Manual For Control of Infectious Diseases in Amphibian Survival Assurance Colonies and

284 Pages·2017·5.86 MB·English

by  Onnie Byers

A Manual For Control of Infectious Diseases in
Amphibian Survival Assurance Colonies and
Reintroduction Programs

Version 2.0: updated 2017

Based on:
Proceedings from a Workshop:
16–18 February 2009
San Diego Zoo
Cover photos courtesy of Allan Pessier and Ron Holt.

A contribution of the IUCN/SSC Conservation Breeding Specialist Group in collaboration
with Amphibian Ark, San Diego Zoo, and Zoo Atlanta

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analysis of issues related to conservation, and believes that reports of these
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© Copyright CBSG 2017

The designation of geographical entities in this book, and the presentation of the
material, do not imply the expression of any opinion whatsoever on the part of IUCN
concerning the legal status of any country, territory, or area, or of its authorities, or
concerning the delimitation of its frontiers or boundaries.

Pessier, A.P. and J.R. Mendelson III (eds.). 2017. A Manual for Control of Infectious
Diseases in Amphibian Survival Assurance Colonies and Reintroduction Programs, Ver.
2.0. IUCN/SSC Conservation Breeding Specialist Group: Apple Valley, MN.

An electronic version of this report can be downloaded at www.cbsg.org
<http://www.cbsg.org/> .
This project was supported by grant LG-25-08-0066 from the Institute of Museum and
Library Services. Any views, findings, conclusions or recommendations expressed in this
publication do not necessarily represent those of the Institute of Museum and Library
Services.
A MANUAL FOR CONTROL OF INFECTIOUS DISEASES IN AMPHIBIAN
SURVIVAL ASSURANCE COLONIES AND REINTRODUCTION
PROGRAMS

EDITORS: ALLAN P. PESSIER & JOSEPH R. MENDELSON III

PROCEEDINGS FROM A WORKSHOP:
16–18 FEBRUARY 2009
Zoological Society of San Diego

Organized by:
Allan P. Pessier, DVM, Dipl. ACVP
San Diego Zoo
&
Joseph R. Mendelson III, PhD
Zoo Atlanta

Facilitated by:
Onnie Byers, PhD
IUCN Conservation Breeding Specialist Group

Updated March 2017 by:

Allan P. Pessier, DVM, Dipl. ACVP
San Diego Zoo
&
Joseph R. Mendelson III, PhD
Zoo Atlanta

Benjamin Tapley, MSc
ZSL London Zoo

Matt Goetz
Durrell Wildlife Conservation Trust
Table of Contents

1. INTRODUCTION………………………………………………………….………………………………….……….. 8

2. LIST OF CONTRIBUTORS………………………………………………….………………………….….……….. 10

3. RISK ASSESSMENT………………………………………………………….….……………………….…………. 11
3.0 INTRODUCTION…………………………………………….………………..………….………….. 11
3.1 GENERAL RECOMMENDATIONS FOR REINTRODUCTION AND
TRANSLOCATION PROGRAMS………………………… .………………………….………… 11
3.2 DISEASE RISK MITIGATION FOR CAPTIVE BREEDING AND REINTRODUCTION
PROGRAMS…………………………….………………………………………..….………….. 13
3.3 TRANSLOCATION……………………………………….…………………………….……………. 19
3.4 REFERENCES……………………………………..…………………………………….…………… 20

4. BIOSECURITY………………………………………………………………………………………….……………. 21
4.0 INTRODUCTION……………………………………………………..………………….…………… 21
4.1 WHAT IS "BIOSECURITY"?............................................................................. 22
4.2 BIOSECURITY DEFINITIONS…………………………………… ………….…………….….…… 23
4.3 COSMOPOLITAN COLLECTIONS AND RISKS OF DISEASE FOR WILD AMPHIBIANS… ... 23
4.4 LEVELS OF BIOSECURITY…………………………………………………………………..…..… 24
4.5 SUMMARY OF BEST PRACTICES AND ISOLATION SECURITY LEVELS……………….….... 30
4.6 BIOSECURITY PRACTICES……………………………………………………………………….… 33
4.7 STAFF TRAINING AND IMPLEMENTATION OF BIOSECURITY PRACTICES………………… 35
4.8 LONG-TERM ISOLATION OF AMPHIBIANS DESTINED FOR REINTRODUCTION
TO THE WILD……………………………………………………………….…………….…….. 35
4.9 FACILITIES FOR LONG-TERM ISOLATION……………………………………………….….…. 36
4.10 HUSBANDRY STAFF HYGIENE AND PROTECTIVE CLOTHING……………………….….…. 37
4.11 HUSBANDRY ROUTINES……………………………………………………………………….... 39
4.12 ENCLOSURE SANITATION……………………………………………………………………...… 41
4.13 EQUIPMENT, ENCLOSURE SUBSTRATES AND DECORATION…………………….….....… 41
4.14 WASTEWATER DISPOSAL…………………………………………………………………..….… 42
4.15 SOLID WASTE DISPOSAL……………………………………………………………….………… 44
4.16 SOURCES OF WATER………………………………………………………………………….….. 45
4.17 SOURCES OF FOOD……………………………………………………………………………...… 45
4.18 FACILITY DESIGN AND BUILDING SPECIFICATIONS………………………………….………. 46
4.19 REFERENCES……………………………………………………………………………….……..… 48

5. CLEANING AND DISINFECTION…………………………………………………………………..……………. 50
5.0 INTRODUCTION…………………………………………………………………………………….… 50
5.1 PRINCIPLES OF CLEANING & DISINFECTION………………………………..……………….…. 51
5.2 CHEMICAL DISINFECTANTS………………………………………………………..…………….… 53
5.3 NON-CHEMICAL METHODS OF DISINFECTION………………………………………………... 56
5.4 CLEANING AND DISINFECTION OF ANIMAL ENCLOSURES……………………………..……. 58
5.5 DISINFECTION OF EQUIPMENT AND TOOLS……………………………………….…………... 59
5.6 DISINFECTION AND STERILIZATION OF SUBSTRATES AND CAGE FURNITURE.…………... 59
5.7 DISINFECTION OF LIVE PLANTS………………………………………………………….……….. 61
5.8 DISINFECTION AND STERILIZATION OF WATER…………………………………….……….... 61
5.9 DISINFECTION AND BIOSECURITY IN THE FIELD……………………………………..………… 62
5.10 TABLE OF DISINFECTANTS…………………………………………………………….…….……. 66
5. 11 REFERENCES…………………………………………………………………………….….…….... 67

6. QUARANTINE…………………………………………………………………………………………..…….…... 70
6.0 INTRODUCTION………………………………………………………………………..……………. 70
6.1 GOALS AND DEVELOPMENT OF QUARANTINE PROGRAMS……………………..……...... 70
6.2 RISK ASSESSMENT IN AMPHIBIAN QUARANTINE………………………………..…………... 72
6.3 QUARANTINE FACILITY AND QUARANTINE STAFF CONSIDERATIONS………..…….….… 73
6.4 ANIMAL HUSBANDRY CONSIDERATIONS IN QUARANTINE……………….…..…… ……... 75
6.5 LENGTH OF THE QUARANTINE PERIOD……………………………………….……..….…..... 79
6.6 MEDICAL CONSIDERATIONS IN QUARANTINE…………………………………………...…… 81
6.7 LABORATORY TESTING AND DISEASE TREATMENT IN QUARANTINE…………….…..….. 84
6.8 NECROPSY……………………………………………………………………………..……..……... 85
6.9 TESTING AND TREATMENT FOR INTERNAL PARASITES………………………..…….…...…. 86
6.10 TESTING AND TREATMENT FOR AMPHIBIAN CHYTRID FUNGI……..….….….………….. 88
6.11 TESTING FOR RANAVIRUS…………………………………………………………....…….……. 93
6.12 CLINICAL PATHOLOGY (HEMATOLOGY AND CLINICAL BIOCHEMISTRY)……..…………. 96
6.13 OTHER INFECTIOUS DISEASES ENCOUNTERED IN QUARANTINE……………..….….…….97
6.14 REFERENCES…………………………………………………………………………………….….. 101

7. DIAGNOSTIC TESTING………………………………………………………………………………………….…105
7.0 INTRODUCTION…………………………………………………….……………………….….…… 106
7.1 DISEASE SURVEILLANCE………………………………………….…………………………..…… 103
7.2 EVALUATION OF LABORATORIES THAT PERFORM MOLECULAR
DIAGNOSTIC TESTING………………………………………….……………..………..……. 110
7.3 DIAGNOSTIC TESTING FOR AMPHIBIAN CHYTRID FUNGI (BATRACHOCHYTRIUM
DENDROBATIDIS AND BATRACHOCHYTRIUM SALAMANDRIVORANS)………….. …. 115
7.4 RANAVIRUS INFECTION…………………………………………………….…..……….……….. 132
7.5 PERKINSUS-LIKE PROTOZOAL DISEASE OF RANID FROGS…………………..…….………. 142
7.6 DISEASE REPORTING AND PARTICIPATION IN DISEASE DATABASES.………..….………. 145
7.7 TESTING AND SURVEILLANCE FOR FECAL PARASITES………………….…..…..…..…….… 146
7.8 HEMATOLOGY AND SERUM BIOCHEMISTRY……………………………….…….…………... 155
7.9 LABORATORIES THAT ACCEPT AMPHIBIAN SAMPLES………………..….…….…………... 157
7.10 REFERENCES…………………………………………………………..……………….……………159

8. TREATMENT AND CONTROL OF DISEASES……………………………………..……………….…..……... 167
8.0 INTRODUCTION………………………………………………………………………….……….…..167
8.1 TREATMENT AND CONTROL METHODS FOR AMPHIBIAN CHYTRID FUNGI…..………….168
8.2 CONTROL OF RANAVIRUS INFECTIONS…………………………………………………..….... .180
8.3 CREATING SPECIFIC-PATHOGEN-FREE AMPHIBIAN POPULATIONS……….…………….. 182
8.4 PARASITE MONITORING AND TREATMENT………………………………….…………..…… 186
8.5 ELECTROLYTE FORMULAS…………………………………………………………..………..….. 190
8.6 EUTHANASIA…………………………………………………………………………..……….….. 190
8.7 REFERENCES…………………………………………………………………………..…….……… 191

9. NECROPSY…………………………………………………………………………………………………..……..195
9.0 INTRODUCTION………………………………………………………………………….…………. 195
9.1 NECROPSY PREPARATION…………………………………………….…………..……………… 196
9.2 MATERIALS NEEDED…………………………………………………..………………………….. 196
9.3 PERFORMING THE NECROPSY……………………………………….………………………….. 197
9.4 ANATOMICAL NOTES FOR AMPHIBIAN NECROPSIES………….…………….……………… 201
9.5 SAMPLE COLLECTION DURING MORTALITY EVENTS……………….……..……………….. 204
9.6 SHIPMENT OF SAMPLES………………………………………………………..….……………… 206
9.7 REFERENCES………………………………………………………………..…….…………………. 206

APPENDIX 1: METHODS TO INDIVIDUALLY IDENTIFY THE AMPHIBIAN SPECIMEN……………….…. 208
APPENDIX 2: WASTEWATER TREATMENT......................................................................... 227
APPENDIX 3: ISOLATED AMPHIBIAN ROOMS AT OMAHA’S HENRY DOORLY ZOO…………...……. 231
APPENDIX 4 DISEASE RISK ANALYSIS AND HEALTJ SURVEILLANCE FOR THE SPECIES RECOVERY
PROGRAMME ……………..……………………………………………………………………………..…………240
CHAPTER 1

INTRODUCTION

The International Union for the Conservation of Nature (IUCN) has estimated
that 30% of all amphibian species are threatened with extinction, and at least 484
species are Critically Endangered (http://www.iucnredlist.org). The major contributing
factor of the most drastic amphibian population declines is the disease chytridiomycosis
caused by amphibian chytrid fungi, specifically Batrachochytrium dendrobatidis and
Batrachochytrium salamandrivorans. These fungi, disseminated by anthropogenic
means, can reduce amphibian biodiversity at new locations in alarmingly short periods
of time. Thus, understanding and controlling infectious diseases such as
chytridiomycosis have become a major focus of both in situ and ex situ amphibian
conservation efforts worldwide.
To formulate a response to the crisis of global amphibian extinctions, the Species
Survival Commission (SSC) of the IUCN convened an international group of leading
amphibian biologists at an Amphibian Conservation Summit resulting in the Amphibian
Conservation Action Plan (ACAP; this document is available for download at
http://www.amphibians.org). The ACAP concluded that the amphibian extinction crisis
“requires a global response at an unprecedented scale from governments, corporations,
civil society and the scientific community.” The ACAP calls for creation of survival
assurance colonies that bring representatives of critically endangered amphibian species
into captivity for safekeeping. Although remediation of factors contributing to species
extinction is preferable to captivity for endangered animals, the rapidity of amphibian
declines makes it likely that innumerable species will be lost long before suitable
solutions are discovered. Therefore, the critical role of survival assurance colonies is to
preserve the option of re-introducing species to their native habitat at a later time. The
global effort to develop such programs is coordinated by the Amphibian Ark, a joint
effort of the World Association of Zoos and Aquariums (WAZA), the IUCN/SSC,
Conservation Breeding Specialist Group (CBSG) and Amphibian Specialist Group
(www.amphibianark.org). The Amphibian Ark estimates 500 amphibian species require
immediate ex situ intervention through establishment of survival assurance colonies.
This is a massive undertaking that calls upon the resources of a wide variety of
institutions including zoos, aquariums, botanical gardens, natural history museums,
governmental agencies, and universities. Yet, as institutions prepare for the “call to
action” from the Amphibian Ark and IUCN, there is a global reality that the field of
amphibian husbandry in general is still very much in its infancy and expertise in
amphibian disease control is limited to a very few institutions worldwide.
In response to urgent needs created by the rapid expansion of amphibian
survival assurance colonies worldwide, guidelines for Best Practices for permanent
quarantine and facility biosecurity were drafted as part of the CBSG/WAZA Amphibian
Ex Situ Conservation Planning Workshop held in El Valle, Panama, in 2006 (proceedings
available at http://www.amphibianark.org/downloads.htm). In addition, the final report
of the Panama workshop outlined additionally needed Action Steps to develop medical
protocols for health screening, prophylactic treatment of important infectious diseases
and disease surveillance in captive amphibian collections. Elements of these Action
Steps have already emerged since 2006 amidst a flood of new information in the
primary literature on amphibian disease testing, disease control protocols, and the
development of creative approaches to meeting strict biosecurity guidelines within
existing ex situ institutional facilities and culture. The present manual aims to formally
address these Action Steps by bringing together recommendations from husbandry
experts and veterinarians with expertise in amphibian medicine and disease risk
assessment to consolidate new information into simple consensus-based protocols that
can be easily adapted by the wide variety of institutions that maintain captive
amphibians.
This manual is intended to serve as a “one-stop shopping” resource for basic
consensus based recommendations concerning amphibian quarantine, necropsy, facility
biosecurity, facility hygiene & disease treatment, and disease screening & surveillance.
These are to be viewed as guidelines and recommendations and not as mandates or
requirements; no simple manual can take into account the specifics of individual
programs and needs. Importantly, this manual is not intended as a replacement for a
complete text on amphibian veterinary medicine (e.g., Amphibian Medicine and Captive
Husbandry, by K. M. Wright & B. R. Whitaker, Malabar Publ., 2001). Similarly, although
the topics of nutrition and basic husbandry are discussed throughout, this manual is
intended merely as a complement to existing texts on amphibian husbandry, such as the
Amphibian Husbandry Resource Guide, Edition 1.0 (Association of Zoos and Aquariums
Publ., 2008) or species-specific protocols such as that prepared for the Panamanian
golden frog (http://www.ranadorada.org/species-info.html). This manual is intended to
be a detailed technical resource for veterinarians and senior staff in ex situ or
conservation programs, to guide them as they develop new or additional amphibian
programs, design routine screening protocols or encounter infectious disease issues in
current collections, or move animals among collections or reintroduce them into the
wild. This manual will be iterative, with subsequent editions made available following an
additional workshop planned for 2011, and occasionally thereafter.
We hope users from all backgrounds find this manual to be a useful primary
resource and, importantly, a convenient portal into the primary literature on amphibian
veterinary science and diseases. The amphibians of the world are in need of
considerable assistance to avoid additional extinctions, and we hope this manual
expedites some of that needed assistance.

Allan P. Pessier
Joseph R. Mendelson III
25 December, 2009
CHAPTER 2

LIST OF PARTICIPANTS

The following persons contributed directly to the final content that appears in
this version of the manual. Most of them participated in the workshop, held at San Diego
Zoo in February, 2009, and added important comments to drafts of various chapters as
they neared completion.

Diane Barber, Fort Worth Zoo
Eric Baitchman; Zoo New England
Don Boyer, San Diego Zoo
Onnie Byers, IUCN Conservation Breeding Specialist Group
Leigh Ann Clayton, National Aquarium Baltimore
Josh Cook, University of Georgia
Ann Duncan, Detroit Zoo
Ryan DeVoe, North Carolina Zoo
Shannon Ferrell, Fort Worth Zoo
Ron Gagliardo, Amphibian Ark & Zoo Atlanta
Michael Garner, Northwest ZooPath
Richard Gibson, Amphibian Ark & Chester Zoo
Andrea Johnson, San Diego Zoo
Megan Jones, San Diego Zoo
Kim Lovich, San Diego Zoo
Bob Mailloux, Sandfire Dragon Ranch
Denise McAloose, Wildlife Conservation Society-Bronx Zoo
Joseph Mendelson, Zoo Atlanta
Pat Morris, San Diego Zoo
Isamara Navarrete, San Diego Zoo
R. Andrew Odum, Toledo Zoo
Brad Lock, Zoo Atlanta
Samuel Rivera, Zoo Atlanta
Allan Pessier, San Diego Zoo
Jennifer Pramuk, Wildlife Conservation Society-Bronx Zoo
Michael Ready, Sandfire Dragon Ranch
Oliver Ryder, San Diego Zoo
Danna Schock, University of Calgary
Mark Schrenzel, San Diego Zoo
Lee Skerratt, James Cook University
Edythe Sonntag, Michigan State University
Simone Vitali, Perth Zoo
Carmel Witte, San Diego Zoo
Kevin Wright, Arizona Exotic Animal Hospital
Kevin Zippel, Amphibian Ark