Mark L Wildhaber Karlie K Ditter Zachary D Beaman Kendell R Bennett Tyler J Grant 20251202 tabular digital data Reston, VA U.S. Geological Survey Authors Open Researcher and Contributor Id (ORCID) are as follows: Mark L Wildhaber: 0000-0002-6538-9083; Karlie K. Ditter: 0000-0001-8970-2022; Zachary D. Beaman: 0000-0001-9649-1585; Kendell R. Bennett: 0000-0001-6081-7002; Tyler J. Grant: 0000-0003-3965-3503 https://doi.org/10.5066/P13JZPPR Using juvenile individuals of four species of carp (Family: Cyprinidae), we tested the potential for two amino acids and one commercial bait stimulus to elicit an extracellular electro-olfactory response within the naris using gelatin-based electrodes. The amino acids alanine and lysine were tested on grass carp (Ctenopharyngodon idella), bighead carp (Hypophthalmichthys nobilis), silver carp (H. molitrix) and black carp (Mylopharyngodon piceus) to provide a comparison among compounds both within and among species. Each row in the electro-olfactogram (EOG) data set represents exposure to a single stimulant in a single fish. Each individual fish was exposed to all three stimulus solutions sequentially in a random order, representing a trial; each individual was a subject for exactly one trial. The response metric was the measured response peak, in millivolts, corrected by subtracting the mean baseline during exposure to well water 60 seconds prior to the stimulant response. The baseline-corrected, absolute value of the response to ultrapure, deionized (UDI or Nanopure) water immediately preceding and following a trial is also included. Other relevant factors that could influence measured individual response, such as nominal concentration of stimulants, fish mass, holding tank temperature, and flow rate of water and solutions over the naris are included. Identifying data, namely date of trial and an individual subject ID (equivalent to a trial ID), are also included. Expanding on the EOG study, we conducted behavior trials on the same four species of invasive carp in a laboratory setting between the dates of 07 July 2024 to 06 November 2024. The feeding stimulant and amino acids alanine and lysine were again used, as well as a fish food solution, to analyze how exposure may impact fish behavior, with a specific focus on avoidance and attraction. Each trial involved two individuals of the same species being exposed to one of the stimulant solutions and well water in a long exposure chamber which drained in the center to maintain a delineation between amino acid and the control water. For each combination of species (4 total) and compound solution (4 total), we performed 10 replicates, totaling 160 trials with each trial utilizing two fish (320 individuals). The tank and fish were recorded from overhead video cameras and fish movement and position within the tank determined using tracking software. After the software provided the initial tracking analysis, additional manual tracking was done to address any errors that occurred due to fish overlap. Data reported includes cumulative duration in percent of time spent on either side of the tank that would help indicate either attractant or avoidance behavior. Other relevant factors such as water temperatures of holding tanks, temperatures of the behavior tank before and after trials and individual fish length and mass were also included. In support of management efforts to control and eradicate grass carp, bighead carp, silver carp, and black carp from U.S. waterways, a variety of research has been done on feed additives and compounds such as food cues (e.g, amino acids), pheromones, and alarm-eliciting compounds that could be used to attract invasive carp as a means for aggregating them for easier harvest. These data were collected in order to establish the electro-olfactory and behavioral responses of these species to the selected compound solutions of interest in a laboratory setting This studies could be used to inform the design of additional behavioral trials in a field setting to determine if introduction of various solutions to serve as attractants or deterrents influenced fish behavior in a way that could be leveraged by managers to control invasive carp species. 20240221 20241106 observed Complete None planned Columbia Environmental Research Center -92.284312 -92.274450 38.912612 38.910784 ISO 19115 Topic Category biota USGS Thesaurus fish invasive species animal behavior National Agricultural Library Thesaurus Amino acids olfactory receptors Ctenopharyngodon Hypophthalmichthys Mylopharyngodon USGS Metadata Identifier USGS:67d18dd4d34ecfe34cc8845a None Columbia Environmental Research Center None Ctenopharyngodon idella Hypophthalmichthys nobilis Hypophthalmichthys moltrix Mylopharyngodon piceus Kingdom Animalia animals Subkingdom Bilateria triploblasts Infrakingdom Deuterostomia Phylum Chordata chordates Subphylum Vertebrata vertebrates Infraphylum Gnathostomata Superclass Actinopterygii ray-finned fishes spiny rayed fishes Class Teleostei Superorder Ostariophysi Order Cypriniformes minnows suckers Superfamily Cyprinoidea Family Cyprinidae carps minnows carps and minnows Genus Ctenopharyngodon grass carps Species Ctenopharyngodon idella grass carp silver orfe TSN: 163537 Genus Hypophthalmichthys bighead carps Species Hypophthalmichthys nobilis bighead carp TSN: 163692 Species Hypophthalmichthys molitrix silver carp TSN: 163691 Genus Mylopharyngodon Species Mylopharyngodon piceus TSN: 639618 None. Please see 'Distribution Info' for details. It is requested that the authors and the USGS Columbia Environmental Research Center be cited for any subsequent publications that reference this dataset. Users are advised to read the dataset's metadata thoroughly to understand appropriate use and data limitations. Mark L Wildhaber USGS - SOUTHEAST REGION Research Ecologist mailing and physical

Columbia Environmental Res Ctr,CERC - Tech Center

Columbia MO 65201 573-876-1847 mwildhaber@usgs.gov This project was funded by the US Department of Interior, US Geological Survey, Columbia Environmental Research Center (CERC) Windows 11 version 23H2, build 22631.3880; notepad++ 64-bit x64 version 8.6.9 Lab materials and instruments were checked, adjusted, or calibrated on varying schedules appropriate to the material or instrument. For electro-olfactogram trials, prepared electrodes were stored in 3 molar KCl solution at between 8 degrees Celsius and 10 degrees Celsius. Electrodes were checked before and after trials for bubbles; bubbles did not automatically exclude data if the baseline was stable and the UDI water response had a value lower than -4 mV. Amino acid and feeding stimulant solutions were remade monthly and stored at 8-10 degrees Celsius when not in use. Solutions were brought to room temperature (19-20 degrees Celsius) prior to testing. Several quality control methods were employed immediately prior to and during data collection trials. A trial comprised a single fish exposure to UDI water at least once at the start, exposures with the two amino acids and feeding stimulant, and exposure to UDI water at the end. The first UDI water exposure was used to test electrode positioning and conductivity. If the measured response to the first UDI water was larger (i.e., smaller in magnitude) than -4 mV, electrodes were readjusted or replaced, and an additional UDI water exposure was used to test the new configuration. Before transitioning from the pre-trial UDI water exposure(s) and the stimulant trial, two criteria needed to be met: 1) a minimum 2.5 min after the previous exposure and 2) at least 1 min of stable baseline without excessive signal noise immediately prior the stimulant exposure. After a trial was completed, the fish was revived to ensure it had been alive throughout the duration of the test. After collection, data were assessed manually for quality by visual inspection of plots of raw data in R (i.e., trial data with extracellular electrical values recorded every 0.05 s) and observation notes taken during trials (i.e., death of fish, records of bubbles in the tubing delivering stimulant solutions). Criteria for low-quality data were 1) noise unrelated to the signal response (e.g., bubbles in the solution delivered for an exposure, change in electrode connectivity or conductivity), 2) a UDI water response prior to the trial with an absolute value less than 4 mV, 3) any repeat exposures of a stimulant solution within a trial, and 4) any trial where the fish was found dead at the end. Low quality data were excluded from this data set. For behavior trials, recorded videos were processed and initially tracked, then trials were interpolated to fill in missing data samples. Manual tracking was done to all trials to address any errors that occurred due to fish overlap to allow for calculation of cumulative duration percent of time spent in zone. After manual tracking concluded, a final check of the videos was done to ensure any errors were corrected. The data for EOG match the details provided, and the values fall within expected ranges. Transcription errors in hand-recorded values (e.g., fish mass, flow rate) were checked for and corrected. For behavior chambers all proportions were checked and fall between 0 and 1 as expected. For EOG, data which met any of the criteria for low quality, as described above in the Attribute Accuracy Report, were excluded from the data set. For behavior data, many other metrics were calculated by EthoVision that were not useful for this study and could not effectively be quality controlled and thus are not included in the data. We did not include data from the acclimation period, described in the methods because this was considered periods where the fish were acclimating. Otherwise, data set is considered complete for the information presented, as described in the abstract. Users are advised to read the rest of the metadata record carefully for additional details. No formal positional accuracy tests were conducted No formal positional accuracy tests were conducted Mark L. Wildhaber Benjamin M. West Karlie K. Ditter Alex S. Peterson Robin D. Calfee Zachary D. Beaman 20230623 publication Fishes vol. 8, issue 7 n/a MDPI AG ppg. 334 https://doi.org/10.3390/fishes8070334 Digital and/or Hardcopy 20230623 publication date Wildhaber et al. 2023 Description of procedures Mark L. Wildhaber Zachary D. Beaman Karlie K. Ditter Benjamin M. West 20241022 publication n/a Wiley https://doi.org/10.1111/jfb.15964 Digital and/or Hardcopy 2024 publication date Wildhaber et al 2024 Description of procedures Electro-olfactogram (EOG) Trials - Test compounds: Amino acids alanine, lysine, and a feeding stimulant (Impulse Amino Acid Feeding Stimulant, BioSource Baits, 422 W Roosevelt Rd., Broadview, IL 60155) were tested on grass, bighead, silver, and black carp. Alanine was chosen based on previous EOG studies as our reference amino acid and we continue to use it here (Wildhaber et al. 2023). Lysine and the feeding stimulant were chosen to evaluate their potential as an attractant. The standard for EOG studies is to compare relative responses to stimulus solutions (Wildhaber et al. 2023). This is done to allow for comparisons across multiple studies with different species and stimuli. Concentrations of 0.0001 molar amino acid solutions were based on previous EOG and behavior studies conducted by Wildhaber et al. (2023, 2024). The concentration of the feeding stimulant was made by diluting the original stock solution to 0.001 dilution of the original solution using well water. 2024 Electro-olfactogram (EOG) Trials - Test organisms: Each fish selected for EOG was exposed to a random sequence of alanine, lysine, and the feeding stimulant, and with ultra-pure deionized water before and after solution exposure. Organisms were sourced from existing cultures at Columbia Environmental Research Center. Twelve juvenile individuals per species were tested: mass and length ranges were 2.5 to 7.43 grams and 43.9 to 90.45 millimeters for grass carp, 1.64 to 3.85 grams and 54.37 to 74.28 millimeters for bighead carp, 2.6 to 8.55 grams and 67.05 to 104.09 millimeters for silver carp, 3.01 to 12.83 grams and 67.61 to 118.48 millimeters for black carp. Laboratory trials for all four species ran from 21 Feb 2024 to 17 Sept 2024 2024 Electro-olfactogram (EOG) Trials - Fish preparation and electrode placement: The procedure used in this study followed Wildhaber et al. (2023). In summary, glass electrodes created from micropipettes and filled with sodium chloride (NaCl) and infused gelatin were used to measure electrical response of fish nares to chemical stimuli. This was done using a differential amplifier (Dagan EX1 Differential Amplifier with a 4001 Dagan Headstage) to collect the data (Dagan Corporation 2855 Park Ave Minneapolis, Minnesota USA). The software used to record the electrical response was LabChart 8 software (ADI Instruments PowerLab 4/35 1994-2019, v8.1.16 12/12/2019). The negative control was the use of fish culture water (i.e., well water). Appropriate electrode placement on the olfactory rosette of a fish was determined through the application of ultrapure deionized (UDI) water (derived from well water) using criteria described below. Amino acids at 0.0001 Molar concentration were tested on grass, bighead, silver and black carp between 21 February 2024 and 17 September 2024. For each trial, buffered tricaine mesylate (MS-222, 300 milligrams per liter) solution was used to anesthetize a fish and the nasal septum of the fish was removed to expose the right olfactory rosette. The fish was then placed in a PVC holding tray for the duration of a trial covered with a wet paper towel. To maintain an anesthetized state and provide oxygen during a trial, a tube was inserted into the fish’s mouth that delivered a continuous flow of 140 milligrams per liter MS-222 in well water. A separate tube was placed over the right naris of the fish through which a solution was continuously delivered over the right nares of the fish. The solution being delivered was either well water, UDI water, or a solution containing an olfactory stimulant. One electrode was placed so that it had proximate but not quite direct contact with the olfactory rosette of the exposed right naris. The second or ground electrode approximately roughly 1 millimeter from the exposed olfactory rosette and touching the skin of the fish. An additional ground-source grounding wire was clipped across the caudal peduncle perpendicular to the lateral line. 2024 Electro-olfactogram (EOG) Trials - Acclimation, exposure and measurement procedure: The exposed olfactory rosette was first acclimated to well water through delivery of well water over the rosette for a minimum of 5 min. After this initial 5 min period, a stable, flat EOG signal for a minimum of 60 seconds with limited noise was used as the criteria for application of a stimulus solution and provided a comparative 60-second negative control signal. Approximately one milliliter of an olfactory stimulant (i.e., amino acid, feeding stimulant, or UDI water) was then delivered to the olfactory rosette over a period of 3-seconds. The olfactory rosette was given at least 2.5 min to recover from exposure to a stimulus solution through exposure to well water. Flow rate of solution over the olfactory rosette was maintained at rates similar to previous studies to ensure comparability. During a trial a fish was exposed to UDI water, then up to three stimulus solutions, followed by UDI water again. The initial UDI exposure was used to test for proper electrode positioning. An initial UDI less than 4 millivolts was used as to determine if the electrodes needed repositioning. For a trial to proceed beyond initial UDI exposure, a minimum of 180 seconds had to occur where there was limited noise and a stable baseline immediately prior to the olfactory stimulant exposure. To ensure a fish was alive throughout a trial, a fish was revived in clean well water after a trial. After a trial was complete and the fish tested for life, the fish was euthanized with buffered MS-222. After being euthanized, the length and weight of the fish were recorded. 2024 Electro-olfactogram (EOG) Trials - data processing and filtering: To control for fine-scale volatility in EOG measurements, we used a moving average with a window of five measurements. A gross EOG response in millivolts was calculated using the moving average maximum value following within 10 seconds of the estimated delivery time. Baseline response before exposure was calculated as mean millivolt response to well water for the 60 seconds prior to the estimated delivery time. The Measured Millivolt Response (MMR) was estimated by subtracting the base response from the gross response. Data were assessed manually for usable EOG recordings using visual plot inspections. Data were included in analyses if all of the following criteria were met: 1) If there was no noise unrelated to the signal response, 2) there was a prior absolute 4mV or greater UDI response, and 3) the fish could be revived after the trial. 2024 Behavior Tanks - Test compounds: Lysine, alanine, and a feeding stimulant were used for this study, along with species-specific fish food solutions, to further progress our understanding of carp behavior when exposed olfactory stimulants. A series of different fish food mixtures was used based on the volume of water of the holding tanks and the type of food fed to certain species of carp. This method allowed us to reach a nominal concentration that for fish food mixtures that would be comparable to the amino acid concentrations. All fish food solutions were made by mixing each of the three different food combinations in 250 milliliters of well water for 30 minutes, afterwards we left the mixture to settle for 24 hours before decanting, and then filtering through grade 615 filter paper. Concentration of both amino acid solutions was nominally 0.0000714 molar. 2024 Behavior Tanks - Exposure chamber design: We used two functionally identical, independent behavior tanks constructed from acrylic tubes (length 152.4 cm, diameter 15.87 cm, water depth roughly 8.57 cm). Behavior chamber design is fully detailed in Wildhaber et al. 2024. All trials were recorded using a local network video recorder (Wisenet XRN-1610SA; Hanwha Vision: Seongnam, South Korea) with one video camera (Wisenet QNO-7080R; Hanwha Vision: Seongnam, South Korea) mounted directly above each behavior tank. To create a system free from external stimuli that could interfere with trials, the tanks were visually and acoustically isolated from the outside environment using foam insulation sheathing board. On both the front and back top side of a tank, there are two openings to introduce individual fish. Heated well water entered the front and back ends of each tank through top openings at 2000 milliliters per minute (mL/min). Throughout all trials, temperatures ranged between 19.6 and 20.3 degrees Celsius. The outflow of water from each tank was controlled by needle valves set, cumulatively, at 4000 mL/min to keep a stable level of water inside the tanks. A small tube was attached to the front and back top openings next to the inflow needle valves that supplied the olfactory stimulants. In each trial, only one side of the tank, randomly selected, received a stimulus. The end of the tank that was not receiving an olfactory stimulus received only well water. All amino acids were mixed 24 hours before the start of a trial and used within two weeks. A mass balance spreadsheet was used to determine how many grams of amino acid was needed to be dissolved into 350 mL local well water, delivered at a rate of 10 mL/min in concert with 2000 mL/min of well water to the stimulus side of a tank was needed to nominally maintain the concentration on one side of the tank during the compound exposure period of the trial. A calibrated peristaltic pump (Masterflex L/S Standard Digital Pump Systems; Avantor, Radnor, Pennsylvania, USA) delivered either a olfactory stimulus or well water through a tube specific to each end on a tank at a rate of 10 mL/min. 2024 Behavior Tanks - Trial procedures: Each trial comprised 4 periods of time. The first period, (acclimation period) was 30 minutes long and included fish introduction along with time allowed for fish acclimation to the tank without the presence of an olfactory stimulus. Two juvenile individuals of a species were introduced into a tank. Using two individuals ensured that behavior patterns in the experimental tanks where consistent to the patterns in the holding tank. The second period (control) lasted 20 minutes and had no stimulus. The third period ran for 5 minutes to allow for addition of the stimulus and establishment of full coverage on the side of the tank to which it was introduced (olfactory stimulant introduction). The last period lasted 20 minutes. During this final period, the stimulus concentration on the side of the tank into which it was introduced was nominally kept at the target concentration (olfactory stimulant exposure). The fish behavior was tracked throughout the control, olfactory stimulant introduction, and olfactory stimulant exposure periods. A series of quality checks on the system was performed before and after each trial. A 10-minute flow test was conducted using the peristaltic pump with well water to test the pumps accuracy before a trial. If the flow rate from the pump differed from the target of 10 mL/min by plus or minus 0.2 mL/min or more, the pump was recalibrated and the 10-minute test was repeated. After the pump test, a 10-minute visual dye test was conducted to ensure the treatment solution would only be present on the treatment side and was fully removed by the center drains. At the end of a trial, we calculated the volume of stimulus solution delivered to ensure it was within 10 mL of the target delivery of the 250 milliliters. At the end of a trial, an additional 10-minute pump flow test as well as a 10-minute visual dye test were repeated to ensure nominal concentrations had been achieved, which was the case for all trials. Also, tank water temperature was taken before and after each trial to ensure consistency. 2024 Behavior Tanks - Trial video processing and data analysis: Videos of the trials were analyzed at 5 samples/second through the video tracking and analysis software, or VTAS (Social Interaction Module within EthoVision XT 17; Noldus: Wageningen, The Netherlands). A template created in the VTAS was made for both tanks to standardize processing across all trials. The arena consisted of a space in which the fish were able to freely swim around in each tank. The arena was broken into two equally sized zones, the front and back zones. Detection settings were created in the VTAS for each of the species of carp being used. Across all trials detection settings varied, as fish color and size often played a key role in how the settings were generated. The VTAS’s automated setup was used first to get the settings closer to what would be needed to track the fish, then additional fine tuning of the settings were made. Frame weight was kept between 1-10, as this was the recommended range from the VTAS’s help manual. For subject identification under detection settings was always set to Unmarked subjects as we were not using tagged or colored markers on the fish for assisted identification. Under Smoothing, video pixel smoothing was set to Low, dropped frames correction was set to “On”, and track noise reduction was set to Off. Under Subject Contour both contour erosion and contour dilation were checked and was always set as “Dilate first, then erode”, however the number for both will vairy depending on the species of carp used. Subject Size would always need fine tuning due to the different size fish per trial. Lastly before running the Acquisition, the Trial List was created and where the video file, arena setting, trial control settings, and the detection settings are all complied in a specified order so that the VTAS will pull the correct info to run initial tracking. Following the VTAS initial tracking analysis, additional manual tracking was done to eliminate any non-fish locations. After video files were processed using the VTAS, we exported calculated data to a spreadsheet containing percentage of a period spent in each half of the tank for each individual fish for each period during a trial. 2024 EOG_Results.csv Comma Separated Value (CSV) file containing data from electro-olfactogram trials. This table provides the date, carp species, stimulant and its concentration, flow rate, and holding tank water temperature, and individual fish mass. Each row represents a single stimulant exposure. Trials, each involving a single individual and three stimulus exposures, are denoted by the subject_id column. Each stimulant exposure has a corresponding baseline-adjusted measured electro-olfactory response and a pre-trial response to ultrapure, deionized (UDI) water. Producer Defined subject_id A unique numeric identifier for an individual fish. Producer Defined The identifier is a shorthand for the species (BLC, GSC, BHC, SVC) and a 6 digit date formatted MMDDYY. On days where multiple trials with the same species were conducted, these IDs end with an underscore and additional numbering (_01, _02, etc.) date A representation of time in which the smallest unit of measure is a day. The value is expressed in YYYYMMDD. It represents the date upon which trials were conducted. Producer Defined 20240221 20240917 YYYYMMDD species The common name of the test organism. A common name is a name other than a scientific name that is commonly used to refer to a species or other taxon. Producer Defined Black Carp Mylopharyngodon piceus Producer defined Grass Carp Ctenopharyngodon idella Producer defined Silver Carp Hypophthalmichthys molitrix Producer defined Bighead Carp Hypophthalmichthys nobilis Producer defined compname Name of the stimulus used for a trial. All amino acids are L-enantiomers and were exposed at 0.0001 molar Producer Defined lysine The stimulus used was a lysine solution at 0.001 M Producer defined alanine The stimulus used was a alanine solution at 0.001 M Producer defined impulse The solution used as a stimulus contained a dilution of a feeding stimulant product known as Impulse Producer defined response_mV The maximum extracellular electrical response of subject's naris recorded within 10 seconds following delivery of a stimulus, measured in millivolts and adjusted by subtracting the baseline electrical signal 60 seconds prior to stimulation. A running average was calculated for a moving window of five measurements across the response spike. The maximum running average was used as the response variable. Producer Defined 0.13125 9.7325 millivolts UDIbeg Absolute value of the lowest (i.e., most negative) extracellular electrical response of naris recorded within 10 seconds following delivery of ultrapure deionized (Nanopure) water at the beginning of a trial, measured in millivolts and adjusted by subtracting the baseline electrical signal 60 seconds prior to UDI water stimulation. This is specifically the value for the UDI water stimulation immediately prior to solution exposures and is therefore the same for rows with the same subject_id. Producer Defined 1.1925 84.28875 millivolts UDIend Absolute value of the lowest (i.e., most negative) extracellular electrical response of naris recorded within 10 seconds following delivery of ultrapure deionized (Nanopure) water at the end of a trial, measured in millivolts and adjusted by subtracting the baseline electrical signal 60 seconds prior to UDI water stimulation. This is specifically the value for the UDI water stimulation immediately following solution exposures and is therefore the same for rows with the same subject_id. Producer Defined 0.07875 48.52125 millivolts flow_rate The inverse flow rate of water and stimulus solutions going over the naris of a subject, measured as mL/s, measured prior to applying any stimulus. Producer Defined 3.41 8.94 milliliters per second beaker _temp_before The degree or intensity of heat or cold measured in the water of the beaker where test subjects were kept after being removed from their holding tanks, prior to start of trial, expressed in degrees Celsius Producer Defined NA No Data Producer defined 19.2 21.9 degrees Celsius cwf_temp_before The degree or intensity of heat or cold measured in the water that was constantly flowing over test subjects during the trials, measured before the start of a trial, expressed in degrees Celsius Producer Defined NA No Data Producer defined 19.9 24.5 beaker _temp_after The degree or intensity of heat or cold measured in the water of the beaker where test subjects were kept after being removed from their holding tanks, at the end of a trial, expressed in degrees Celsius Producer Defined NA No Data Producer defined 20.2 21.9 degrees Celsius cwf_temp_after The degree or intensity of heat or cold measured in the water that was constantly flowing over test subjects during the trials, measured at the end of a trial, expressed in degrees Celsius Producer Defined 20.4 24.6 degrees Celsius mass_g The wet mass of the test subject following euthanasia immediately after the trial, expressed in grams Producer Defined 1.64 12.83 grams length_mm The length of the test subject measured with electronic calipers following euthansia immediately after the trial, expressed in millimeters. Producer Defined 43.9 118.48 millimeters Behavior_Results.csv Comma Separated Value (CSV) file containing data on where the fish were positioned during behavior trials. Each row represents a period of time in a trial that was conducted on two individuals of the same species. The olfactory stimulant delivered during the trial, initial positioning of fish and compound within the tank, manual track editing status, and the averaged values of proportion of time are included in the dataset. Producer Defined trial_id A unique numeric identifier for a trial. Each trial involves two individual fish of the same species in the same tank exposed to the same olfactory stimulant. This is a key attribute linking data between tables. Producer Defined Trials were assigned sequential numbers. species The common name of the test organism. A common name is a name other than a scientific name that is commonly used to refer to a species or other taxon. Producer Defined grass carp Ctenopharyngodon idella Producer defined silver carp Hypophthalmichthys molitrix Producer defined bighead carp Hypophthalmichthys nobilis Producer defined black carp Mylopharyngodon piceus Producer defined compname Name of the stimulus used for a trial. All amino acids are L-enantiomers. Producer Defined Impulse The solution used as a stimulus contained a dilution of a commercial bait stimulus product known as Impulse Producer defined FishFood The solution contained a decant made from species specific fish food Producer defined Alanine The stimulus used was a alanine solution at 0.001 M Producer defined Lysine The stimulus used was a lysine solution at 0.001 M Producer defined date A representation of time in which the smallest unit of measure is a day. The value is expressed in YYYYMMDD. It represents the date upon which a trial was conducted. Producer Defined 20240722 20241106 YYYYMMDD fishplace If viewing the trial tank from above, the latitudinal half of the trial tank to which the two fish were introduced at the start of the trial. Halves were of equal dimensions and were divided by a line parallel to the shorter walls of the tank. There was no physical divider or line between halves. For each tank, the definitions 'front' and 'back' are consistent across trials and attributes. Producer Defined Identifers of latitudinal halves as front and back. For each tank, the definitions 'front' and 'back' are consistent across trials and attributes. compplace If viewing the trial tank from above, the half of the trial tank to which the olfactory stimulant was introduced during the exposure periods of the trial. Halves were of equal dimensions and were divided by a line parallel to the shorter walls of the tank. There was no physical divider or line between halves. For each tank, the definitions 'front' and 'back' are consistent across trials and attributes. Producer Defined Identifers of latitudinal halves as front and back. For each tank, the definitions 'front' and 'back' are consistent across trials and attributes. tracked A binary indicator of whether the trial video was edited with manual tracking of individuals in Ethovision. Producer Defined track_edited The video was edited with manual tracking of individuals Producer defined period An abbreviated name for each recorded period of a trial. Each trial had three recorded and tracked periods that were 20 minutes each. Producer Defined comp_acclimation The 5 minute period between the control pre-exposure and compound exposure periods when compound is being added to the exposure side of the tank and is expected to be reaching nominal concentrations. Producer defined comp_exposure Compound exposure period. The final period of the trial where the compound was expected to be present in one half of the tank at the nominal concentration. This period was twenty minutes in duration and was initiated 5 minutes after addition of the compound began, i.e. at the end of the comp_acclimation period. Producer defined control_preexp Pre-exposure control period. The twenty minute pre-exposure period which occurred approximately 30 minutes after the introduction of the fish. No compound is present in the tank during this period and it is intended to serve as a control. Producer defined prop_time The proportion of the period spent in the half of the tank where the compound was delivered. Producer Defined 0.0 1.0 unitless proportion Behavior_FishSize.csv Comma Separated Value (CSV) file containing size data for fish utilized in the behavior trials. Each row represents a period of time in a trial that was conducted on two individuals of the same species. A unique identifier for each fish, fish species, length of fish, and mass of fish are included in the dataset. Producer Defined trial_id A unique numeric identifier for a trial. Each trial involves two individual fish of the same species in the same tank exposed to the same olfactory stimulant. This is a key attribute linking data between tables. Producer Defined Trials were assigned sequential numbers. indiv_id A unique numeric identifier for a fish used in a trial. Producer Defined Numbers were assigned to fish sequentially. species The common name of the test organism. A common name is a name other than a scientific name that is commonly used to refer to a species or other taxon. Producer Defined grass carp Ctenopharyngodon idella Producer defined silver carp Hypophthalmichthys molitrix Producer defined bighead carp Hypophthalmichthys nobilis Producer defined black carp Mylopharyngodon piceus Producer defined length_mm The total length, from snout to tail, of the fish, measured at the end of the trial, reported in millimeters. Numeric. Producer Defined 41.7 116.85 millimeters mass_g The wet mass of the whole body of the fish, measured in grams at the conclusion of the trial Producer Defined 0.9 12.41 grams Behavior_ChamberTemperatures.csv Comma Separated Value (CSV) file containing temperature data for the trials. Each row represents a period of time in a trial that was conducted on two individuals of the same species. The fish species, temperature of the holding tank, temperature of the trial tank at the start of the experiment, temperature of the trial tank at the end of the experiment, and tank identification are included in the dataset. Producer Defined trial_id A unique numeric identifier for a trial. Each trial involves two individual fish of the same species in the same tank exposed to the same olfactory stimulant. This is a key attribute linking data between tables. Producer Defined Trials were assigned sequential numbers. date A representation of time in which the smallest unit of measure is a day. The value is expressed in YYYYMMDD. It represents the date upon which a trial was conducted. Producer Defined 20240722 20241106 YYYYMMDD species The common name of the test organism. A common name is a name other than a scientific name that is commonly used to refer to a species or other taxon. Producer Defined grass carp Ctenopharyngodon idella Producer defined silver carp Hypophthalmichthys molitrix Producer defined bighead carp Hypophthalmichthys nobilis Producer defined black carp Mylopharyngodon piceus Producer defined hold_tank_temp The degree or intensity of heat or cold measured in the tank where the fish were being held prior to taking part in the trial, expressed in degrees Celsius. Numeric. Producer Defined 18.7 20.7 degrees Celsius trial_tank_temp_start The degree or intensity of heat or cold measured in the testing tank at the start of the trial, expressed in degrees Celsius. Numeric. Producer Defined NA No Data. The temperature was not measured. Producer defined 19.6 20.8 degrees Celsius trial_tank_temp_end The degree or intensity of heat or cold measured in the testing tank at the end of the trial, expressed in degrees Celsius. Numeric. Producer Defined NA No Data. The temperature was not measured. Producer defined 19.1 20.3 degrees Celsius tank An identifier for the tank where the trail was run. Producer Defined The two trial tanks were identified as blue and yellow (yel). These are meaningless identifiers assigned to distinguish between the two exposure systems. This data release is made up of four files containing tabular data in CSV format. Electro-olfactogram data, including response, fish, and water data are contained in a single CSV file named EOG_Results.csv Data for behavior trials are spit between 3 tables. Behavior_results contains cumulative position data for each trial. Behavior_FishSize contains length and mass data for the fish used in the trials. Behavior_ChamberTemperatures contains temperature data for the holding and trial tanks. Data within these three tables are linked through the attribute 'trial_id' Wildhaber et al. 2025 GS ScienceBase U.S. Geological Survey mailing and physical

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