Fish Quarantine: Current Practices in Public Zoos and

Aquaria
Authors: Catherine A. Hadfield, and Leigh A. Clayton
Source: Journal of Zoo and Wildlife Medicine, 42(4) : 641-650
Published By: American Association of Zoo Veterinarians
URL: https://doi.org/10.1638/2011-0034.1

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 Journal of Zoo and Wildlife Medicine 42(4): 641–650, 2011
Copyright 2011 by American Association of Zoo Veterinarians

FISH QUARANTINE: CURRENT PRACTICES IN PUBLIC ZOOS

AND AQUARIA

Catherine A. Hadfield, M.A., Vet.M.B., M.R.C.V.S., and Leigh A. Clayton, D.V.M., Dipl. A.B.V.P.
(Avian)

Abstract: The primary goal of quarantine is to reduce the risk of introducing infectious diseases into established
collections. Fish quarantine is inherently complex because of the variety of species, environmental requirements,
and facilities. To examine current practices, questionnaires were submitted to 60 public zoos and aquaria,
predominantly in North America. Questions reviewed system type (closed, flow-through), quarantine length,
diagnostics, treatments, and cleaning and disinfection. Forty-two of the 60 institutions responded. Most
institutions had separate quarantine protocols for freshwater teleosts, marine teleosts, and elasmobranchs.
Ninety-five percent of institutions had a minimum quarantine period of 30 days or more. Sixty-four percent of
institutions used isolated areas for some or all of their fish quarantine. Twenty-five percent had designated fish
quarantine staff. All institutions used regular visual examinations to assess animal health. Fifty-four percent of
the institutions carried out routine hands-on diagnostics on some fish; this was more common for elasmobranchs
than teleosts. All institutions carried out necropsies on mortalities. Fifteen percent of institutions performed
histopathology on almost all fresh mortalities; 54% percent performed histopathology on less than 10% of
mortalities. Prophylactic treatments were common in closed systems, in particular, formalin immersion for
teleosts, freshwater dips and copper sulfate immersion for marine teleosts, and praziquantel immersion for marine
teleosts and elasmobranchs. Institutions using dips generally did so at the start or end of quarantine.
Fenbendazole- and praziquantel-medicated foods were used commonly in teleosts, but dosages varied greatly.
Cleaning and disinfection of systems and equipment increased in response to known pathogens. These results can
be used to compare and discuss fish quarantine practices at display facilities in order to improve quarantine
success.
Key words: Copper sulfate, fish quarantine, formalin, praziquantel, salinity.

INTRODUCTION allowing monitoring, diagnostics, and treatments

Most public zoos and aquaria acquire fish from
a variety of sources including from the wild or
from large commercial facilities.11,13 Infectious
diseases are common in these fish, particularly
following environmental stressors such as trans-
port.17,22,26 These diseases can lead to high mor-
tality, especially ectoparasitic protozoa in teleosts
(e.g., Ichthyophthirius, Cryptocaryon, and Amyloodi-
nium) and monogeneans in teleosts and elasmo-
branchs (e.g., Gyrodactylus).22,26 Several diseases
are associated with chronic morbidity and loss of
aesthetics (e.g., Neobenedinia, leeches, and lym-
phocystis).22,26 If these diseases are introduced
into large exhibits, options for monitoring, diag-
nostics, and treatment can be severely limited by
accessibility, species composition, system vol-
ume, and life-support equipment.10,25 In some
cases, control of a pathogen in an exhibit may be
impossible. Quarantine is used to reduce the risk
of introducing infectious diseases into exhibits by
From the National Aquarium, 501 E. Pratt Street,
Baltimore, Maryland 21202, USA. Correspondence
should be directed to Dr. Hadfield (khadfield@
aqua.org).
while fish are isolated from collection animals.
Quarantine also allows animals to adjust to new
environmental conditions while under close ob-
servation and allows the collection of baseline
data.
The Association of Zoos and Aquariums
(AZA) recognizes that quarantine standards for
other zoo and aquarium animals cannot always be
applied to fish, as there is enormous variability in
what is effective and practical for a given institu-
tion, depending on the animals acquired, their
environmental requirements, and the facilities
available.4 Fish quarantine recommendations
have been published elsewhere.7,8,11–13,15,26,28,29 Two
papers have reported quarantine protocols in
specific institutions (National Aquarium, Balti-
more, and Oceanário de Lisboa); however, there
has been no review of current practices.6,29
This paper summarizes the results of a survey
of fish quarantine protocols at public zoos and
aquaria. The goal is not to recommend a specific
protocol, as the advantages and disadvantages will
depend on the species acquired and the facilities
available. However, it is essential to understand
the current practices in the industry in order to
compare and discuss quarantine protocols within

641

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 642 JOURNAL OF ZOO AND WILDLIFE MEDICINE

institutions. These discussions can help improve
fish quarantine practices and reduce morbidity
and mortalities in both quarantine and collection
animals.
MATERIALS AND METHODS
Between November 2008 and February 2009,
60 questionnaires were submitted to curatorial or
veterinary staff at large zoos and aquaria, pre-
dominantly in North America, with some in
Europe and Asia. The questionnaire included
questions on system type (closed or flow-
through), quarantine length, diagnostics, treat-
ments (including dosage regimes), and cleaning
and disinfection. Of the 18 questions, seven were
closed and 11 were open-ended. The open-ended
questions were used where categories were hard
to establish before the survey was carried out, and
where comments were encouraged (e.g., diagnos-
tic testing, treatment protocols). Where clarifica-
tion of answers was needed, and where dosages
were out of the expected range, follow-up was
carried out by electronic mail. Institutions with
predominantly flow-through systems were ana-
lyzed separately. Treatments were considered
routine if they were part of the quarantine
protocols and used prophylactically on most
members of a group. Some treatments were not
part of the standard quarantine but were used
commonly in response to specific pathogens or
situations; these were analyzed separately. Im-
mersion treatments were designated as dips if
they were 5 min or less, and baths if longer than 5
min. Where answers were incomplete or the
question was not applicable (e.g., freshwater dips
for institutions with only freshwater systems), the
institutions were excluded. The number of com-
plete responses to each question was reported as
the n-number. Bivariate analyses (nonparametric
Spearman correlation coefficients [SCC]) were
used to assess relationships between the number
of fish acquired and the availability of dedicated
space or staff. Other statistics were univariate.
RESULTS
Forty-two of the 60 institutions responded
(70% return rate), although not all questions were
answered by every institution. Several institutions
also submitted detailed quarantine protocols.
Most institutions were AZA accredited (74%,
31/42), including three of the eight international
institutions. Most institutions acquired more than
500 fish per year (59%, 19/32), and many acquired
more than 2,000 fish per year (28%, 9/32). Thirty-
eight of the responding institutions used closed
(recirculating) systems (90%, 38/42), and the
remaining four institutions had predominantly
flow-through systems (10%, 4/42).
Flow-through systems receive a continuous
supply of new water (e.g., local seawater). Of the
four institutions with predominantly flow-through
systems, two maintained designated fish quaran-
tine areas physically isolated from exhibit or
backup (holding) areas, with designated staff,
and used a minimum quarantine period of 30
days. Two institutions used backup areas and
exhibits for fish quarantine and used a quarantine
period of less than 30 days. These two institutions
reported seeing the same diseases on exhibit and
in quarantine because of exposure to waterborne
pathogens through the incoming water. In all four
institutions with flow-through systems, hands-on
examinations were rare. No medications were
used routinely; the only routine treatments were
saltwater or freshwater dips.
Closed systems rely on filtration and pumps to
recirculate water and maintain suitable oxygena-
tion and water chemistry. Each closed system is
independent, so quarantine can be used more
effectively to reduce the risk of introducing
infectious diseases. The following results are from
institutions with closed systems only.
Most institutions with closed systems had
designated fish quarantine areas physically isolat-
ed from exhibit or backup areas (64%, 23/36).
Some institutions used only these areas (19%, 7/
36), but many would also quarantine fish in
backup areas (44%, 16/36). The remaining insti-
tutions had no isolated fish quarantine and
quarantined all fish in backup areas (36%, 13/
36). The use of isolated quarantine areas was not
correlated with the number of fish acquired
(Spearman correlation coefficient 0.22, P ¼ 0.27).
Many institutions had designated staff for fish
quarantine (25%, 9/36). Institutions that acquired
less than 500 fish per year did not have designated
staff. Institutions that received more fish were
significantly more likely to have dedicated quar-
antine staff (Spearman correlation coefficient
0.49, P ¼ 0.009).
Some institutions reported never quarantining
fish within an exhibit system (19%, 6/32). The
remaining institutions would rarely or occasion-
ally quarantine fish on exhibit (81%, 26/32). This
was often because an exhibit system was empty,
and was more common with small, isolated
systems and signature species. Another reason
for quarantine on exhibit was a lack of suitable
quarantine space in backup (e.g., for cold-water

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 HADFIELD AND CLAYTON—FISH QUARANTINE PRACTICES
Table 1. Quarantine protocols for fish in public zoos and aquaria with closed systems.a
Fish quarantine groups
No fish protocols
General fish protocols
FW teleosts, SW teleosts, elasmobranchs
FW teleosts (cold water/warm water), SW teleosts
(cold water/warm water), elasmobranchs
FW teleosts (cold water/warm water), SW teleosts
(cold water/warm water), syngnathids, elasmobranchs
FW teleosts, SW teleosts, syngnathids, elasmobranchs
Other
a
FW, freshwater; SW, marine.
species). Prophylactic dips for ectoparasites were
sometimes used prior to starting quarantine on
exhibit.
All institutions aimed to quarantine teleosts
separately from elasmobranchs and invertebrates.
However, many reported mixing these groups on
rare occasions (e.g., teleosts with live rock or
sponges that were important food sources) (41%,
13/32). Most institutions either never used gravel
in their quarantine tanks or only used gravel
where it was considered necessary for the species
(e.g., garden eels, small wrasses) (69%, 20/29).
Some had gravel in most or all of their quarantine
tanks (14%, 4/29).
Protocols: Most institutions had separate quar-
antine protocols for freshwater teleosts, saltwater
teleosts, and elasmobranchs (66%, 25/38) (Table
1). Several of these also had separate protocols for
cold-water species, although the differences be-
tween cold-water and warm-water protocols var-
ied among institutions (e.g., three institutions
used a longer quarantine period with cold-water
fish, and five did not use copper sulfate on cold-
water fish). All institutions tailored their actual
quarantine plans to accommodate known or
suspected species’ sensitivities. Syngnathids in
particular were considered ‘‘sensitive’’ by most
institutions, and some institutions had separate
written protocols for this group; these were
similar among institutions (e.g., no copper sulfate
therapy, no nitrofurans, no or low-dosage forma-
lin, protocols for oral dosing using brine shrimp,
Artemia) (18%, 9/38). Most institutions main-
tained a written list of species with suspected
sensitivities to medications (63%, 22/35). Institu-
tions that had the same quarantine protocols for
all fish, or had no specific quarantine protocols
for fish, were mostly those that acquired few fish
(,500 per year) or that acquired only local species
with similar environmental requirements.
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643
% of institutions (n ¼ 38)
5
16
32
16
13
5
13
Duration of fish quarantine: Almost all institu-
tions aimed for a minimum quarantine period of
30 days or more (95%, 35/37). The minimum
quarantine was often longer for cold-water fish
(e.g., 45–60 days). Many institutions reported that
fish were often in quarantine for longer than the
minimum because of the duration of prophylactic
treatments or medical concerns. The longest
minimum quarantine period was 90 days, which
applied to all animals in one institution, and to all
elasmobranchs and to teleosts not treated with
copper sulfate or formalin in a second institution.
Several institutions shortened the quarantine
period for logistic reasons in specific cases,
especially for larger elasmobranchs.
Diagnostics: All institutions used regular visual
examinations to assess animal health. Many
institutions carried out routine hands-on diagnos-
tics on teleosts (41%, 15/37). These usually
consisted of examinations, skin scrapes, and gill
biopsies. For small teleosts, a subset of the group
was typically sampled (e.g., 5–10%), whereas most
larger teleosts were sampled as individuals. Most
institutions carried out routine hands-on diagnos-
tics on elasmobranchs (54%, 20/37), including
examinations, skin scrapes, gill endoscopy/biop-
sies, morphometrics, diagnostic imaging (radiog-
raphy, ultrasonography), blood work, blood
culture, fecal samples, coelomic aspirates, and
individual identification (e.g., microchips). The
remaining institutions did not run routine diag-
nostics but carried out diagnostics as necessary if
there were disease concerns (46%, 17/37).
All institutions performed necropsies on mor-
talities that occurred during quarantine, and
several reported that moribund fish in quarantine
were euthanized for diagnostics. One institution
reported sampling up to 2% of individuals from
less valuable groups for diagnostic necropsies.
Histopathology was less common: some reported
 644 JOURNAL OF ZOO AND WILDLIFE MEDICINE
running histopathology on all mortalities that
were fresh enough, although only a subset of a
group was submitted in a mass mortality event
(15%, 4/26); most reported running histopathol-
ogy on less than 10% of mortalities, usually when
there was no final diagnosis based on history,
water quality, or gross necropsy (54%, 14/26); and
one did not use histopathology (4%, 1/26).
Diagnostics were generally carried out by
veterinary staff (veterinarians or veterinary tech-
nicians) and students (53%), husbandry staff
alone (10%), or a combination (37%, 11/30).
Treatments: Almost all institutions with closed
systems routinely used medications during quar-
antine (94%, 34/36). The most commonly report-
ed medications, frequency of use, and dosage
regimes are listed in Table 2. Several institutions
used prophylactic treatments more commonly for
fish that came from large distributors and for fish
that were going into large exhibits. Treatments
tended to be more conservative for animals
acquired from other zoos and aquaria. Other than
dips done on arrival and mild salinity changes, all
institutions waited 3 to 7 days before starting
prophylactic treatments.
Most institutions used salinity changes in
quarantine, including freshwater dips, low-dose
hypersalinity baths, and saltwater dips (Table 2).
Protocols often included provisions to maintain
adequate temperature and pH during dips and
baths. Some institutions also used dips diagnos-
tically, looking for ectoparasites in the water post-
treatment (18%, 6/34). Institutions that used low-
dose hypersalinity baths treated almost all fresh-
water fish, but some fish were excluded from dips
(e.g., tetras and mormyrids were excluded from
saltwater dips).
Formalin was a common treatment for teleosts
and some elasmobranchs (Table 2). Dosage and
duration varied, with higher doses associated with
shorter treatment duration (Table 2). The highest
doses were often used on entry and/or exit from
quarantine. Most protocols required increased
aeration of the water during formalin treatment.
Some protocols required filters to be isolated
during treatment. Groups of fish that were known
or suspected to be sensitive to formalin were
excluded from treatment or dosed at a lower
concentration (e.g., syngnathids, scaleless fish,
and soft, freshwater fish). Of the 23 institutions
using formalin, 4% used an assay to measure drug
levels in the water, and 9% routinely used
formalin with malachite green.
Copper sulfate treatment was common for
marine teleosts (Table 2). This treatment was
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not used in freshwater systems or where alkalinity
was below 50 ppm. Copper levels were increased
slowly over 3 to 7 days, and were assayed at least
once a day and redosed as needed to maintain
therapeutic levels. Groups of fish that were known
or suspected to be sensitive to ionized copper
therapy were sometimes excluded from treatment
(e.g., syngnathids, juvenile tropical marine fish,
some cold-water fish). Alternatives included sa-
linity changes, formalin, chloroquine, or chelated
copper sulfate, e.g., Copper Safe (Mardel, Virbac
Animal Health Inc., Fort Worth, Texas 76137,
USA, or Sentry AQ, Sergeant’s Pet Care Products,
Inc., Omaha, Nebraska 68130, USA) or Cupr-
amine (Seachem Laboratories, Madison, Georgia
30650, USA). Some institutions either extended
quarantine or carried out further diagnostics if
copper treatment was not possible.
Chloroquine was rarely part of the routine
quarantine but was used commonly in teleosts,
either as an alternative to copper sulfate therapy
or to specifically treat Cryptocaryon, Ichthyophthir-
ius, or Amyloodinium (Table 2). Protocols often
required the system to be darkened through
immersion treatment. Of the 12 institutions using
chloroquine, 25% routinely used an assay to
measure drug levels in water.
Organophosphates were not part of the routine
quarantine for fish but were used commonly in
response to specific parasites that were resistant
to other medications (e.g., copepods, isopods,
leeches, and some monogeneans) (Table 2). Or-
ganophosphates were also used by three institu-
tions as an alternative to praziquantel baths in
elasmobranchs, although praziquantel was pre-
ferred where possible (8%, 3/37). Almost all of
the institutions that reported using organophos-
phates in quarantine used trichlorfon (92%, 12/
13). No assays were used. Lufenuron and diflu-
benzuron were occasionally used as alternatives.
Fenbendazole was often used in teleosts (Table
2). It was almost always dosed orally using
gelatin-based foods (e.g., Mazuri aquatic gels,
Purina Mills, St. Louis, Missouri 63166, USA).
Oral doses of gelatin-based foods varied widely
(Table 2). For larger fish, fenbendazole was
sometimes dosed at the individual’s body weight
and fed in prey items (Table 2). For syngnathids,
fenbendazole was often soaked into Artemia; dose
varied widely (Table 2). Several institutions
reported possible sensitivities in temperate, ben-
thic freshwater species. Levamisole was reported
as a potential alternative for sensitive species and
was dosed either orally or by immersion.
 HADFIELD AND CLAYTON—FISH QUARANTINE PRACTICES 645

Table 2. Common fish quarantine medications reported by institutions with closed systems.a

Predominant
Medication n % Institutions treatment groups Common dosage regimes

Freshwater dip 38 71 (teleosts), 19 Marine teleosts, some 0 ppt for up to 5 min, often at
(elasmobranchs) marine elasmobranchs the start and/or end of
quarantine
Saltwater dip 35 20 Freshwater teleosts 28–35 ppt for up to 5 min,
often at the start and/or end
of quarantine
Low-dose 35 37 Freshwater teleosts 3–5 ppt long-term, e.g., 7–30
hypersalinity bath days or for the duration of
quarantine
Formalin bath 36 64 Mostly teleosts 12.5 mg/L 7 days for 3–4 tx
(freshwater, brackish, 20–25 mg/L 1–2 days every 2–7
and/or marine) days for 3–5 tx
100–250 mg/L 5–60 min
Copper sulfate bath 37 61 Marine teleosts only. Not Slow increase to 0.15–0.25 mg/
used in elasmobranchs L ionized copper for 14–30
days
Chloroquine bath 37 5 (routine use), 27 Teleosts only 10–15 mg/L, e.g., 10–30 days,
(common use) or 7 days repeated every 7
days for 2–4 tx, or 1 day
repeated every 14 days for 2
tx
Organophosphate 37 0 (routine use), 36 Some teleosts and 0.25–0.75 mg/L 1–7 days for 2–
bath (common use) elasmobranchs 4 tx
Fenbendazole oral 36 56 (teleosts), 22 Teleosts and some Gel: 25–10,000 mg/kg of food
(elasmobranchs) elasmobranchs weight. Median 2,000 mg/kg;
at 2% food intake ¼ 40 mg/
kg of body weight
In prey: 25–50 mg/kg body
weight
Soaked into Artemia: 5–2,500
mg/kg wet weight for 15–30
min
Fed once or twice daily for 1–5
days, often repeated in 7–14
days
Metronidazole bath 36 11 Some teleosts 6.8–15 mg/L 5–8 hr
Metronidazole oral 36 28 Some teleosts Gel: 1,000–18,750 mg/kg food
weight. Median 6,250 mg/kg;
at 2% food intake ¼ 125 mg/
kg of body weight
In prey: 25–100 mg/kg body
weight
Fed once or twice daily for 5
days, sometimes repeated in
7–14 days
Praziquantel bath 36 75 Elasmobranchs, marine 2 mg/L 1–7 days, sometimes
teleosts repeated every 3–14 days for
2–3 tx; up to 5–10 mg/L 3 hr
Praziquantel oral 36 44 Some teleosts Gel: 50–12,000 mg/kg food
weight. Median 4,000 mg/kg;
at 2% food intake ¼ 80 mg/
kg of body weight
Fed once daily for 3 days, often
repeated in 7–14 days
Nitrofuran bath 36 25 Some teleosts Dependent on formulation
a
Tx, treatments.

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 646 JOURNAL OF ZOO AND WILDLIFE MEDICINE
Metronidazole was used less commonly (Table
2). Oral doses (usually in gelatin-based foods)
varied widely. For larger fish, metronidazole was
sometimes fed in prey items.
Praziquantel use was common, particularly
immersion treatment for elasmobranchs and ma-
rine teleosts (Table 2). Higher-dose baths were
often used on entry and/or exit from quarantine.
Several institutions reported using lower doses in
syngnathids or excluding them from treatment.
Praziquantel is relatively insoluble in water;
methods of application included initial dissolu-
tion in ethyl alcohol, or using a fine mesh to
contain the drug as it dissolved. Of the 29
institutions using praziquantel as a bath, 3% used
an assay to measure drug levels in water. Oral
routes (usually in gelatin-based foods) were used
mostly for teleosts or for specimens in larger
quarantine systems, and the doses again varied
widely (Table 2).
Many institutions used nitrofurans, which in-
cluded various combinations of nitrofurazone,
furazolidone, and methylene blue (Table 2). These
antibacterials were typically added to the water at
the start of quarantine and left in for 5 to 10 days.
Most institutions using this medication did not
use it in syngnathids. Routine use of other
antibiotics was rare: two institutions used oxolinic
acid or oxytetracycline in teleosts at the start of
quarantine (6%, 2/36). Most institutions used
antibiotics only in response to bacterial skin
disease or signs of systemic inflammation (e.g.,
hyperemia).
Two institutions routinely used formalin-inac-
tivated vibriosis vaccines in syngnathids (6%, 2/
36), and two institutions (6%; 2/36) used DC-
DHA Selco (Inve Aquaculture, Salt Lake City,
Utah 84104, USA) disinfection of Artemia for
syngnathids to reduce Vibrio spp.
Cleaning and disinfection: Following bath treat-
ments, most institutions routinely used activated
carbon to adsorb medication (58%, 19/33). Of
these 19 institutions, 84% disposed of the carbon
in municipal waste, 5% by incineration, and 11%
used a professional contractor.
Half of the institutions used ultraviolet disin-
fection on some of their quarantine systems (52%,
14/27). Some institutions used ozone disinfection
on a few of their quarantine systems or treated
water with ozone after use in quarantine systems
(22%, 6/27). Many institutions did not use any
ultraviolet or ozone disinfection in quarantine
(44%, 12/27).
For equipment used in quarantine systems, most
institutions reported using bleach or other chlorine
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disinfection followed by drying (74%, 25/34).
Other products included peroxygen compounds,
iodophors, quaternary ammonium compounds,
biguanides, alcohols, polymeric compounds, or
freshwater for equipment from saltwater systems.
Cleaning and disinfection between quarantine
groups varied. If there had been no concerns
during the quarantine period, most institutions
made no changes to the system. Many institutions
would keep the system free of fish for a period of
time, although this was often an effect of batching
shipments. Some institutions would drain and dry
the system or use bleach or other chlorine
disinfection. If there had been infectious disease
concerns during quarantine, a variety of disinfec-
tants were used, as well as temperature or salinity
changes, tailored to the disease(s) involved.
DISCUSSION
Forty-two institutions responded to this survey.
Although this represents only a sample of the
zoos and aquaria that display fish, most were
AZA-accredited and most of the larger aquaria
were included, so the results should be indicative
of current industry practices.
Isolation is an essential component of quaran-
tine. A separate area with designated staff is
ideal.4,29 Although designated staff were more
common in institutions that acquired more fish,
separate quarantine space was not correlated with
the number of fish acquired. Most institutions
also quarantined fish in backup areas. Although
these systems are separate, their close proximity
to systems with collection animals may increase
the risk of cross-contamination through splash-
ing, aerosolization, and fomites.22 Many institu-
tions rarely or occasionally quarantined fish on
exhibit. With empty exhibits, there is no direct
exposure of collection animals, but the exhibit
design may limit monitoring, access, and treat-
ment options. However, quarantine of fish with an
existing population was reported for exhibit
reasons (e.g., signature species, poorly stocked
tanks) or because suitable space was not available
in backup. This does increase the risk of exposure
of collection animals to novel pathogens and can
limit monitoring, access, and treatment options.
Teleosts, elasmobranchs, and invertebrates
were held separately whenever possible. This
allows a wider range of treatment options, as
several medications used in teleosts are potential-
ly toxic to some invertebrates and elasmobranchs
(e.g., copper sulfate, organophosphates).22,26
There was great variability in quarantine pro-
tocols among institutions, which is likely due to
 HADFIELD AND CLAYTON—FISH QUARANTINE PRACTICES
Table 3. Aspects of fish quarantine plans common
to more than 50% of institutions with closed systems (n
¼ 36).
Fish group Aspects of quarantine plans
All fish groups Minimum 30 days isolation
Regular visual examinations
Necropsies on all mortalities,
histopathology on some
Freshwater Formalin, if tolerated, repeated
teleosts Fenbendazole medicated feed
Diagnostics if morbidity or
mortalities seen
Marine teleosts Freshwater dips on entry and exit
Copper sulfate long-term bath, if
tolerated
Formalin, if tolerated, repeated
Praziquantel bath
Fenbendazole medicated feed
Diagnostics if morbidity or
mortalities seen
Elasmobranchs Praziquantel bath
Hands-on examination with
diagnostics
the variety of species acquired and the facilities
available. However, several management and
treatment options were used by more than 50%
of the institutions with closed systems (Table 3).
Most institutions used different protocols for
freshwater teleosts, marine teleosts, and elasmo-
branchs. Many also had separate protocols for
cold-water fish and tropical fish, although the
differences varied by institution. Many had sepa-
rate protocols for syngnathids, which generally
included less medication and, in some cases,
vibriosis vaccines and disinfection of foods.
The AZA recognizes that a 30-day quarantine is
an adequate standard for fish but that this period
may need to be shortened or extended for certain
species or diseases.4 Ninety-five percent of all
institutions with closed systems used a minimum
fish quarantine period of 30 days or more. This is
similar to a survey of 21 aquaria in Europe, where
86% of institutions used a quarantine period of
more than 30 days (I. Boerman, unpubl. data). In
this current study, quarantine duration was often
extended to allow for treatments and was often
longer for cold-water fish, in which pathogens and
clinical signs are slower to present.22 Some
institutions reported situations where practical
limitations required a quarantine period of less
than 30 days, particularly for large sharks.
All institutions used visual examinations to
assess animal health and carried out diagnostics
on live fish if there was significant morbidity.
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647
Routine sampling of a subset of each population
is recommended in the literature and was carried
out by half of the institutions.12,13 Common
diagnostics in teleosts focused on surveying
populations for ectoparasites, which are a com-
mon cause of acute morbidity and mortalities.
Common diagnostics in elasmobranchs were
more intensive, as these animals are often man-
aged as individuals.
All institutions used necropsies to identify
potential pathogens. This differs from a European
survey in which only 47% of aquaria reported
carrying out necropsies on fish in quarantine (I.
Boerman, unpubl. data). Histopathology was less
common. This was probably associated with the
prevalence of infectious diseases that can be
identified at gross necropsy, as well as the cost
and time required for histopathologic evalua-
tion.22,26,31
Prophylactic treatments were common in closed
systems. Several institutions reported using pro-
phylactic treatments more aggressively with fish
that were wild caught or were obtained from large
distributors. These animals are more likely to carry
common and/or potentially serious patho-
gens.4,11,13,22 Prophylactic treatments were also used
more aggressively in fish going into large exhibits
because subsequent treatment would be limit-
ed.4,11,13 Because treatments can have adverse
effects on the animals and the life-support system,
and because toxicity data are limited, most insti-
tutions maintained lists of potentially sensitive
species to help guide treatment decisions.
The most common ectoparasite treatments
consisted of salinity changes, copper sulfate,
formalin, and praziquantel. Salinity changes in-
cluded freshwater dips for marine fish and low-
dose hypersalinity baths for freshwater fish. These
were the only treatments used routinely by
institutions with flow-through systems. Salinity
treatments have several practical advantages,
including cost, availability, and easy disposal,
and they are well tolerated by most species.22,26
Copper sulfate was commonly used for marine
teleosts. It can be toxic to many species but when
used correctly is an effective treatment for
ectoparasitic ciliates and dinoflagellates.16,22,23,26
Institutions used a slow increase to therapy, and
drug levels were measured at least daily. Most
protocols offered alternatives for species that may
be sensitive. Chloroquine was also used as an
antiprotozoal treatment, rather than as an anti-
bacterial, and has the benefit of a wider therapeu-
tic index than copper.22
 648 JOURNAL OF ZOO AND WILDLIFE MEDICINE
Formalin was used mostly for teleosts, with
higher doses associated with shorter treatment
duration. Supplemental aeration was used be-
cause of the decrease in dissolved oxygen seen
with formalin treatment.3 Some of the bath
treatments lasted several days; assays are lacking
but would be useful to look at how the drug level
changes over time. Formalin is approved by the
United States Food and Drug Administration
(FDA) for use in food fish but is toxic to some
species and is a potential carcinogen.9,14 Malachite
green was used as an antiprotozoal with formalin
in a few institutions. However, because of toxicity
and human health concerns, malachite green has
been placed under high enforcement priority by
the FDA and should be avoided where possible.
Praziquantel was commonly used as a bath
treatment for elasmobranchs and marine teleosts,
primarily to control monogeneans. Although
praziquantel is relatively expensive, it has a wide
therapeutic index and is effective against adult
monogeneans.15,22,26 Assays were not common, and
because drug levels will likely be affected by
method of dissolution, water chemistry, and life-
support systems, it would be useful to investigate
drug concentration in the water.
Endoparasite treatments were less common and
included oral praziquantel and fenbendazole.
There was enormous variation in the drug doses
in the medicated gelatin-based foods. Possible
reasons include a lack of pharmacokinetic data
and wide ranges in the literature: a review of three
commonly used formularies shows wide ranges
for oral praziquantel (4,000–20,000 mg/kg of food
weight) and oral metronidazole (5,000–40,000
mg/kg of food weight).15,22,26 It is also possible
that historic quarantine protocols are continued
as long as no adverse effects are seen. Of
particular concern is the wide dose range seen
with fenbendazole, as there are increasing reports
of fenbendazole toxicity, including toxicity in
sharks at 25 mg/kg orally.20,21 Oral medications
should be dosed based on estimated food intake
and available pharmacokinetic and efficacy data.11
Antibacterial nitrofurans were used by 25% of
institutions immediately post-transport. Nitrofu-
rans are used to reduce bacterial load in the water,
as damage to the skin and mucus is common
following transport.17,22,26 However, immersion is
the route of administration, and both human
contact and disposal are of concern.22,26 In the
United States, nitrofurans have been placed under
high enforcement priority by the FDA and should
be avoided where possible.
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Antibacterial treatment of Artemia and vibriosis
vaccines were reported specifically for syngna-
thids. Vibriosis is a common cause of morbidity
and mortalities in syngnathids, especially follow-
ing skin trauma and stress.2,5,18
Medication use and disposal should follow all
relevant legislation. In the United States, few
drugs are approved for use in fish, but drugs
approved in other major food-producing animals
can be used in confined populations of nonfood
fish under the care of a veterinarian, when
necessary. This applies to most medications
discussed in this paper, apart from malachite
green and nitrofurans.
Disinfection of water can be used to reduce
pathogen loads. Ultraviolet filtration can provide
safe and effective disinfection if used appropri-
ately.24,27 Although ultraviolet filtration was the
most common form of disinfection, it was used
only on a subset of quarantine systems in less than
half of the institutions, probably because of the
cost. Ozone can provide effective disinfection for
fish systems.27 Because of potential adverse ef-
fects, ozone production must be tightly controlled
and by-products monitored closely.27 Ozone was
not commonly used on quarantine systems,
probably because the variable bioload and small
volume of many systems increase the risk of toxic
effects, and ozone disinfection is both technically
demanding and expensive. Two institutions used
ozone to treat water coming out of quarantine
systems, which may help reduce the bacterial and
viral load in the water before it is discharged or
used in other systems.19
Activated carbon was used commonly to ad-
sorb in-water medications.1 The carbon was
predominantly disposed of in municipal waste.
The total volume of carbon is likely to be small,
particularly when compared to aquaculture, but
disposal of both medicated water and carbon
should be considered when using immersion
treatments.
Cleaning and disinfection varied significantly,
although chlorine-based compounds were most
commonly reported. Cleaning and disinfection
was increased in the face of specific disease
concerns to reduce the risk to subsequent popu-
lations.29,30
This questionnaire did not look at effectiveness
of fish quarantine protocols; it was assumed that
institutions used protocols that provided ade-
quate results for the commonly acquired species.
It would, however, be interesting to look at
morbidity and mortality in relation to the quar-
antine protocols. Such a study would need to
 HADFIELD AND CLAYTON—FISH QUARANTINE PRACTICES
incorporate information on source, shipping,
species, and environmental parameters (e.g.,
stocking density, temperature ranges, and water
quality).
CONCLUSIONS
Despite the diversity of species and facilities,
there were many similarities in fish quarantine
protocols at public zoos and aquaria with closed
systems. These included a minimum quarantine
period of 30 days, visual examinations, necropsies
on mortalities, and prophylactic treatments for
ectoparasites. The study also showed some of the
challenges encountered when working with di-
verse species; no single quarantine protocol could
be applied to all institutions. The information
from this survey can be used to compare and
discuss fish quarantine practices and protocols at
display facilities, in order to improve quarantine
success.
Acknowledgments: The following institutions
generously provided information on their fish
quarantine procedures: Adventure Aquarium,
New Jersey (USA); Alaska Sealife Center, Alaska
(USA); Aquarium of Niagara, New York (USA);
Audubon Aquarium of the Americas, Louisiana
(USA); Calvert Marine Museum, Maryland
(USA); Chicago Zoological Society, Brookfield
Zoo, Illinois (USA); Cincinnati Zoo and Botanical
Garden, Ohio (USA); Cleveland Metroparks Zoo,
Ohio (USA); Columbus Zoo and Aquarium, Ohio
(USA); Dallas Aquarium, Fair Park, Texas
(USA); Denver Zoological Gardens, Colorado
(USA); Disney’s Animal Programs, Florida
(USA); Florida Aquarium, Florida (USA); In-
dianapolis Zoological Society, Indiana (USA);
John G. Shedd Aquarium, Illinois (USA); Mon-
terey Bay Aquarium, California (USA); Mote
Marine Laboratory and Aquarium, Florida
(USA); Mystic Aquarium, Connecticut (USA);
National Aquarium, Maryland/Washington, D.C.
(USA); New England Aquarium, Massachusetts
(USA); Newport Aquarium, Kentucky (USA);
North Carolina Aquaria—Fort Fisher, Pine Knoll
Shores, Roanoke Island, North Carolina (USA);
Oklahoma City Zoological Park, Oklahoma
(USA); Omaha’s Henry Doorly Zoo, Nebraska
(USA); Oregon Zoo, Oregon (USA); SeaWorld
Orlando, Florida (USA); SeaWorld San Antonio,
Texas (USA); SeaWorld San Diego, California
(USA); Smithsonian National Zoological Park,
District of Columbia (USA); South Carolina
Aquarium, South Carolina (USA); Steinhart
Aquarium, California Academy of Sciences, Cal-
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649
ifornia (USA); Texas State Aquarium, Texas
(USA); Wildlife Conservation Society, New York
Aquarium, New York (USA); World Aquarium,
Missouri (USA); Aquarium du Québec, Canada;
Bermuda Aquarium, Bermuda; Bristol Zoo Gar-
dens, England; Oceanário de Lisboa, Portugal;
Underwater World, Singapore; Toronto Zoo,
Canada; Vancouver Aquarium Marine Science
Centre, Canada; Zoological Society of London,
England.
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Received for publication 15 February 2011