Journal of
Applied Ichthyology
J. Appl. Ichthyol. 26 (2010), 612–613
 2010 Blackwell Verlag, Berlin
ISSN 0175–8659
Short Communication
Evaluation of PIT-tagging in an endangered Iberian cyprinid
By D. Pires, F. Ribeiro and M. F. Magalhães
Universidade de Lisboa, Faculdade de Cieˆncias, Lisboa, Portugal, Departamento Biologia Animal ⁄ Centro de Biologia Ambiental
Introduction
Tagging has been increasingly used in freshwater fisheries
science to gain information on fish growth, movement and
behaviour and for mark-recapture population assessments
(Lucas and Baras, 2000; Gibbons and Andrews, 2004).
However, effects of tagging on fish are still equivocal, with
studies reporting adverse effects on survival, growth and
behaviour (e.g. Armstrong and Rawlings, 1993; Knights and
Lasee, 1996; Jepsen et al., 2002) contrasted by those showing
no adverse effects (e.g. Connors et al., 2002; Acolas et al.,
2007; Newby et al., 2007). Therefore, thorough evaluations of
tagging reliability are needed for previously unstudied species
to assure fish welfare and unbiased data collection (Lucas and
Baras, 2000; Gibbons and Andrews, 2004). This is especially
critical in rare and endangered species, as adverse tagging
effects can strongly impact declining populations and result in
poor conservation prescriptions.
Passive Integrated Transponders (PITs) are among the best
candidates for uniquely marking small, endangered fish. PIT
tags are small, have millions of unique alphanumeric codes and
unlimited functional life, and allow repeated non-destructive
sampling (Gibbons and Andrews, 2004). Recent studies have
documented the suitability of PIT-tagging for small fish (e.g.
Skov et al., 2005; Ruetz et al., 2006; Acolas et al., 2007;
Knaepkens et al., 2007; Bolland et al., 2009), although evi-
dence for endangered cyprinids remains scarce (but see Ward
et al., 2008). This study evaluated the reliability of PIT-tagging
for the Mira chub Squalius torgalensis (Coelho, Bogutskaya,
Rodrigues and Collares-Pereira), a critically endangered cyp-
rinid endemic to the Iberian Peninsula. Specific objectives were
to analyse the retention of PIT tags and their short-term effects
on survival, growth, and swimming performance of the Mira
chub.
Materials and methods
Fish were captured in the wild (Torgal stream, SW Portugal)
by electrofishing and transported in aerated vats to the
experiment site (University of Lisbon). Fish injured by
sampling, in poor condition or with diseases were discarded.
Overall, 80 fish, 66–124 mm fork length, were stocked into
aerated aquaria (50 · 30 · 25 cm), with gravel bottoms and
filtered water. The length range used was that typically
encountered in the wild and which appeared acceptable for
PIT-tagging based on results in other small fish (Acolas et al.,
2007; Ward et al., 2008). Fish were acclimated to experimental
conditions for 7 days. Aquaria were under ambient tempera-
ture and photoperiod conditions in an isolated room. Fish
U.S. Copyright Clearance Centre Code Statement: 0175–8659/2010/2604–0612$15.00/0
Received: August 27, 2009
Accepted: November 22, 2009
doi: 10.1111/j.1439-0426.2010.01464.x
were fed a daily ration of commercial dry food, using
automatic feeders.
Three treatments were considered: control, sham-tagging,
and PIT-tagging. Treatments were tested on groups of four fish
per aquarium, in six aquaria (i.e. three treatments · six
replicates · four fish). Fish were randomly assigned to treat-
ments, and aquarium effects were assumed to be null. All fish
were anaesthetised using clove oil, as this has minimal
behavioural and physiological impacts (Anderson et al.,
1997). Initial fork length (mm) and total weight (0.1 g) were
recorded. PIT tags (Trovan ID-100A, 2.12 · 11.5 mm) were
inserted into the body cavity of fish through an incision made
just off the midventral line of the body, anterior to the vent,
using a sharpened needle (Ruetz et al., 2006). Sham-tagging
involved the surgical procedure but no transponder implant.
Incisions were not sutured but clinical grade cyanocaricle
adhesive (3M Vetbond) was applied to close the wound.
Tag-to-body weight ratio ranged between 0.4% and 2.9% in
air. After treatment, fish were allowed to recover until they
regained balance and swimming, and then returned to the
experimental aquaria.
Fish were held for observation for 30 days. Aquaria were
inspected for mortalities and tag retention throughout the
experiment. Fish were scanned for tag presence and function
using a handheld detector (Trovan LID 571, EID Aalten
B.V.). Lost tags on the bottom of the aquaria were manually
removed. Dead fish and lost tags were not replaced. At days 1,
3, 7, and 30 (D1, D3, D7 and D30), fish were video recorded
for 30 min, between 9.00 and 11.30 h. Swimming performance
was considered as the number of cells in a reference grid
(16 · 9 cm mesh) crossed by each fish per 30 s, in scans taken
at 5, 15, 25 min of record. Finally, all fish were measured and
weighted at D30.
Contingency analysis was used to compare cumulative
mortalities per treatment. One-way analysis of variance was
used to test for variation in length and weight among
treatments, and repeated anova was performed on swimming
counts to test for variation depending on treatment and time.
Prior to analysis data were log10-transformed to comply with
anova assumptions of normality and homecedasticity (Zar,
1999). Significance of statistical testing was assessed at
P < 0.05.
Results
Survival was not different among treatments (92–100%;
v2 = 0.000, P = 1.000), with only two PIT-tagged (65 and
76 mm) and one sham-tagged fish (85 mm) lost in D1, D6, and
PIT-tagging in an endangered cyprinid
Table 1 Comparison of traits of Mira chub Squalius torgalensis in control, sham-tagging and PIT-tagging treatments, at 0, 1, 3, 7, and 30 days
post-marking. Values are mean ± standard deviation
Fork length (mm)
Day post-marking D0 D30
Treatment Control 81.3 ± 12.8 81.5 ± 12.6
Sham-tagging 80.8 ± 13.5 80.8 ± 13.6
PIT-tagging 81.3 ± 13.9 81.3 ± 13.9
D15, respectively. Length and weight at D30 showed no
variation among treatments (F = 0.030, P = 0.968 and
F = 0.223, P = 0.801, respectively). Swimming performance
did not differ among treatments (F = 0.989, P = 0.409) or
over time (F = 2.528, P = 0.078), with tagged fish reducing
swimming in D1 and D3 only (Table 1). Tag retention was
80%, with losses occurring from D7 to D29 in fish 66–75 mm.
Conclusions
PIT tags showed little or no short-term adverse effects on Mira
chub survival, growth or swimming performance. PIT-tagging
did not markedly increase mortality or decrease chub growth,
consistent with evidence for other small fish (e.g. Ruetz et al.,
2006; Acolas et al., 2007; Ward et al., 2008; Bolland et al.,
2009). Likewise, PIT-tagging had minimal impact on swim-
ming performance, similar to what has been reported for
bullhead (Knaepkens et al., 2007) and juvenile rainbow trout
(Newby et al., 2007). Finally, tag retention was comparable to
that found in other experimental studies (Acolas et al., 2007;
Ward et al., 2008; Bolland et al., 2009).
These findings suggest that PIT tags were effective on Mira
chub, and should be considered when selecting a method for
uniquely marking other Iberian cyprinids. Specifically, PIT
tags appeared a promising alternative to Floy and Visible
Implant tags, traditionally used with Iberian cyprinids, which
are plagued by high tag-to-body size ratios and poor retention
(see Prenda and Granado-Lorencio, 1994; Aparicio and
Sostoa, 1999). Nevertheless, additional research on the long-
term effects and reliability of PIT tags under natural condi-
tions is needed prior to their broad-scale use in behaviour and
population assessments in small, endangered Iberian cyprinids.
Acknowledgements
This study was conducted under permits of the Portuguese
Forestry Authority (10 ⁄ 2008) and the Institute for Nature
Conservation and Biodiversity (316 ⁄ 2007). Funding was
provided by the Portuguese Science Foundation
(PPCDT ⁄ BIA-BDE ⁄ 56272 ⁄ 2004), which also supported D.
F. Pires (SFRH ⁄ BD ⁄ 21861 ⁄ 2005) and F. Ribeiro
(SFRH ⁄ BPD ⁄ 46761 ⁄ 2008). Thanks are due to Margarida
Corado for assistance in the experiment, and Dietrich Schnack
for helpful comments on a draft of the manuscript.
References
Acolas, M.L.; Roussel, J.M.; Lebel, J.M.; Baglinière, J.L., 2007:
Laboratory experiment on survival, growth and tag retention
613
Total weight (g) Swimming performance (Nr. of cells crossed per 30 s)
D0 D30 D1 D3 D7 D30
7.4 ± 4.3 7.2 ± 3.9 9.5 ± 5.6 7.8 ± 5.1 8.3 ± 6.0 8.5 ± 5.5
7.2 ± 4.7 6.9 ± 4.3 10.0 ± 9.6 8.9 ± 4.4 12.4 ± 5.9 5.2 ± 1.7
7.3 ± 4.6 6.7 ± 4.2 3.1 ± 1.2 4.2 ± 2.3 7.6 ± 2.9 5.5 ± 2.2
following PIT injection into the body cavity of juvenile brown
trout (Salmo trutta). Fish. Res. 86, 280–284.
Anderson, W.G.; Kinley, R.S.; Colavecchia, M., 1997: The use of clove
oil as an anesthetic for rainbow trout and its effects on swimming
performance. North Am. J. Fish. Manag. 17, 301–307.
Aparicio, E.; Sostoa, A., 1999: Field evaluation of visible implant tag
retention by Barbus haasi. J. Appl. Ichthyol. 15, 199–200.
Armstrong, J.D.; Rawlings, C.E., 1993: The effect of intragastric
transmitters on feeding behaviour of Atlantic salmon, Salmo
salar, parr during autumn. J. Fish Biol. 43, 646–648.
Bolland, J.D.; Cowx, I.G.; Lucas, M.C., 2009: Evaluation of VIE and
PIT tagging methods for juvenile cyprinid fishes. J. Appl.
Ichthyol. 25, 381–386.
Connors, K.B.; Scruton, D.; Brown, J.A.; McKinley, R.S., 2002: The
effects of surgically-implanted dummy radio transmitters on the
behaviour of wild Atlantic salmon smolts. Hydrobiologia 483,
231–237.
Gibbons, J.W.; Andrews, K.M., 2004: PIT tagging: simple technology
at its best. Bioscience 54, 447–454.
Jepsen, N.; Koed, A.; Thorstad, E.B.; Baras, E., 2002: Surgical
implantation of telemetry transmitters in fish: how much have we
learned? Hydrobiologia 483, 239–248.
Knaepkens, G.; Maerten, E.; Tudorache, C.; De Boeck, G.; Eens, M.,
2007: Evaluation of passive integrated transponder tags for
marking the bullhead (Cottus gobio), a small benthic freshwater
fish: effects on survival, growth and swimming capacity. Ecol.
Freshw. Fish. 16, 404–409.
Knights, B.C.; Lasee, B.A., 1996: Effects of implanted transmitters on
adult bluegills at two temperatures. Trans. Am. Fish. Soc. 125,
440–449.
Lucas, M.C.; Baras, E., 2000: Methods for studying spatial behaviour
of freshwater fishes in the natural environment. Fish Fish. 1,
283–316.
Newby, N.C.; Binder, T.R.; Stevens, E.D., 2007: Passive Integrated
Transponder (PIT) tagging did not negatively affect the short-
term feeding behavior or swimming performance of juvenile
rainbow trout. Trans. Am. Fish. Soc. 136, 341–345.
Prenda, J.; Granado-Lorencio, C. 1994: Winter home range and
habitat quality of three fish species (Cyprinidade) in a Mediter-
ranean river. Doñana. Acta vertebr. 21, 61–77 (in Spanish).
Ruetz, C.R., III; Earl, B.M.; Kohler, S.L., 2006: Evaluating Passive
Integrated Transponder tags for marking mottled sculpins: effects
on growth and mortality. Trans. Am. Fish. Soc. 135, 1456–1461.
Skov, C.; Brodersen, J.; Brönmark, C.; Hansson, L.A.; Hertonsson, P.;
Nilsson, P.A., 2005: Evaluation of PIT-tagging in cyprinids. J.
Fish Biol. 67, 1195–1201.
Ward, D.L.; Childs, M.R.; Persons, W.R., 2008: PIT tag retention and
tag induced mortality in juvenile bonytail and Gila chub. Fish.
Manag. Ecol. 15, 159–161.
Zar, J.H., 1999: Biostatistical Analysis. Prentice Hall International
Inc., Upper Saddle River, NJ.
AuthorÕs address: Daniel Pires, Departamento de Biologia Animal,
Faculdade de Ciências, Universidade de Lisboa, C2,
Piso 2, Campo Grande, P-1749-016 Lisboa,
Portugal
E-mail: dfcpires@fc.ul.pt