S109491942100030X

Managing the H ealth of
C a p t i v e G ro u p s o f R e p t i l e s
and Amphibians
Trent Charles van Zanten, BSc (Hons), DVMa,*,
Shane Craig Simpson, BVSc (Hons), GCM(VP), CMAVAb
KEYWORDS
Amphibian Diagnostics Examination Fecal Husbandry Nutrition
Quarantine Reptile
KEY POINTS
The health of large herpetological collections is heavily centered on maintaining species-
appropriate environmental parameters and husbandry techniques.
Dietary provisions for each species must be evidence-based, provide adequate macronu-
trients and micronutrients, and use clean and fresh ingredients provided in a practical
manner.
Reducing disease transmission relies on careful collection planning, experienced staff,
controlled entry into the collection, and awareness of potential zoonoses.
Quarantine is focused on clearing pathogens through diagnostic testing rather than hold-
ing animals for a specified period.
Veterinarians are required to assess new acquisitions, perform routine diagnostics, treat
medical cases, and provide guidance on husbandry and collection management.
INTRODUCTION
Reptiles and amphibians have been maintained in large collections with increasing fre-
quency over the last 20 years, which is due to their popularity in the pet trade,
increased study of their wild ecology, recognition of the need for species-focused
conservation programs, and the need to supply the meat and skin trade with
captive-bred animals and eliminate the demand for wild stock. Herpetological collec-
tions can range from private keepers who house several of their favorite species at
home, to zoos exhibiting hundreds of animals from a diverse range of species, as
a
Conservation, Research and Veterinary Services, Wildlife Reserves Singapore, Jurong Bird
Park, 2 Jurong Hill, Singapore 628925; b The Unusual Pet Vets, 210 Karingal Drive, Frankston,
Victoria 3199, Australia
* Corresponding author.
E-mail address: [email protected]
Vet Clin Exot Anim 24 (2021) 609–645

https://doi.org/10.1016/j.cvex.2021.05.005 vetexotic.theclinics.com
1094-9194/21/ª 2021 Elsevier Inc. All rights reserved.
610 Trent Van Zanten & Shane Simpson
well as to a farming operation rearing several thousand American bullfrogs for

slaughter. In all scenarios, a linking factor in the success and health of herptiles is
providing species-appropriate environmental conditions, nutrition, and husbandry
scaled to the size and demands of the collection. Clinicians and keepers face similar
challenges when managing herpetological captives: each taxon is reliant on specific
conditions that can be challenging to consistently provide, their progression of dis-
ease is often slow and insidious, they display limited signs of illness, and many un-
knowns remain regarding their optimal husbandry, disease prevention, and
treatment. Veterinary wellness programs catered by experienced clinicians have the
most value in large collections; however, these principles can be applied equally to
smaller menageries frequently encountered in private practice. Veterinarians manag-
ing large herpetological collections must have a sound understanding of what influ-
ences the health of a reptile or amphibian (“herptile”), the physiology and husbandry
of the species kept, how to assist with collection management decisions, and imple-
ment disease surveillance protocols for new acquisitions and long-term captives.
THE TRIFECTA OF HERPETOLOGICAL HEALTH
Maintaining the health of a herpetological collection is dependent on 3 separate but

related factors: the condition of the animals, their captive environment, and the
management techniques they experience (Fig. 1). For ectothermic species where
internal homeostasis depends on external physical factors,1,2 environmental and
husbandry conditions have more impact than animal-related factors. Under optimal
conditions, most species are incredibly hardy. Thus, maintaining the health of a
large collection shifts from focusing on individual animals during acquisition to
ensuring appropriate care and conditions for the species after entering the
collection.3,4
SOURCING THE COLLECTION
Selecting healthy foundation stock is essential to any animal enterprise and an

important point of contact for the client and veterinarian. New additions should be
sourced from reputable breeders/dealers and preferably physically assessed before
purchase. Captive-bred stock are preferred; wild-caught animals should be avoided
because of the infectious disease risk they pose to the existing collection, poor wel-
fare during capture and transportation, low rates of acclimation to captivity, and the
impact of unsustainable harvesting on wild populations.5,6 Rare exceptions include
sourcing species unavailable from other institutions where the need for captive repo-
pulation is identified by a taxon-advisory group,7 and appropriate permits and docu-
mentation for the process are obtained. Evidence demonstrates that wild-caught
herptiles suffer far greater mortalities than captive-bred animals and cause greater
difficulty and disappointment for surveyed keepers.4,8 Wild-caught herptiles also
represent considerable biosecurity and zoonotic risks to existing collections, local
wildlife, and keepers by introducing exotic pathogens directly or through foreign
parasitic vectors.9,10 The importation of wild-caught animals has declined signifi-
cantly ( 70% over a 12-year period) because of increased regulation and CITES pro-
tection of coveted species, while the importation of “ranched” animals (hatched and
sold from wild-caught adults) from Africa, Asia, and South America has seen a 50-
fold increase in recent years.11 Ranching systems can be readily overexploited as
a front for wild-caught operations because of limited oversight and trade
regulation.11,12
Managing the Health of Captive Groups of Reptiles 611
Fig. 1. Representation of the relative importance and interactions between animal, environ-
ment, and management factors contributing to the health of individual animals and the sus-
tainability of a herpetological collection.
ENVIRONMENTAL CONDITIONS AND HUSBANDRY

Enclosure Size, Design, and Construction
Enclosure design should suit the species’ needs and purpose of the collection. Pets
and animals exhibited for education are often afforded more space and naturalistic
furnishings aimed at appealing aesthetics and providing environmental stimulation.
By contrast, specimens kept for research, breeding, and commercial industries are
generally given more spartan conditions to maximize cleanliness, ease of mainte-
nance, and the number of animals per unit area.13,14 Neither approach is inappro-
priate; however, all animals must receive adequate space and sanitary conditions to
fulfill their physiologic and behavioral needs. A range of commercial designs appro-
priate for most small to medium-sized species are available; however, customized en-
closures are generally more affordable and appropriate for large reptiles and
commercial operations.
Traditionally reptiles have been housed in comparatively small enclosures under the
perception that their low metabolic rate and activity inferred low space requirements,
recommending floor dimensions (with body length, measured snout to tail, repre-
sented by BL) as little as 0.5 BL 0.33 BL for snakes and 2 BL 1 BL for lizards.15
While animals may survive in such accommodations, recent investigations into the
behavior of wild herptiles indicate that such dimensions may be inadequate,16
compromising their welfare by restricting normal movement and activities. Such re-
strictions are particularly relevant to large athletic species such as monitor lizards
and diurnal elapids/colubrids, indicating species-specific spatial requirements. Previ-
ous publications have suggested successful maintenance areas (SMAs) and ideal
maintenance areas (IMAs) in terms of floor space for reptiles and amphibians based
on BL, described in Table 1.13 For arboreal species, height must also be considered.
SMA floorplans provide minimum dimensions geared toward breeders/wholesalers/
research conditions, while the larger IMA floorplans are intended for hobbyists/
zoologic institutions—the area between these values indicating appropriate space
adjusted for species and size. Although larger enclosures generally improve welfare,
reduce self-inflicted injuries, and bolster reproductive activity—juveniles (particularly
snakes) are prone to anorexia and poor body condition in open spaces due to stress
from exposure and inexperience thermoregulating resources in artificial
environments.17
Rectangular enclosures are common because they are inherently strong and stack-
able, providing wide thermal gradients, few corners for animals to collide with or
abrade their snouts, and improved viewing.18 Square, round, and oval enclosures
are also successful, particularly for aquatic species. Enclosure height must prevent
terrestrial specimens from escaping or traumatizing themselves and allow arboreal an-
imals to climb, perch, and move vertically, which can be afforded by making the
longest horizontal dimension to a vertical one.18 Construction materials can include
plastic, glass, plexiglass, steel, melamine coated wood, fiberglass, concrete, or any
combination thereof. Materials selection depends on indoor versus outdoor enclosure
Table 1
Comparison of successful maintenance area (SMA) and ideal maintenance area (IMA)
recommendations for various herpetological taxa5,11,19
Species SMA IMA

Terrestrial snakes 0.6 m2/m BL 1.2 m2/m BL
3
Arboreal snakes 0.6 m /m BL 1.2 m3/m BL
2
Terrestrial lizards 0.2 m /0.1 m BL 0.5 m2/0.1 m BL
3
Arboreal lizards 0.2 m /0.1 m BL 0.5 m3/0.1 m BL
2
Tortoises and semiaquatic turtles 0.2 m /0.1 m BL 0.5 m2/0.1 m BL
3
Purely aquatic turtles 0.2 m /0.1 m BL 0.5 m3/0.1 m BL
2
Crocodilians 0.2 m /0.1 m BL 0.5 m2/0.1 m BL
3
Terrestrial amphibians 0.2 m /0.1 m BL 0.5 m3/0.1 m BL
3
Arboreal amphibians 0.2 m /0.1 m BL 0.5 m3/0.1 m BL
Abbreviation: BL, body length from snout to tail.

placement, local climate, maintenance of specific temperature and humidity, security,

ease of disinfection, providing nonabrasive surfaces, enrichment requirements, and
human viewing.19 Ventilation should be adequate to prevent microbial overgrowth in
stagnant air, which can be achieved using air vents or mesh screens on opposite sides
of the enclosure to create cross-current airflow.20 Adequate ventilation is difficult to
achieve in traditional glass aquariums without significant modification.
Outdoor housing is possible where the local climate reflects an animal’s native
range or can be similarly approximated with modification (eg, greenhouses in
temperate regions to stabilize higher temperatures for crocodilians). Outdoor enclo-
sures (typically open-topped pits or enclosed aviaries) are often larger than indoor
setups and can afford more naturalistic furnishings to create conditions conducive
to group housing and incite natural behaviors that improve breeding success
(Fig. 2).13 Increased reproductive activity, reduced ongoing expenses, and space
for group housing make outdoor systems are very popular with large-scale hobbyists
Fig. 2. Outdoor housing for reptiles. (A) Lockable outdoor housing for temperate elapids
and blue-tongued lizards made from converted rabbit hutches. (B) Converted aviary for
temporary reptile housing with a supplemental heat source. (C) Outdoor breeding enclosure
for chameleons with supplemental heating and water dripper system. (D) Outdoor breeding
facility for semiaquatic turtles within a protective aviary to prevent predation.
and commercial industries.3 “Wild” conditions bring about “wild” problems—enclo-

sures require predator proofing, and access of noxious pests such as rodents, metal-
dehyde baited snails, or cane toads (Rhinella marina) should be prevented.21,22
Weather extremes, inappropriate population dynamics (competition, bullying, starva-
tion), and parasitism can have a greater impact in poorly managed outdoor enclosures
than either wild or indoor conditions. Setups must be secure and protected against
natural disasters. Most problems can be managed or eliminated with careful enclosure
design, making outdoor accommodation optimal for large collections and displays.
All facilities should have hospital housing for unwell animals separate from the main
collection and quarantine facilities where possible. Hospital enclosures prioritize
cleaning and monitoring the occupant. Thus, they should be minimalistic (water
bowl, newspaper or water substrate, hide, heat and light source, perch for arboreal
species) and made from easily disinfected surfaces (eg, HDPE plastic).13,23 Racking
systems designed for breeding snakes/lizards are ideal for this situation; however,
any suitably sized plastic tub with adequate ventilation holes can be used, providing
they can maintain the temperature and with humidity requirements needed for
recovery.19
Temperature Regulation and Heating

Ectothermy dictates that herptiles rely on environmental temperatures for physiologic
and behavioral functions. These body temperatures are typically maintained within a
narrow range above ambient temperature through behavioral thermoregulation.1,24
Many amphibians experience thermoregulatory constraints from preventing evapora-
tive water loss at higher temperatures, selecting cooler preferred body temperatures
(PBTs) with higher moisture content.24–26 The measurement of PBT and establishment
of a preferred optimal temperature range (POTR) maintain the PBT has been evaluated
in many species using combined analyses of temperatures during field studies,
coelomic temperature logging, thermal shuttle boxes, peak temperature survival
times, physiologic processing, and protein denaturation studies.27–29 Swaths of pub-
lished references for commonly available species are available describing tempera-
ture ranges in captivity. For species where the POTR is unknown, inferences are
made based on temperatures within their wild distribution, altitude, habitat, and
ecological niche as well as data from similar species. Understanding a species’ unique
microclimate requirements is essential, as temperatures can vary significantly among
fossorial, aquatic, terrestrial, and arboreal strata in the same location.30 These temper-
atures are best provided as a gradient from the upper optimal temperature (UPOT) to
lower optimal temperature (LPOT) ends of the animal’s POTR either horizontally or
vertically, depending on the animal’s ecology.13 Providing a uniform constant temper-
ature can reduce growth and inhibit normal behavior patterns in Leopard geckos and
other species.31 Diurnal heliothermic species such as monitors and most agamids are
more reliant on basking than ambient temperature to achieve PBT (relying on radiant
heat sources), and larger species can maintain a core PBT under cooler ambient con-
ditions when provided a warm basking site or direct sunlight because of thermal
inertia.32,33 By contrast, thigmothermic or poikilothermic species are more dependent
on ambient temperature and conduction of heat (often from ventrally derived heat
sources).34 Temperature gradients are created based on the species and scale of
operation. Sunlight, heat lights, radiant panels, heat cord, aquarium heaters, pool
heating, and greenhouses have all been successfully used. Smaller collections hous-
ing a range of species may utilize individually controlled enclosures, while background
room heating or cooling to keep ambient conditions at the LPOT combined with a
small enclosure basking site can improve power efficiency in large establishments
and provides supplemental heat if enclosure heating fails.3 Semiaquatic species often
require heated water and basking sites to adequately thermoregulate.30,35 By
contrast, depending on the surrounding climate, most Urodeles and herptiles with
ranges of high latitude/altitude may require aquarium chillers or air-conditioning to
maintain their lower POTR.36,37 Animals housed outdoors must be afforded shaded
or sheltered areas to prevent overheating.
Daily and seasonal temperature variations affect the diel activity and reproductive
patterns of many species, which should be reflected in captivity. Variation exists be-
tween even closely related taxa regarding circadian temperature preferences;
nocturnal Australian geckos do not display differences in thermoregulation between
day and night,38 while tokay geckos demonstrate peaked metabolism and seek
warmer temperatures during the night.39 Most diurnal reptiles reduce thermoregula-
tory efforts and utilize cooler temperatures at night.38 This nightly temperature drop
is recommended when simulating environmental conditions to promote natural phys-
iologic behavior and development in zoologic settings; however, for meat and skin
production systems, maintaining temperatures at the UPOT together with increased
feeding is utilized to accelerate growth rates in juveniles prior to slaughter.40,41 Tem-
perature changes over the course of the year to induce seasonal dormancy in nontrop-
ical reptiles are natural processes and recommended in healthy animals because of
correlations with reduced rates of preovulatory dystocia, hepatic disease, and obesity
when practiced.42,43 Gonadal development, sex steroid/sperm production, follicular
cycling, and breeding behavior in many herptiles (particularly those from temperate
or subtropical climates) are associated with seasonal temperature variation upregulat-
ing hypothalamic–pituitary–gonadal axis activity and thyroid hormone production as
well as directly stimulating gonadal cells.44–46 These seasonal changes often improve
the breeding success of captive situations and can be artificially manipulated to in-
crease clutch number and minimize nonproductive periods in some species.47,48
When artificial heat sources are used, the heat output should be regulated via a ther-
mostat to prevent excessive heat load, and animals should be prevented from directly
accessing heat sources with physical barriers or careful placement of heaters out of
reach from occupants to prevent thermal burns.49 Recording of temperatures at
different locations in all enclosures should be performed regularly to confirm the
appropriate thermal gradient.13 Electrical elements near water or with risk of breakage
should employ ground fault circuit interrupters to prevent electrocution, and an unin-
terrupted power supply should be considered to reduce the effects of power outages
on the collection.
Lighting and Photoperiod

Providing ultraviolet (UV) lighting and regular photoperiod creates great discussion in
herpetocultural industries. Light serves important physiologic functions in herptiles.
Visible light (400–750 nm) permits vision, thermoregulation, and immune function
and influences seasonal reproductive activity and other neuroendocrine responses
through circadian melatonin production.50,51 UV-A (320–400 nm) composes the visible
light range for many reptiles and affects social interactions between conspecifics.30,52
UV-B (290–320 nm) is required for cutaneous vitamin D3 synthesis, contributes to cal-
cium metabolism, and can stimulate appetite.50 Wild reptiles and amphibians are
exposed to a daily and seasonal photoperiod in addition to variable visible/UV light
exposure each day depending on shade/cover, activity, elevation, and the time of
year. UV and visible light exposure thus depends on species ecology.53 Many
nocturnal geckos and frogs demonstrate basking behavior for short periods during
the morning/evening and experimentally can maintain higher systemic vitamin D3
concentrations when given access to UV-B radiation.54–56 Herpetoculturists agree

that diurnal reptiles (agamids/iguanids, monitors, chelonians, skinks, some frogs)
require UV-B to avoid syndromes of metabolic bone disease, but many argue
nocturnal species (snakes, geckos, crocodilians, most amphibians) do not require arti-
ficial light supplementation to thrive in captivity. Lighting is forgone because of added
expense and maintenance. Although these animals can survive without light provi-
sions, increasing evidence demonstrates improved health, development, and welfare
when light sources are added. Corn snakes demonstrate normal behavior patterns
and increased vitamin D3 synthesis when provided with UV-B,57 crocodiles and
caiman experience better growth and improved immune function when exposed to
natural light,40,58 and frogs raised without UV-B and given only oral vitamin D3 supple-
mentation have reduced bone density compared with wild counterparts.59 In Panther
chameleons, adequate maternal UV-B provision reduced the incidence of full-term
dead embryos in clutches,60 and reduced rates of postovulatory dystocia are
observed when females are supplied UV-B to facilitate calcium metabolism for ovipo-
sition.13,47 Further research is required to investigate the light requirements in a greater
range of species, but all evidence indicates that providing captive reptiles and am-
phibians full-spectrum lighting should be recommended because of the potential ben-
efits for their health, welfare, reproductive function, and behavior.
The light intensity, wavelength, and duration of exposure required are dependent on
species ecology. Herptiles of higher latitudes should experience seasonal variations in
day length and light intensity compared with equatorial species, where photoperiod
light exposure is more consistent. In some chelonians, a period of shortened day
length is demonstrated to trigger reproductive behavior, and seasonal photoperiod
(and temperature) shifts are recommended as an adjunct breeding stimulus.61,62
The UV requirements for over 250 species have been determined based on observa-
tions in their natural range, microhabitat, basking behavior, and measured ranges of
UV exposure and temperature requirements, categorizing animals into 4 Ferguson
zones according to the UV index (UVI).53,63 Examples of species from this list are
included in Table 2, and requirements for species not yet determined can be esti-
mated from related and ecologically similar taxa on the list.53 Not all related species
are alike in terms of preferred UVI: 2 chameleons from Madagascar (Calumma brevi-
corne and C nasutum) are Ferguson zone 1 reptiles, whereas C parsonii is a Ferguson
zone 3 species requiring much higher exposure.53,64
A wide range of UV-B lighting options exist, and suitable choices for each Ferguson
zone are provided in Table 2. These lights vary considerably in output wavelength and
intensity allowing keepers to select an appropriate option for their species. Mercury
vapor and metal halide lamps and can produce considerable heat, and this should
be factored into the selection process if other heat sources are to be used. Any
mesh placed between the light and the animal can reduce the visible and UV light
output by as much as 50%, and glass/most plastics between the light filter almost
all UV-B and UV-A, preventing access by the animal.50 Conversely, aluminum reflec-
tors can increase the irradiance of a lamp 218% compared with its standard output,
increasing the functionality of low output lamps for higher Ferguson zones.53 UV pro-
duction should be measured monthly in-situ with a quality UVI meter, and most lights
require replacement every 6 to 12 months from progressive UV output decay through
solarization—even if the light is apparently working.20
Natural light should be used wherever possible to provide the greatest wavelength
range and intensity and is ideal for facilities where the climate is appropriate for the
species—for both reduced cost to the keeper and optimized light exposure to the an-
imal. Herptiles kept outdoors should be offered both exposed and shaded areas
Table 2
Ferguson zone divisions, UV index (UVI) estimates, and appropriate methods of provision for
various herpetological taxa in captivity52,62
UVI Maximum
Ferguson Zone Recorded Method of UV
Zone Characteristics Range UVI Provisiona Example Species
1 Crepuscular/ 0–0.7 0.6–1.4 Shade method Fire salamander,
shade dweller blue dart frog,
spiny hill turtle,
emerald tree
boa, crocodile
skink
2 Partial sun/ 0.7–1.0 1.1–3.0 Shade method (in White’s tree frog,
occasional large cages, eastern box
basker gentle sunbeam turtle, western
can be used) hognose snake,
angle-headed
dragon sp,
spectacled
caiman
3 Open/partial 1.0–2.6 2.9–7.4 Sunbeam method Amazon milk frog,
sun basker leopard tortoise,
diamond
python,
Argentine tegu
4 “Midday” 2.6–3.5 4.5–9.5 Sunbeam method Galapagos
open sun tortoise,
basker Komodo
dragon,
chuckwalla,
Cunningham’s
skink
a
Shade method: a lower UV level over the entire enclosure / fluorescent tube ideal sunbeam
method: higher UVI focused on basking site; lower background UV level / mercury vapor, metal
halide, or T5-HO fluorescent tube.
providing an ultraviolet gradient. Indoor-housed specimens can be placed outside for

short periods (15–30 minutes) on a regular basis ensuring these setups be monitored,
well ventilated, and partially shaded; glass/plastic can create dangerous heat build-up
in confined spaces. Further investigation is required into the minimum time and fre-
quency of sunlight exposure required for maintaining normal systemic vitamin D
concentrations.
Care must be taken with the placement of enclosure lighting to adhere to recom-
mended distances from animals and prevent direct access to the light, particularly
with mercury vapor and metal halide lamps, where the risk of burn injuries is high; how-
ever, even fluorescent tubes can be dangerous if broken by strong lizards or snakes.49
Some lights produce very short wavelength UV-B and UV-C (below 290 nm), both of
which are toxic to epithelial cells at close range and can result in photokeratoconjunc-
tivitis, severe photodermatitis, and death with prolonged exposure.65,66
Humidity and Water

The sensible and insensible fluid loss of captive herptiles must be compensated for to
prevent dehydration. Drinking water alone is insufficient without creating an
appropriate microclimate to replace cutaneous and respiratory losses.67 Water bal-

ance is essential to kidney health, ecdysis, growth, reproduction, and digestion.33
Establishing a humidity gradient that replicates an animal’s native habitat can be chal-
lenging because of the inverse relationship between humidity and ventilation. Rainfor-
est and semiaquatic species are provided high (70%–100%) relative humidity (RH)
through limited enclosure ventilation, a moisture-laden substrate, and a substantial
water source—combined, this creates a stagnant environment that promotes bacterial
and fungal overgrowth—predisposing inhabitants to respiratory and dermatologic in-
fections.68 The accessory saccular lung lobe extending from the cervical trachea of
some chameleons is suspected to predispose chameleons to pneumonia in poorly
ventilated environments.69,70 Amphibians have high requirements for humidity owing
to their permeable skin; however, stagnant conditions with excessive environmental
pathogens will result in bacterial, fungal, or algal dermatosepticemia.71,72 Better
means for providing rainforest species high humidity involve creating humid microcli-
mates using hides filled with moist sphagnum, substrates with drainage layers to
remove excess moisture, timed misting systems to allow cages to periodically dry
out, and IV-line drip systems as a water source for arboreal species.13 If very high hu-
midity is required (eg, tree boas, dendrobatids, Abronia, Tribolonotus), increased
moisture can be paired with increased ventilation using screen top/sided enclosures
or axial fans to increase airflow.49 The RH of entire rooms can be raised using vapor-
izers or humidifiers, allowing features like drip systems in each enclosure to be used
without excessively wetting every surface. For arid-adapted species, a lower RH
(20%–60%) can be provided by limiting access to water over the course of the
week and using nonabsorbent or desiccating substrates. Even arid animals that
require humid microclimates periodically can suffer dysecdysis without a humid
retreat or brief misting approaching shedding.73 Regularly soaking tortoises and
providing a higher RH to juveniles during periods of growth is demonstrated to have
a greater impact on preventing pyramiding than diet, lighting, and temperature.74
Water should be kept clean, preferably being replaced daily except for large, filtered
sources. Arboreal herptiles can be provided elevated bowls or misting, terrestrial spe-
cies can be given water bowls or puddles, and aquatic species can be given water
sources large enough to swim in and maintain PBT. Water receptacles must be appro-
priately sized (ideally large enough for soaking), readily disinfected, and shaped to
permit escape and avoid accidental drowning. Dechlorination by aging tap water or
adding dechlorinator is recommended for amphibians and aquatic reptiles, and the
use of reverse osmosis/deionized (RO/DI) water with reconditioning salts will remove
toxins, impurities, and infectious agents (including Batrachochytrium).75 RO water also
appears to reduce the incidence of spindly leg syndrome in Harlequin toads, the
reason for this is unknown.76 Aquatic systems can be supplied via constant influx-
efflux of fresh, clean water (open systems) or the periodically changing a percentage
of the total water volume (semiclosed systems); the former being a challenge to main-
tain except in the largest facilities.75 Appropriate biological filtration, temperature,
aeration, water changing, and water testing (for pH, ammonia, nitrite, nitrate, GH,
KH) should be provided and factored into the maintenance of aquatic semiclosed
systems.
Substrate and Furnishings

Substrate choice is a major determinant of collection health and is decided by the pur-
pose of the collection and the species kept. In large facilities, substrates that are easily
cleaned take priority over aesthetics and enrichment; however, providing for behav-
ioral needs like burrowing, foraging, and hiding and furnishing a comfortable surface
with enough traction for locomotion are required. Quarantine, laboratory, and farming
facilities may elect no substrate at all for frogs, crocodiles, lizards, or chelonians in
plastic, fiberglass, or smooth concrete pens, with the transition from land to water sur-
faces provided by sloping the tub, using plumbed drainage systems for continuous
cleaning.4,19,41 Snakes and lizards in intensive industries are maintained on news-
paper, butcher paper, or wood shavings such as aspen or cypress, which facilitates
hiding and burrowing of these animals.77 In zoos or private collections, shredded/
ground coconut husk, washed river sand, sterilized soil, or sphagnum moss are excel-
lent choices when paired with appropriate maintenance. Cedar and treated pine prod-
ucts contain a resin that is potentially toxic to herptiles affecting the brain, liver, and
respiratory tract and should not be used.77,78 Similarly, substrates that are dusty or
form a powder should be avoided because of irritation of the airways, eyes, and
scales.13 Crushed walnut or pecan shells cause impactions when ingested and pro-
mote Aspergillus spp spore growth and are not recommended.13,20 Artificial compact-
ing sands, fine gravel, and clay-based cat litters should also not be used because of
the risk of gastrointestinal impaction.49 Stones larger than the animal’s head are safe
but are difficult to keep clean. Moisture-retentive substrates affect the microclimates
within the enclosure and minimize insensible water loss by increasing both local and
enclosure-wide humidity, and this is particularly important to fossorial species or juve-
nile reptiles with a high surface area/volume ratio, where chronic dehydration from the
use of desiccating substrates can cause mortalities.4,79,80 Persistently wet, warm sub-
strate without top-down ventilation to replace the warm moist air with cool, dry air will
promote rapid bacterial overgrowth; therefore, moisture-retentive substrates are not
recommended for closed plastic tubs or racking systems without adequate top venti-
lation.13,18,79 Bioactive substrates utilizing a “cleanup crew” of nitrifying bacteria,
fungi, plants, and invertebrate detritivores (eg, springtails and isopods) to break
down fecal and food waste have become incredibly popular as they reduce patho-
genic bacteria in the environment, stabilize humidity and reduce stress to the animal
through less frequent cleaning.79,81 Bioactive systems require a deep substrate
base (bottom drainage layer, mesh separation, nutrient-rich/aerated loam, and often
a surface cover of leaf litter/stones) in addition to live plants, natural furnishings,
and regular water supply to thrive, and depend on a stable population of “janitors”
to aerate the soil, preventing saturation, impaction, and toxic anaerobic condi-
tions.13,82 Such arrangements are very successful and have revolutionized keeping
dendrobatids; however, their initial expense and ongoing maintenance can prove
challenging to apply to large-scale production industries. Additionally, during an infec-
tious disease outbreak, the substrate must be discarded and replaced regularly, which
is expensive and time-consuming.
Minimalist enclosure furnishings should include a dark hide for cover, a water bowl,
a haul-out area for semiaquatics, and a basking surface for “sun-worshippers.” Ani-
mals without hiding places demonstrate overt stress and higher cortisol levels and
are prone to illness through stress-induced immunosuppression.14,83–85 The “bare-
minimum” approach to herpetoculture should be considered only in quarantine,
research, or large-scale production facilities. Enrichment options such as ground
cover, wood/branches, rocks, live plants, and artificial backgrounds should be consid-
ered for all captive herptiles to fulfill normal ecological activities and simulate their nat-
ural habitat (Fig. 3). Arboreal species have an innate need to climb, perch, and be off
the ground and fail to thrive without these provisions.17 Furnishings should be fixed in
position to prevent collapse and injury to the animal. Natural furnishings from wild
sources require disinfection prior to use—wood and rocks can be scrubbed with dilute
bleach, and soil and leaf litter may be sterilized by baking in a dry oven.13,79 Doing so
Fig. 3. Naturalistic enclosures. (A) Larger enclosures afford more opportunities for cohabita-
tion, increased enrichment opportunities, and improved aesthetics for onlookers. (B) Arid-
themed setup incorporating multiple focused basking sites and multiple feeding opportu-
nities for inhabitants. (C) Naturalistic housing for venomous species should include trap
boxes, as shown in the rear of the image. (D) Dart frog vivarium with copious growth of
live plants over a bioactive substrate.
removes parasites, fungi, bacteria, and viruses that may be potentially pathogenic,
permitting recolonization with beneficial microbiota.
DIETARY REQUIREMENTS
The nutritional needs for reptiles and amphibians is highly understudied; thus, dietary
inadequacy remains a leading cause of disease in herpetoculture.13 Captive diets
should always be based on studies of wild observations, yet this can be both chal-
lenging and conflicting—1 study in wild adult bearded dragons identified animal mat-
ter constituting 61% DM (dry matter) of the total diet, whereas a second demonstrated
that insects are comprised in only 10% of DM.86,87 Food items should be freshly pre-
pared; however, freezing or partial cooking of foods is acceptable and may release
some nutrients locked in plant cell walls.13,88 Frozen food items should be disposed
of within 6 months if not used, and uneaten food should be removed from enclosures
within 6 hours to reduce ingestion of rancid material. Ontogenetic shifts in dietary re-
quirements are common and should be accounted for when feeding juveniles versus
adults.89 Despite the psychological stimulation it provides, feeding live vertebrate prey
is not considered acceptable, and hunting behavior can be replicated with forceps or
tong feeding; additionally, prey items can seriously wound disinterested predators
when confined together.90 Food is a potential vector for parasites, bacteria, fungi,
and viruses. Animal prey should be frozen for 4 weeks and plant matter thoroughly
washed before feeding, wild-caught food items should be avoided, and transferal of
unwanted items from 1 enclosure to the next should never be practiced.68,88 Prepara-
tion of carnivorous and herbivorous diets should be performed separately to reduce
potential cross-contamination of pathogens like Salmonella, Campylobacter, and
Escherichia coli.91 The vitamin and mineral requirements of most herptiles remain un-
determined, and overprovision can be toxic; however, supplementation in captivity is
required in most cases for insectivores, omnivores, and less so, herbivores. Some
general rules apply:
1. Herpetocultural supplements have little oversight, and composition varies greatly.
Many available options are not complete multivitamins.88 Selection is based on
careful analysis of labeling and species needs. Human multivitamins are not recom-
mended. Ca and vitamin supplements should not be premixed.
2. Calcium is required in almost all cases but should not be supplied with a
phosphorus-based salt (negates the calcium supplement) or iron and zinc (Ca re-
duces absorption). Ca supplements containing phosphorus require a Ca:P>20:1
to overcome the poor Ca:P of commercially available insects.88
3. Not all species metabolize dietary vitamin D efficiently, and many iguanids and cha-
meleons preferentially synthesize vitamin D when provided with ample UV-B
radiation.60,92,93
4. Vitamin A is frequently not provided in supplements but traded for a stable precur-
sor b-carotene, which many reptiles cannot convert.88
Commercially formulated diets are increasingly popular; however, these also suffer
poor oversight in their production, and buyers must carefully assess options to ensure
guaranteed analysis and appropriate nutrient analysis on a DM basis from the product
label.94 Formulated diets have great potential in herpetoculture, and continued
research into species-specific requirements for complete nutrition and maximal
growth is required.
The known requirements for captive herptiles are summarized:
Insectivores
This group includes nearly all amphibians, most lizards, some turtles, and a few
snakes.88 Specific nutritional requirements are unknown for most species and are
extrapolated from NRC requirements for laboratory insectivores.95 Larval insects
(except silkworms) contain excessive fat, but all invertebrates have adequate protein
to support growth.95,96 Commercially raised insects are severely Ca-deficient,
whereas annelids and crustaceans have positive (yet still inadequate) Ca/P ratios.97
Ca and vitamins A/D require supplementation for captive insectivores.95,97 Gut-
loading insects with a quality dry, nutrient-dense, Ca-fortified diet containing greater
than 8% Ca for 12 hours to 5 days prior to feeding can overcome these deficits98,99
and should be augmented through the use of regular calcium and multivitamin dusting
of feeders, though this is less effective for aquatic animals and inadequate after 4 to
5 hours if not eaten.100,101 Improved growth and nutrient availability are obtained
from feeding a variety of invertebrates instead of a monoculture diet.88,102 The risk of

pesticide ingestion in animals fed wild invertebrate sources is exceedingly rare. All in-
sectivores should be provided with UV-B to promote vitamin D uptake. Fireflies are
toxic to many reptiles and amphibians and should never be fed.103
Carnivores
Carnivores include most snakes, all crocodilians, some lizards and amphibians, and
many aquatic turtles.88 Carnivorous reptile nutritional requirements are based on lab-
oratory analyses of mammalian predators that require high protein (25%–60% DM),
high fat (30%–60% DM) and low carbohydrates (<10% DM) with a species-
appropriate amino acid profile.104,105 Dietary deficiencies are rare when fed whole
prey, as most choices represent a balanced nutrient base.88 Prey should be fed a
commercial formulated diet; not seed/scraps/dog food creating poor nutritional qual-
ity. Muscle and viscera has a low Ca/P ratio and should not be fed exclusively.68 Very
specific dietary requirements (eg, ophiophagy in King cobras) can either be accommo-
dated in captivity or attempts can be made to convert to more conventional diets
without apparent ill effect.106 Some fish are thiamine and vitamin E deficient after
freezing because of activation of thiaminase and oxidation of lipids, warranting sup-
plementation.107 Feeding a range of fish species will circumvent this. Crocodile farms
commonly use by-products of other animal industries, which is acceptable for animals
to be culled but not for long-term use.108
Omnivores
Includes some lizards (eg, bearded dragons, sailfin dragons, blue-tongued skinks) and
turtles (sliders, box turtles, sea turtles, Australian short-necked turtles). Variable,
adaptable nutritional requirements of 15% to 40% protein, 5% to 40% fat, 20% to
75% carbohydrates DMB.109 Fruits should be avoided because of low overall nutri-
tional content, otherwise a mix of plant, insect and animal material can be offered in
addition to a large proportion of commercial dragon/turtle pelleted diet, with calcium
and multivitamin supplementation required less frequently than for insectivores. The
juveniles of omnivorous species typically consume far more insect and animal protein
than adults.88
Herbivores
Herbivores include most iguanids, some agamids, monkey-tailed skinks, some sea
turtles, and most tortoises. They require a dietary composition of approximately
15% to 35% protein, greater than 10% fat, 50% to 75% carbohydrates (containing
15%–45% crude fiber) DMB.109 Herbivore diets are challenging to accommodate in
captivity. Reptile herbivores are hindgut fermenters receiving proteins, vitamins, and
volatile fatty acids for digestion from symbiotic gut flora.110 All herbivores require
appropriate UV-B to synthesize vitamin D and allow dietary Ca absorption.88,93 Natural
diets are very diverse and offering variety in captivity is more important than 1 “best
choice” as few captive options are similar to what these animals naturally feed on.88
Salad greens and vegetables are typically too low in fiber, Ca, trace minerals/vitamins
and high in simple carbohydrates; fed in large volumes these predispose herbivores to
hepatic lipidosis and intestinal dysbiosis.88,111 It is difficult to provide a balanced diet
with only greens, vegetables, and fruit, and providing Ca/vitamin supplements with
green-foods often remains insufficient.92 Commercial herbivore diets vary greatly in
suitability, but several brands closely match the needs of herbivorous chelonians/liz-
ards and can account for 40% to 50% of the diet. Timothy and grass hays closely
match the nutrient profile of wild desert tortoises, and together with fresh grasses
and native vegetation should contribute 40% (as fed) to tortoise diets.112,113 Fresh
greens, vegetables, flowers, and weeds can be comprised in 20% of the chelonian
diet and in 50% of the iguanid diet.92 When fed predominantly balanced commercial
diets, limited Ca/vitamin supplementation is required for herbivores.88
CONSIDERATIONS REGARDING STRESS AND WELFARE IN HERPETOLOGICAL

COLLECTIONS
Animals under human care should be afforded the 5 domains of animal welfare (phys-
ical related factors such as adequate/balanced nutrition and water, a comfortable
physical environment, maintenance of a healthy/fit physical state, the ability to ex-
press rewarding behaviors, and opportunities to maximize positive experiences while
minimizing aversive ones) to ensure physical and mental wellbeing, regardless of their
reason for captivity.114,115 At a minimum, for herptiles this translates to having choice
when accessing required environmental parameters within their enclosure, providing
food/water based on their natural history, and providing enrichment through features
in their setup or interactions with conspecifics where appropriate.81 Regular rear-
rangement of furnishings, creating social complexity through cohabitation, training
and interaction with caretakers, and controlled low level “stressors” (eg, male combat,
seasonal temperature cycling, seasonal dietary restriction) add intrinsic psychological
value for many taxa and should be accounted for (Fig. 4).81,116 Naturalistic exhibits
frequently support these requirements; however, basic setups that are species-
appropriate and well maintained can provide good welfare (Fig. 5).117 Reptile welfare
and cognition studies are identifying many inconsistencies and inadequacies in previ-
ously accepted mantra. Space provisions for captive reptiles are currently being ques-
tioned: snakes are frequently denied the opportunity for rectilinear motion or
positioning in small enclosures, and monitor lizards with extensive wild ranges and
active diurnal behavior are routinely confined to cages twice their total length.16,118
There is a movement away from keeping reptiles in small, barren setups toward hous-
ing them in ways that better fulfill their enrichment and cognitive needs.119 Another
contended issue is stocking density and communal housing. For commercially farmed
species (crocodiles, alligators, soft-shelled turtles, bullfrogs) ideal stocking rates are
known,41,120,121 but these can rarely be applied to similar animals (eg, snapping turtles
are more aggressive than soft-shelled turtles, increased territoriality of dendrobatids
compared with bullfrogs).5,122 Many species in the pet trade, zoos, and conservation
programs have no published data on stocking density and may be subjected to unnec-
essary social stress in captivity. Breeders must balance providing for these physio-
logic and psychological needs with maximizing animals per unit area for peak
production. Clinicians must be able to recognize signs of stress in herptiles and poten-
tial stressors and advise on how these can be controlled.81
PLANNING THE COLLECTION
Successful herpetological enterprises require clear definable goals and adequate

curatorial input to ensure such targets are attained. Institutions should budget for pro-
posed financial inputs/outputs to prevent overexpenditure and compromised welfare.
Thought should be given to maintaining population sustainability and counteracting
unexpected losses or repopulation costs. Zoologic and conservation facilities must
select species based on regional biodiversity, conservation and educational value,
and are guided by the needs and recommendations of governing zoologic organiza-
tions to promote genetic diversity, meet population targets, repatriation goals, and
prevent surplus breeding.3,123 Where species recovery or reintroduction is the primary
Fig. 4. Enrichment and training. (A) Young Komodo dragon taken on a controlled walk
around a zoo for regular enrichment. (B) Raised nest box for egg deposition and hiding
in arboreal reptiles. (C) Artificial humid tunnels for amphibians. (D) Acclimation and training
of an African Dwarf crocodile (bottom left of image) to transport container prior to
shipping.
output, planning is required to maintain genetic diversity; however, for taxa threatened
with imminent extinction, increasing the global metapopulation takes priority.
Controlled short-term inbreeding may be required, with few long-term consequences
for wild populations based on genetic analysis of previously fragmented popula-
tions.124,125 Skilled, educated, experienced staff are invaluable to provide daily
assessment of both animals and equipment, and less experienced team-members
should receive ongoing opportunities for continuous education.3 Staff dedicated to
particular roles (food preparation, general husbandry, quarantine) are preferred and
reduce disease transmission versus 1 staff member performing all these tasks.
Facility design heavily influences husbandry and maintenance (Fig. 6). Species from
different biogeographical regions (temperate, tropical, arid) can be maintained in
separate rooms that are temperature, photoperiod, and humidity controlled, allowing
background regulation of conditions corresponding to each climatic zone, with indi-
vidual cages offering species-specific microclimates more effectively.3 Utilizing
Fig. 5. Racking systems. (A 1 B) Racking design at the Author’s facility for housing Tiliqua
skinks; each tub provides ample space, basking, UV, digging, hiding, and foraging opportu-
nities required for this species. (C) Modified racking system for elapids with cutout viewing
panels to allow inspection prior to enclosure access. (D) Racking system to house nocturnal
lizards.
natural sunlight with minimal filtering provides an accessible, beneficial and aesthetic
background light source in enclosures.125 Water supply, electrical access, adequate
storage, and waste disposal in all rooms will greatly improve workflow.13 Colder loca-
tions require insulated or heated flooring to prevent the “heat-sink” effect on concrete
enclosures at ground level, which can negatively affect thermoregulation and basking
behavior.3
Interspecific and intraspecific cohabitation of herptiles is possible. Inappropriate
cohabitation is stressful for captives, and pairing cannibalistic or territorial species
can be unacceptably dangerous.81 Successful cohabitation requires increased space
(adding half the minimum enclosure dimensions for each additional animal),18 multiple
additional resources such as basking sites/hides/feed bowls/perches (n11 resources
at a minimum, where n 5 number of animals), providing visual barriers, closely moni-
toring the group (with fortnightly weight/condition scoring) and removing problem in-
dividuals.3,125,126 Specimens are matched on size and behavioral traits, and species
Fig. 6. Enclosure design features. (A) Sand and charcoal filtration system for aquatic display
located external to building to improve access and reduce heat accumulation. (B) RO/DI
filtration system for amphibian water supply to remove contaminants and stabilize water
conditions. (C) Dual air-conditioning system running at alternate times to reduce overload
on a single unit and act as a backup in the event of failure. (D 1 E) Example of drainage,
separation, substrate and surface layering in a bioactive substrate, and modification of a
bioactive media during medical treatment using a sponge for easy cleaning and replace-
ment. (F) Multilayered water draining to prevent fluid accumulation in substrates of a semi-
aquatic setup. (G) Guard poles around natural tree trunks in outdoor setup allowing
controlled access to climbing resources while preventing escape. (H) Quarantine trap box
design for venomous species. (I) modified bucket-drip system for simple, cost-effective water
supply and humidity control in a chameleon breeding facility.
capable of predating on another should not be paired. Males often cannot be

cohabited because of territorial aggression, and although male–female and female–
female groups are often successful, this is not always the case (eg, most female cha-
meleons are intolerant of males outside breeding and will not accept other females
except in very large cages).69 Species from different geographic locations should
not be paired because of increased pathogen susceptibility in an immunologically
naı̈ve population.13 Some species asymptomatically carry organisms that cause
disease in others, such as chelonians and crocodilians harboring commensal proto-

zoa like Entamoeba causing severe gastrointestinal disease in squamates—mixing
of these taxa (particularly snakes with chelonians) should be avoided (Fig. 7).111
Maintaining accurate records of an individual’s identification, physical state, prior
history, husbandry conditions, daily observations, reproductive events, and medical
interventions is crucial in large collections.127 Individual identifiers (microchips, photo-
graphs, scale/toe cuttings, cage cards, scute tags) are recommended for all animals.
Records should be stored electronically with duplicate copies maintained off-site,3
and this can be done using excel spreadsheets or through an online database service
for animal collections.128
REDUCING DISEASE TRANSMISSION
Different collections have differing biosecurity requirements and disease risks. In

closed collections with no new animals entering the collection under any means, dis-
ease prevention is the focus of biosecurity and veterinary work. By comparison, open
collections (most operations, particularly wholesalers and retailers) focus on detecting
new diseases, although prevention must still be considered.4,129 Written biosecurity
protocols should be available to all staff and reviewed regularly for currency. Daily
workflow should ensure vulnerable groups (neonates, delicate species) are fed/
cleaned first, followed by mature/hardy animals; those with medical conditions are
handled last.3 Feeding and cleaning facilities should be physically separated, and
gloves worn/changed between cages (washing hands thoroughly between animals
is acceptable). Each room should contain separate, labeled, nontransferable equip-
ment. Footbaths between large enclosures (or at a minimum between quarantine
and hospital) is recommended; however, these must be well maintained to prevent
inactivation of disinfectant with organics.3,4,41 Daily, weekly, monthly, quarterly, and
annual cleaning schedules should be developed; these requirements vary but at a
minimum, feces, food waste and other unsanitary materials should be removed daily,
and soiled substrate or furnishings that cannot be disinfected disposed of.130 Disin-
fecting bioactive substrates will kill beneficial microorganisms; instead feces can be
removed and substrate changed in sections if needed. Naturalistic enclosures should
be reserved for collections (or parts thereof) where infectious disease prevalence is
low, as clearing pathogenic organisms established in these enclosures is diffi-
cult.13,18,79 Organic matter must be removed from enclosures using detergent and
hot water for effective disinfection.130 Cleaning items should be disposable and not
reused between enclosures. Disinfectants should be chosen based on disease risk
analysis and requirements; chlorhexidine, iodophors, and phenolic compounds
have demonstrated fatal toxicities when in contact with amphibians, and chlorhexidine
bathing has previously caused 2 turtle mortalities.130–133 Quaternary ammonium com-
pounds such as F10 represent a safe choice for most situations and have a broad
effective spectrum against bacteria (may be limited against Pseudomonas), bacterial
spores, mycobacterium, fungi, and enveloped and nonenveloped viruses.130,131,134 All
disinfectants require a contact time of 10 to 30 minutes depending on the compound,
and some are more effective when not rinsed off after use.130
Stress can be reduced by minimizing unwanted disruptions, keeping changes to
enclosure furnishings/keepers/routines to a minimum, creating appropriate social
groups, and providing optimal microclimates and nutrition. These steps will help pre-
vent chronic corticosterone elevation, which negatively affects growth, reproduction,
and immune function.81 Pest species that can pose a direct threat to captives, intro-
duce diseases, or consume captive diets include birds, rodents, insects, small
Fig. 7. Multispecies enclosures. (A) Species that naturally co-occur and use similar habitats in
the wild (such as this land mullet and tiger snake) and are of similar size make appropriate
cohabitants; however, housing snakes with other species always presents some degree of
risk. (B) Large outdoor enclosure cohousing Tomistoma and Siamang; both species naturally
never share the same space in these enclosures and have adequate provisions to avoid each
other. (C) Birds can be safely cohoused with some reptiles, such as tortoises, with few chan-
ces of predation or disease spread. (D) Squamates and chelonians should not be cohabited
in enclosures unless the health status of both species is thoroughly investigated prior to
introduction to reduce disease transmission.
carnivores and wild reptiles/amphibians.3,135 Establishing an integrated pest manage-

ment program targeting indirect suppressive control measures (eliminate desirable
pest resources and improve sanitation in animal facilities) is the cornerstone of pest
control, with direct suppression (baiting, repellents, removal) being used adjunc-
tively.135 Routine epidemiologic evaluation of disease events will aid the control of
future outbreaks, identify areas of inadequate husbandry, and provide guidelines for
collection management. Doing so requires evaluation of husbandry notes, veterinary
records, pathology reports, collection plans, and enclosure designs on a regular ba-
sis.136 Facilities housing high densities of reptiles and amphibians should practice
strict personal hygiene to minimize zoonotic disease potential. A list of zoonotic
genera linked to herptile-transmitted cases is provided in Table 3.137,138 Preventing

zoonotic infections entails prohibiting immunocompromised individuals from working
with reptiles, avoiding using live foods, keeping human kitchen areas away from ani-
mal facilities, washing hands and wearing gloves during maintenance, draining water
into toilets instead of the sink, using face-masks/shields when cleaning, and screening
for zoonotic pathogens as a part of collection health management.9,138,139
QUARANTINE
Quarantine is essential to prevent the entry of infectious diseases into collections,

monitor recovery after shipment, and allow animals to adjust to new husbandry mea-
sures. Stressed reptiles are more susceptible to infectious disease and increase path-
ogen shedding; thus, quarantine is required to avert this increased transmission
risk.140,141 Quarantine facilities should be physically separated from the main collec-
tion, utilize a separate ventilation system, and ideally employ separate staff (at a min-
imum, tend to quarantined animals on alternate days to or after working with the main
collection).141,142 Enclosures should accommodate the needs of the animal and mini-
mize stress during acclimation, containing minimal furnishings that can be either dis-
infected or disposed of.140 Newspaper, water, no substrate, or thin replaceable
sponge (for amphibians) are all ideal substrate choices.19 Dedicated quarantine equip-
ment should not leave the facility. Accurate health and husbandry records before and
after transfer from isolation are essential and often required as a part of international
standards for zoologic and farming industries.125,142
Disease risk analysis (DRA) involves estimating the current presence, exposure, and
consequences of introducing a pathogen to a collection and should be performed for
all major infectious diseases for the species being acquired and for other species
currently held in the collection.141,143 Steps to performing DRA include (1) hazard iden-
tification, (2) risk assessment, (3) risk management, and (4) risk communication.143 To
complete this, the animal’s origins, medical records, disease pathophysiology, recom-
mended transmission controls, and ways the disease risk can be communicated to
relevant parties should be identified. A list of pathogens to be considered for DRA
in herpetocultural facilities is provided in Table 4.141 The preexistence of these path-
ogens in a collection, and the function of those animals can greatly influence the de-
cision matrix of a DRA. Where the collection is aimed at displaying animals and not
focused on breeding and distribution, preexisting chronic viral or bacterial disorders
Table 3
Zoonotic pathogens causing disease in humans with confirmed or suspected links to reptile/
amphibian-transmitted cases136–138
Bacteria Salmonella, Aeromonas, Plesiomonas, Campylobacter, Clostridium,

Mycobacterium, Coxiella burnetii, Leptospira, Chlamydia, Rickettsia,
Borrelia,
Fungi Entomophthorales, Metarhizium, Beauveria
Viruses Western equine encephalitis virus, Eastern equine encephalitis virus, West
Nile virus
Protozoa Sarcocystis, Cryptosporidia
Pentatomidae Armillifer, Kiricephalus, Porocephalus, Raillietiella sp.
Helminths Spirometra, Diphyllobothrium, Trichinella, Gnathostoma, Alaria,
Echinostoma,
Arthropods Ticks (Spreading vector-borne diseases), trombiculid mites, snake mites
Table 4
Pathogens to be considered in disease risk analyses for different herpetological species140
Amphibians Chelonians Crocodilians Lizards Snakes

Ranavirus Herpesvirus Pox Virus Adenovirus Paramyxovirus
Chytridiomycosis Iridovirus West Nile Virus Nidovirus Reptarenavirus
Chlamydiosis Picornavirus Chlamydiosis Nannizziopsis 1 Nidovirus
Helminthiasis Amebiasis Mycoplasmosis Paranannizziopsis Bornavirus
Helminthiasis Helminthiasis Cryptosporidiosis Adenovirus
Intranuclear Helminthiasis Ophidiomyces
Coccidiosis Mites Cryptosporidiosis
Mycoplasmosis Ticks Helminthiasis
Ticks Amebiasis
Mites
Ticks
that can otherwise be managed on a case-by-case basis without euthanasia of

asymptomatic individuals may preclude the need to screen for these organisms during
quarantine or the elimination of specimens testing positive. Conversely, a refuge pop-
ulation maintained in a conservation program would typically aim to remain free of in-
fectious diseases, necessitating screening for all major pathogens during quarantine
and euthanasia of confirmed positives for contagious disorders.
Quarantine protocols entail a sequence of key disease-rule outs. Entering quaran-
tine, all specimens must be thoroughly examined, their reference weights recorded,
and their obvious disorders identified and addressed on arrival.140 Permanent identi-
fication should be established on entry. Transport boxes should be discarded or dis-
infected prior to reuse.142 Animals are ideally quarantined individually to allow better
assessment of behavior, appetite, and fecal output.140 Any problems identified in
quarantine require extension of the predetermined quarantine period until the condi-
tion is resolved.141 Ectoparasites must be screened for on multiple occasions; mites
are easily missed, and ticks, which represent clinically significant vectors of disease,
are commonly shipped from the country of origin on wild-caught or outdoor-housed
animals.140,144,145 Internal parasite screening (direct smear, flotation, and sedimenta-
tion) should not be performed on arrival—instead, waiting 1 to 2 weeks for animals to
begin feeding and shedding more parasites after the stress of transport results in bet-
ter detection.141,146 A second fecal exam is performed 2 to 4 weeks later. Wellness
bloodwork, fungal skin cytology, viral PCR, serologic testing, radiographs, or ultra-
sound can be taken as required, keeping the contact time with quarantining animals
to a minimum. PCR for snake viruses (particularly in python/boid collections) and
Batrachochytrium in amphibians are valuable during quarantine because of their
high sensitivity/specificity.147,148 PCR and serologic testing of some diseases (Crypto-
sporidium, Mycoplasma) have limited value in healthy animals not currently shedding,
and negative serologic results will not exclude early infections.141 Many novel patho-
gens cannot be tested for, warranting a consistent quarantine structure for every ship-
ment. The quarantine period depends greatly on the DRA of incoming animals; wild-
caught specimens require longer a quarantine and more rigorous testing than animals
with well-documented health records and no identified disease risks.149 Thus, quaran-
tine periods can range from as little as 14 to 90 days, with 3 months being standard in
most collections.141 Sunshinevirus and tortoise picornavirus can be shed from asymp-
tomatic animals for several years, indicating that we cannot “out-quarantine” some
viral diseases; clearing these pathogens requires testing prior to entering a collec-
tion.147,150 Ideally, no new animals should enter quarantine until the previous batch
has completed its duration, and an “all-in/all-out” policy is recommended if new ani-
mals must enter beforehand—quarantine for all animals is extended until the last
group has completed its duration.141,142
VETERINARY INVOLVEMENT: WHY, WHERE, WHEN, AND HOW OFTEN?
Veterinarians have a role in collection management when acquiring and sexing new
stock, during quarantine, and for annual health assessment, reproductive assess-
ment, disease treatment, and end-of-life care. All new animals require physical
assessment and history collection pertaining to prior husbandry, diet, their origins,
breeding, and medical history.142 During quarantine, veterinarians perform diagnos-
tics and analyze samples to identify infectious agents or investigate other health con-
cerns identified at initial examination.
Annual health assessment is ideally performed on all animals in the collection, and
this is often timed prior to the onset of breeding season (or immediately before bruma-
tion) to assist with management decisions through the year.129,151 Annual health
assessment may not be feasible when working with thousands of animals or
dangerous species (Fig. 8).129 In these cases, examining a percentage of the collec-
tion can be planned annually based on specimen value, population size, the disease
prevalence, and the sensitivity/specificity of diagnostic tests using epidemiologic ser-
vices such as Epitools to calculate the number of samples required to diagnose and
control a disease.152 Performing annual examinations more or less frequently depends
on the collection’s disease status, the species held, and the purpose for which they
are kept.4 Annual examinations can include bloodwork, reproductive ultrasound, ra-
diographs for periodontal disease/progressive conditions, fecal parasite, fungal, and
viral screening.129 Ultrasonographic reproductive evaluation is sometimes
required.4,47 Veterinarians may be hired to develop health management programs
requiring regular visitation or permanent contracts; in other cases, they may be called
on only to treat clinical cases and assess new acquisitions.4
Euthanasia and quality-of-life evaluations are critical to the humane treatment of all
animals. Veterinarians must ensure that slaughter practices in farming operations are
appropriate and follow current standards. Multimodal (2-step or 3-step) euthanasia or
slaughter techniques are recommended entailing inducing anesthesia, chemical/
physical euthanasia, followed by pithing or freezing for assurance of death.153 Interna-
tional euthanasia guidelines through the AVMA and Swiss FVO are available for ani-
mals kept for pet/conservation/research purposes and those bred for food.153–155
Currently, no standardized quality-of-life assessment exists for geriatric or chronically
ill herptiles, and some facilities adopt scoring systems used for mammalian species as
a basis for the decision to euthanize (Douay G, personal communication, 2020). Such
decisions are based on the animal medical status, deterioration in behavior, and the
level of long-term care required. Necropsies should be performed in all specimens af-
ter death or euthanasia, preferably within 24 to 48 hours of death if refrigerated.156
Postmortem sample submission is case-based, and histopathology on all gross ab-
normalities is recommended at a minimum; however, a definitive diagnosis is rarely
obtained from gross examination, and specific ancillary testing is generally
required.156
PREVENTIVE HEALTH
Diagnostic tests are essential when managing the health of large herpetological col-
lections. Protocols should be customized for each facility, as differing geography
and species kept will ultimately affect the recommended testing. A variety of
Fig. 8. Saltwater crocodile farming. (A) Incubator room for crocodile eggs is temperature-
controlled and humidity-controlled and requires complete disinfection prior to entry. (B)
Breeding pen design for adult crocodiles. (C) Crocodile facilities commonly raise hatchlings
in water tanks in total darkness to reduce stress and competition without the provision of
any light. (D) Grow-out pens for subadult crocodiles prior to slaughter.
diagnostics are available, with the challenge being selecting the correct tests to
perform, with the correct time and frequency to perform them. Testing can be divided
into 2 broad groups, screening diagnostics and targeted diagnostics.
SCREENING DIAGNOSTICS
These tests are not designed to look for a specific problem but rather to check animals
for a range of potential issues. The results of these tests may then indicate whether
further investigation is required.
Fecal Testing
The collection and analysis of fecal samples for internal parasites should be consid-
ered routine, if not mandatory, for all reptiles and amphibians entering a collection
and during quarantine. Because of the intermittent shedding of many parasites and
their eggs, serial sampling should be conducted to ensure the animals are clear before
release into the main collection. The actual frequency and duration that fecal testing is
performed will depend on the species being tested, the known parasitic pathogens for
that species, the housing method (ie, the likelihood of reinfestation), and the original
source of the animal (eg, captive-bred vs wild-caught). As a rule, ascarids, strongyles,
cestodes, trematodes, pentastomids, acanthocephalans (more so amphibians),
microsporidia (Cryptosporidium), and Entamoeba are always pathogenic and should
be cleared from the collection. Fecal samples must be as fresh as possible, free
from contaminants, and not dry when examined.111 Gross examination of the sample
can be conducted, noting the volume, color, and consistency and the presence of
blood, mucous, or obvious parasites. Direct fecal smears should be made and exam-
ined shortly after obtaining the sample unless it has been refrigerated or is large
enough such that the center of the sample remains moist. Fecal smears are particu-
larly useful at detecting protozoans and motile bacteria. Just as with small-animal
medicine, fecal flotation is an important diagnostic tool. This technique is useful to
detect parasites such as coccidia, nematodes, cestodes, and pentastomids; sedi-
mentation can be used to detect trematodes.157,158 Fecal cytology, particularly using
acid-fast stains to detect Cryptosporidia, can be used in collections where this is a
known concern. Many “parasites” found on fecal examinations may in fact constitute
normal gastrointestinal flora of the reptile or amphibian. The decision to treat is often
subjective and based on an appreciation for the normal flora seen in each species and
the presence of clinical signs attributable to the parasite’s presence. Fecal culture as a
screening test in herptiles is of limited value, particularly in relation to the presence of
Salmonella sp. All reptiles should be considered as potentially carrying Salmonella,
and a positive culture should therefore not necessarily be interpreted as that animal
harboring a pathogen.
Blood Testing
The routine analysis of hematological and biochemical data is extremely useful as a
screening tool to detect illness. Sample collection in the animal’s normal environment,
after it has had a chance to settle (not at import/purchase), will reduce stress leukocy-
tosis. There can be considerable variation in blood values between individuals
depending on their physiologic state, so routine blood screening allows the develop-
ment of baseline values for individual animals that can be used as comparisons with
later samples. Additionally, normal reference ranges may not be readily available for
the species in question. Hematological assessment can range from packed cell vol-
ume measurement (often all that is available in small patients) to complete blood
counts based on examination of prepared blood smears. Total white cells count
can be conducted along with cell morphology and assessment for the presence of
blood parasites such as Hemogregaria, assorted microfilaria, and bacterial/viral inclu-
sions.147 Biochemical analysis is most useful in clinically ill animals, though establish-
ing baseline data for animals shortly after entry into collections can be invaluable.
PCR and Viral Screening

Several key viral diseases can be prevented from entering a collection via appropriate
testing while animals are in quarantine. Examples of these infections include para-
myxoviruses (eg, Ferlavirus, Sunshinevirus), Bornavirus, Reptarenavirus, Nidovirus,
Adenovirus, Herpesvirus, and Ranavirus; most of which require oro-cloacal swabs
or fresh-frozen tissues.147 It is vital that the appropriate tests for the species in ques-
tion be performed. Testudidae herpesvirus and mycoplasmosis of chelonians can be
screened using PCR or virus isolation (for herpesvirus); however, false negatives are
common in clinically well animals, and given their high prevalence in captive popula-
tions, results are unlikely to add value unless required for export/conservation/ex situ
release purposes or when clinical cases are frequent.159 When combined with endo-
scopic gastric/intestinal mucosal biopsy or lavage samples, Cryptosporidium can be
identified readily via PCR and differentiated from species derived from prey items.157
Chytridiomycosis in amphibians is readily identified from skin swab PCR and is recom-
mended for all recent acquisitions; however, screening via cytologic examination of
stained/unstained skin smears may be preferred for annual assessment.148 The fungal
pathogens Ophidiomyces and Nannizziopsis sp. can be screened for using PCR from
lesions on the arrival of new imports (particularly bearded dragons 1 snakes collected
from the wild in the United States).160 Systemic amebiasis caused by Entamoeba inva-
dens and intranuclear coccidiosis and can be screened for via qPCR; however, sensi-
tivity is poor (trichrome staining is more effective), and given that Entamoeba is a
commensal organism in many chelonians, annual screening for this pathogen is nor-
mally required only where other reptiles are housed with or alongside chelonians,
particularly in collections with previous clinical cases.157
TARGETED DIAGNOSTICS
These tests are used when an animal develops clinical signs of disease or when
assessing for a specific condition.
Diagnostic Imaging
Various diagnostic imaging modalities can be used to assess animals with clinical dis-
ease or incorporated into the annual health assessment for many species. Ultrasonog-
raphy can be incorporated into routine reproductive evaluations on breeding
operations, particularly for female snakes, varanids, agamids, and chelonians. It is
used to evaluate follicular size prior to pairing animals, detect ovulation, confirm the
formation of developed eggs/embryos, and assess reproductive pathology, all of
which assist reproductive decision-making over the course of a season.161 Radio-
graphs can be used for several screening purposes. Geriatric animals can be
assessed for spinal and long bone abnormalities annually. Dental radiography can
be performed regularly on lizards with acrodont dentition to assess for the develop-
ment of periodontal disease (though CT provides greater detail for lesion severity). Ra-
diographs can provide postoviposition evaluation in chelonians (not applicable to
snakes/most lizards because of lower calcification of shells) and for assessment of uri-
nary calculi in tortoises in regions where high silicates predispose this disorder.162
Serology
Serologic evaluation is infrequently used as a stand-alone diagnostic in reptiles either
as a component of annual health assessment in collections or as a part of import
screening. Evidence of an immune response on its own is not enough to indicate
active infection and requires either quantification of increased immune response or
pairing with evidence of the pathogen’s presence with the animal (eg, PCR). Excep-
tions to this are conditions that persistently infect animals, such as chelonian myco-
plasmosis and testudinid herpesvirus—ELISA serology for Mycoplasma agassizii
and M testudineum may detect subclinical, intermittent shedders, and chronically
infected animals.159 Similar reports exist for ophidian paramyxovirus in snakes; how-
ever, limited availability and poor reliability (a negative result does not exclude early
infection, requiring second-round testing for the 6-week to 9-week period to develop
a twofold increase in antibody titers) means these techniques are rarely used in
quarantine assessment.141 May be considered in extremely valuable collections as a

means of annual screening for such chronic disorders.
Endoscopy
Cryptosporidia can be detected via endoscopic biopsy of gastric mucosa, or alter-
nately from gastric lavage samples (followed by subsequent PCR, or acid-fast staining
and histology) and is more sensitive than fecal cytology, but could be time-consuming
for large imports and should be considered only for at-risk species or specimens
showing GI abnormalities at the time of acquisition.157 In populations considered at
risk of mycobacteriosis, chlamydiosis, or other hepato-centric infections, coelioscopic
hepatic biopsy followed by tissue-PCR is considered a sensitive method of screening
for these disorders and may be considered for use in large collections as a part of
annual screening and prior to transfer between exhibits containing prior positive cases
or to other facilities.163
Fecal Testing
Just as fecal testing is an important screening diagnostic; it can be just as important as
a targeted diagnostic when investigating clinical disease. Those tests already dis-
cussed above can be performed should the clinical signs warrant them.
Blood Testing
Bloodwork is critical to investigating disease in most species. In reptiles and amphib-
ians, having baseline values to compare with disease-state samples is critical because
of their individual variations.
THE FUTURE OF PREVENTIVE MEDICINE
Prophylactic medicine is a developing area in exotic animal management and con-

tinues to evolve as new diseases (and technologies to manage them) are continuously
discovered. There is a tremendous scope for the development of new techniques and
preventive health programs in herpetoculture. However, prophylactic management
should still focus on optimizing husbandry practices for the health of the species being
kept.
Vaccination is currently being explored in some herpetocultural industries, and an
inactivated vaccine is currently in production against West Nile Virus in crocodil-
ians—this product is licensed for use in the USA for alligators from 1 month in
age.164 Once vaccinated, uninfected animals can become PCR positive and cannot
be differentiated from those that are truly infected.165 An autogenous poxvirus vaccine
developed from skin lesions in crocodilians sped healing and reduced scar formation
in affected animals.164 A similar autogenous vaccine has been used to manage Myco-
plasma crocodyli outbreaks in Nile crocodiles; however, a trial using an inactivated
vaccine proved unsuccessful.166,167 Vaccine development has been investigated for
ophidian Ferlavirus; however, to date, a positive antibody response has been inade-
quate and commercial options are not viable or recommended.164 Similarly, an inac-
tivated herpesvirus vaccine for use in chelonians showed no antibody response when
delivered subcutaneously.164
SUMMARY
The cornerstone of effective preventive health in large herpetological collections is an

in-depth knowledge of the ecology and husbandry of the species being kept, followed
by designing effective screening and control measures for potential pathogens. When
this is understood and provided for, reptiles and amphibians can prove hardy, long-
lived captives and breed readily. Developing health programs requires trust and
open communication between the veterinarian and the client; without this, most pro-
tocols will not be maintained. As more is learned about emerging reptile and
amphibian disease, preventive health represents an important area of research and
development for clinicians, presenting new opportunities for its application in a variety
of collections when adhering to these key principles.
CLINICS CARE POINTS
The environmental conditions under which a reptile or amphibian collection is maintained,

followed by the management techniques used to care for the animals, collectively have a far
greater impact on the success and health of captive herptiles than the disease status of
individual animals. Detailed knowledge of the natural history and subsequent application to
the husbandry of the species being kept will significantly reduce the incidence of illness in
the collection.
The understanding of welfare in captive reptiles and amphibians is continually evolving, and
a transition away from bare, cramped, “sterile” housing to larger and more naturalistic
setups (even at a large scale) is currently in effect. Bioactive enclosures come with their own
set of challenges, and veterinarians should be aware of these potential problems and how to
mitigate them.
Herptile nutrition can be generally subdivided into carnivore, insectivore, omnivore, and
herbivore diets, with carnivores being the least complicated and herbivores often proving
the most challenging to successfully establish long term. Limited regulatory oversight in the
production of formulated reptile diets means a careful review of the product label is
required prior to using them in large collections, despite their potential perks and ease of
use.
Quarantine alone is insufficient to prevent the introduction of pathogens to an established
collection, yet the decision to test for every known disorder on an acquisition of specimens is
not realistic. Performing a DRA on all incoming animals encourages appropriate disease
screening based on species, place of origin, and consequences of pathogen introduction and
will result in optimized decisions on quarantine duration and lab expenditure.
Large collections will benefit greatly from the development of an annual health calendar for
veterinary involvement under which individual health assessment, assistance with husbandry
planning, and decisions on screening and targeted diagnostics can be made for all specimens
based on epidemiologic principles.
DISCLOSURE
The authors have nothing to disclose.
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Managing the H ealth of

Vet Clin Exot Anim 24 (2021) 609–645

well as to a farming operation rearing several thousand American bullfrogs for

THE TRIFECTA OF HERPETOLOGICAL HEALTH

Maintaining the health of a herpetological collection is dependent on 3 separate but

SOURCING THE COLLECTION

Selecting healthy foundation stock is essential to any animal enterprise and an

ENVIRONMENTAL CONDITIONS AND HUSBANDRY

Species SMA IMA

Abbreviation: BL, body length from snout to tail.

placement, local climate, maintenance of specific temperature and humidity, security,

and commercial industries.3 “Wild” conditions bring about “wild” problems—enclo-

Temperature Regulation and Heating

Lighting and Photoperiod

concentrations when given access to UV-B radiation.54–56 Herpetoculturists agree

providing an ultraviolet gradient. Indoor-housed specimens can be placed outside for

Humidity and Water

appropriate microclimate to replace cutaneous and respiratory losses.67 Water bal-

Substrate and Furnishings

from feeding a variety of invertebrates instead of a monoculture diet.88,102 The risk of

CONSIDERATIONS REGARDING STRESS AND WELFARE IN HERPETOLOGICAL

PLANNING THE COLLECTION

Successful herpetological enterprises require clear definable goals and adequate

capable of predating on another should not be paired. Males often cannot be

disease in others, such as chelonians and crocodilians harboring commensal proto-

REDUCING DISEASE TRANSMISSION

Different collections have differing biosecurity requirements and disease risks. In

carnivores and wild reptiles/amphibians.3,135 Establishing an integrated pest manage-

genera linked to herptile-transmitted cases is provided in Table 3.137,138 Preventing

Quarantine is essential to prevent the entry of infectious diseases into collections,

Bacteria Salmonella, Aeromonas, Plesiomonas, Campylobacter, Clostridium,

Amphibians Chelonians Crocodilians Lizards Snakes

that can otherwise be managed on a case-by-case basis without euthanasia of

VETERINARY INVOLVEMENT: WHY, WHERE, WHEN, AND HOW OFTEN?

PCR and Viral Screening

quarantine assessment.141 May be considered in extremely valuable collections as a

THE FUTURE OF PREVENTIVE MEDICINE

Prophylactic medicine is a developing area in exotic animal management and con-

The cornerstone of effective preventive health in large herpetological collections is an

CLINICS CARE POINTS

 The environmental conditions under which a reptile or amphibian collection is maintained,

The authors have nothing to disclose.

1. Pough FH. The advantages of ectothermy for tetrapods. Am Naturalist 1980;

64. Edmonds D, Razaiarimahefa T, Kessler E, et al. Natural exposure to ultraviolet-B

84. Warwick C. Reptilian ethology in captivity: observations of some problems and

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The environmental conditions under which a reptile or amphibian collection is maintained,