{"Bibliographic":{"Title":"A study to determine the biological feasibility of a new fish tagging system, 1985-1986","Authors":"","Publication date":"1986","Publisher":""},"Administrative":{"Date created":"08-16-2023","Language":"English","Rights":"CC 0","Size":"0000120490"},"Pages":["mahnken\nU.S Department of Energy\nA Study to Determine the Biological\nBonneville Power Administration\nDivision of Fish & Wildlife\nFeasibility of a New Fish Tagging System\nCoastal Zone and Estuarine\nStudies Division\nNorthwest and Alaska\nFisheries Center\nSH\nNational Marine Fisheries Service\n153\nP51\nDecember 1986\n1985-86\nAnnual\nReport\n1985","This report was funded by the Bonneville Power Administration (BPA), U.S.\nDepartment of Energy, as part of BPA's program to protect, mitigate, and\nenhance fish and wildlife affected by the development and operation of\nhydroelectric facilities on the Columbia River and its tributaries. The views\nin this report are the author's and do not necessarily represent the views of\nBPA.\nFor copies of this report, write:\nBonneville Power Administration\nDivision of Fish and Wildlife\nPublic Information Officer - PJ\nP.O. Box 3621\nPortland, OR 97208","SH\n51453\n153\npsl\nP51\n1985-86\nA STUDY TO DETERMINE THE BIOLOGICAL\nFEASIBILITY OF A NEW FISH TAGGING SYSTEM, 1985-1986\nby\nEarl F. Prentice\nDonn L. Park\nThomas A. Flagg\nand\nScott McCutcheon\nCoastal Zone and Estuarine Studies Division\nNorthwest and Alaska Fisheries Center\nNational Marine Fisheries Service\nNational Oceanic and Atmospheric Administration\n2725 Montlake Boulevard East\nSeattle, Washington 98112\nFunded by\nDepartment of Energy\nBonneville Power Administration\nDivision of Fish and Wildlife\nContract Number DE-AI79-84BP11982\nProject Number 83-319\nDecember 1986","ABSTRACT\nAn ongoing cooperative project between the Bonneville Power\nAdministration and the National Marine Fisheries Service was initiated in 1983\nto evaluate the technical and biological feasibility of adapting a new\nidentification system to salmonids. The system is based upon the passive\nintegrated transponder (PIT) tag. This report discusses the work completed in\n1985 and is divided into laboratory and field studies. All studies were\nconducted with the tag implanted into the body cavity of the test fish via a\n12-gauge hypodermic needle.\nLaboratory studies with juvenile chinook salmon and steelhead showed no\nadverse effect of the tag on growth or survival. Once the tag was established\nin the body cavity, its location was found to be consistent over time.\nBehavioral tests showed no significant effect of the tag on opercular rate,\ntail beat frequency, stamina, or post fatigue survival on juvenile\nsteelhead. Active swimming did not affect tag retention in steelhead. Tests\nrevealed a minimum size threshold for tag retention in juvenile steelhead at\n8.5 g before acceptable tag retention levels were achieved. No effect on\ngrowth or survival was observed for juvenile chinook salmon or steelhead.\nThe polypropylene encapsulated tags had an unacceptable failure rate due\nto moisture contacting the tag's electronic circuitry. The use\nof\npolypropylene encapsulated PIT tags was not recommended. The tag manufacturer\nnow produces the tag encapsulated in glass--which should provide significant\nimprovements in tag longevity and tag retention.\nNo evidence of infection due to tagging procedures was observed in tagged\nfish. Nevertheless, it was demonstrated that the PIT tag and tagging\napparatus could be disinfected against Aeromonas salmonicida by exposure to a\n50% or stronger solution of ethanol for a minimum of 1 minute.","Maturing Atlantic salmon were PIT tagged. In males, tag retention was\n100% prior to and after spawning. Females had 100% tag retention prior to\nspawning and 83% retention after multiple hand strippings. Lost tags\naccompanied the egg mass during strippings and were easily detected in the\nspawning bucket.\nAll field tests using juvenile salmonids were conducted at McNary Dam,\nwhereas tests using adult fish were conducted at Bonneville Dam. The PIT tag\nmonitoring equipment is described and discussed. The tag monitoring equipment\nshowed a high degree of reliability, efficiency, and accuracy. During the\n6-month testing period, tag reading efficiency exceeded 90%, and tag reading\naccuracy for juvenile chinook salmon was 100%. Only two minor equipment\nfailures occurred during the testing period.\nField studies used migrant spring and fall chinook salmon; no significant\neffects of the tag on survival could be determined when compared to\ntraditional tagging and marking methods. No significant difference was\nobserved in the recovery rate between branded and PIT tagged juvenile fall\nchinook salmon released into McNary reservoir and recovered at the dam. The\nPIT tag data were acquired with 90% fewer PIT tagged fish being released than\nbranded fish and a 33-fold reduction in the number of tagged fish being\nphysically handled to recover the data. Adult steelhead were successfully PIT\ntagged and automatically interrogated as they passed through a PIT tag monitor\ninstalled on a Denil fish ladder. It was concluded that a PIT tag monitor for\nadults can be installed at any location that can accommodate a coded wire tag\nmonitor.\nFuture work related to PIT tag systems development is described and\ndiscussed.","CONTENTS\nPAGE\nINTRODUCTION\n1\nPART I:\nLABORATORY STUDIES\n3\nStudy 1: Comparison Between Functional and Sham PIT Tags\n3\nIntroduction\n3\nMethods and Materials\n3\nResults and Discussion\n5\nStudy 2: PIT Tag Longevity\n11\nIntroduction\n11\nMethods and Materials\n11\nResults and Discussion\n12\nStudy 3: PIT Tag Effect on Locomotive Ability\n16\nIntroduction\n16\nMethods and Materials\n17\nResults and Discussion\n21\nSwimming Stamina\n21\nStride Efficiency\n22\nOpercular Beat Rate\n27\nPost-Test Survival and Tag Retention\n31\nStudy 4: Serial Tagging to Determine Minimum Fish Size for Tagging\nIntroduction\n33\nMethods and Materials\n34\nResults and Discussion\n34\nStudy 5: Tag Placement in Adult Salmon\n37\nIntroduction\n37","PAGE\nMethods and Materials\n38\nResults and Discussion\n38\nStudy 6: Sterilizati on Technique for Tagging Equipment\n41\nConclusions and Recommendations\n41\nPART II:\nFIELD STUDIES\n43\nStudy 1: Evaluate Juvenile PIT Tag Monitor Reliability\n43\nIntroduction\n43\nMethods and Materials\n43\nResults and Discussion\n45\nStudy 2: Evaluate Tag Reading Efficiency of the Juvenile\nPIT Tag Monitor\n48\nIntroduction\n48\nMethods and Materials\n49\nTest 1\n49\nTest 2\n50\nTest 3\n50\nResults and Discussion\n51\nTest 1\n51\nTest 2\n51\nTest 3\n53\nStudy 3: Comparison of the PIT Tag to Traditional Tagging and\nMarking Methods\n55\nIntroduction\n55\nMethods and Materials\n55\nResults and Discussion\n56","PAGE\nStudy 4: McNary Reservoir Release\n58\nIntroduction\n58\nMethods and Materials\n58\nResults and Discussion\n60\nStudy 5: Monitoring PIT Tags in Adult Fish\n62\nIntroduction\n62\nMethods and Materials\n63\nResults and Discussion\n65\nConclusions and Recommendations\n67\nPART III: SYSTEMS DEVELOPMENT\n69\nStudy 1: PIT Tag Injection Devices\n69\nStudy 2: Quality Control Monitor for Tagging\n69\nStudy 3: Hatchery Release Monitor\n71\nStudy 4: Design and Placement of Future Monitoring Systems\n72\nConclusions and Recommendations\n76\nACKNOWLEDGMENTS\n77\nLITERATURE CITED\n78\nAPPENDIX A-- -Preliminary Investigation of the Inactivation of\nAeromonas salmonicida, a Fish Pathogen\n80\nAPPENDIX B--Budget Information\n88","INTRODUCTION\nA cooperative program between the National Marine Fisheries Service\n(NMFS) and the Bonneville Power Administration (BPA) to evaluate the technical\nand biological feasibility of the passive integrated transponder (PIT) tag for\nsalmonid research has been under way since 1983. The PIT tag is being\ndeveloped as a research and management tool for monitoring the movements of\njuvenile and adult salmonids in the Columbia River Basin. Preliminary results\nshow that fish injected with this tag can be automatically recognized by\ndetecting/recording devices strategically located within the collection\nfacilities at hydroelectric dams. The PIT tag is an electronic tag 10 mm long\nby 2.1 mm in diameter that can be coded with one of 35 billion unique codes.\nThe tag can be automatically detected and decoded in situ--eliminating the\nneed to sacrifice, anesthetize, handle, or restrain fish during data\nretrieval.\nIn 1983 and 1984, juvenile and adult salmon were injected with sham\n(non-functional) PIT tags to determine suitable anatomical areas for tag\nplacement, develop tag injection techniques, and determine the effect of the\ntag on growth and survival. The body cavity was selected as the best area for\ntag placement for most applications from a biological and social standpoint.\nFrom 1984 to 1985, work continued to evaluate the effect of the tag on\ngrowth and survival of juvenile fish and to further refine the tagging\ntechnique. Functional PIT tags were used in studies for the first time.\nPrototype juvenile and adult PIT tag monitoring systems were evaluated in\nfield tests. Tag decoding efficiency averaged 90.5% for four different tests\nusing juvenile fish and 94.4% in tests using adult fish. Tag reading accuracy\nwas 100% for all tests.","2\nThis report covers the work conducted under the 1985 to 1986 work plan\nand is divided into three parts. Each of these studies concentrate on\ndifferent developmental aspects for the PIT tag. The species of fish used in\nthese studies varies, and was governed both by availability and\napplicability. The Laboratory Studies (Part I) focus on tag retention,\nreliability, and effects on behavior. This study establishes minimum fish\nsize criteria for tagging with the polypropylene encapsulated tag. The Field\nStudies (Part II) evaluate the PIT tag monitors and compare the PIT tag to the\ntraditional tagging and marking methods. Systems Development (Part III)\nfocuses on design and quality control measures needed to develop the PIT tag\nfor use in large scale studies.","3\nPART I: LABORATORY STUDIES\nStudy 1: Comparison Between Functional and Sham PIT Tags\nIntroduction\nAll laboratory tests through 1984 used sham, non-functional, tags. The\nsham tags were the same size and shape as functional tags and had the same\nexternal coating. These tests defined an acceptable anatomical area for tag\nplacement (intraperitoneally near the mid-ventral line and posterior of the\npectoral fins) and resulted in techniques for implanting the tag. The\nobjective of the 1985 study was to compare results obtained from fish injected\nwith sham tags to those injected with functional tags.\nMethods and Materials\nThe study was conducted at the University of Washington's Big Beef Creek\nResearch Station. Juvenile fall chinook salmon, Oncorhynchus tshawytscha,\nwere initially maintained in 2.4-m diameter tanks with running fresh water\n(surface water). Standard husbandry practices were followed in maintaining\nthe fish. Fish were randomly selected from the main population on 15 April\n1985 to establish five groups: functional tag, functional tag sacrifice, sham\ntag, sham tag sacrifice, and control. At the time the groups were established\nand at the termination of the study (20 August), a sub-sample of 10 fish from\neach group was weighed (+ 0.5 g) and measured (+ 3.0 mm) . The number\nof\nreplicates and number of fish per replicate are shown in Table 1.\nThe PIT tags and sham tags were injected into the body cavity of the fish\nusing a 12-gauge hypodermic needle. The control fish were handled, but not\ninjected with the hypodermic needle. During tag insertion, the needle was\nangled in a posterior direction, 2 to 3 mm to either side of the mid-ventral","4\nTable 1.--Test plan for Part I, Studies 1 and 2 using fall chinook salmon.\nRearing\nNumber of\nNumber of fish\nStudy\nTreatment\nreplicates\nper replicate\narea\n1\nControl\nFresh water\n6\n100\n1\nFunctional tag\nFresh water\n6\n100\n1\nFunctional tag sacrifice\nFresh water\n2\n100\n1\nSham tag\nFresh water\n6\n100\n1\nSham tag sacrifice\nFresh water\n2\n100\n2\nControl\nSeawater\n1\n300\n2\nFunctional tag\nSeawater\n1\n300","5\nline at the posterior end of the pectoral fins. Tagging methods are still\nbeing developed and automated, however, they generally followed methods\ndescribed by Prentice et al. 1985. At tagging, a single tag was loaded into\nthe barrel of a needle and, upon needle insertion into the fish, the tag was\nreleased via a push-rod attached to the plunger of the hypodermic syringe.\nTag location within the body cavity as well as tissue response to the tag were\ndetermined by examining fish that died or were sacrificed. The first two\nsacrifice groups were terminated and examined on 25 and 29 May, the third on\n19 July, and the fourth group on 20 August. All fish that died during the\nstudy were examined for tag retention and cause of death. At the termination\nof testing, all tagged fish were sacrificed and examined for tag location and\ntissue response to the tag.\nResults and Discussion\nNo significant difference (P<0.05) in length or weight was seen between\nreplicates within a treatment or between treatments at the start of the\nstudy. Similarly (P<0.05), growth rates were not different at the end of the\nstudy (127 elapsed days). These results are similar to that previously\nreported (Prentice et al. 1984 and 1985), suggesting the PIT tag does not\nsuppress growth.\nTag retention (sham and functional) was poor ranging from 58 to 93% at\n127 days (Table 2) No explanation can be given for the one sham tag\nreplicate with only a 58% tag retention, whereas the next lowest tag retention\nvalue was 74%. The overall percentages (combined replicates) of tag retention\nfor sham and functionally tagged fish were 80 and 86%, respectively.\nTag retention among the sacrificial groups was also poor. The first sham\ngroup was sacrificed on Day 40 of the test with a 5% tag loss. The second\nsham group was sacrificed on Day 97 showing a 25% tag loss. Similar high and","functional\nPercent\ntags\n96\n84\n95\n98\n99\n99\n90\n95\n-\n-\n-\n-\n-\n-\n-\n-\n-\n-\n-\n-\n-\n-\nretention\nPercent\ntag\n76\n90\n92\n89\n82\n89\n85\n93\n83\n74\n93\n85\n88\n58\n95\n75\n-\n-\n-\n-\n-\n-\nEnding weight\n(g)\n5.85\n3.15\n2.20\n4.17\n1.76\n6.55\n5.23\n3.50\n3.76\n5.65\n2.49\n5.33\n1.48\n3.41\n3.94\n2.63\n2.50\n3.18\n4.47\n4.61\n3.35\nSD\n-\n21.3\nAve.\n16.8\n21.3\n16.9\n27.4\n27.5\n20.3\n25.6\n24.3\n15.4\n22.5\n17.1\n22.7\n15.6\n15.3\n19.9\n24.5\n24.5\n18.5\n(g)\n9.8\n9.3\n-\nStarting weight\n(g)\n1.01\n0.70\n0.86\n0.97\n1.13\n0.91\n0.93\n0.76\n0.67\n1.08\n0.18\n0.65\n0.86\n0.87\n1.02\n0.62\n0.65\n0.72\n0.85\n0.90\n0.99\n0.69\nSD\nAve.\n(g)\n4.7\n4.2\n4.3\n5.4\n4.5\n4.6\n5.1\n4.8\n5.1\n4.6\n4.4\n4.7\n4.3\n4.8\n4.6\n4.5\n4.4\n4.6\n4.5\n4.6\n4.6\n3.9\nEnding length\nTable 2. Summary of data comparing functional and sham tagged fish to control fish.\n5.98\n(mm)\n6.56\n6.61\n7.32\n4.28\n11.80\n8.59\n10.40\n6.00\n5.69\n4.83\n6.89\n5.04\n7.09\n6.96\n5.79\n5.59\n5.85\n7.01\n6.89\n5.08\n6.22\nSD\nAve.\n(mm)\n130\n118\n103\n127\n120\n140\n139\n125\n131\n130\n114\n126\n94\n114\n128\n114\n114\n130\n121\n129\n95\n116\nStarting length\n(mm)\n5.29\n4.99\n4.37\n5.04\n5.66\n5.23\n5.03\n4.44\n2.58\n6.03\n4.65\n3.78\n5.70\n3.43\n4.40\n2.95\n2.74\n2.91\n5.06\n4.69\n5.51\n6.46\nSD\nThe experiment was extended 30 days after the sham sacrifice.\nAve.\n(mm)\n78\n76\n76\n79\n81\n79\n79\n79\n80\n78\n77\n80\n78\n81\n80\n79\n77\n77\n77\n78\n78\n78\nsurvival\nStarting Percent\nSample size varies due to availability of fish.\n98\n97\n97\n100\n99\n100\n99\n99\n96\n98\n95\n98\n99\n99\n99\n100\n97\n89\n99\n95\n96\n99\nnumber\n692\n100\n100\n100\n100\n100\n100\n100\n100\n100\n100\n100\n100\n100\n103\n100\n100\n100\n100\n100\n100\n100\nperiod\n(days)\nTest\n127-\n127\n127\n127\n127\n127\n127\n127\n127\n127\n127\n127\n127\n45\n127\n127\n127\n127\n127\n127\n40\n97\nReplicate\n1\n2\n3\n4\n5\n6\n1\n2\n3\n4\n5\n6\n1\n2\n1\n2\n3\n4\n5\n6\n1\n2\nSham sacrifice\nPIT sacrifice\nSham tagged\nPIT tagged\nTreatment\nControl\n1/\n2","7\nvariable tag loss occurred with the functional PIT tag sacrificial groups. At\nthe end of 45 days, 15% of the tags were lost from the first sacrificial\ngroup, whereas by the end of 127 days, only 7% were lost from the second\nsacrificial group. No good explanation can be offered for the variability in\ntag retention between replicates, but the small size of the fish (about 80 mm)\nat tagging was thought to be a factor controlling tag retention.\nDuring the experiments, tags were observed protruding through the body\nwall (Table 3)\nSome fish that had shed tags showed two scars in the\nabdominal region, one from the tagging needle and a second scar presumed to be\nwhere the tag exited the body cavity.\nThe exact mechanism by which the tag is rejected is unknown. It is\nprobable, however, that the peritoneal cavity of small fish is very limited in\nits capacity to accommodate a proportionally large foreign body such as the\nPIT tag. If at the time the tag is injected into the body cavity of small\nfish, it does not lie nearly parallel to the body axis, but lies at some\nangle, the body organs (intestine, gut, etc.) may exert pressure against the\ntag. Since there would be limited room in the peritoneal cavity under these\nconditions for the tag to reorient itself, the force against the tag could\ncause the tag to be pushed through the body wall. This condition could be\nfurther aggravated if there was a slight tissue reaction to the encapsulating\nmaterial or to the rough edges that occasionally occur on the tag . We have\nseen very few incidences of tissue reaction to the tag, however, such\nreactions may be short term and/or difficult to detect. The above condition\nmay not occur in large fish with peritoneal cavities of greater volume.\nA large number of functional tags failed to operate after 127 days of\ntesting. The percent of functional tags ranged from 84 to 99% (Table 2).\nWithin the sacrificial group, 10% of the tags failed within 45 days. The tag\nmanufacturer was made aware of the failure problem and is in the process of\nmodifying production techniques to increase the tag's longevity.","8\nTable 3. Summary of wound condition after tagging and tag location within\nthe body cavity of juvenile fall chinook salmon over time with a\ndescription of wound condition and tag location codes.\nDays post tagging\nCode\n40-45\n97\n127\nWound code a /\nPercent fish within a classification code\nb\nA\n7.3\n0\n0.6\nb\n0\n0.2\nB\n8.3\nb\nC\n84.4\n100.0\n99.2\nc/\nTag location code\nd\n/\n3.9\nA\n2.1\n0\nd\n86.5\n69.1\n83.3\nB\nd\n4.4\n1.0\nC\n0.0\nd\n5.2\n25.0\n6.9\nD\nd\nE\n6.3\n1.5\n4.9\nA = An open wound.\nB = A wound that is closed by a thin membrane and is\nhealing; at times a slight red or pinkish coloration\nis noticeable in the area of the wound.\n= A wound completely healed and may or may not be\nC\nnoticeable by the presence of a scar. There is no\nred or pink coloration in the area of the wound.\nb/ Percentage based on data from the combined sham and\nfunctional PIT tagged groups examined from Days 40-45.\nc/ A = Tag located between the pyloric caeca and mid-gut.\nB = Tag located near the abdominal musculature and often\nembedded in the posterior area of pyloric caeca near\nthe spleen or in the adipose tissue at the posterior\narea of the pyloric caeca.\nC = Tag found in an area other than those noted;\ngenerally between the mid-gut and air bladder or\nbetween the liver and pyloric caeca.\nD = No tag present.\nE = Tag partially protruding through abdominal wall.\nd / Percentages based only on the sham sacrificial group\nexamined on Day 40.","9\nSurvival was high among all groups, ranging from 89 to 100% (Table 2).\nControl fish showed a slightly (but not significantly) higher survival (97 to\n100%) than sham tagged (89 to 100%) or functionally tagged fish (95 to 99%)\nThe difference in survival between the control group and the other two\ntreatment groups was attributed to initial tagging mortality. Initial tagging\nmortality was from perforation of the intestine or laceration of the kidney\nwith the tagging needle at the time of tagging. Fish suffering such injuries\ndied within the first 4 days after tagging. All other mortalities among test\nand control fish were attributed to bacterial kidney disease or bacterial gill\ndisease.\nNo correlation was seen between tag retention and survival (r=0.030,\nP<0.05) among any test group (Table 2). The passing of the tag through the\nbody wall did not cause an increase in mortality. No infection or other\ndisease problems were visually observed among fish that were rejecting or had\nrejected their tag.\nTag wound condition and tag placement were documented for fish in four\nsacrificial groups (two sham and two functionally tagged groups) (Table 3)\nNearly 85% of the fish examined (n=195), regardless of treatment, showed the\ntagging wound to be completely healed with only a scar indicating the area of\nneedle insertion by Days 40 and 45. During this same period, 7.3% of the fish\nshowed an open wound and 8.3% showed a wound that was closed but slightly\ndiscolored. All fish (n=99) sacrificed after 90 days showed the wound to be\ncompletely healed. At the termination of the study (127 days), 102 fish from\na functional sacrifice group were examined, and 99.2% of the fish had\ncompletely healed wounds, 0.6% showed open wounds, and 0.2% had wounds that\nwere closed but slightly discolored.","10\nIn a previous study, data for juvenile steelhead Salmo gairdneri, showed\nthat after 30 days, all tagging wounds were completely healed (Prentice et al.\n1985). The fish used in that study were larger than the fall chinook salmon\nused in the present study. A second difference between the studies was that\nthe number of fish used per observation was limited (n=6) in the earlier work,\nthus the precision of the estimate is not comparable to the present study. In\nspite of the slight difference in results between the two studies, it is our\nopinion that no problem exists from the tagging. To date there has been no\nevidence of infection or excessive mortality resulting from PIT tagging fish.\nTag location within the body cavity was consistent regardless of the\ntreatment (sham or functional PIT tag) or time observed (Table 3)\nThe\nmajority of the tags were observed near the abdominal musculature either\nembedded in the posterior area of the pyloric caeca near the spleen or in the\nadipose tissue at the posterior area of the pyloric caeca. These results are\nconsistent with those obtained in a previous study, where 96% of the tags were\nfound in similar locations (Prentice et al. 1985).\nTag retention was a problem among both the test replicates and\nsacrificial groups regardless of treatment. Tag loss occurred throughout the\nstudy and showed signs of continuing by the presence of tags protruding from\nthe body wall. Close examination of these fish did not reveal where the tags\nmay have been within the body cavity prior to their migration through the\nabdominal wall.","11\nStudy 2: PIT Tag Longevity\nIntroduction\nThe only information pertaining to the longevity of the functional PIT\ntag is from the tag manufacturer who thoroughly tests the tags under\nlaboratory conditions. Field testing is necessary, however, to provide\nvaluable information, unobtainable in the laboratory, that is needed to design\nstudies and interpret their results. The objective of the study was to\ndetermine, under field conditions, the longevity of functional tags placed in\njuvenile salmon.\nMethods and Materials\nJuvenile fall chinook salmon were obtained from the same populations\nutilized in Study 1. On 2 April 1985, two 300-fish test groups were\nestablished at Big Beef Creek: one control and one functional tag group\n(Table 1). Tags were injected into the body cavity of fish as previously\ndescribed. All fish in each test group were weighed (+ 0.5 g) and measured\n(+ 3.0 mm) at the time the test groups were established. The identification\nnumber of each fish was recorded. The two test groups were maintained in\nseparate tanks in fresh water until smolted.\nAt the time of smoltification, as determined by visual observations, all\nfish were transported to the NMFS Manchester Marine Experimental Station near\nManchester, WA, (5 May) vaccinated against Vibrio sp.; and acclimated to\nseawater over a 5-day period. All fish in each test group were counted and\nthe presence of the functional tag verified prior to placement in seawater.\nThe PIT tag and control groups were maintained in separate seawater net -\npens. Standard husbandry practices were followed for the duration of the\nstudy. All dead fish were examined for cause of death, and the presence of","12\nthe tag was verified if applicable. Additional observations as to tag\npresence and functionality took place on 6 March, 21 August, and 5 November,\n1986. At termination of the study on 6 March 1986, all fish were measured,\nand a subsample of 25 fish from each treatment was weighed.\nResults and Discussion\nA total of 35 days elapsed from the time the fish were tagged to the time\nthey were transferred to seawater (Table 4) . During that period, two tagged\nfish died of kidney damage that occurred during tagging. Four control fish\ndied during freshwater rearing; one from jumping from a rearing tank and the\nother three from unknown causes. During seawater culture (306 days), a total\nof 9 tagged fish and 16 control fish died. The cause of death was bacterial\nkidney disease.\nNo significant difference (P<0.05) in growth between control fish and\ntagged fish was observed during 341 days of rearing (Table 4 and Figs. 1\nand 2). The mean starting fork lengths of control and tagged fish were\n70.0 mm + 3.8 (SD) and 69.8 mm + 3.8 (SD), respectively. After 341 days, the\nmean lengths were 254 mm + 26.0 (SD) for control fish and 256 mm + 24.8 (SD)\nfor tagged fish.\nTag longevity was poor. A total of 40 tags out of the initial 300 failed\n(13.3%) after 341 days in fish (Table 4). The nonfunctional tags were\nreturned to the manufacturer for inspection. They concluded that body fluids\nentered the tag through the ends of improperly sealed tags. At the time the\ntags are manufactured, they are pressure tested to several atmospheres using a\nleak indicator. It was discovered however, that micro-openings occur\noccasionally in the end seal of the tag. These openings closed under pressure\ntesting, and the defective tags were not detected. However, under normal","functional\nb Thirteen fish were missing from the group due to predators. The percent mortality is calculated on the basis of accounted for mortality.\nPercent\n/\n98.3\nTable 4.--Summary of growth, survival, and tag retention and longevity information for PIT tagged and control fall chinook salmon cultured\ntags\n98.7\n96.3\n92.4\n-\n-\n-\n-\na / The number of fish at the start of the period has been adjusted for mortalities, missing fish, fish with no tag, and fish with non-\nretention\nPercent\n91.7\n93.7\n100.0\n100.0\ntag\n-\n-\n-\n-\nEnding length\n7.00\n(mm)\n24.78\n9.61\n8.27\n4.83\n9.96\n17.72\n25.95\nSD\nMean\n(mm)\n79.0\n139.6\n202.0\n256.3\n82.4\n139.0\n198.0\n254.0\nStarting length\n(mm)\n3.77\n7.00\n9.61\n18.27\n3.80\n4.83\n9.96\n17.72\nSD\nMean\n(mm)\n70.0\n79.0\n139.0\n202.0\n70.0\n82.0\n139.0\n198.0\nEnding weight\n1.62\n64.49\n1.11\n68.45\n(g)\nSD\n-\n-\n-\n-\n.\nMean\n5.6\n225.1\n6.2\n247.5\n-\n-\n-\n-\n(g)\nOver an elapsed period of 341 days, 40 tags out 300 (13.3%) failed.\nStarting weight\n0.64\n1.62\n0.62\n1.11\n(g)\nSD\n-\n-\n-\n-\nMean\n(g)\n7.5\n5.6\n3.6\n6.2\n-\n-\n-\n-\nsurvival\nPercent\n99.0\n98.5\n98.3\n97.2\n98.7\n97.0\n97.6\n97.9\nof fish\nEnding\nnumber\n261 b\n297\n264\n233\n209\n296\n287\n280\nStarting\nnumber\nof fish\n300\n268\n237\n215\n300\n296\n287\n280\nfor 341 days.\nPeriod\n(days)\n0-35\n36-106\n107-217\n218-341\n0-35\n36-106\n107-217\n218-341\nfunctional tags.\nPIT tagged\nTreatment\nControl\n/\nc","14\n270\nT\n240\n210\n180\n150\n120\n90\nI\nStandard error\nSeawater entry\n60\nControl group\nPIT tagged group\n30\n0\n0\n50\n100\n150\n200\n250\n300\n350\nDays\nFigure 1. 6 -- Comparison of weight change of PIT tagged and control fish over\ntime.","15\n270\n250\n230\n210\n190\n170\n150\n130\n110\nI\nStandard error\nSeawater entry\n90\nControl group\nPIT tagged group\n70\n0\n50\n100\n150\n200\n250\n300\n350\nDays\nFigure 2. @ --Comparison of length change of PIT tagged and control fish over\ntime.","16\nconditions, capillary action drew fluids into the tag and caused shorting of\nthe electronic circuitry. The manufacturer of the tag will be providing tags\nwith a glass enclosure in 1986. This change in manufacturing should eliminate\nleakage problems and substantially increase tag longevity.\nTag retention initially was poor. In the first 35 days of culture, 8.3%\nof the tags (25 tags) were not retained within the body cavity. During the\nnext 107 days of rearing, an additional 6.3% of the tags were rejected. Tag\nrejection, however, was zero during the following 234 days. The increase in\nfish size during the last 234 days of the study may have accounted for the\nimproved tag retention.\nThe tag rejection process did not jeopardize the survival of the fish.\nDuring the 341 days of culture, 17 tagged fish died (vs 26 control fish), ,\nwhile 42 tags were rejected. The exact mechanism of tag rejection remains\nunknown.\nStudy 3: PIT Tag Effect on Locomotive Ability\nIntroduction\nBoth internal and external ultrasonic telemetry tags have been shown to\nadversely effect the fishes swimming ability and respiratory rate (McCleave\nand Stred 1975; Lewis and Muntz 1984) and, therefore, could potentially alter\nmigratory ability. Though the PIT tag is only about 3% of the volume of\ncommonly used juvenile radio telemetry tags (Monan 1985), there is concern\nthat swimming performance could be affected. The present study evaluated the\nphysiological/behavioral effects of the PIT tag on locomotive ability for two\nsize ranges of steelhead, these tests are ongoing and will eventually include\nother size ranges of steelhead and chinook salmon. Locomotive performance was","17\nevaluated by assessing swimming stamina, tail beat (swimming) proficiency, and\nrespiratory rates.\nMethods and Materials\nTwo size ranges of steelhead were evaluated in the present study:\nfingerling fish tested in July 1985 averaged 83 mm + 8 (SD) in length and\n6.5 g + 1.8 (SD) in weight. In October 1985, juvenile steelhead were tested,\nthese fish averaged 112 mm + 9 (SD) in length and 17.2 g + 4.4 (SD). At\ntesting, random samples (n=200) were removed from the main population and\nintraperitoneally tagged with the PIT tag using procedures described by\nPrentice et al. (1984). A control, non-tagged, group (n=200) was also\nestablished from the main population at this time. Swimming performance tests\nwere conducted on Days 0 (same day as tagging) 1, 2, 3, 4, 7, 9, 11, 14, 17,\n21, and 25, following tagging, with 12 tagged and 4 control fish tested each\nday.\nSwimming tests were conducted in a modified version of the Blaska\nrespirometer-stamina chamber described by Smith and Newcomb (1970)\n(Fig. 3). These chambers were divided into multiple compartments to allow the\nsimultaneous testing of four fish. Each test chamber was equipped with an\nelectrified screen at the downstream end, assuring maximum fish performance.\nIn these tests, fish were individually anesthetized [tricaine methane-\nsulfonate (MS-222)], weighed (+ 0.1 g) and fork length measured (+ 1 mm), and\nthen placed into a test compartment. After a 1-h recovery period, the initial\nwater velocity was set at 1.5 body lengths per second (1/s) and increased 0.5\n1/s every 15 minutes until all the fish reached fatigue (i.e., could no longer\nhold position in the current and remained impinged against the electrified\nscreen).","13\n10\nEnd View\n12\n16\nside and end views. For loading, the chamber is tilted, partially\nfilled with water, and end plate and vane are removed. Fish are\nplaced in the test compartments, vane and end plate are replaced,\nis\nproduced with motor driven propeller and varied via motor speed\ncontroller. Direction of water flow is toward propeller in inner\ntube, water is turned at the end plate, and returned through the\n11\nFigure 3. . -- Diagram of modified Blaska respirometer-stamina - chamber, showing\nand chamber is filled with water and leveled. Water flow\n15\n15. Axle for tilting chamber\nspace between the inner and outer tubes (see arrows). .\n13. End plate (removable\n16. Compartment divider\n10. Test compartment\n11. Removable vane\nfor fish loading)\nSide View\n12. Outflow\n14. Inflow\n10\n9\n8\n14\n7\n6\n1. Variable speed control\n7. Outer tube (plexiglas)\n8. Inner tube (plexiglas)\n9. Electrified screen\n5\nM\n3. Tachometer\n4\n5. End plate\n6. Propeller\n3\n2. Motor\n4. Pulley\n2\n1","19\nIn these studies, the step-wise 1/s value was based on the mean length of\nthe four fish in the chamber. The swimming speed of each fish was calculated\nfrom the relationship of the mean length of the fish in the chamber, and\nlength of each individual fish, to the water flow within the chamber by the\nformula:\nwhere:\nSp\n= swimming speed of individual fish in body lengths\nper second (1/s)\n1 = mean length of the four fish (mm)\nlii = length of the individual fish (mm)\nV = water velocity in the chamber (1/s)\nIndividual swimming speed was corrected for the effects of solid blocking\n(for any fish whose size was greater than 10% of the cross-sectional area of\nits swimming compartment) using the formula of Bell and Terhune (1970):\nVE = Vt\nwhere: Vf = effective velocity (1/s)\nVt = average velocity through the empty test section (1/s)\nA = maximum cross-sectional area of the object in the test\nsection (mm2)\nAt = test section area (mm2)\nA swimming stamina profile (U-critical) was established for each group,\nusing the swimming speed at fatigue and the time of fatigue as an integrated\ntime/velocity measure of impingment, by the methods described by Beamish\n(1978):","20\nU-critical = U + (ti/tii x U11)\nwhere: U-critical = critical swimming speed (1/s)\nU = highest velocity maintained for the prescribed period (1/s)\nUii = velocity increment (1/s)\nti = time (in minutes) fish swims at fatigue (impingment)\nvelocity\ntii = prescribed period of swimming (in minutes)\nSwimming (tail beat) proficiency was determined for all tested fish by\ndocumenting the number of tail beats per minute over the range of swimming\nspeeds using a video camera with a superimposed stop watch function.\nRespiratory rate was determined by documenting the number of opercular beats\nper minute.\nTail-beat frequency (TBF) and opercular beat rate (OBR) per minute were\nmonitored using a video camera. Data were recorded with fish maintaining\nposition in the central portion of the swimming tunnel and not moving relative\nto the video recording equipment. The TBF and OBR were documented two to\nthree times throughout each 15-minute velocity increment. Stride length\n(distance traveled per tail beat) was calculated by the formula:\nSL = Sp/TBF\nwhere: SL = stride length\nSp = swimming speed of individual fish in body lengths per\nsecond (1/s)\nTBF = tail beat frequency, complete cycles per minute","21\nStride efficiency (number of tail beats per minute required to maintain a\nunit swimming speed of one body length per second) was calculated for each\nwater velocity increment from the tail beat frequency data by the formula:\nSE = TBF/Sp\nwhere: SE = stride efficiency\nTBF = tail beat frequency, complete cycles per minute\nSp\n= swimming speed of individual fish in body lengths per\nsecond (1/s)\nAll tested fish (tagged and control) were held for 14 days post-test to\nestablish stress survival profiles. These fish were fed daily, and the\npopulations were inspected regularly to document mortality. At the end of the\n14-day holding period, all fish were examined to determine tag retention.\nThe swimming stamina data, stride efficiency data, and respiratory rate\ndata were compared between tagged and control fish, and between post-tag\ntesting dates, using the non-parametric Mann-Whitney test.\nSwimming\nproficiency profiles for tagged and control fish were calculated using\nstandard regression techniques. All data analyses followed the methods of\nSokal and Rohlf (1981).\nResults and Discussion\nSwimming Stamina. -- Changes in swimming stamina levels have proven to be a\nreliable indicator of significant stressors in fish (Beamish 1978; Flagg\n1981). Depressions in swimming stamina levels have been noted in teleost fish\nupon exposure to many stressors, including both external and internal\ntelemetry tags (McCleave and Stred 1975; Lewis and Muntz 1984). The present\nstudy indicates that neither the act of tagging nor the presence of the PIT\ntag is a significant stress to steelhead, as measured by swimming stamina\ntests.","22\nThe Mann-Whitney statistical tests indicate that the PIT tag does not\ncompromise the swimming stamina (U-critical) of steelhead. Fish were tested\nduring Days 0-25 post-tag and there were no statistical differences (P<0.01)\nbetween tagged and control fish at any test day (post-tag) for either\nfingerling (Table 5) or juvenile (Table 6) steelhead. The swimming stamina of\nPIT tagged and control fish varied slightly between test days (Figs. 4 and 5), ,\nhowever, no trend is evident, and the data suggest that a swimming stamina\nlevel (U-critical) of 4.6-5.2 body lengths per second is representative of the\nfish used in this study (Tables 5 and 6). This swimming stamina level is\nwithin limits documented by other authors (Beamish 1978) and indicates that\nthe PIT tagged steelhead in these studies had good locomotive ability.\nStride Efficiency Measures of tail beat frequency have been used by\nresearchers to document changes in physiological condition of fish (Beamish\n1978; Stevens 1979; Flagg and Smith 1982). Recently, Lewis and Muntz (1984)\nshowed that external ultrasonic tagging adversely affects the tail beat\nfrequency of rainbow trout, Salmo gairdneri. However in our tests, the PIT\ntag did not affect the tail beat efficiency of steelhead. These data suggest\nthat tagging and the presence of the PIT tag are not significant physiological\nimpairments to steelhead.\nStride efficiency (number of tail beats per minute required to maintain a\nunit swimming speed of one body length per second) was documented as a\ncomparative measure of propulsive efficiency. The Mann-Whitney statistical\ntests showed there were no statistical differences (P<0.01) between test (PIT\ntagged) and control (non-tagged) fish at any post-test day (0-25) for either\nsize range of steelhead tested (Tables 5 and 6).","23\nTable 5. Stride efficiency, opercular beat rate, and swimming stamina of\nPIT tagged and control fingerling steelhead (6.5 g average)\nTest\nStride\nOpercular\nSwimming\nB/\nb/\nb/\nday\nefficiency\nbeat rate\nstamina\npost\na/\n-\ntag\nGroup\nSE\nSE\nSE\nX\nX\nX\n0\nT\n86.7\n6.1\n133.3\n5.4\n4.4\n0.19\nC\n74.3\n10.0\n131.0\n8.3\n4.8\n0.43\n1\nT\n94.1\n4.3\n145.1\n2.9\n4.2\n0.07\nC\n93.8\n6.8\n145.1\n5.2\n4.1\n0.09\n2\nT\n88.0\n3.3\n150.5\n3.8\n4.6\n0.19\nC\n83.4\n4.4\n146.3\n5.6\n5.2\n0.25\n3\nT\n95.3\n4.0\n154.0\n2.7\n5.7\n0.18\nC\n89.1\n7.6\n155.4\n6.2\n5.3\n0.35\n4\nT\n101.6\n4.2\n144.3\n3.5\n4.9\n0.33\nC\n102.0\n8.6\n147.3\n6.7\n5.9\n0.55\n7\nT\n89.9\n3.5\n150.8\n3.0\n5.4\n0.16\nC\n84.2\n6.4\n144.6\n4.9\n5.4\n0.31\n9\nT\n95.8\n3.1\n139.9\n3.5\n5.2\n0.20\nC\n99.4\n6.1\n126.6\n4.9\n4.9\n0.00\n11\nT\n95.5\n3.4\n143.5\n2.6\n5.3\n0.15\nC\n105.3\n8.6\n144.5\n5.5\n5.2\n0.40\n14\nT\n100.3\n3.7\n141.9\n3.4\n5.3\n0.20\nC\n97.7\n5.3\n141.8\n7.1\n5.4\n0.35\n17\nT\n102.9\n4.2\n143.0\n3.3\n5.1\n0.16\nC\n108.6\n8.0\n143.4\n3.9\n4.9\n0.90\n21\nT\n93.6\n3.0\n136.0\n3.5\n5.8\n0.09\nC\n95.3\n4.7\n136.9\n5.6\n5.6\n0.00\n25\nT\n103.5\n4.2\n143.0\n3.4\n5.3\n0.30\nc\nC\n102.3\n5.4\n150.0\n6.4\n-\ntagged\n95.6\n143.8\n5.1\nX\nX control\n94.6\n142.7\n5.2\nT = PIT tagged, n = 12 tagged fish tested each day\nC = control, n = 4 control fish tested each day\nb\nX = mean\nSE = standard error\n= significantly different, P<0.01; (note: there were no statistical\n*\ndifferences noted in these data)\n/\nNo data due to equipment malfunction.\nc","24\nTable 6. -Stride efficiency, opercular beat rate, and swimming stamina of\nPIT tagged and control juvenile steelhead (17.2 g average)\nTest\nStride\nOpercular\nSwimming\nB/\nb/\nb/\nday\nefficiency\nbeat rate\nstamina\npost\na/\n-\n-\ntag\nGroup\nSE\nSE\nSE\nX\nX\nX\n0\nT\n105.8\n2.8\n153.8*\n1.6\n4.0\n0.09\nC\n92.4\n6.9\n139.3*\n4.3\n3.9\n0.20\n1\nT\n105.5\n2.9\n150.4\n2.6\n4.1\n0.14\nC\n98.8\n4.5\n153.0\n3.0\n4.5\n0.22\n2\nT\n100.2\n3.1\n149.5\n3.7\n4.2\n0.26\nC\n103.9\n7.8\n151.4\n4.3\n4.9\n0.09\n3\nT\n97.5\n3.0\n145.2\n2.7\n4.7\n0.19\nC\n100.6\n4.5\n148.6\n2.4\n4.8\n0.03\n4\nT\n105.5\n3.7\n145.5\n1.9\n4.5\n0.15\nC\n92.6\n4.7\n147.0\n5.4\n4.3\n0.62\n7\nT\n90.7\n3.1\n144.7\n3.2\n4.9\n0.14\nC\n93.4\n5.2\n147.0\n6.6\n4.4\n0.54\n9\nT\n100.7\n3.2\n145.1\n2.3\n4.8\n0.01\nC\n96.3\n4.1\n154.1\n6.1\n4.9\n0.13\n11\nT\n102.4\n3.1\n152.1\n3.0\n4.7\n0.21\nC\n93.9\n4.2\n146.3\n6.2\n5.2\n0.15\n14\nT\n94.4\n2.4\n145.2\n3.1\n4.9\n0.03\nC\n89.6\n4.2\n152.3\n5.1\n5.1\n0.28\n17\nT\n104.1\n3.9\n142.2\n3.3\n4.7\n0.18\nC\n89.2\n4.6\n141.0\n6.8\n4.9\n0.10\n21\nT\n98.2\n2.8\n149.5\n2.6\n4.8\n0.08\nC\n97.1\n4.9\n152.7\n4.0\n4.9\n0.07\n25\nT\n97.2\n2.5\n151.6\n2.7\n4.6\n0.13\nC\n98.7\n5.5\n148.5\n6.1\n4.6\n0.19\n-\ntagged\n100.2\n147.9\n4.6\nX\nX control\n95.5\n148.4\n4.7\nT = PIT tagged, n = 12 tagged fish tested each day\na\nC = control, n = 4 control fish tested each day\nb X = mean\nSE = standard error\n*\n= significantly different, P<0.01","25\n7\n6\n5\n4\nPIT tagged\n3\nControl\n1 2 3 4 7 9 11 14 17 21 25\n0\n0\nDays post-tag\nFigure 4. -- Mean fingerling swimming stamina (U-critical) of PIT tagged and\npost-tag. steelhead (6.5 g average) trout during control Days 0-25\nBrackets indicate + one standard error.","26\n7\n6\n5\n-\n4\nPIT tagged\nControl\n3\n0 1 2 3 4 7 9 11 14 17 21\n0\n25\nDays post-tag\nMean juvenile tag. swimming steelhead stamina (17.2 (U-critical) of PIT tagged\nFigure\n5.\nBrackets indicate + one g average) standard trout during and Days control 0-25\n-\npost-\nerror.","27\nStride efficiency varied slightly between test days, and on Day 0 was\nreduced, although not significantly (P<0.01), from control levels for both\nfingerling and juvenile fish. In addition, the control fish were slightly,\nbut not significantly (P<0.01), more stride efficient throughout the tests\n(Tables 5 and 6; Figs. 6 and 7). However, this advantage varied between test\ndays, and no clear trend was evident--suggesting that a stride efficiency of\n94.6-100.2 tb/l/s is representative of fish used in this study. The results\nof this test suggest that interperitoneally tagging with the PIT tag does not\naffect the stride efficiency of steelhead.\nOpercular Beat Rate.--Changes in respiratory metabolism have also been\nused by researchers to document changes in the physiological condition of\nfish. Lewis and Muntz (1984) showed that external ultrasonic tags raise the\nrespiratory (opercular beat) rate, and the authors suggested that these type\ntags cause physiological compromises in rainbow trout. In the present study,\nOBR was documented as a comparative measure of respiratory efficiency. The\ndata suggest that the PIT type tags do not physiologically compromise\nsteelhead.\nIn the tests on fingerling steelhead, OBR exhibited an unexplained\nprogressive increase during the first 4 days (for both test and control fish),\nand subsequently, peaked and stabilized (Table 5 and Fig. 8). However, the\nMann-Whitney statistical tests indicated there were no statistical differences\n(P<0.01) between the PIT-tagged and control fish (fingerling steelhead) at any\ntest day in this series of tests (Table 5). Therefore, it seems probable that\nsome external environmental influence caused the variations in OBR level noted\nin tests on fingerling steelhead.","28\n115\n110\n100\n90\n80\n70\nPIT tagged\n60\nControl\n0 1 2 3 4 7 9 11 14 17 21\n0\n25\nDays post-tag\n6. . -- Mean steelhead stride (6.5 efficiency g of PIT tagged and control\nFigure\nBrackets indicate + one average) standard trout error. during Days fingerling 0-25 post-tag.","29\n120\n110\nT\n100\n90\n80\n70\nPIT tagged\n60\nControl\n0\n17\n21\n25\n0\n1\n2\n3\n4\n7\n9\n11\n14\nDays post-tag\nFigure 7. . -- Mean stride efficiency of PIT tagged and control juvenile steelhead\n(17.2 g average) trout during Days 0-25 post-tag. Brackets\nindicate + one standard error.","30\n160\n150\n140\n130\n120\n110\nPIT tagged\nControl\n0\n0\n1\n2\n3\n4\n7\n9\n11\n14\n17\n21\n25\nDays post-tag\nFigure 8. - Mean opercular beat rate of PIT tagged and control fingerling\nsteelhead (6.5 g average) trout during Days 0-25 post-tag.\nBrackets indicate + one standard error.","31\nIn the tests on juvenile steelhead, control OBR was significantly\n(P=0.01) reduced from that of PIT-tagged fish at Day 0 post-tag. However, by\nDay 1 post-tagging, control OBR had increased to that of the PIT-tagged fish,\nand there were no further significant differences (P<0.01) between test and\ncontrol fish for the remainder of the test series (Table 6 and Fig 9). Since\ncontrol OBR increased to equal the PIT-tagged fish by Day 1 post-tagging, the\nsignificance of the lower OBR for control fish at Day 0 is unclear.\nHowever, since only one of the test days (out of 24 observations) showed\na statistical difference from controls, it is apparent that neither tagging\nnor the presence of the PIT tag normally compromise the respiratory efficiency\nof steelhead. The data suggest that an OBR of 140-150 is most commonly\nrepresentative of (swimming) steelhead (Tables 5 and 6).\nPost-Test Survival and Tag Retention The effects of tagging on fish can\nvary due to tag type, size, and placement. Recent tagging/survival studies\nusing juvenile salmonids indicated that the PIT tag has excellent (up to 99%)\nretention and does not adversely affect survival (Prentice et al. 1985).\nHowever, the potential interactions of tagging and stress have not been fully\ndocumented. Severe exercise, such as swimming to fatigue, is a stress that\nhas the potential to induce trauma (possibly causing tag rejection) or even\ndeath (Black 1958; Beamish 1978; Flagg et al. 1983).\nIn the present study, all fish were held 14 days after their stamina\ntest, and survival and tag retention were documented to assess whether the act\nof tagging and/or the presence of the PIT tag were detrimental to fish\nencountering a severe secondary stress (e.g., swimming to fatigue).\nNeither the act of tagging nor the presence of the PIT tag had any effect\non the fishes post-stress (fatigue test) survival. Of the 414 steelhead","32\n160\n150\n140\n130\n120\n110\nPIT tagged\nControl\n0\n0\n1\n2\n3\n4\n7\n9\n11\n14\n17\n21\n25\nDays post-tag\nFigure 9. -- -Mean opercular beat rate of PIT tagged and control juvenile\nsteelhead ( 17.2 g average) trout during Days 0-25 post-tag.\nBrackets indicate + one standard error. Asterisk (*) indicates\nsignificant (tagged VS. control) difference (P<0.01).","33\nsurveyed during these series of tests, none of 312 PIT-tagged nor 102 control\nfish died ( (100% survival). In addition, PIT tag retention was 100%. At the\ntermination of the 14-day holding, all PIT-tagged fish were sacrificed and\nnecropsies performed to determine tissue reaction to the tags. No adverse\ntissue reaction and no tag migration were noted.\nThus, it appears that the PIT tag does not impact the fish's ability to\nsurvive severe secondary stress (e.g., swimming to fatigue). It also appears\nthat this type of severe stress (even during the first few days post-tag) does\nnot compromise tag retention.\nThis study indicates that the PIT tag does not compromise the swimming\nefficiency, swimming stamina, or respiratory rate of either fingerling or\njuvenile steelhead. In addition, this study supports previous work showing\nexcellent PIT tag retention and survivability.\nHowever, the full\nphysiological/behavioral effect of the PIT tag on smolting or migrating fish\nis still not known. During the 1986 season, these type tests will continue in\nthe hat chery using smolting steelhead and three size ranges of chinook\nsalmon. In addition, locomotion tests will be conducted on migrating spring\nchinook salmon, fall chinook salmon, and steelhead at McNary Dam.\nStudy 4: Serial Tagging to Determine Minimum Fish Size for Tagging\nIntroduction\nPIT tag retention in juvenile fish has been variable (Prentice et al.\n1984 and 1985). In 1985, we conducted a study to compare the functional PIT\ntag to sham tags (see Part I, Study 1 of this report), and tag loss varied\nbetween 7 and 42%. Similarly, a study to determine tag longevity (Part 1,\nStudy 2) showed a high tag rejection rate (8.3% within the first 35 days).","34\nThe objective of this study was to evaluate the relationship between fish\nsize at tagging and tag retention. The criteria for successful tagging was\n96% or greater tag retention over a 45-day period. The study was not outlined\nin our 1984-85 BPA Work Plan, but was conducted after the results of Studies 1\nand 2 of this report were available.\nMethods and Materials\nJuvenile steelhead were used for the study. The population was\nmaintained in a 2.4-m diameter tank with running fresh water; standard\nhusbandry practices were followed. The study was conducted at the Big Beef\nCreek Research Station. Fish were randomly selected from the main population\nto establish eight test groups, each consisting of two replicates of 150 fish\neach (Table 7). Each replicate was maintained in a 1.2-m diameter tank with\nrunning fresh water. One test group was established about every 14 days\nbetween 1 August and 23 October 1985. Thirty-two days elapsed between the\nestablishment of the seventh and eighth test groups.\nAll fish were injected with functional PIT tags in a manner similar to\nthat described previously. Fifty fish in each replicate were weighed to the\nnearest 0.5 g, and all fish were measured to the nearest 3.0 mm (fork length)\nat the start and end of the study (45 elapsed days). Each test tank was\nexamined for rejected tags at 1- to 3-day intervals. All fish were sacrificed\nat the end of the study and examined for tag presence.\nResults and Discussion\nAll data for the study are summarized in Table 7. The data presented are\nfor combined replicates since there were no apparent differences between\nreplicates for weight, length, number of tags rejected, or survival.","Table 7.--Summary of serial tagging study to determine the minimum fish size for optimal tag retention and survival.\n/ Summary data are for combined replicates since no significant difference was seen between replicates for growth,\nretention\nPercent\n100.0\ntag\n91.7\n86.4\n95.0\n94.0\n98.0\n98.0\n99.7\nsurvival\nPercent\n96.3\n98.3\n98.0\n99.7\n98.3\n100.0\n99.0\n99.3\nEnding length\n(mm)\n10.1\n7.9\n9.1\n6.7\n5.3\n8.5\n7.9\n11.9\nSD\nMean\n(mm)\n84.1\n83.6\n89.5\n92.7\n100.6\n123.0\n121.5\n138.0\nb / Group 3 fish were from a separate population than all other test groups.\nEnding weight\nSD\n(g)\n2.5\n2.0\n2.6\n2.5\n3.6\n5.3\n4.4\n8.0\nMean\n(g)\n8.5\n7.6\n8.8\n10.5\n13.4\n23.5\n22.6\n32.8\nStarting length\n(mm)\n4.3\n4.7\n4.9\n4.7\n6.8\n7.6\n13.6\n10.6\nSD\nMean\n(mm)\n63.9\n69.4\n71.8\n75.9\n85.0\n92.9\n103.7\n128.6\nStarting weight\n(g)\n0.7\n1.2\n1.0\n1.2\n2.1\n2.8\n4.4\n6.9\nSD\nsurvival, or tag retention.\nMean\n(g)\n3.3\n4.3\n4.3\n6.1\n8.5\n10.8\n15.8\n28.0\na/\nNumber\nfish\nof\n300\n300\n301\n301\n302\n301\n300\n301\nGroup\nb/\n3°\n2\n1\n4\n5\n6\n7\n8\na","36\nOur criteria for successful tag retention was 96% over a 45-day period.\nThis criteria was achieved only with fish within the fifth through the eighth\ntest groups. The mean weight and length of fish within the fifth group was\n8.5 g + 2.1 (SD) and 85 mm + 6.8 (SD). The poorest tag retention was observed\nin the first test group (13.6% tag rejection). Tag rejection occurred on the\nfirst day after tagging and continued throughout the 45 days of testing in\nGroups 1 through 4. The majority of the tags were rejected between Days 13\nand 30. The few tags that were rejected from Groups 5 through 8 occurred\nafter Day 26, with the exception being two tags rejected from Group 7 on\nDay 8. The exact number of tags rejected during a specific period was\ndifficult to ascertain. Once tags were rejected, fish had a tendency to\ninjest them and were capable of passing them through the intestinal tract at a\nlater date. All fish within each test group were sacrificed at the end of 45\ndays, and the presence or absence of the tag within the body cavity was\nconfirmed. Upon examination of the fish, we found up to four tags in the\nstomach of one fish and several other fish that had injested one or two\ntags. How many fish had injested tags and passed them prior to the\ntermination of the study is unknown.\nSurvival was high between test groups, ranging from 96.3 to 100%\n(Table 7). The lowest survival was in the first test group, which had the\nsmallest fish. Damage to the intestinal tract from the tagging needle\naccounted for a number of the initial (first 4 days) mortalities among the\nfish. This was especially true with the smaller fish.\nNo clear relationship was seen between survival and tag loss if the\npercent survival for each group of fish is compared to the percent tag\nrejection (Table 7). Thus, there appears to be no severe adverse effect to","37\nthe fish during the tag rejection process. The exact mechanism or reason for\ntag rejection is unknown at this time. We have observed fish with either a\nscar or a partially protruding tag through the abdominal wall; no infection or\nother adverse tissue reaction to the tag could be observed in such fish. We\nhave further observed fish with protruding tags. If these fish are left in\nthe population, they will continue to grow and survive.\nPresently the minimum size fish that meets our criteria for successful\ntag retention weighs 8.5 g + 2.1 (SD) and measures 85 mm + 2.1 (SD). . High\nsurvival (greater than 96%), however, can be achieved with fish much smaller\nthan the above size restriction. The process of rejecting the tag does not\nappear to compromise the health or survival of the fish. The mechanism or\nreason for tag rejection between various size groups of fish is unknown.\nModification to the tag's encapsulation material from polypropylene to glass\nand altering tagging procedures slightly may improve tag retention.\nStudy 5: Tag Placement in Adult Salmon\nIntroduction\nNumerous morphological and physiological changes take place as a salmon\nmatures. These changes may alter the response of a fish to foreign material\nsuch as a PIT tag within its body cavity. For instance, since wound healing\nability may be impaired in maturing fish, tag implantation may subject the\nfish to infection and thus increase the chance for tag loss through an open\nwound or cause premature death. Furthermore, the questions of whether a tag\nplaced in the body cavity would cause internal damage to eggs and whether a\ntag would be retained during spawning need to be answered. The objective of\nthis study, therefore, was to obtain information on wound healing, tag\nretention, and tag effect during spawning in maturing adult salmon.","38\nMethods and Materials\nThe study was conducted at the Manchester Marine Experimental Station and\nthe Northwest and Alaska Fisheries Center (NWAFC) in Seattle, Washington. A\ntotal of 84 maturing female and male Atlantic salmon, Salmo salar, were used\nin the study. The fish were reared to near maturity in seawater net-pens at\nManchester.\nAll fish were PIT tagged intraperitoneally on 15 October 1985. Initially\nthe tag was injected through the abdominal musculature about 3 to 5 cm\nanterior to the pelvic girdle along the mid-ventral line. This procedure was\nsubsequently modified by moving the injection point about 1 to 2 cm to either\nside of the mid-ventral line. Tag insertion was made with a 12-gauge needle\nand a modified hypodermic syringe.\nThe fish were divided into two groups. One group consisted of 10 males\nand 33 females retained in seawater until spawning. The second group\nconsisted of 11 males and 30 females transported to fresh water at the\nNWAFC. All fish were weighed to the nearest 100 g and measured (fork length)\nto the nearest 1 cm. Fish weight ranged from 2,500 to 10,000 g, and lengths\nranged from 61 to 80 cm. All fish were examined for wound healing, readiness\nto spawn, and general condition on 18, 21, 22, 23, 25, and 29 October and\n4 November. The study was terminated on 5 November. When fish were\ndetermined to be ripe, eggs were collected by squeezing the peritoneal cavity\nby hand (stripping). All fish were lightly anesthetized (MS-222) for spawning\nand scanned for tag code using a hand-held scanning unit. Individuals that\nspawned were subjected to 3 to 4 strippings.\nResults and Discussion\nDuring the study, no adverse reaction by the tissue to the tag was\nnoted. A11 tagging wounds were closed and healing on the first day of","39\nobservation (3 days post tagging). No infection or discoloration was noted in\nthe area of the tagging wound.\nThree fish in the seawater group were removed from the study immediately\nafter tagging when severe external bleeding was noted in the area of the tag\nwound. The bleeding problem was eliminated in subsequent tagging by moving\nthe tag injection site about 1 to 2 cm to either side of the mid-ventral line\nthereby avoiding the ventral artery. We recommend this change in tagging\nprocedure for all size ranges of fish. The distance from the mid-ventral line\nshould vary, however, with the size of fish being tagged.\nAll 21 males matured, and milt was collected from each fish. A total of\n48 females were spawned from the population of 60 fish in the study\n(Table 8). At the termination of the study (5 November), 12 fish had not yet\nripened.\nOverall, there was 100% tag retention among male fish and 83% among\nfemales. Four tags were passed during the first stripping and four during the\nsecond to fourth strippings (Table 8). There was no clear relationship\nbetween tag retention among freshwater or seawater test groups. No adverse\neffects could be noted to the eggs from the tag's presence. When a tag was\npassed, it was easily observed among the eggs.\nAll fish were easily identified with one or two scans of a portable tag\ndetector using lightly anesthetized fish. During the observation periods, the\nfish were placed in a 1.2-m diameter tank and guided to the tank's side where\ntag detection was accomplished from the exterior of the tank without removing\nthe fish from the water.","40\nTable 8 . -- --Spawning dates and PIT tag rejection for Atlantic salmon females.\nDate\nNo. females\nCumulative\nNo. tags\nspawned\nspawned per date\nno. spawned\nnot retained\n1a/\n21 Oct\n21\n21\n22 Oct\n4\n25\n0\n23 Oct\n7\n32\n0\n2b/\n25 Oct\n7\n39\n3c/\n29 Oct\n3\n42\n2d/\n4 Nov\n6\n48\na/\nOne tag not retained during first. stripping.\nb\n/ One tag not retained during third and fourth stripping.\nc/\nOne tag not retained during first, second, and fourth stripping.\n/\nd\nTwo tags not retained during first stripping.","41\nStudy 6: Sterilization Technique for Tagging Equipment\nPresently, the PIT tag is injected into the fish's body cavity using a\n12-gauge hypodermic needle attached to a modified syringe. The same unit\n(needle and syringe) is used for consecutive fish. This procedure has not\nresulted in any documented disease transfer from fish to fish; however, the\nfish used in the tests were healthy. To reduce the potential of transferring\ndiseases from fish to fish via the tagging apparatus, a practical means of\ndisinfection is needed. Battelle Northwest, Sequim Marine Laboratory, was\ncontracted to evaluate the problem and to provide recommendations. Their\nreport is presented in Appendix A.\nConclusions and Recommendations\n1. The presence of the PIT tag within the body cavity of juvenile fall\nchinook salmon and steelhead will not significantly (P<0.05) affect growth or\nsurvival.\n2. Tag retention can be expected to be high (tag retention of 96% or\ngreater over 45 days) for juvenile steelhead weighing more than 8.5 g and\nmeasuring greater than 85 mm.\n3. The exact mechanism for tag rejection in juvenile fall chinook salmon\nand steelhead is unknown but may be primarily mechanical and, in part, related\nto fish size.\nThere is no correlation between survival and tag rejection for\n4.\njuvenile fall chinook salmon and steelhead.\n5. Tag location in juvenile fall chinook salmon is consistent (greater\nthan 90%) within the body cavity over time, suggesting that once the tag is\nestablished within the cavity it remains stationary.","42\n6. Even though no infection in the area of the tagging wound was noted,\nand survival of tagged fish was not significantly different from control fish,\nwe recommend that both the tags and tagging apparatus be disinfected (when\npractical) to reduce the chance for disease transmission from fish to fish.\n7. Tag longevity was poor with up to 8.3% of the tags failing to\nfunction after 35 days due primarily to liquids entering the tag through\nfaulty end seals on the polypropylene capsule. We do not recommend the use of\nthe polypropolene encapsulated PIT tags at this time. We believe however,\nthat this problem will be overcome by the introduction of glass encapsulated\ntags in 1986 1 /\n8. The PIT tag does not have a significant effect on the opercular rate,\ntail beat frequency, stamina, and post fatigue survival of fingerling or\njuvenile steelhead.\nActive swimming does not affect tag retention in fingerling or\n9.\njuvenile steelhead (100% retention over 14 days in all tests). The PIT\ntag\nwill not significantly affect locomotive ability of juvenile steelhead in the\nsize range tested.\n10. The PIT tag can be injected safely into maturing adult salmon\nwithout jeopardizing their health, survival, and egg or sperm viability.\n1/\nPreliminary 1986 data show that by encapsulating the tags in glass, tag\nlongevity and retention are greatly improved.","43\n11. The PIT tag is retained within the body cavity of adult female salmon\nat a high rate even after multiple hand-strippings.\n12. We recommend that until additional laboratory and field tests are\nconducted and the data analyzed, that a cautious approach be taken in the use\nof the PIT tag, even though all the information to date is encouraging.\nPremature use of the tag may give biased results stemming from a lack of\nunderstanding of the technical limitations of the tag and monitoring system\nand an incomplete understanding of the biological ramifications of injecting\nthe tag into fish. We believe that if test results continue to be as\nencouraging as they are, the tag should be ready for use in the field by 1987.\nPART II: FIELD STUDIES\nStudy 1: Evaluate Juvenile PIT Tag Monitor Reliability\nIntroduction\nThe objective of the study was to determine the reliablity of juvenile\nPIT tag monitoring equipment installed at McNary Dam during the 1985 field\nseason. The continuous operation of the equipment is essential not only to\nensure the accuracy and reliability of the collected data but also to\ndetermine areas for design improvement.\nMethods and Materials\nThe study was conducted at McNary Dam on the Columbia River near\nUmatilla, Oregon. Two juvenile PIT tag monitors were installed directly on\nthe fish discharge ports of the juvenile wet separator (Fig. 10) Water\nvelocity through the monitors was up to 0.3 m/sec. Monitor A was 147.3 cm\nlong by 20.3 cm high by 30.5 cm wide and had three monitoring loops.\nMonitor B was 122.0 cm long by 20.3 cm high by 30.5 cm wide and had two\nmonitoring loops. Both monitors were made of clear PVC and had a plastic","44\nDewatering area\n(perforated\nplater)\nTo raceway\nor river\nSubsample\nCoil\nCoil\nCoil\ngate\nC\nB\nA\nFlow\nA Side\nMonitor A\nMonitor B\nFlow\nB Side\nCoil Coil\nB\nA\nSubsample\ngate\nTo raceway or river\nFigure 10. Location of juvenile salmon PIT tag monitors at McNary Dam during\n1985.","45\nshield to protect the loops from weather. Monitor A was operated by a triple\nexcitor and power supply mounted in a single housing and wired directly to the\nloops. Monitor B was operated by a dual excitor and power supply mounted and\nwired as Monitor A. The excitors of Monitors A and B were connected to\nindividual controller units, printers, and computers (Fig. 11).\nTo evaluate the reliability of the electronic components of the PIT tag\nmonitoring system, all equipment except the printer and computer were left\ncontinuously in an operational mode from 27 April to 20 July 1985. The\nequipment was again activated from 4 August to 28 September 1985. During the\nactive period, a total of 16 tests (8 tests per monitor) were conducted to\ndetermine monitor tag reading reliability.\nThe tests were conducted on a monthly basis from April to September 1985\n(Table 9). Each test consisted of releasing neutrally buoyant plastic fishing\nbobbers (5.8 cm long by 2.5 cm diameter) containing a functional PIT tag. The\nnumber of bobbers release per test ranged from 8 to 204 (Table 9). The bobbers\nwere released into the entrance of each monitor and recovered upon their exit\nfor reuse.\nResults and Discussion\nThe prototype juvenile PIT tag monitoring equipment performed well during\nthe 1985 field season with only two electronic equipment problems. The\nmonitoring equipment was turned off on 20 July while a leak in a section of\nthe\nflume was repaired. Monitor A malfunctioned during power-up\non\n4 August. Two controller cards within the controller malfunctioned, and two\ncapacitors failed within the power supply. The failure of the capacitors\nprobably caused the controller cards to malfunction. All repairs were made in\nthe field within 1 hour.","46\nMONITOR A\n30.5 cm\n147.3 cm\n20.3 cm\nFlow\nC\nA\nB\nCoils\nMONITOR B\n30.5 cm\n122.0 cm\n20.3 cm\nFlow\nB\nA\nCoils\nPower\nExcitor\nExcitor\nPower\nsupply\nassembly\nsupply\nassembly\nController\nController\nPrinter\nComputer\nComputer\nPrinter\nFigure 11. -- Diagram of the juvenile salmon PIT tag monitoring system at McNary\nDam during 1985.","47\nTable 9 .--Summary of reliability tests conducted at McNary Dam in 1985.\nNumber\nNumber\nTotal\nNumber of\nPercent\nNumber\nof\nof bobbers\nnumber of\nbobbers\nbobbers\nreading\nTest date\nMonitor\ntrials\nper trial\nbobbers\nnot read\nread\nerrors\n4/29\n2\nA\n51\n4\n204\n4\n98\n4\n98\n0\nB\n50\n4\n200\n5/10\nA\n52\n3\n156\n0\n100\n0\n4\n98\n0\nB\n49\n4\n196\n5/22\n0\nA\n51\n3 to 4\n184\n7\n96\n5\n98\n0\nB\n51\n4\n204\n5/28\nA\n26\n4\n104\n3\n97\n0\n3\n97\n0\nB\n23\n4\n92\n6/5\n4\n104\n100\n0\nA\n26\n0\nB\n27\n4\n108\n1\n99\n0\n7/17\n4\n8\n0\n100\n0\nA\n2\n8\n0\n100\n0\nB\n2\n4\n98\n8/2\n0\nA\n10\n4\n40\n1\n42\n0\n100\n0\nB\n12\n3 to 4\n9/25\n91\n0\nA\n28\n4\n112\n10\nB\n32\n4\n128\n0\n100\n0\n97\n246\n2\nTotal\n3 to 4\n912\n25\nA\n17\nTotal\n246\n3 to 4\n978\n98\n0\nB","48\nMonitor A again malfunctioned sometime after 5 August. One of the three\nmonitoring loops failed which caused the detuning of the remaining two\nloops. We estimated that a 6 to 7% decrease in tag reading ability resulted\nfrom this failure and detuning (Table 9).\nResults of the monthly tag reading tests are shown in Table 9. A total\nof 912 tags were passed through Monitor A, and 978 tags through Monitor B\nduring 8 tests per monitor. Out of 1,890 tags, two tags were misread. No\nexplanation can be given for the two reading errors. No other misreadings\nwere experienced in any other study conducted during 1985 using PIT tags.\nOverall reading efficiency for all tests was 97 and 98% for Monitors A and B,\nrespectively. The slight difference in overall tag reading efficiency between\nMonitors A and B was due to the detuning of the detector loops on Monitor A as\npreviously discussed.\nThe overall results of the reliability tests suggest that the PIT tag\nmonitoring equipment can withstand the rigors of field operation over an\nextended time. The results of the tag reading tests with the bobbers showed a\nhigh degree of reliability in reading efficiency, and the results were similar\nto those obtained with fish. This suggests that the bobbers used in these\ntests are a dependable substitute for fish in determining monitor reliability.\nStudy 2: Evaluate Tag Reading Efficiency of the Juvenile PIT Tag Monitor\nIntroduction\nJuvenile PIT tag monitors were evaluated for tag reading efficiency under\nsimulated field conditions in 1984 (Prentice et al. 1985). Results showed a\nmean reading efficiency of 90.5%. However, a question remained whether this\nlevel of reading efficiency could be obtained under actual test conditions in\nthe field. This study was designed to answer that question.","49\nMethods and Materials\nTwo juvenile PIT tag monitors installed on the wet separator at McNary\nDam on the Columbia River were evaluated. The monitors are described in\nPart II, Study 1 of this report (Fig. 10). Three tests were conducted, two\nusing juvenile migrant spring chinook salmon and one with migrant fall chinook\nsalmon.\nTest --Juvenile migrant spring chinook salmon used in the study were\nrandomly collected from the juvenile salmon collection and inspection facility\nat McNary Dam on 8 May 1985. At the facility, a subsample of fish passing\nthrough the juvenile collection system was diverted into an inspection room\nwhere they were dipnetted; anesthetized; and inspected for fin clips,\ndescaling, injuries, species composition, and brands. Only fish showing\nlimited scale loss and no previous marks, tags, or injuries were used in the\nstudy. The fish were PIT tagged in the same manner as previously described.\nTwenty-five groups of fish, 20 fish per group, were tagged, measured to the\nnearest 3 mm (fork length), and recorded on a computer file and printer. The\nfish ranged in length from 95 to 215 mm and averaged 147 mm. Each group of\nfish was held in a 132-liter holding container receiving a continuous supply\nof aerated ambient river water.\nThe fish were held between 20 and 25 h prior to their release directly\ninto the wet separator (Fig. 10). Prior to release, each group was examined\nfor tag loss and mortality. All mortalities were replaced with fish from the\n25th group of fish. The individual code and length of the replacement fish\nwere substituted for the removed mortalities, thus all release groups had 20\nfish. Two groups of fish were released into the wet separator at 30-min\nintervals, one in the A side, the other in the B side (Fig. 10).","50\nAll fish were allowed to pass through the wet separator on their own\nvolition. All PIT tagged fish were interrogated, and PIT tag codes were\nrecorded automatically using the systems previously described. The code of\neach PIT tagged fish, monitor, detection loop, date of passage, and time of\npassage (hour, minute, and second) were recorded into a computer and printer\nfile.\nTest 2.--At the termination of the study, comparing the PIT tag to\ntraditional tagging and marking methods (Part II, Study 3), all surviving PIT\ntagged fish within each of four test groups were retained. On 3 June 1985,\nadditional fish obtained from the inspection facility were tagged and added to\neach of the four groups as needed to adjust the total number of fish per group\nto 26. A fifth group of 20 fish was tagged as replacement fish for any\nsubsequent mortalities. All fish handling, holding, releasing (two releases\nof two groups per release), and tag monitoring were conducted in a manner\nsimilar to Test 1.\nTest .--Juvenile migrant fall chinook salmon ranging in length from 85\nto 160 mm were used in the test. The fish were obtained from two sources.\nGroups 1 through 13 were obtained from the subsample as were the fish in\nTest 1. These fish had up to 24 h of rest prior to being handled. Low\nnumbers of fish in the subsample made it necessary to obtain the needed fish\nfor Groups 14 through 24 from a raceway system. Many fish from the raceway\ndid not have an opportunity to recover from stress resulting from their\npassage through the dam's collection facility before being handled for\ntagging. After tagging, all test groups were held for 24 h. The rest of the\nmethods and materials, number, and size of the test groups were all similar to\nTest 1; the main differences between Tests 1 and 3 were the species used, time\nof year, and prevailing environmental conditions.","51\nResults and Discussion\nTest 1.--A total of 480 PIT tagged spring chinook salmon were released\ninto the wet separator; 9 fish were not detected for an overall tag reading\nefficiency of 98.1%. All of the tags that were detected were read correctly\n(100% reading accuracy).\nThe elapsed time for spring chinook salmon to exit the wet separator in\nTest 1 ranged from 16 seconds to 36 h 27 min (Fig. 12). Eighty-one percent of\nthe fish were detected within the first 30 min after release, 9% in the next\n30 min, and 5% within the following 60 min. Two fish resided in the wet\nseparator for extended periods: 20 h 13 min and 36 h 27 min. No explanation\ncan be given for the long residence time for these two fish; however, this\nphenomenon has been observed previously (Park et al. 1984).\nBased upon our 1984 work, our criteria for acceptable tag reading\nefficiency was 90% with 99% reading accuracy (Prentice et al. 1985). The\nresults of this test far exceeded that criteria.\nTest 2.--The -- results obtained in Test 2 were similar to that of Test 1.\nOverall tag reading efficiency for Test 2 was 97.1% (3 fish were not detected\nout of 104 fish released). All tags that were detected were correctly decoded\n(100% tag reading accuracy).\nPassage time of PIT tagged fish out of the wet separator was similar to\nthat for Test 1 (Fig. 12). Within the first 30 min, 74% of the fish exited\nthe system, an additional 11% passed through the system in the next 30 min,\nand 12% within the following 60 min. No fish remained in the separator longer\nthan 3 h and 44 min.","52\n90\n80\n70\n60\n50\n40\n30\nTest I\nTest II\n20\n10\n2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 35 36\n0\n1\nTime (hours)\n12. exiting Percentage the of PIT tagged spring chinook salmon detected while\nFigure\nMcNary Dam wet separator.","53\nTest 3.--Post tagging mortality was different between the two sources of\nfish. Fish from the subsample (Groups 1 through 13) showed a 0.38% mortality\n(1 fish died), whereas fish from the raceway showed a 4.1% mortality (11 fish\ndied). Overall 24-h post tagging mortality was 2.2%. Raceway mortality (non-\ntagged fish) during the same period was 1.7%. The difference in mortality\nbetween the two sources of fish likely indicates the effect of stressing a\nfish twice within a short period without sufficient recovery time.\nOverall tag reading efficiency was 92.5%, with all tags being read\ncorrectly (100% tag reading accuracy). We believe, however, that the tag\nreading efficiency was affected by fish dying within the wet separator. Tag\nreading efficiency was different between the two sources of fish: Group 1\nthrough 13 (n=260), 95.4% and Groups 14 through 24 (n=220), 89.1%.\nWe believe that the difference in mortality between the two sources of\nfish continued after release into the wet separator. Since the residence time\nfor the fall chinook salmon in the wet separator was long (Fig. 13), there was\na high probability for mortality to occur. After death, a fish would have\ndecayed rapidly and lost its tag in the 20° to 21°C water present during the\ntest. Tags lost in this manner would not be available for detection but would\ndrop through the wet separator's perforated floor.\nThe time for fall chinook salmon to exit the wet separator was much\ndifferent than for spring chinook salmon in Tests 1 and 2 (Figs. 12 and 13).\nWithin the first 30 min, 16.1% of the fish in Test 3 exited the separator\ncompared to 81 and 74% for fish in Tests 1 and 2, respectively. Similar\ndifferences were seen in exit times during the next 30 min, with only 0.2% of\nthe fish in Tests 3 exiting in this test compared to 8.9 and 10.9% in Tests 1\nand 2, respectively. Within the first 24 h in Test 1 and 2, 99.8 and 100%,","54\nFirst 24 hour passage\n18\n16\n14\n12\n10\n8\n6\n4\n2\n0\n1\n2\n3\n4\n5\n6\n7\n8\n9\n10\n11\n12\n13\n14\n15\n16\n17\n18\n19\n20\n21\n22\n23\n24\nTime (hours)\nSubsequent passage\n16\n14\n12\n10\n8\n6\n4\n2\n0\n23\n25\n27\n1\n3\n5\n7\n9\n11\n13\n15\n17\n19\n21\n29\n31\n33\n35\n37\n39\n41\n43\n45\n47\n49\n51\n53\nTime (days)\nFigure 13. Percentage of PIT tagged fall chinook salmon detected while\nexiting the McNary Dam wet separator in the first 24 h and\nsubsequent days.","55\nrespectively, exited the wet separator, whereas in Test 3 only 67.3% exited in\nthe same period. No definitive explanation can be given for the long\nresidence time in the wet separator.\nStudy 3: Comparison of the PIT Tag to\nTraditional Tagging and Marking Methods\nIntroduction\nBranding and coded wire tags (CWT) have traditionally been used as means\nof identifying groups of fish on the Columbia River. Often fish must be\nrandomly collected at dams during periods of elevated water temperatures and\nthen branded and/or tagged. Although marking fish during these conditions is\nstressful to salmonids and normally should be avoided, situations often\nnecessitate such an approach. The objective of Study 3 was to compare the\nsurvival of fish injected with PIT tags to survival of fish tagged and marked\nby traditional methods. If no adverse effects to marking or tagging were seen\nunder these harsh field conditions, it is unlikely that severe problems would\nresult under more favorable conditions.\nMethods and Materials\nThe comparative study between traditional methods of marking and tagging\nand marking with the PIT tag was conducted at McNary Dam. Outmigrating fall\nchinook salmon collected from the juvenile collection and inspection facility\nwere used in the study. The fish ranged in fork length from 104 to 181 mm.\nThe study was conducted from 21 May to 9 June 1985.\nThe survival of PIT tagged fish was compared to that of control fish\n(handled, but not tagged or marked), CWT, CWT and branded, and branded fish.\nTraditional tagging and branding techniques were used in the study. All","56\ntreatments were combined and held as four replicate groups since each\ntreatment could be recognized by its identifying tag or mark (Table 10). .\nTwenty-five fish per treatment for a total of 125 fish per group were used in\nthe study. The fish were held for 14 days in four knotless nylon nets\nsuspended within a raceway receiving a continuous supply of untreated ambient\nriver water. The fish were examined daily for mortality. The data were\nanalyzed for differences using the G2 statistic at the P=0.05 level (Sokal and\nRohlf 1981).\nResults and Discussion\nNo statistical difference (G2=6.14 df=4, probability 0.19) between the\nsurvival of fish injected with the PIT tag and other treatment groups was\nshown at the end of 14 days of holding (Table 10). During the first 7 days of\nholding, only one control and one PIT tag fish died out of the 500 fish in the\nstudy. A total of 4 control, 13 PIT tagged, 6 branded, 8 CWT, and 7 CWT plus\nbranded fish died during the 14 days of holding. At the termination of the\nstudy, two control and two CWT fish were heavily infected with a fungus and\nwould probably not have survived an additional 1 to 2 days. The condition of\nall fish in the test groups was rapidly deteriorating at the end of the\n14 days of holding.\nAll dead fish were usually examined for cause of death. The fish\nexamined showed descaling and fungus infection in the caudal area. No signs\nof disease or fungus were seen on live or dead fish in the vicinity of the\nwound made by the injection needle. All PIT tagged fish showed complete\nclosure of the injection wound.\nThe holding of migrant fall chinook salmon captured at a collection\nfacility during the late part of the run and during a period of elevated water","57\nTable 10. - Summary of survival data comparing PIT tagged fish and\ntraditionally marked and or tagged fish after 14 days of holding.\nStarting\nDead\nEnding\nReplicate\n(n)\n(n)\n(n)\nTreatment\nControl\nI\n25\n0\n25\nPIT tag\n25\n5\n20\nBrand\n25\n2\n23\nCWT\n25\n2\n23\nCWT + brand\n25\n1\n24\nII\nControl\n25\n2\n23\nPIT tag\n25\n2\n23\nBrand\n25\n0\n25\nCWT\n25\n3\n22\nCWT + brand\n25\n3\n22\nIII\nControl\n25\n0\n25\nPIT tag\n25\n5\n20\n23\nBrand\n25\n2\nCWT\n25\n0\n25\nCWT + brand\n25\n0\n25\n2\n23\nIV\nControl\n25\nPIT tag\n25\n1\n24\nBrand\n25\n2\n23\nCWT\n25\n3\n22\nCWT + brand\n25\n3\n22","58\ntemperature is a stressful situation. It is believed, however, since no\nadverse effect of the PIT tag to survival was seen under these conditions,\nthat under more favorable conditions of capture, tagging, and holding, the PIT\ntag would not create any severe problems to migrant fall chinook salmon.\nStudy 4: McNary Reservoir Release\nIntroduction\nThe 1985 workplan did not include a reservoir release study, however,\nbased on the encouraging results of our planned 1985 field tests, we felt that\na reservoir release would provide valuable information for future planning\npurposes. A test plan was prepared and approved by BPA and the Columbia River\nFish\nPassage Committee. The objective of the study was to compare the\ncollection ratio of freeze branded fish to PIT tagged fish at the McNary Dam\njuvenile fish collection facility.\nMethods and Materials\nTesting was conducted from 7 August to 26 September 1985 at McNary Dam.\nA total of 4,400 juvenile outmigrant fall chinook salmon ranging in fork\nlength from 90 to 172 mm were marked and tagged over a 5-day period. Each day\na replicate consisting of 880 fall chinook salmon was randomly sampled from\nthe juvenile collection facility. No weak, highly descaled, or previously\nmarked fish or species other than fall chinook salmon were used in the\nstudy. of the 880 fish, 80 fish were randomly subsampled, injected with PIT\ntags, and measured. The remaining 800 fish were marked with a freeze brand\n(Park and Ebel 1974), and the upper caudal fin was clipped 2 / All fish were\ntransferred via flowing water to a 1,800-liter transport tank located on a\n2\nFreeze brands are difficult to read until about 4 days after marking, thus\na upper caudal clip is generally used by researchers as a flag whenever brands\nare expected to be read prior to 4 days.","59\ntruck. Brands were changed for each replicate (daily), and each PIT tagged\nfish had an individual code. Both PIT tagged and branded fish were held\ntogether in the truck transport tank for 24 h with flow through water prior to\nbeing transported to McNary Yacht Harbor at Hat Rock, Oregon, 11 km upstream\nfrom McNary Dam. The fish were transferred from the truck via gravity flow\nthrough a hose to a barge containing a transport tank receiving a continuous\nsupply of river water. The fish were then barged to the main river channel\nand released. Prior to release, all dead fish were collected for tag and mark\nobservation comparisons.\nPIT tag detection was performed by two automatic monitoring systems\nlocated on the wet separator at McNary Dam (see Part II, Study 1 for a\ndescription). The tag monitor systems required no handling of fish and\nautomatically stored tag codes and time of tagged-fish passage through the\ndetectors on computer files and a printer. The monitor systems were\npositioned to interrogate 100% of the fish passing through the juvenile\ncollection facility (Fig. 10).\nBranded fish were monitored by NMFS personnel at the juvenile salmon\ncollection and inspection facility at McNary Dam. A subsample of the fish\nexiting the wet separator was diverted to an inspection room; the subsample\ndiversion gates were located downstream from the PIT tag monitors (Fig. 10).\nThe gates were operated by a timer system which allowed sampling for 1.4 min,\n3 times per hour or 7% of the time fish passed out of the separator. The\nsubsampled fish were dipnetted; anesthetized; and inspected for fin clips,\ndescaling, injuries, and brands. The subsampled fish were then diverted to a\nraceway for transport downstream. The study was terminated when\nthe\ncollection system shut down for the season on 26 September 1985.","60\nResults and Discussion\nResults of the reservoir release comparative study are summarized in\nTable 11. No statistical difference was observed (P<0.001) between the\nrecovery of branded and PIT tagged fish. The total number of PIT tagged fall\nchinook salmon detected exiting from the collection facility was 64 (16%).\nThis represented 100% of the PIT tagged fish that were guided and passed\nthrough the collection facility at McNary Dam. The 758 branded fish (19%) is\nan estimate. The estimate is based upon expanding the actual number of fish\nobserved in the subsample (53) by a factor of 14.3 to adjust for the\nsubsampling rate.\nIn all, 13,239 fish were handled for branding and brand sampling to\nobtain the 53 fish in the subsample. To obtain statistically equal data, only\n400 fish were handled during PIT tag marking, and an estimated 138,926 fish\nwere passively monitored. Therefore, 97% more fish were handled to obtain\nbrand information in comparison to PIT tag data. This handling difference\nequates to a ratio of 33:1. In addition, 99% of the fish sampled for the\nbrand evaluation during this testing period were non-branded and were\nunnecessarily stressed.\nPost branding mortality (24-h) was slightly higher among branded fish\nthan the PIT tagged fish,--2 vs 1.5%. The water temperature at the time of\ntagging ranged between 20° and 21°C. The branded fish, as noted, received a\nsmall caudal clip as a marker. The combination of clipping the caudal fin and\nhigh water temperature may explain the mortalities that occurred prior to\nrelease of the fish. Upon recapture, several of the branded fish showed\ndeterioration of the caudal fin in the clipped area. We do not believe this\nfactor biased the data, however in future studies, we will avoid using any fin\nclip under adverse environmental conditions.","deviation\nStandard\nThe expanded value is based upon adjusting the actual observed number of fish in the subsample\n(%)\n+9\n+4\n/ No expansion factor is required since the number of fish observed represents 100% of the PIT\nobserved\nPercent\n19\n16\n11. -Recovery of branded and PIT tagged fall chinook salmon at McNary Dam.\nnumber fish\nb/\nobserved\nExpanded\n64 c/\n758\nnumber fish\nobserved\nActual\n53\n64\ntagged fish passing through the collection facility.\nhandled\nTotal\n13,239\n400\nfish\n14.3 to adjust for the subsampling rate.\na/ All data are for combined replicates.\nPre-release\nmortality\n2.3\n1.5\n(%)\n.\nnumber of\nTotal\nfish\n4,000\n400\nTreatment a/\nPIT tagged\nTable\nBrand\nby\nb/\nC","62\nThe initial comparison between the PIT tag and brand showed very\nencouraging results, with the PIT tag being considered a more statistically\nreliable marking method than marking with brands. Also, significantly fewer\nfish were stressed during the marking and sampling procedures with the PIT\ntag.\nWe recommend that further testing be conducted, as outlined in our 1986\nworkplan, using: (1) releases of steelhead, spring chinook salmon, and fall\nchinook salmon; (2) releases made at both inriver sites as well as from\nhatcheries; and (3) monitoring conducted at both Lower Granite and McNary\nDams. If results for 1986 are as conclusive as those we have seen in 1985, we\ncould recommend the use of the PIT tag as a tool for obtaining data to address\nsome of the problems on the Columbia River system in 1987.\nStudy 5: Monitoring PIT Tags in Adult Fish\nIntroduction\nThe PIT tag has significant potential as a tool to identify adult fish\nreturning to a river system. The tag can either be: (1) placed in smolts\nresulting in data being recovered during their outmigration at dams equipped\nwith automatic tag monitors and again, when as adults, they pass monitors on\ntheir upstream spawning migration or (2) placed in adults at some point on\ntheir spawning migration, with data subsequently recovered as in (1) above.\nThe former use may replace current CWT or freeze branding techniques. The\nlatter use would complement radio-tracking and CWT/freeze branding studies\nwhere research is needed on adult losses, migration delays, stock\nidentifications, and fall-back problems at dams or other migratory obstacles.\nIf the PIT tag is to have broad application for research, detection and\nautomatic data recording must be assured under a variety of field","63\nconditions. Therefore, our objectives were to: (1) evaluate the feasibility\nof monitoring PIT-tagged adult salmonids in a variety of situations applicable\nto Columbia River dams and (2) assess the accuracy and reliability of the PIT\ntag detector system when used with adult salmonids.\nThe 1985 PIT tag studies expanded the 1984 research by: (1) conducting\nthe research at an existing CWT trapping station instead of a simulated site,\n(2) modifying the detection system to provide more power and thus increasing\ntag reading efficiency, (3) improving the PIT tag quality, (4) increasing\ndetection by using a tandem detection system (multiple loops), and (5) adding\nadditional testing on the use of a PIT jaw tag.\nMethods and Materials\nSince this phase of testing was to be under actual field conditions, an\nexisting adult trap was necessary for a testing site. The interim fish trap\nlocated at the north shore fish ladder at Bonneville Dam was chosen due to its\nproximity to the newly completed fish-collecting facility and because this\nexisting trap could be used without interfering with normal fish passage\n(Fig. 14) 3/ Two modifications to the interim trap were necessary: (1) a\nscreen was installed in the approach channel from the fish ladder, providing a\nclosed system and (2) a 2.7-m long section of the flume was removed\nimmediately below a magnetic CWT detector located at the exit of a Denil fish\nladder. This flume section was replaced with two PIT tag detectors joined end\nto end (Fig. 14). Each detector consisted of a 1.2-m long section of 30-cm\n3/\nThe interim fish trap was constructed upon the completion of the\nBonneville Second Powerhouse in 1981 to provide a north shore adult fish trap\nduring the interim time before the completion of the north shore fish\ncollection facility.","64\nComputer\nController\nPrinter\nPower\nsupplies\nEntrance\nEnclosure\nchannel\nscreen\nHolding\npool\nPower\ncables\nExit\nchannel\nDenil\nfish\nData\nladder\ncables\nFish ladder\nFalsewein\nCWT\ndetector\nTandem PIT tag\ndetectors with\nexcitors\nTrap\nFigure 14. -- -Bonneville Dam interim fish trap and testing facility, 1985.","65\ndiameter PVC pipe containing two detector loops shielded with 4.8-mm thick\naluminum. A dual excitor was located inside the shielded box, and the power\nsupply, controller, computer, and printer were located in a mobile office\nstationed 100 feet away. The PIT tags used for this test were improved by the\nmanufacturer to provide more range than those used in the 1984 study.\nTesting was conducted from 11 to 19 July 1985, using adult steelhead\nranging in fork length from 51 to 82 cm. Steelhead entering the new trapping\nfacility were diverted directly into an anesthetic tank containing 40 ppm\nMS-222. The anesthetized fish from all 10 replicates (10 fish per replicate)\nwere then internally tagged with PIT tags (Prentice et al. 1985). For\nReplicates 1 and 3, the fish were also tagged with PIT jaw tags (Prentice et\nal. 1985). All fish were measured and placed into a 568-liter transport\ncontainer. After recovering from the anesthetic, the fish were transported to\nthe interim trap and released (water-to-water) into the holding area. The\ntime of release, length of fish, and PIT tag number were entered on the\ncomputer to create a release file. The holding pool had only one exit, the\n6.7-m long Denil fish ladder used as an approach to the magnetic CWT detector,\nthe PIT tag detector, and the holding trap. Codes from the PIT tags were read\nautomatically as the fish passed through the tunnel at flow velocities up to\n0.3 m/second. These data along with the passage time were simultaneously\nplaced on hard copy and floppy disk for storage.\nResults and Discussion\nResults of tests conducted under actual field conditions with the\nautomatic detection system for PIT-tagged adult salmonids are summarized in\nTable 12. Detection efficiency ranged from 90 to 100%, with an average\ndetection of 98%. These results should be representative of fish tagged\ninternally as juveniles and detected upon returning as adults.","66\nTable 12. -- Detection of PIT tags placed in adult steelhead at working coded wire\ntrapping facility on Bonneville Dam, July 1985.\nMean\nMean\npassage\nRelease\nNo. fish\nNo. fish\nlength\ntime\nDetection\nReplicate\ndate/time\nreleased\ndetected\n(mm)\n(h)\n(%)\n1,\n11 Jul - 1100\n10\n10\n625\n9.75\n100\n2\n12 Jul - 0900\n10\n10\n645\n18.32\n100\n3\nb\n13 Jul - 0925\n10\n10\n698\n8.56\n90\n4\n13 Jul - 1358\n10\n9\n636\n2.39\n90\n5\n14 Jul - 1009\n10\n10\n656\n12.28\n100\n6\n14 Jul - 1104\n10\n10\n672\n4.96\n100\n7\n15 Jul - 0840\n10\n10\n627\n3.12\n100\n8\n16 Jul - 0924\n10\n10\n627\n8.16\n100\n9\n16 Jul - 1434\n10\n10\n637\n16.60\n100\n10\n17 Jul - 0842\n10\n10\n613\n4.70\n100\nTotals\n100\n98\nAve.\n644\n8.88\n98\na\nReplicates 1 and 3 were double tagged with PIT internal tag and PIT jaw tag.\nIn\nboth cases, jaw tag data are identical to that shown for internal.\nb\nAll internal PIT tags were detected, however, for replicate three, one PIT jaw\ntag was not detected.","67\nIn some instances, the PIT tag may be used to obtain adult information\nonly. In this case, the fish could be externally tagged. In the two\nreplicates where the fish were double tagged with both an internal PIT tag and\na PIT jaw tag, both methods of tagging performed equally, with a mean\ndetection of 95% and each non-detections occurring on separate fish.\nOne of the primary goals of any research or management activity, where\nliving organisms will be returned to the environment, is to reduce handling\nstress. After testing the PIT tag on adult salmonids, we believe this\nobjective was met. In fact, the primary advantage of this system is the\nability to recover data (read tags) from moving fish, thus totally eliminating\nadditional handling stress to that fish and other fish which would be trapped\nin the sampling process. Furthermore, the 98% detection rate achieved during\nthe test of the adult PIT tag system exceeded the design criteria of 95%\ndetection. For these reasons, we feel that this system could be used at\nexisting CWT trapping facilities to increase data collection as well as\nenhance the quality of the data and fish collected.\nThe performance of the PIT jaw tag was equal to the internal PIT tag,\nsuggesting that the PIT jaw tag could be a viable method of tagging adult\nsalmonids when returns from non-automated sources are necessary (i.e.,\ncommercial or sport fisheries).\nConclusions and Recommendations\n1. The PIT tag monitors can be installed at dams and give consistent and\nreliable results. We recommend that a minimum of two independent double loop\nassemblies be used wherever PIT tags are to be remotely detected, and one\ncontroller, exiter, and power supply be maintained in a convenient location to\nserve as an emergency replacement unit in case of a component failure.","68\n2. The PIT tag can be read efficiently and accurately in juvenile fall\nand spring chinook salmon that are moving up to 0.3m/sec as they pass\nvolitionally through a PIT tag detection system.\nThe PIT tag does not significantly impair survival of juvenile\n3.\nmigrant fall chinook salmon compared to the survival of traditionally tagged\nand marked fish.\n4. Based on branded and PIT tagged juvenile fall chinook salmon released\nin McNary reservoir being collected at the McNary juvenile fish collection\nfacility in the same ratio, PIT tagged fish behave and survive in a manner\nsimilar to fish traditionally marked.\n5. The use of the PIT tag, in many types of juvenile salmon studies\ncould reduce the number of test fish required by up to 90% and reduce stress\nto the fish by only requiring the fish to be handled at the time of tagging.\nAll data collection can be automatic without handling the fish or restraining\ntheir passage.\n6. Adult steelhead migrants can be successfully PIT tagged and\nautomatically interrogated as they volitionally pass through a PIT tag\ndetection system installed on a Denil fish ladder.\n7. With properly installed tag detection equipment, PIT tag reading\nefficiency for adult migrant steelhead can be expected to be greater than 95%\nwith 100% accuracy.\n8. The PIT tag detection system for adult salmonids can be used at\nexisting coded wire tag trapping facilities with minimal revision.\n9. The use of the PIT tag with adult migrant fish can increase data\ncollection and enhance the quality of the data collected.","69\nPART III: SYSTEMS DEVELOPMENT\nStudy 1: PIT Tag Injection Devices\nPIT tags are presently injected into fish with a modified hypodermic\nsyringe and needle. Each injector is loaded by hand, requiring a tag to be\nmanually inserted into the needle. This procedure has been satisfactory for\ntest purposes requiring small numbers of fish, however, as greater numbers of\nfish are tagged, a more efficient means of placing the tag in the needle is\nrequired. Complicating the design of a tagging system is a self-imposed\nrequirement that both the needle and tag be disinfected prior to use.\nPresently, several designs for a tagging system that meet our requirements are\nunder evaluation. The final design and implementation of an automatic tag\ninjection system must wait until the tag manufacturer decides upon a packaging\nsystem (tags in a strip, cartridge, etc).\nStudy 2: Quality Control Monitor For Tagging\nAt the time a fish is PIT tagged, every assurance must be made that the\ntag injected into the fish is functional and can be interrogated and the data\nrecorded. Furthermore, since each fish can be identified by a unique\nidentification number, individual information such as length and/or weight can\nbe recorded and associated with the identification number at the time of\ntagging. Figure 15 shows a quality controlled tagging system to be evaluated\nin 1986. The system will consist of two similar tagging stations. Each\nstation will have a 150-cm diameter tag detection loop, a tag monitor, an\nelectronic measuring board and balance, and a controlled fish release area.\nThe components of the two stations are connected to a multiplexer, computer,\nand printer. The tagging procedure at a station would require a number of\nsteps. A fish would be removed from an anesthetic tank and injected with a","Anesthetic tank\nTAGGING STATION II (similar to Station I)\nPIT tag data scanner and tag reading coil\nElectronic measuring board\nFish release monitor\nElectronic balance\nFigure 15.--Conceptual drawing of a quality control system for tagging.\nPIT tag\nscanner\ndata\nPrinter\nPIT tag\nreading\ncoil\nTAGGING STATION I\nMultiplexer\nComputer\nElectronic\nmeasuring\nboard\nModum\nElectronic\nbalance\nControl lights\nFish with\nFish release monitor\nno tags\nTagged\nfish","71\nPIT tag. While holding the fish in hand, the fish would be placed through the\ntag detection loop. A message would appear on the tag monitor's screen if the\ntag was successfully read and entered into the computer. The operator would\nthen place the fish on the electronic measuring board and touch a stylus to\nthe fork of the tail to obtain fork length. The fish would then be weighed on\nan electronic balance. The data from the measuring board and balance would be\nentered into the computer automatically. If all data\nentered\nwere\nsuccessfully, a green light would show and a rubber gate would open allowing\nthe operator to release the fish. All data would be automatically entered on\ncomputer files and a hard copy made. If for some reason not all the data\nentered the computer, a red light would show on the tagging console and a\nrubber gate over a repeat exit would open. The two stations could be operated\nsimutaneously since the multiplexer acts as a controller and a buffer for the\nsystem.\nTo date, not all the components have been linked together and fully\ntested. However, we have individually tested the tag detection loop, tag\nmonitor, electronic balance and measuring board, multiplexer, computer, and\nprinter. Actual field testing of the system awaits the 1986 field season.\nThe design of this system has been reviewed by U.S. Fish and Wildlife Service\npersonnel associated with fish tagging.\nStudy 3: Hatchery Release Monitor\nMortality and tag loss may occur between the time fish are tagged and\nreleased at a hatchery. Therefore, it is essential to know the actual\nidentification of each fish at the time of release so that tags that are no\nlonger a part of the study can be eliminated from the data base.","72\nMonitoring tagged fish at time of release from a hatchery is challenging,\nsince the highest concentration of tagged to non-tagged fish will occur within\na hatchery rearing system when all the fish will be released within a short\ntime. Under these conditions, precautions are needed to reduce the likelihood\nof two tagged fish entering a monitoring loop simultaneously to prevent\nreading error. Furthermore, the monitoring system must be designed to rapidly\nmonitor fish without stress.\nDesign work was completed on a hatchery monitor under the 1985-86\nworkplan (Fig. 16). The monitor consists of four pipes measuring 10.2 cm in\ndiameter by 61.0 cm long. Each pipe is equipped with two PIT tag monitoring\nloops connected to tag monitoring equipment. All of the monitors are\nconnected to a computer and printer. As each PIT-tagged fish passes through a\nmonitor, its number will be recorded automatically on a computer file and be\nprinted. After the release, the release file will be compared to the file\ncreated at the time of tagging and missing fish will be noted. The release\nmonitor will be tested at Dworshak National Fish Hatchery in March 1986.\nStudy 4: Design and Placement of Future Monitoring Systems\nThe results obtained during the 1985 field season at McNary Dam provided\nvaluable insight into the future design and placement of juvenile monitoring\nequipment at collector dams. Initial monitor design and placement made it\ndifficult to clean the orifices on the wet separator and, thus, could\npotentially increase debris problems within the fish collection system.\nSuggested modifications include narrowing the monitor entrance and adding a\ndewatering section. In addition, it was determined that a series of two\nmonitors (with two detector coils each) per flume should provide optimal PIT\ntag reading efficiency. Based upon this experience, an improved new PIT tag","(4 each)\n(2 each)\n(1 each)\n(1 each)\n(2 each)\nTag monitoring equipment\n(4 each)\nPower supplies\nTag excitors\nMultiplexer\nControllers\nComputer\nPrinter\ndiameter PVC\n10.2 cm\nRELEASE GATE WITH PIT TAG MONITORS\nFigure 16. .--Hatchery PIT tag release monitor system.\n61.0 cm\n45.7 cm\nRaceway release gate\nand PIT tag monitor\n73.7 cm\nHATCHERY PIT TAG RELEASE MONITOR\nsystem\nCrowder\nscreen","74\nSubsemple\nPit tag monitors\nTo raceway or river\nE and F\nPit tag monitors\nC and D\nWet separator\nFlow\nFlow\nDewater\nsections\nPit tag monitors A and B\nPorosity control\n.\nFigure 17. . -- Proposed location of juvenile PIT tag monitors at McNary Dam","PIT tag monitors C and D\nRaceways\nFigure 18. -Location of juvenile PIT tag monitors at Lower Granite Dam.\nInclined\nseparator\nUpwell\nscreen\nWet\nSample\ntank\nmonitors\nA and B\nPIT tag\nPIT tag monitors E and F\nRaceways","76\nmonitoring system will be installed at McNary Dam for the 1986 field season\n(Fig. 17). This unit should improve operational efficiency by lowering debris\nproblems in the system. A similar system will be installed at Lower Granite\nDam (Fig. 18). The U.S. Army Corps of Engineers and the Fish Passage\nCommittee have approved the design and installation of the new monitors at\nboth dams.\nA tag monitor system has also been designed for Little Goose Dam. A\nseries of controlled tests incorporating both the PIT tag monitors and fish\ncounters working in close proximity to one another must be completed before\nthe design is available for review.\nConclusions and Recommendations\n1. As soon as the PIT-tag manufacturer decides on final tag design and\npackaging, we recommend a semi-automatic or automatic tag injection system be\ndeveloped to reduce the time required to tag a population of fish.\n2. We recommend that a PIT tag tagging station and quality control\nsystem be designed and fully tested in 1986. Such a system should be designed\non the same principle as that used for CWT.\n3. We recommend that the system to monitor PIT-tagged fish leaving\nhatchery raceways be evaluated in 1986.\n4. We recommend that an improved PIT tag detection system be installed\nat McNary Dam to overcome the potential debris problem that existed at the wet\nseparator in 1985.\n5. PIT tag detection systems can be installed at Lower Granite Dam\nwithout major modifications to the existing system; we recommend that such a\nsystem be installed in 1986.","77\nACKNOWLEDGMENTS\nSupport for this research came from the region's electrical rate payers\nthrough the Bonneville Power Administration.\nSpecial thanks is given to David Cross for his technical support. We\nalso thank Lee Ferguson, Carol Ranck, and their crews for the support given\nduring the project. Thanks is also given to Brad Ebby (COE) and his crew, for\nwithout their assistance the field studies could not have proceeded.","78\nLITERATURE CITED\nBeamish, F. W. H.\n1978. Swimming capacity. In: W. S. Hoar and D. J. Randall, editors.\nFish Physiology 7. Academic Press Inc., New York, N.Y., p. 135.\nBell, W. H. and L. D. Terhune.\n1970. Water tunnel design for fisheries research. Fish. Res. Board\nCan., Tech. Rep. 195:1-69.\nBlack, E. C.\n1958. Hyperactivity as a lethal factor in fish. J. Fish. Res. Board\nCan., 15:573-586.\nFlagg, T. A.\n1981. Swimming stamina and survival related to swimming fatigue in\nresponse to direct seawater entry during the parr-smolt transformation\nof coho salmon (Oncorhynchus kisutch). M. Sci. Thesis., University of\nWashington. 58 pp.\nFlagg, T. A. and L. S. Smith.\n1982. Changes in swimming behavior and stamina during smolting of coho\nsalmon. Salmon and trout migratory behavior symposium, E. L. Brannon\nand E. 0. Salo, editors. June 1981. University of Washington press.\np. 191-195.\nFlagg, T. A., E. F. Prentice, and L. S. Smith.\n1983. Swimming stamina and survival following direct seawater entry\nduring parr-smolt transformation of coho salmon (Oncorhynchus\nkisutch). Aquaculture 32:383-396.\nLewis, A. E. and W. R. A. Muntz.\n1984. The effects of external ultrasonic tagging on the swimming\nperformance of rainbow trout, Salmo gairdneri Richardson. J. Fish.\nBiol. 25:577-585.\nMcCleave, J. D. and K. A. Stred.\n1975. Effect of dummy transmitters on stamina of Atlantic salmon (Salmo\nsalar) smolts. J. Fish. Res. Board Can. 32:559-563.\nMonan, Gerald E.\n1985. Advances in tagging and tracking hat chery salmonids; coded wire\ntags, multiple-coded and miniature radio tags, and the passive\nintegrated transpounder tag. P. 33-37. U.S. Dep. Commer. , NOAA Tech.\nRep. NMFS 27.\nPark, Donn L. and Wesley J. Ebel.\n1974. Marking fishes and invertebrates. II. Brand size and configuration\nin relation to long-term retention on steelhead trout and chinook\nsalmon. Mar. Fish. Rev. , Vol. 36 No. 7. e pp. 1-6.","79\nPark, D L. G. Matthews, J. R. Smith, T. E. Ruehle, J. R. Harmon, and\nS. Achord.\n1984. Evaluation of transportation of juvenile salmonids and related\nresearch on the Columbia and Snake River, 1983. U.S. Dep. of Commer.,\nNatl. Oceanic Atmos. Admin., Natl. Mar. Fish. Serv., Northwest and\nAlaska Fish. Cent., Seattle, WA. 58 p. plus Appendix (Report to U.S.\nArmy Corps of Engineers, Contract DACW68-78-C-0051).\nPrentice, E. P., D. L. Park, and C. W. Sims.\n1984. A study to determine the biological feasibility of a new fish\ntagging system. U.S. Dep. of Commer., Natl. Oceanic and Atmos. Admin.,\nNatl. Marine Fish. Serv., Northwest and Alaska Fish. Cent., Seattle,\nWA.\n38 p. (Report to Bonneville Power Administration, Contract\nDE-A179-83BP11982, Project 83-19).\nPrentice, Earl F., Carl W. Sims, and Donn L. Park.\n1985. A study to determine the biological feasibility of a new fish\ntagging system. U.S. Dep. of Commer. Natl. Oceanic Atmos. Admin.,\nNatl. Mar. Fish. Serv., Northwest and Alaska Fish. Cent., Seattle, WA.\n36 P. plus Appendixes (Report to Bonneville Power Administration,\nContract DE-A179-83BP11982, Project 83-19).\nSmith, L. S. and T. W. Newcomb.\n1970. A modified version of the Blaska respiromenter and exercise chamber\nfor large fish. J. Fish. Res. Board Can. 27:1331-1336.\nSokal, R. R. and F. J. Rohlf.\n1981. Biometry. W. H. Freeman and Co., San Francisco, California.\nStevens, E. D.\n1979. The effects of temperature on tail-beat frequency of fish swimming\nat constant velocity. Can. J. Zool. 57:1628-1635.\nWedemeyer, G. A., N. C. Nelson, and W. T. Yasutake.\n1979. \"Potentials and Limits for the Use of Ozone as a Fish Disease\nControl Agent.\" In Ozone: Science and Engineering Vol. 1, pp. 295-318,\nPergamon Press Ltd.","80\nAPPENDIX A\nFinal Report\nPRELIMINARY INVESTIGATION OF THE\nINACTIVATION OF AEROMONAS SALMONICIDA,\nA FISH PATHOGEN\nR. A. Elston\nBattelle/Marine Research Laboratory\nSequim, Washington\nNovember 1986\nPrepared for the\nNational Oceanic and Atmospheric Administration\nunder a Related Services Agreement\nwith the U.S. Department of Energy\nContract DE-AC06-76RLO 1830\nPacific Northwest Laboratory\nRichland, Washington 99352","81\nINTRODUCTION\nIn order to provide some preliminary indication of the ability of a\nsterilizing agent to inactivate a common fish pathogen, initial studies\ndescribed here were conducted. These studies were in support of a tagging\nprogram in which electronic fish tags (PIT tags) were used to mark salmonid\nfishes from a variety of Columbia River Basin stocks. The studies resulted\nfrom a concern that the repeated use of fish tagging injectors could serve\nas a vector for fish pathogens. It was realized that an exhaustive\ninvestigation of sterilizing agents on various pathogens of differing degrees\nof sensitivity to the sterilants was beyond the scope of the effort here.\nThus the results provided here utilizing a relatively sensitive indicator\nand easy to detect bacterium provide a guideline for the minimal conditions\nwhich should be used in the maintenance of sterilizing solutions. Further\nextensive work with a variety of pathogens such as Renibacterium salmoninarum\nand infectious pancreatic necrosis virus would be required to definitively\nestablish the efficacy of the concentrations of ethanol used here or other\nsterilants for their inactivation.","82\nMETHODS\nInjectors used for intraperitoneal injection of fish were obtained from\nthe National Marine Fisheries Service (NMFS) as well as the tagging devices\n(PIT tags). . Injector tips were dipped in sterile petroleum jelly prior to\nthe test in order to simulate conditions of actual use in the field. An\nisolate of Aeromonas salmonicida was also obtained from the NMFS.\nBacterial suspensions of A. salmonicida were prepared by inoculating\ntryptic soy broth with a loopful of the isolate. Density of 18 to 24 hour\ncultures and an approximation of cell concentration was made by measuring\noptical density at a wavelength of 620 nm. Sterilized tag injectors were\ndipped into the bacteriological broth (to as depth of about 1 cm) containing\nbetween 1 X 10 6 and 1 X 107 organisms per ml. The tags were expelled after\nthe devices were withdrawn from the broth and placed in the sterilizing\nsolutions for the appropriate test time. Untreated controls were given a\nsimilar immersion in the bacterial broth but were not subjected to the dip\nin the test sterilizing solutions.\nFollowing the sterilizing treatments, the injectors were swabbed with\nsterile cotton tip applicators which were then used to qualitatively\ninoculate tryptic soy agar plates. Plates were incubated for up to 3 days\nand examined for the presence or absence of bacteriological growth. A\nseries of four experiments was conducted to determine the minimal\nconcentration of ethanol which would completely sterilize all test injectors.","83\nRESULTS\nA summary of the test results is given in Table 1. Preliminary\nexperiments suggested that a concentration of as low as 30% ethanol would\ninactivate the bacterium. Further experimentation (Experiments 3 and 4,\nTable 1) with 30% and 50% ethanol indicated that the lower concentration\n(30%) was not effective in inactivation (only 2/10 test samples were\ninactivated) but that 50% ethanol was effective in inactivating 10/10 test\nsamples. The inactivation occurred after one minute of exposure to the\nethanol solution. The first experiments with small sample sizes had\nsuggested that exposure of the contaminated injectors to the sterilant\nresulted in sterilization within one minute although several injectors\nwere tested with a five minute treatment in the sterilant. The results\nthus indicate that for Aeromonas salmonicida or for microorganisms of similar\nsensitivity to ethanol that a one minute exposure of the PIT tag injecting\ndevices in 50% ethanol is sufficient to kill the bacteria.","84\nTABLE 1. Inactivation of Aeromonas salmonicida with an ethyl aclohol rinse.\nPROPORTION OF POSITIVE BACTERIOLOGICAL PLATES\nConcentration\nof Alcohol in\nExperiment\nSterilizing\nDuration of Treatment*\nNumber\nSolution\n1 Minute\n5 Minutes\n1\n90%\n0/2\n0/2\n50%\n0/2\n0/2\nUntreated Control\n2/2\n2/2\n2\n50%\n0/2\n0/2\n30%\n0/2\n0/2\n10%\n2/2\n2/2\nUntreated Control\n2/2\n2/2\n3\n30%\n8/10\n-\nUntreated Control\n10/10\n-\n4\n50%\n0/10\n-\nUntreated Control\n10/10\n-\n*Proportion of total plates with bacterial growth for each indicated\ntreatment.","85\nDISCUSSION\nIt must be noted that the results presented here cannot be applied to\nother microorganisms which may not have the same sensitivity to ethanol.\nFor example. the cell wall of the gram positive fish pathogen, Renibacterium\nsalmoninarum, could render it more resistant to the treatments which were\neffective for Aeromonas salmonicida. This possibility can only be verified\nby further testing.\nWedemeyer et al. 1979, found that A. salmonicida was more resistant to\nboth chlorine and ozone treatment for inactivation than was the etiologic\nagent of enteric redmouth disease (ERM), Yersinia ruckeri. A concentration\nof 0.05 mg\\L inactivated Y. ruckeri 30s while a concentration of 0.1 mg\\L\nfor 30s was required to inactivate A. salmonicida. The inactivation of\ninfectious hematopoietic necrosis virus (IHNV) in hard lake water required\nchlorine at 0.5 mg/L for 10 minutes or 1.0 mg/L for 30s. Under similar\nconditions, 0.7 mg/L chlorine destroyed infectious pancreatic necrosis\nvirus (IPNV) within 2 minutes. These values may provide some indication\nof the relative resistance of the microorganisms to inactivating agents\nbut can not be assumed to be directly proportional to the sensitivity of\nthe same microorganisms to ethanol since the mechanism of inactivation may\nbe different.\nOne important component of the approach to the control of diseases\nthrough the use of tagging equipment is to determine which diseases are\nknown or considered to be probable to exist in a given watershed. Obviously,\nif infectious agents which are potentially more resistant to a given method","86\nof inactivation are not present in a particular drainage, then these agents\nwould not be considered in the inactivation of fish handling equipment.","87\nREFERENCES\nWedemeyer, G. A. , N. C. Nelson, and W. T. Yasutake. 1979. \"Potentials\nand Limits for the Use of Ozone as a Fish Disease Control Agent.\" = In Ozone:\nScience and Engineering Vol. 1, pp. 295-318. Pergamon Press Ltd.","88\nAPPENDIX B\nBudget Information","89\nBUDGET INFORMATION\nA. Summary of expenditures\nPersonnel Services and Benefits\n87.9K\nTravel & Transportation of Persons\n9.3K\nTransportation of Things\n5.5K\nRent, Communications & Utilities\n0\nPrinting & Reproduction\n0.1K\nContract & Other Services\n7.5K\nSupplies & Materials\n280.2K\n276.3K\nEquipment\n0\nGrants\nSupport Cost (Including DOC ovhd.)\n33.6K\nTOTAL\n693.7K\nB. Major items purchased\n1. PIT tags (50,000)--Contract 85-ABC-00182\n2. PIT tag monitoring systems for juvenile migrants at Lower Granite and\nMcNary Dams--Contract 50-ABNF-6-0048.","FIGURES\nFigure 1. -- -Compari of weight change of PIT tagged and control fish over\ntime.\nFigure 2.--Comparison of length change of PIT tagged and control fish over\ntime.\nFigure 3. -- Di agram of modified Blaska respirometer-stamina chamber, showing\nside and end views. For loading, the chamber is tilted, partially\nfilled with water, and end plate and vane are removed. Fish are\nplaced in the test compartments, vane and end plate are replaced,\nand chamber is filled with water and leveled. Water flow is\nproduced with motor driven propeller and varied via motor speed\ncontroller. Direction of water flow is toward propeller in inner\ntube, water is turned at the end plate, and returned through the\nspace between the inner and outer tubes (see arrows).\nFigure 4. -- -Mean swimming stamina (U-critical) of PIT tagged and control\nfingerling steelhead (6.5 g average) trout during Days 0-25\npost-tag. Brackets indicate + one standard error.\nFigure 5. .--Mean swimming stamina (U-critical) of PIT tagged and control\njuvenile steelhead (17.2 g average) trout during Days 0-25 post-\ntag. Brackets indicate + one standard error.\nFigure 6.--Mean stride efficiency of PIT tagged and control fingerling\nsteelhead (6.5 g average) trout during Days 0-25 post-tag.\nBrackets indicate + one standard error.\nFigure\n7.--Mean stride efficiency of PIT tagged and control juvenile steelhead\n(17.2 g average) trout during Days 0-25 post-tag. Brackets\nindicate + one standard error.\nFigure 8.--Mean opercular beat rate of PIT tagged and control fingerling\nsteelhead (6.5 g average) trout during Days 0-25 post-tag.\nBrackets indicate + one standard error.\nFigure .--Mean opercular beat rate of PIT tagged and control juvenile\nsteelhead (17.2 g average) trout during Days 0-25 post-tag.\nBrackets indicate + one standard error. Asterisk (*) indicates\nsignificant (tagged VS. control) difference (P<0.01).\nFigure 10.--Location -- of juvenile salmon PIT tag monitors at McNary Dam during\n1985.\nFigure 11.--Diagram of the juvenile salmon PIT tag monitoring system at McNary\nDam during 1985.","Figure 12. . .--Percentage of PIT tagged spring chinook salmon detected while\nexiting the McNary Dam wet separator.\nFigure 13. . -- Percentage of PIT tagged fall chinook salmon detected while\nexiting the McNary Dam wet separator in the first 24 h and\nsubsequent days.\nFigure 14. . -- Bonneville Dam interim fish trap and testing facility, 1985.\nFigure 15 . --Conceptual drawing of a quality control system for tagging.\nFigure 16. . -- --Hatchery PIT tag release monitor system.\nFigure 17 .--Proposed . location of juvenile PIT tag monitors at McNary Dam.\nFigure 18.--Location of juvenile PIT tag monitors at Lower Granite Dam."]}