Progress 01/12/16 to 09/30/20
Outputs
Target Audience:State and federal agencies that batch mark fish. Aquaculture producers that might batch mark fish. Researchers that need to batch mark fish as part of their experimental protocols. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?We reported these results to the USFWS ADAP Program What do you plan to do during the next reporting period to accomplish the goals?This is a final report
Impacts
What was accomplished under these goals?
Objectives 1&2. We conducted range-finding and focused salinity tolerance tests to determine the level of salt that crappies (both Black Crappie and White Crappie) can handle for periods of 3.5 minutes. The final focused salinity tolerance tests were conducted at five treatment levels (1%-5%) salt concentrations, with 3 replicates per treatment level. We monitored immediate and 24-h mortalities for Black Crappie and White Crappie. Black Crappie experienced no immediate mortality at any concentration and no 24-h mortality at treatment levels up to 4%. The 24-h mortality for Black Crappie was 10% at a salt concentration of 5%. White Crappie experienced no immediate or 24-h mortalities at salt concentrations below 3%, but immediate and 24-h mortalities averaged about 13% at treatment concentrations of 3% and 4%. Objective 3. We determined an appropriate dunking density tolerated by crappies for a period of 20 minutes. We tested five treatment densities ranging from 0.016 to 0.250 kg/L, with 3 replicates per treatment level. We monitored immediate and 24-h mortality rates for both crappie species. Black Crappie had no immediate or 24-h mortality at dunking densities below 0.125 kg/L. Immediate and 24-h mortalities for Black Crappie at the highest dunking density (0.250 kg/L) averaged less than 5%. White Crappie exhibited immediate mortalities that increased from 4% at 0.016 kg/L to 20% at 0.250 kg/L. The 24-h mortalities generally mirrored the immediate mortalities at all treatment levels. The protocol utilized to mark crappies was conducted on batches of crappies held at a dunking density of 0.250 kg/L. The protocol included 3.5 min at 4% salt concentration as the osmotic induction for the calcein mark, followed by an approximately 10 sec rinse in clean water. The second step was a static immersion in a 1% calcein bath for 7 minutes, followed by another rinse in clean water. Fish were held overnight to determine initial marking efficacy and marking mortality. We marked approximately 96,500 2.6-g Black Crappie and 86,100 3.0-g White Crappie at two state fish hatcheries. The process took 35 man hours to mark Black Crappie and 50 man hours to mark White Crappie. The 24-h mortality for both marking effots was less than 0.5%. Efficacy of the calcein mark was 100%. Fish were graded using a mark quality scale (0-3), based on the requirements outlined in INAD #10-987T. We scored mark quality on the pectoral fin ray, scales, and isthmus. Black Crappie scores averaged 3 on all three body parts at the time of marking. Likewise, White Crappie mark quality averaged 3 on the pectoral fin ray, scales, and isthmus at the time of marking. Objective 4. Black and White Crappies are being held outdoor in 3,700-L blue polytanks. Pond water is continuously supplied to the tanks and returned to an earthen pond. Crappie are feeding on zooplankton naturally occurring in the earthen pond. Crappies are examined every 1-2 months and scored according to the aforementioned mark quality scheme. At 142 d post marking, mark quality on the pectoral fin ray, scales, and isthmus averaged 2.7 to 3.0 for Black Crappie. At 129 d post marking, mark quality on the pectoral fin ray, scales, and isthmus averaged 2.0-3.0 for White Crappie. Crappie are now being fed a combination of Fathead Minnow and Golden Shiner. Accuracy of detecting external calcein marks was determined separately for each crappie species through another blind mark detection test. Specimens used in the second blind test included 30 live calcein marked crappies combined with 30 live unmarked crappies from the same hatchery produced cohort. One individual set up the test and a second individual determined which crappies were marked with calcein. Calcein marks were assessed by seeking local darkness under the cover of 3-mm thick black plastic sheeting. The NIGHTSEA (Lexington, Massachusetts) optics VG-3 barrier filter glasses were worn, and UV light was directed upon the specimen. Accuracy of detecting external calcein marks was defined as the number of specimens scored correctly divided by the sample size expressed as a percentage. Accuracy of detecting calcein marks on sagittal otoliths was determined in a blind test for each species. Specimens used in the blind test included calcein marked Black Crappies (n = 10) and White Crappies (n = 10), and unmarked White Crappies (n = 10). Accuracy of detecting calcein marks on otoliths was defined as the number of specimens scored correctly divided by the sample size, and expressed as a percentage. Two readers independently determined which of the 30 samples were calcein marked, further noting whether the right, left, or both otoliths displayed the calcein mark. One reader was familiar with observing calcein marks on otoliths, while the second reader was in no way familiar with observing calcein marks on otoliths. The longevity of external calcein marks was estimated 11 times over a ~400-d period for subsamples of ~450 White Crappies and ~450 Black Crappies. These crappies were held in 3,789-L outdoor circular pools at UAPB. Experimental pools were allowed to experience a natural photoperiod. Water was pumped from an earthen pond through the pools and returned to the pond. Crappies initially fed on naturally occurring zooplankton. Later, Western Mosquitofish Gambusia affinis were fed to crappies at a rate of 10% of body weight per week. The external calcein mark on six body parts of each specimen was graded using the ordinal scale provided by the USFWS calcein INAD protocol. Ordinal mark scores ranged from three, which described a readily visible bright green mark, to two, which describes a clearly visible green mark, to one, a dimly visible dull green mark. A score of zero denoted that no mark could be seen. The body regions graded included those specified by the INAD and included the isthmus as the additional body region. One trained reader made estimates of calcein mark quality over the duration of this study. Mean mark quality of each body part was plotted to display variability of the mark through time. The longevity of calcein marks on otoliths was estimated for subsamples (n = 10) of both species from the experimental blue pools at ~500-d post-marking. The longevity of calcein marks on otoliths was reported as the maximum number of days post-marking when the calcein mark was still visible. The ordinal scale used to grade external calcein marks was applied to grade the quality of calcein marks on the sagittal otoliths of 20 known calcein marked crappies. We referred to these scores if a known calcein marked crappie was scored incorrectly. This information was helpful in describing the level of experience necessary to identify calcein marks on otoliths. ?
Publications
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Progress 10/01/18 to 09/30/19
Outputs
Target Audience:State and federal agencies that batch mark fish. Aquaculture producers that might batch mark fish. Researchers that need to batch mark fish as part of their experimental protocols. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?
Nothing Reported
How have the results been disseminated to communities of interest?We reported these results to the USFWS ADAP Program What do you plan to do during the next reporting period to accomplish the goals?
Nothing Reported
Impacts
What was accomplished under these goals?
We attempted the osmotic induction and calcein marking with fingerling largemouth bass during the summer 2018. This work was conducted under an agreement with the U.S. Fish and Wildlife Service (INAD #10-987). Prior work with crappies suggested osmotic induction with a 4% salt bath for 3.5 min followed by a 0.5% calcein solution for 7 min produced lasting external marks on another centrarchid. However, this procedure resulted in 100% mortality within 48 h of marking. During summer 2019, a series of trials were conducted to determine the reason for complete mortality using an established protocol. We again proceeded to mark fish using a 4% salt bath for 3.5 min followed by a 0.5% calcein solution for 7 min. As during the previous year, this procedure resulted in 100% mortality within 48 h of marking. Fish were stocked into outdoor tanks in this and every subsequent trial unless otherwise specified. A second trial was conducted with graded levels of salt (0%, 2%, and 4%) followed by a 7-min bath in a 0.5% calcein solution. Calcein solutions in this and all subsequently-described trials were always 0.5%. Mortality (48-h) was 100% in all treatment except the control, which experienced no mortality. A third trial with graded levels of salt (0%, 2%, and 4%) and a sham 7-min bath (no calcein) resulted in no mortalities after 48 h. A fourth trial with 2% salt and graded periods (3, 5, and 7 min) of 0.5% calcein bath resulted in 80% mortality after 48 h in all treatments and no mortalities in the control. We speculated that our calcein was contaminated and ordered fresh calcein. A fifth trial was conducted with 2% salt and 3.5-min bath in 5% calcein or 3.5-min in a sham bath. These fish were held in indoor tanks for 48 h and mortality in the treatment and control were both zero. A sixth trial was conducted with 2% salt and graded periods (3, 5, and 7 min) of 0.5% calcein bath or a 7-min sham. Fish were stocked directly outside. All treatments experienced 100% mortality, while the control experience none. A seventh trial was conducted with 2% salt and a 3-min 0.5% calcein bath or 3-min sham. Half the treatment and control replicates were stocked directly outside, while half the treatment and control replicates were stocked inside. All treatments and replicates experienced no mortalities after 48 h, except the treatments stocked directly outside, which experienced 100% mortality. A final trial was conducted with graded levels of salt (0%, 2%, and 4%) and a 3-min bath in 0.5% calcein. The control for this trial was 0% salt and 3-min sham calcein bath. All fish were held inside for 48 h. No mortalities occurred in any treatment except the 0% salt which experienced 10% mortality. After 48 h, fish were moved to outside tanks. Within 48 h, mortalities ranged from 60% to 80% in treatments, while the control experienced 10% mortality. A protocol for calcein marking of fingerling Largemouth Bass has not been established.
Publications
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Progress 10/01/17 to 09/30/18
Outputs
Target Audience:State and federal agencies that batch mark fish. Aquaculture producers that might batch mark fish. Researchers that need to batch mark fish as part of their experimental protocols. Changes/Problems:Calcein marking of Largemouth Bass following protocols for crappie (another centrarchid species) were unsuccessful. Mortality was 100% and no further studies could be conducted this year, because of size requirements of the USFWS INAD for calcein (fish < 1 g/fish). What opportunities for training and professional development has the project provided?One technician participated during this reporting period. How have the results been disseminated to communities of interest?We reported these results to the USFWS ADAP Program What do you plan to do during the next reporting period to accomplish the goals?We will conduct a series of experiements designed to pinpoint the hurdle to success, including changing osmotic induction salt concentrations and duration of calcein bath.
Impacts
What was accomplished under these goals?
We attempted the osmotic induction and calcein marking with fingerling largemouth bass during the summer 2018. This work was conducted under an agreement with the U.S. Fish and Wildlife Service (INAD #10-987). Prior work with crappies suggested osmotic induction with a 4% salt bath for 3.5 min followed by a 0.5% calcein solution for 7 min produced lasting external marks on another centrarchid. However, this procedure resulted in 100% mortality within 48 h of marking.
Publications
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Progress 10/01/16 to 09/30/17
Outputs
Target Audience:State and federal agencies that batch mark fish. Aquaculture producers that might batch mark fish. Researchers that need to batch mark fish as part of their experimental protocols. Changes/Problems:Based on our success, we intend to expand the project to include osmotic induction and calcein marking of Largemouth Bass. What opportunities for training and professional development has the project provided?On graduate student, one Research Associate, and one technician participated during this reporting period How have the results been disseminated to communities of interest?We have presented these results at professional meetings at the state and national levels. What do you plan to do during the next reporting period to accomplish the goals?We have presented these results at professional meetings at the state and national levels.
Impacts
What was accomplished under these goals?
Objectives 1-3 met during previous reporting period. Objective 4. Black and White Crappies are being held outdoor in 3,700-L blue polytanks. Pond water is continuously supplied to the tanks and returned to an earthen pond. Crappie are feeding on zooplankton naturally occurring in the earthen pond. Crappies are examined every 1-2 months and scored according to the aforementioned mark quality scheme. At 142 d post marking, mark quality on the pectoral fin ray, scales, and isthmus averaged 2.7 to 3.0 for Black Crappie. At 129 d post marking, mark quality on the pectoral fin ray, scales, and isthmus averaged 2.0-3.0 for White Crappie. Crappie are now being fed a combination of Fathead Minnow and Golden Shiner. Accuracy of detecting external calcein marks was determined separately for each crappie species through another blind mark detection test. Specimens used in the second blind test included 30 live calcein marked crappies combined with 30 live unmarked crappies from the same hatchery produced cohort. One individual set up the test and a second individual determined which crappies were marked with calcein. Calcein marks were assessed by seeking local darkness under the cover of 3-mm thick black plastic sheeting. The NIGHTSEA (Lexington, Massachusetts) optics VG-3 barrier filter glasses were worn, and UV light was directed upon the specimen. Accuracy of detecting external calcein marks was defined as the number of specimens scored correctly divided by the sample size expressed as a percentage. Accuracy of detecting calcein marks on sagittal otoliths was determined in a blind test for each species. Specimens used in the blind test included calcein marked Black Crappies (n = 10) and White Crappies (n = 10), and unmarked White Crappies (n = 10). Accuracy of detecting calcein marks on otoliths was defined as the number of specimens scored correctly divided by the sample size, and expressed as a percentage. Two readers independently determined which of the 30 samples were calcein marked, further noting whether the right, left, or both otoliths displayed the calcein mark. One reader was familiar with observing calcein marks on otoliths, while the second reader was in no way familiar with observing calcein marks on otoliths. The longevity of external calcein marks was estimated 11 times over a ~400-d period for subsamples of ~450 White Crappies and ~450 Black Crappies. These crappies were held in 3,789-L outdoor circular pools at UAPB. Experimental pools were allowed to experience a natural photoperiod. Water was pumped from an earthen pond through the pools and returned to the pond. Crappies initially fed on naturally occurring zooplankton. Later, Western Mosquitofish Gambusia affinis were fed to crappies at a rate of 10% of body weight per week. The external calcein mark on six body parts of each specimen was graded using the ordinal scale provided by the USFWS calcein INAD protocol. Ordinal mark scores ranged from three, which described a readily visible bright green mark, to two, which describes a clearly visible green mark, to one, a dimly visible dull green mark. A score of zero denoted that no mark could be seen. The body regions graded included those specified by the INAD and included the isthmus as the additional body region. One trained reader made estimates of calcein mark quality over the duration of this study. Mean mark quality of each body part was plotted to display variability of the mark through time. The longevity of calcein marks on otoliths was estimated for subsamples (n = 10) of both species from the experimental blue pools at ~500-d post-marking. The longevity of calcein marks on otoliths was reported as the maximum number of days post-marking when the calcein mark was still visible. The ordinal scale used to grade external calcein marks was applied to grade the quality of calcein marks on the sagittal otoliths of 20 known calcein marked crappies. We referred to these scores if a known calcein marked crappie was scored incorrectly. This information was helpful in describing the level of experience necessary to identify calcein marks on otoliths.
Publications
- Type:
Theses/Dissertations
Status:
Accepted
Year Published:
2017
Citation:
Farris, G.F. 2017. Assessment of Arkansas Game and Fish Commission Crappie Stock Enhancements. Masters Thesis. University of Arkansas at Pine Bluff, Pine Bluff, Arkansas. 67 pages.
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Progress 01/12/16 to 09/30/16
Outputs
Target Audience:State and federal agencies that batch mark fish. Aquaculture producers that might batch mark fish. Researchers that need to batch mark fish as part of their experimental protocols. Changes/Problems:
Nothing Reported
What opportunities for training and professional development has the project provided?Two graduate students, one university technician, and 7 state agency hatchery personnel have participated in the marking process and understand the marking protocol. We have presented these results at professional meetings at the state and national levels. How have the results been disseminated to communities of interest?We have presented these results at professional meetings at the state and national levels. What do you plan to do during the next reporting period to accomplish the goals?We will continue to monitor mark persistence in Black Crappie and White Crappie. Based on our success, we intend to expand the project to include osmotic induction and calcein marking of Largemouth Bass.
Impacts
What was accomplished under these goals?
Objectives 1&2. We conducted range-finding and focused salinity tolerance tests to determine the level of salt that crappies (both Black Crappie and White Crappie) can handle for periods of 3.5 minutes. The final focused salinity tolerance tests were conducted at five treatment levels (1%-5%) salt concentrations, with 3 replicates per treatment level. We monitored immediate and 24-h mortalities for Black Crappie and White Crappie. Black Crappie experienced no immediate mortality at any concentration and no 24-h mortality at treatment levels up to 4%. The 24-h mortality for Black Crappie was 10% at a salt concentration of 5%. White Crappie experienced no immediate or24-h mortalities at salt concentrations below3%, but immediate and 24-h mortalities averaged about13% at treatment concentrations of 3% and 4%. Objective 3. We determined an appropriate dunking density tolerated by crappies for a period of 20 minutes. We tested five treatment densities ranging from 0.016 to 0.250 kg/L, with 3 replicates per treatment level. We monitored immediate and 24-h mortality rates for both crappie species. Black Crappie had no immediate or 24-h mortality at dunking densities below 0.125 kg/L. Immediate and 24-h mortalities for Black Crappie at the highest dunking density (0.250 kg/L) averaged less than 5%. White Crappie exhibited immediate mortalities that increased from 4% at 0.016 kg/L to 20% at 0.250 kg/L. The 24-h mortalities generally mirrored the immediate mortalities at all treatment levels. The protocol utilized to mark crappies was conducted on batches of crappies held ata dunking density of 0.250 kg/L. The protocol included 3.5 min at 4% salt concentration as the osmotic induction for the calcein mark, followed by an approximately 10 sec rinse in clean water. The second step was a static immersion in a 1% calcein bathfor 7 minutes, followed byanother rinse in clean water. Fish were held overnight to determine initial marking efficacy and marking mortality. We marked approximately 96,5002.6-g Black Crappie and 86,100 3.0-g White Crappie at two state fish hatcheries. The process took 35 man hours to mark Black Crappie and 50 man hours to mark White Crappie. The 24-h mortality for both marking effots was less than 0.5%. Efficacy of the calcein mark was 100%. Fish were graded using a mark quality scale (0-3), based on the requirements outlined in INAD #10-987T. We scored mark quality on the pectoral fin ray, scales, and isthmus. Black Crappie scores averaged 3 on all three body parts at the time of marking. Likewise, White Crappie mark quality averaged 3 on the pectoral fin ray, scales, and isthmus at the time of marking. Objective 4. Black and White Crappies are being held outdoor in 3,700-L blue polytanks. Pond water is continuously supplied to the tanks and returned to an earthen pond. Crappie are feeding on zooplankton naturally occurring in the earthen pond. Crappies are examined every 1-2 months and scored according to the aforementioned mark quality scheme. At 142 d post marking, mark quality on the pectoral fin ray, scales, and isthmus averaged 2.7 to 3.0 for Black Crappie. At 129 d post marking, mark quality on the pectoral fin ray, scales, and isthmus averaged 2.0-3.0 for White Crappie. Crappie are now being fed a combination of Fathead Minnow and Golden Shiner. We continue to monitor mark persistence.
Publications
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