mesh. CAC was produced by euthanizing a donor bicolor damselfish with a quick blow to the head, making six superficial cuts on the lateral sides of the fish, and rinsing the cuts with 15 mL of treatment water which was added to 20 mL of treatment water. CAC was made immediately following the establishment of successful recording for each individual and one donor fish was used for each experimental fish. Bicolor Damselfish Data.csv: Bicolor Damselfish Data.csv data includes lengths and masses of experimental and donor (sacrificed for CAC) bicolor damselfish. This dataset also includes which fish were included in analysis and treatment (oil or control). If a particular fish was excluded from analysis the reason is listed. Fish ID corresponds to comments in Labchart files. If NA is listed for a fish length or mass it was not collected by the researcher.
Slick oil collected from surface skimming operations during the DWH spill (sample ID: OFS-20100719-Juniper-001 A00884) was used to prepare a high-energy water accommodated fraction (HEWAF) on the day that 24-hour incubations began by mixing 1 g of oil per liter of damselfish culture water at low speed for 30 s in a Waring CB 15 blender (Torrington, CT). The blended oil and seawater mixture was immediately transferred to a glass separatory funnel, allowed to settle for 1 hour, and the lower 90% was drained and retained as the 100% water-accommodated fraction (WAF).
Samples for PAH analysis were collected in 250 mL amber bottles directly from tanks used for static incubations at the beginning and end of 24-hour incubations and were shipped overnight on ice to ALS Environmental (Kelso, WA) for gas chromatography/mass spectrometry—selective ion monitoring (GC/MS—SIM; based on EPA method 8270D).
Instruments:
A non-polarizable Ag-AgCl electrode (Warner Instruments, Hamden, CT), fitted with a 1.5 mm glass capillary tube (Warner Instruments) pulled to a fine tip and filled with 3 M potassium chloride (Sigma Aldrich) was placed on the longest lamellae of the anterior dorsal section of the OE. The output from the recording and reference electrode was differentially amplified 1000x (DP-311, Warner Instruments), filtered (0.1Hz-0.1kHz, 50/60Hz; DP-311, Warner Instruments), digitized and filtered (60 Hz, Power Lab, AD Instruments, Dunedin, NZ), and recorded (Chart Software v. 8.1.3, AD Instruments, Colorado Springs, CO).
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