49th Scientific Symposium of the UJNR Aquaculture Panel
PROGRAM and ABSTRACTS
Theme: Control and Management of Aquaculture Disease
Dates/Times Eastern Time: November 2nd and 3rd, 2021; 7:00 to 9:00 p.m.
Dates/Times Japan Time: November 3rd and 4th, 2021; 8:00 to 10:00 a.m.
Location: Online with the Microsoft Teams link for both days HERE
Aim of the Symposium
The US-Japan Natural Resources (UJNR) Aquaculture Panel is a cooperative research exchange,
between the US’ National Oceanic and Atmospheric Administration (NOAA) and Japan’s
Fisheries Research and Education Agency (FRA), jointly addressing environmental and technicalissues that affect the aquaculture industries of both nations.
The 2021, 49th UJNR Aquaculture Scientific Symposium is the first year of the current threeyear theme of “Control and Management of Aquaculture Disease”. Commercial aquaculture
must manage pests and pathogens that can cause disease and negatively impact animal wellbeing and economic productivity. Investigation and identification of effective controls,
mitigation measures, and treatments of disease is therefore a top industry and government
priority in Japan and the US.
Program
Note: All talks are scheduled for 20 minutes, which includes 15 minutes for presentation and 5
minutes for questions.
Tuesday, November 2, 2021 (Eastern); Wednesday, November 3, 2021 (Japan)
Welcome and Aim of the Symposium
Michael Rust, US Panel Chair, NOAA Fisheries Office of Aquaculture

19:00-19:05 (E)08:00-08:05 (J)Session 1: Disease Control(Moderators: Satoshi Miwa, Caird Rexroad)
Isolation and Characterization of Hirame Aquareovirus (HAqRV):A New Aquareovirus Isolated From Diseased Hirame Paralichthys
olivaceus
1

19:05-19:25 (E)08:05-08:25 (J)Yasuhiko Kawato, Fisheries Technology Institute, FRA
Weissellosis: An Emerging Disease of Farmed Rainbow Trout
Tim Welch, National Center for Cool and Cold Water Aquaculture,
USDA/ARS

19:25-19:45 (E)08:25-08:45 (J)Development of a Method to Quantify Endogenous IFNγ Protein in
Amberjack Species
Yuta Matsuura, Fisheries Technology Institute, FRA

19:45-20:05 (E)08:45-09:05 (J)Use of a Flavobacterium columnare DnaK Recombinant Protein Vaccine
to Guard Against Columnaris Disease in Channel Catfish
Miles Lange, Aquatic Animal Health Research Unit, USDA/ARS

20:05-20:25 (E)
09:05-09:25 (J)α-glucosidase Activity of Nocardia seriolae Isolated From Seawater Fishin Japan
Yoshiko Shimahara, Fisheries Technology Institute, FRA

20:25-20:45 (E)09:25-09:45 (J)Emergent Pathogens in Aquaculture: How Should We Identify Them?
Linda Rhodes, Northwest Fisheries Science Center, NOAA

20:45-21:05 (E)09:45-10:05 (J)Wednesday, November 3, 2021 (Eastern); Thursday, November 4, 2021 (Japan)
Session 2: Education and Disease Management(Moderators: Satoshi Miwa, Luke Gardner)
The Role of National Fisheries University and Studies of Fish Disease
Shinya Yasumoto, National Fisheries University, FRA

19:00-19:20 (E)
08:00-08:20 (J)Aquaculture Medicine in the 21st Century: Training the VeterinaryCommunity to Meet the Needs of a Global Industry
Iskande Larkin, University of Florida

19:20-19:40 (E)
08:20-08:40 (J)Epitope Mapping of Anti-RSIV Monoclonal Antibody Using a Phage
Display Library
Tomokazu Takano, Fisheries Technology Institute, FRA

19:40-20:00 (E)
08:40-09:00 (J)Managing the Threat of Emerging Diseases in Bivalve Mollusc
Aquaculture
Ryan Carnegie, Virginia Institute of Marine Science, College ofWilliam & Mary

20:00-20:20 (E)
09:00-09:20 (J)Advancing Epidemiological Tools for Marine Disease Management –Modeling Dispersion of OsHV-1μvar in Pacific Oyster Populations
Meghana Parikh, Northeast Fisheries Science Center, NOAA

20:20-20:40 (E)
09:20-09:40 (J)2Using a OneHealth Approach to Evaluate How Marine Pathogens andParasites That Are Infectious to Cultured Finfish, Shellfish, andMacroalgae Within the United States May Be Affected by Climate
ChangeMichelle Rub, Northwest Fisheries Science Center, NOAA

20:40-21:00 (E)
09:40-10:00 (J)Science Symposium Closing
Hideaki Aono, Japan Panel Chair, FRA

21:00-21:05 (E)
10:00-10:05 (J)List of Participants
UNITED STATES:
Name
Panel
Dr. Michael Rust
Dr. Luke Gardner
Dr. Caird Rexroad
Mr. Clete Otoshi
Speakers
Dr. Linda Rhodes
Dr. Michelle Rub
Dr. Meghana Parikh
Dr. Miles Lange
Dr. Tim Welch
Dr. Ryan Carnegie
Dr. Iske Larkin
Invited to Attend
Ms. Danielle Blacklock
Mr. David O’Brien
Dr. Seth Theuerkauf
Ms. Marifrancis Moschopoulos
Dr. Janet Whaley
Dr. John Shelley
Dr. Penny Swanson
Dr. William Fairgrieve
Mr. Kellen Parrish
Dr. Kathleen Hartman
Dr. Alicia Marston
Dr. Thomas Waltzek
Dr. Marvirstine Briggs-Fisher

Affiliation
NOAA Fisheries, Office of Aquaculture
California Sea Grant
USDA, ARSNOAA Fisheries, Office of Aquaculture
NOAA Fisheries, NWFSCNOAA Fisheries, NWFSCNOAA Fisheries, NEFSC
USDA, ARS, Aquatic Animal Health Research Unit
USDA, ARS, National Center for Cool and Cold Water
Aquaculture
College of William & Mary, Virginia Institute of Marine
Science
University of Florida, College of Veterinary Medicine
NOAA Fisheries, Office of Aquaculture
NOAA Fisheries, Office of Aquaculture
NOAA Fisheries, Office of Aquaculture
NOAA Fisheries, Office of Aquaculture
NOAA Fisheries, Office of International Affairs andSeafood Inspection and Office of Aquaculture
NOAA Fisheries, Office of International Affairs andSeafood Inspection
NOAA Fisheries, NWFSCNOAA Fisheries, NWFSCNOAA Fisheries, NWFSC
USDA, APHISUSDA, APHISUSDA, APHISUSDA, APHIS
3

Dr. Lori Gustafson
Dr. Christine Ellis
Dr. Tim Leeds
Dr. Gary Richards
Dr. Michael Deshotel
Dr. Brett Dumbauld
Dr. Dina Proestou
Dr. Neil Thompson
Dr. Benjamin LaFrentz
Dr. Ben Beck
Dr. Timothy Sullivan
Dr. Jennifer Matysczak
Dr. Paul Wills

USDA, APHISUSDA, APHISUSDA, ARS
USDA, ARS
USDA, ARS
USDA, ARS
USDA, ARS
USDA, ARS
USDA, ARS
USDA, ARS
USDA, National Institute of Food and Agriculture
FDA, Center for Veterinary Medicine
Florida Atlantic University, Harbor Branch

NOAA Fisheries: National Oceanic and Atmospheric Administration, National Marine FisheriesServiceNWFSC: Northwest Fisheries Science Center
NEFSC: Northeast Fisheries Science Center
USDA: United States Department of Agriculture
ARS: Agricultural Research Service
APHIS: Animal and Plant Health Inspection Service
JAPAN:
Name

Affiliation

Panel
Dr. Aono Hideaki
Dr. Gen Kouichiro
Dr. Ozaki Akiyuki
Dr. Takano Masatsugu
Dr. Oikawa Hiroshi
Arai Daisuke
Dr. Yamano Keisuke
Dr. Nakayasu Chihaya
Dr. Okumua Takuji
Saito Yoho
Dr. Kamaishi Takashi
Dr. Ito Takafumi
Dr. Miwa Satoshi
Ishihi Yuka
Dr. Shibuno Takuro

FRA, FTIFRA, FTI, Planning and Coordination Department
Ministry of Agriculture, Forestry and Fisheries, Fisheries Agency
FRA Headquarters, General Planning and Coordination Department
FRA, FTI, Planning and Coordination Department
FRA, FTI, Planning and Coordination Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA Headquarters, General Planning and Coordination Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Management Department
FRA, FTI, Management Department

Speakers
Dr. Kawato YasuhikoDr. Matsuura Yuta
Dr. Shimahara Yoshiko

FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department

4Dr. Yasumoto Shinya
Dr. Takano Tomokazu

FRA, NFU
FRA, FTI, Aquaculture Research Department

Invited to Attend
Dr. Urawa Shigehiko
Dr. Sakai Takamitsu
Dr. Ishida Noriko
Dr. Matsuyama Tomomasa
Yoshino Tomoaki
Dr. Sato Jun
Dr. Maeda Tomoki
Nakagawa Yukihiro
Dr. Umeda Kousuke
Dr. Kiryu Ikunari
Dr. Yuasa Kei
Dr. Kurobe Tomofumi
Dr. Mekata Toru
Dr. Inada Mari
Dr. Yoshinaga Tomoyoshi
Dr. Itoh Naoki
Dr. Karakawa Nanae
Kawahara Miku
Kuboyama Azusa
Vince Neil Fuertes
Dr. Sano Motohiko
Dr. Hirono Ikuo
Dr. Kondo Hidehiro
Dr. Kato Goshi
Dr. Matsumoto Moe
Lah Lik Ming
Dr. Shirakashi Sho
Dr. Matsuo Kazutoshi
Daikai Takateru
Ota Chiharu
Nobuhisa Tsuchiya
Asato Masaki
Omine Risako

FRA, FRI, Salmon Research Department
FRA, FTI, Planning and Coordination Department
FRA, FTI, Environment and Fisheries Applied Techniques Research
Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
FRA, FTI, Aquaculture Research Department
Graduate School of Agricultural and Life Sciences, The University ofTokyo
Graduate School of Agricultural and Life Sciences, The University ofTokyo
Graduate School of Agricultural and Life Sciences, The University ofTokyo
Graduate School of Agricultural and Life Sciences, The University ofTokyo
Graduate School of Agricultural and Life Sciences, The University ofTokyo
Graduate School of Agricultural and Life Sciences, The University of
Tokyo
Tokyo University of Marine Science and Technology
Tokyo University of Marine Science and Technology
Tokyo University of Marine Science and Technology
Tokyo University of Marine Science and Technology
Tokyo University of Marine Science and Technology
Tokyo University of Marine Science and Technology
Aquaculture Research Institute, Kindai University
Animal Products Safety Division, Ministry of Agriculture, Forestry and
Fisheries
Food Safety Policy Division, Ministry of Agriculture, Forestry and
Fisheries
Food Safety Policy Division, Ministry of Agriculture, Forestry and
Fisheries
International Research Collaboration Division, Agriculture, Forestry and
Fisheries Research Council Secretariat
Fishery and Ocean Technology Center, Okinawa Prefecture
Fishery and Ocean Technology Center, Okinawa Prefecture5

Mizutani Kouta

Ibaraki Prefectural Fisheries Research Institute

FRA: Japan Fisheries Research and Education Agency
FTI: Fisheries Technology Institute
NFU: National Fisheries University
FRI: Fisheries Resource Institute6

Abstracts and Annotated Bibliographies
Abstract Index
Tuesday, November 2, 2021 (Eastern); Wednesday, November 3, 2021 (Japan)
Isolation and Characterization of Hirame Aquareovirus
19:05-19:25 (E);
Y. Kawato(HAqRV): A New Aquareovirus Isolated From
08:05-08:25 (J)Diseased Hirame Paralichthys olivaceus
19:25-19:45 (E);
Weissellosis: An Emerging Disease of Farmed Rainbow
T. Welch
08:25-08:45 (J)Trout
19:45-20:05 (E); Y.
Development of a Method to Quantify Endogenous
08:45-09:05 (J) Matsuura
IFNγ Protein in Amberjack Species
Use of a Flavobacterium columnare DnaK
20:05-20:25 (E);
M. Lange
Recombinant Protein Vaccine to Guard Against
09:05-09:25 (J)Columnaris Disease in Channel Catfish
20:25-20:45 (E); Y.
α-glucosidase Activity of Nocardia seriolae Isolated
09:25-09:45 (J) Shimahara
From Seawater Fish in Japan
20:45-21:05 (E);
Emergent Pathogens in Aquaculture: How Should We
L. Rhodes
09:45-10:05 (J)Identify Them?
Wednesday, November 3, 2021 (Eastern); Thursday, November 4, 2021 (Japan)
19:00-19:20 (E); S.
The Role of National Fisheries University and Studies
08:00-08:20 (J) Yasumotoof Fish Disease
Aquaculture Medicine in the 21st Century: Training the
19:20-19:40 (E);
Veterinary Community to Meet the Needs of a Global
I. Larkin
08:20-08:40 (J)Industry
19:40-20:00 (E);
Epitope Mapping of Anti-RSIV Monoclonal Antibody
T. Takano
08:40-09:00 (J)Using a Phage Display Library
20:00-20:20 (E);Managing the Threat of Emerging Diseases in Bivalve
R. Carnegie
09:00-09:20 (J)Mollusc Aquaculture
Advancing Epidemiological Tools for Marine Disease
20:20-20:40 (E);Management – Modeling Dispersion of OsHV-1μvar in
M. Parikh
09:20-09:40 (J)Pacific Oyster Populations
Using a OneHealth Approach to Evaluate How Marine
Pathogens and Parasites That Are Infectious to Cultured
20:40-21:00 (E);
M. Rub
09:40-10:00 (J)Finfish, Shellfish, and Macroalgae Within the UnitedStates May Be Affected by Climate Change

7

7
9
11
13
15
16

19
21
24
26
28

30

Isolation and Characterization of Hirame Aquareovirus (HAqRV): A New Aquareovirus
Isolated From Diseased Hirame Paralichthys olivaceus.
Yasuhiko Kawato*1, Toru Mekata1, Toyohiro Nishioka2, Ikunari Kiryu1, Takamitsu Sakai3,
Tomoki Maeda4, Satoshi Miwa4, Kanae Koike5, Masahiro Sadakane6, Koh-ichiro Mori3
1

Nansei Field Station, Fisheries Technology Institute, Japan Fisheries Research and Education
Agency, Mie 519-0193, Japan
2
Yashima Field Station, Fisheries Technology Institute, Japan Fisheries Research and Education
Agency, Kagawa 761-0111, Japan.
3
Nagasaki Field Station, Fisheries Technology Institute, Japan Fisheries Research and Education
Agency, Nagasaki 851–2213, Japan.
4
Tamaki Field Station, Fisheries Technology Institute, Japan Fisheries Research and Education
Agency, Mie 519-0423, Japan
5
Technical Center, Hiroshima University, Japan
6
Department of Applied Chemistry, Graduate School of Advanced Science and Engineering,
Hiroshima University, Japan*E-mail: ykawato@affrc.go.jp
Hirame or Japanese flounder (Paralichthys olivaceus) is an important flatfish for aquaculture
in Japan. As a part of stock enhancement effort, large numbers of hirame juveniles are released
annually by public corporations, and hence, a total of over 20 million juveniles are produced every
year in more than 100 hatcheries in Japan. Mass mortalities of hirame larvae or juveniles due toreovirus-like infection have been reported since the 2000s. Currently no effective protection
measures against the disease have been established because the causative agent has not been
identified so far. The goal of present study is to identify the virus causing the mass mortality inhirame juveniles by Koch’s postulates. We successfully isolated a novel Aquareovirus (hirame
aquareovirus: HAqRV) from diseased fish using hirame natural embryo (HINAE) cell line. A
cytopathic effect characterized by aggregation of the virus-infected cells was observed 20 days
after virus inoculation. In electron microscopy, the spherical virion (75 nm in diameter) with multilayered capsid structure was observed. The viral genome consisted of 11 segments, and regions
encoding 7 virion structural proteins (VP1 - VP7) and 5 non-structural proteins (NS1 - NS5) werepredicted. The deduced amino acid sequences of those proteins were highly similar to those of theviruses of genus Aquareovirus. The terminal sequences (5’-GUUUUA and UCAUC-3’) of eachsegment were conserved among the 11 segments and they were identical to those of Aquareovirus
A and G. However, the similarities of the complete genome sequence between the HAqRV andother aquareoviruses were less than 60% at nucleotide level. Phylogenetic analyses based on the
deduced amino acid sequences of VP2 (RNA dependent RNA polymerase), VP7 (outer capsid
protein), and NS5 (fusion associated transmembrane protein) suggest that the HAqRV is notclassified into the known species of Aquareovirus. Pathogenicity of HAqRV was clearly
demonstrated in accordance with Koch’s postulates by by experimental challenge tests using
hirame juveniles at 20 days post hatching. HAqRV had specific tissue tropism to the epitheliumof the digestive tract including hepatocytes and caused syncytia in the infected tissues. The results
suggest that the HAqRV is likely a new Aquareovirus species which is highly virulent for thehirame at early life stages. We believe that the data from this study will significantly contribute todevelop protection measures against HAqRV, which is a big threat to hirame hatcheries in Japan.
8For example, qPCR assay developed in this study can be used for rapid diagnosis and investigating
the route of infection. The cell culture system can be used for validating the disinfectant conditions.
This project is ongoing, and we have been working hard to achieve a goal of development of theprotection measures against the disease and improve hirame production in Japan.
Annotated Bibliography of Key Works
Isshiki, T., Nagano, T., Abe, M., Miyazaki, T. 2003. Histopathological changes probably
associated with a virus in larval and juvenile marbled sole Pleuronectes yokohamae. Fish. Pathol.,
38:143–149.
The seed production of marbled sole Pleuronectes yokohamae has been attempted at a hatchery in
Kagawa Prefecture, Japan since 1989, while a subacute disease with a considerable amount ofmortality occurred in larval and juvenile fish (24-69 days old). The evident histopathologicalfeatures were the formation of syncytia and necrosis in the liver, renal tubules, pancreas, andintestinal epithelium. Electron microscopy of affected hepatocytes revealed formation of inclusion
bodies consisting of crystalline arrays of virus particles (75-80 nm in diameter) whichmorphologically resemble reovirus. Although aquareovirus infection was suspected in the reported
cases, the causative agent could not be identified as virus isolation failed.
Blindheim, S., Nylund, A., Watanabe, K., Plarre, H., Erstad, B., Nylund, S. 2015. A newaquareovirus causing high mortality in farmed Atlantic halibut fry in Norway. Arch. Virol.,
160:91–102.
A new aquareovirus (Atlantic halibut reovirus: AHRV) was isolated from cultured Atlantic halibut
Hippoglossus hippoglossus fry at a facility in Norway where massive mortalities had occurred
during the start-feeding phase. The virus replicated in BF-2 and CHSE-214 cell cultures andproduced syncytia and plaque-like cytopathic effects. AHRV was associated with necrosis of theliver and pancreas, and syncytium formation in these tissues in halibut fry. Transmission electron
microscopy revealed that the viroplasm contained virions, non-enveloped, icosahedral particles
approximately 70 nm in diameter with a double capsid layer, amorphous material, and tubular
structures. The RNA-dependent RNA polymerase (RdRp) gene from the AHRV isolates showed
the highest amino acid sequence identity (80 %) to an isolate belonging to the species Aquareovirus
A. A similar disease of halibut fry has also been described in halibut production facilities in Canada
and Scotland.
Kawato, Y., Mekata, T., Nishioka, T., Kiryu, I., Sakai, T., Maeda, T., Miwa, S., Koike, K.,
Sadakane, M., Mori, KI. 2021. Isolation and characterization of hirame aquareovirus (HAqRV):A new Aquareovirus isolated from diseased hirame Paralichthys olivaceus. Virology, 559:120–130.
We isolated a novel Aquareovirus (hirame aquareovirus: HAqRV) from hirame or Japaneseflounder Paralichthys olivaceus suffering from reovirus-like infection. Complete genome
sequence of the isolated virus suggested that HAqRV is likely a new Aquareovirus species.
Pathogenicity of HAqRV was clearly demonstrated in accordance with Koch's postulates byexperimental infection using hirame juveniles. The present study would contribute to develop
protection measures against the infection with HAqRV, which is a big threat to the hatcheryproduction of Japanese flounder in Japan.

9

Weissellosis: An Emerging Disease of Farmed Rainbow Trout
Timothy J. Welch
National Center for Cool and Cold Water Aquaculture, Agricultural Research Service/U.S.
Department of Agriculture, Kearneysville, WV, 25430., USA.
E-mail: tim.welch@usda.gov
Weisselosis, caused by the gram-positive lactic acid bacterium Weissela ceti, is an emerging
infectious disease of farmed rainbow trout. This disease was first recognized in China in 2007 but
was soon after found in Brazil in 2008, in the United States and Japan both in 2011, and more
recently in Mexico in 2015, Colombia in 2016, Canada in 2017, and Peru in 2018. This disease isof major concern as losses can be as high as 40-80% of production and thus results in significant
loss of animals with coinciding economic impact. Genome sequence analysis of W. ceti strainsfrom the US, Brazilian, and Japanese outbreaks revealed striking genetic similarity between strainsfrom these geographically diverse sites and suggests emergence from a common source. This is
surprising given the geographical distance between these sites and a lack of obvious
epidemiological links between farms and countries. In contrast, the Canadian outbreak strain isdistinct from the US, Japanese and Brazilian isolates and suggests that this strain represents an
independent emergence of this pathogen. Injection vaccination with formalin-killed whole-cell
preparations of W. ceti provides excellent protection against experimental challenge when tested
under laboratory conditions. Recent work has also demonstrated that vaccination induces a robust
IgM antibody response and that serum factors provide protection against experimental challenge,
suggesting a role for specific IgM in vaccine-induced protection. In practice, vaccination has beenhighly successful in controlling this disease in both North Carolina and Canada. Gaining a better
understanding of the origins of this pathogen and the epidemiological pathways that havefacilitated its rapid dissemination to highly disparate locations will be critical for controlling its
further spread.
Annotated Bibliography of Key Works
Welch, TJ, Good, CM. Mortality associated with Weissellosis (Weissella sp.) in USA farmed
rainbow trout: Potential for control by vaccination. Aquaculture. 388-391:122-127.
This was the first report of this emerging pathogen in North Carolina, USA. This paper also
reported the development and validation of an efficacious vaccine against W. ceti. This vaccine
was additionally shown to be effective when mixed with another vaccine used in North Carolina
thus demonstrating that these vaccines can be delivered together minimizing the increased
production costs and negative effects caused by multiple injections. Vaccination has been used tocontrol W. ceti in North Carolina since 2012. Early detection of this pathogen and rapid
development and implementation of this vaccine aided in control efforts and reduced the likelihoodof further pathogen dissemination in the US.
Figueiredo HC, Soares SC, Pereira FL, Dorella FA, Carvalho AF, Teixeira JP, Azevedo VA, Leal
CA. Comparative genome analysis of Weissella ceti, an emerging pathogen of farm-raised rainbow
trout. BMC Genomics. 2015.
This paper presents whole genome comparisons between Brazilian and US strains of W. ceti andwas first to demonstrate the remarkable similarity in the genetic structure of these W. ceti strains10

despite their lack of epidemiological linkage. Several genomic islands and putative virulence genes
were also identified including adhesins, hemolysins, and antibiotic resistance genes. This work
provided important insight into the pathogenesis, evolution, and epidemiology of this emerging
pathogen.
Snyder AK, Hinshaw JM, Welch TJ. Diagnostic tools for rapid detection and quantification of
Weissella ceti NC36 infections in rainbow trout. Lett Appl Microbiol. 2015 Feb; 60(2):103-110.
This paper describes the development and validation of rapid and quantitative PCR-based assays
for the identification of W. ceti from diseased fish and other environmental samples. The assays
were validated in a controlled experimental challenge and in confirming the diagnosis of
Weissellosis in a commercial rainbow trout farm during an outbreak investigation. The assays
allow rapid Weissellosis diagnosis, enhanced pathogen surveillance for epidemiological studies.
Mitomi K, Hoai TD, Nishiki I, Yoshida T. First isolation of Weissella ceti responsible foroutbreaks of weissellosis in farmed rainbow trout in Japan. J Fish Dis. 2018 May; 41(5):847-850.
These authors describe the identification of W. ceti in cultured trout in Japan. It was revealed thatJapanese outbreaks coincided with outbreaks occurring in the US and that the strains were
phenotypically very similar to US strains.

11Development of a Method to Quantify Endogenous IFNγ Protein in Amberjack Species
Yuta Matsuura*, Tomokazu Takano, Tomomasa Matsuyama
Nansei Field Station, Fisheries Technology Institute, Japan Fisheries Research and Education
Agency, Mie 516-0193, Japan*E-mail: matsuura_yuta53@fra.go.jp
Interferon gamma (IFNγ) is a pivotal cytokine that promotes and orchestrates adaptive cellmediated immunity against intracellular pathogens. The capacity of T lymphocytes in mammals
to produce IFNγ has been measured using specific antibodies to analyze cell-mediated immune
responses against infection or immuno-stimulants such as vaccines. However, measurement of fish
IFNγ protein has not been possible due to a lack of research tools. The development of vaccines
for intracellular fish pathogens that are thought to be targeted by cell-mediated immunity has beena difficult task in such a situation. Serious economic losses due to Nocardiosis caused by an
intracellular bacterium “Nocardia seriolae” have occurred in the aquaculture of amberjack species
in Japan. The method to quantify IFNγ of amberjack species is required to develop vaccines for
Nocardiosis.
To develop the method for quantification of IFNγ, we firstly generated antibodies that reactwith endogenous amberjack IFNγ. A monoclonal antibody (mAb) and a polyclonal antibody (pAb)that specifically react with endogenous IFNγ produced by Japanese amberjack Seriola
quinqueradiata and greater amberjack Seriola dumerili were established. We then constructed asandwich enzyme-linked immunosorbent assay (ELISA) for the IFNγ of the species using theantibodies. The sandwich ELISA could detect the endogenous IFNγ at concentrations less than
100 pg/mL in leukocytes culture supernatant without cross-reactivity with another cytokine (IL4/13). Using the ELISA, IFNγ produced by Japanese amberjack leukocytes were analyzed. The
level of IFNγ secreted by stimulated leukocytes isolated from N. seriolae-infected fish wassignificantly higher than that secreted by cells of naïve fish (p<0.05) and was lasted a long time(at least eight months) following the infection. A portion of the fish that survived for eight months
after infection were re-challenged with N. seriolae, and all re-challenged fish survived, whereas
100% of control fish died of the infection. Thus, the fish that had recovered from N. seriolae
infection were considered to have acquired cell-mediated immunity to the organism.
In conclusion, we established a new sandwich ELISA method that enables for the first time
the quantification of IFNγ at the protein level in teleost. Our results obtained using the methodsuggest that the developed method would help evaluate the capacity of cell-mediated immunity torespond against intracellular pathogen infection. Assessment of IFNγ expression at the protein
rather than genetic level could provide information that would enhance understanding of theimmunologic process and contribute to vaccine development.
Annotated Bibliography of Key Works
Matsuura, Y., Takano, T., Matsuyama, T., Sakai, T., Terashima, S., Nakayasu, C. 2020.
Development of a method to quantify endogenous IFNγ protein in amberjack species. Fish
Shellfish Immunol., 107 (A):251-259.
The authors established the method to quantify a cytokine IFNγ, an important marker of cellmediated immune responses in fish for the first time. The established method could contribute to12the vaccine development against Nocardiosis that has caused serious damages in amberjack
farming in Japan.
Matsuura, Y., Takasaki, M., Miyazawa, R., Nakanishi, T. 2017. Stimulatory effects of heat-killedEnterococcus faecalis on cell-mediated immunity in fish. Dev. Comp. Immunol., 74:1-9.
The paper reported that heat-killed lactic acid bacteria Enterococcus faecalis enhanced cellmediated immunity in fish. The heat-killed bacteria triggered the induction of Th1 responses andincreases in CD4+ Th lymphocytes and effector cells including CD8+ lymphocytes andmacrophages. Thus, the heat-killed one is a promising vaccine adjuvant to induce cell-mediatedimmunity in fish and could contribute to the studies of vaccines against Nocardiosis.
Matsumoto, M., Araki, K., Hayashi, K., Takeuchi, K., Shiozaki, K., Suetake, H., Yamamoto M.
2017. Adjuvant effect of recombinant interleukin-12 in the Nocardiosis formalin-killed vaccine of
the amberjack Seriola dumerili. Fish Shellfish Immunol., 67:263-269.
The authors succeeded production of the recombinant interleukin-12 cytokine that is an inducer ofcell-mediated immunity. Furthermore, they demonstrated that the cytokine had an adjuvant effectagainst Nocardiosis. The work could contribute to the development of vaccines against
Nocardiosis.

13

Use of a Flavobacterium columnare DnaK Recombinant Protein Vaccine to Guard Against
Columnaris Disease in Channel Catfish
Miles D. Lange
United States Department of Agriculture, Agricultural Research Service, Aquatic Animal HealthResearch Unit, Auburn, AL, 36832, USA
E-mail: miles.lange@usda.gov
The incidence of different bacterial diseases and their ongoing contribution to on-farm losses
including the cost of therapeutants during the production cycle persists among the U.S. catfish
industry accounting for as much $17 million in overall annual lost revenue. Flavobacterium
columnare causes substantial losses among cultured finfish species. The Gram-negative bacterium
is an opportunistic pathogen that manifests as biofilms on the host’s mucosal surfaces as thedisease progresses. We previously established that the dominant mucosal IgM antibody response
to F. columnare is to the DnaK protein found in the extracellular fraction (Lange et al. 2016).
Recent advances in recombinant protein technology have made the production and subsequenttesting of individual immunogens quite effective. To establish the efficacy of using recombinant
protein technology to develop a new vaccine against columnaris disease, we are reporting on
several vaccine trials using a recombinant F. columnare DnaK protein (rDnaK). In the first study,
three groups of channel catfish were immunized by bath immersion with a live attenuated F.
columnare isolate, rDnaK or sham immunized. After six weeks, an F. columnare laboratory
challenge showed a significant increase in survival in both the live attenuated and rDnaK vaccines
when compared to the non-immunized control. A rDnaK-specific ELISA revealed significant
levels of mucosal IgM antibodies present in the skin of catfish immunized with rDnaK at four- andsix-weeks post immunization. In the second study, three groups of channel catfish were bathimmunized with rDnaK alone or with rDnaK after a brief osmotic shock or sham immunized. In
study two, after six weeks a laboratory challenge with F. columnare was conducted and showed asignificant increase in survival in the rDnaK and in rDnaK with osmotic shock when compared to
the non-immunized control. The rDnaK ELISA demonstrated significant levels of mucosal IgM
antibodies in the skin of catfish groups immunized with rDnaK at six weeks post immunization.
To further understand the processes which have conferred immune protection in the rDnaK group,
we conducted RNA sequencing of skin samples from the non-immunized and rDnaK treatedchannel catfish at one-week and six weeks post immunization. Differential gene expression was
identified between the non-immunized and immunized skin and gene ontology analyses (Lange etal. 2019). We next confirmed and extended these findings by evaluating the efficacy of therecombinant F. columnare DnaK protein vaccine when using different immersion adjuvant
strategies. The results demonstrate significant protection of channel catfish at after 6 to 8 weeks
post vaccination (Lange et al. 2021). Experimental methods to enhance the catfish immune
response to recombinant F. columnare DnaK protein under different strategies continues as this
vaccine remains a promising candidate for experimental trials in a production setting.
Funding: USDA Research Projects #6010-32000-027-00-D and #6028-32000-007-00-D.
Annotated Bibliography of Key Works
Lange, M.D., B.H. Beck, J.D. Brown, B.D. Farmer, L.M. Barnett, and C.D. Webster. 2016.

14

Missing the target: DNAk is a dominant epitope in the humoral immune response of channel
catfish (Ictalurus punctatus) to Flavobacterium columnare: Fish and Shellfish Immunology.
51:170-179.
This paper was the first to demonstrate that an F. columnare 70 kDa extracellular protein,
identified as a DnaK chaperonin, is the dominant antigen during a mucosal IgM response toinfection. This study also demonstrated that mucosal catfish IgM antibodies generated after F.
columnare infection are cross reactive to DnaK proteins identified from other Gram-negative
bacteria.
Lange, M.D., J. Abernathy and B.D. Farmer. 2019. Evaluation of a recombinant Flavobacterium
columnare DnaK protein vaccine as a means of protection against columnaris disease in channel
catfish(Ictaluruspunctatus).
Frontiersin
Immunology.
10:1175
https://doi.org/10.3389/fimmu.2019.01175
This paper was the first to test the efficacy of a new F. columnare recombinant protein vaccine
and demonstrated that protection could be conferred after immunization with a single antigen. The
study also identified adaptive immunity processes using high-throughput RNA sequencing throughwhich protection was likely achieved in the skin post-immunization.
Lange, M.D., J. Abernathy, B.D. Farmer and B.H. Beck. 2021. Use of an immersion adjuvant witha Flavobacterium columnare recombinant protein vaccine in channel
catfish. Fish and
Shellfish Immunology. 117: 136-139.
This paper demonstrated the effectiveness of using a bath immersion formulated adjuvant to reduce
the amount of immunization time required to confer protection against F. columnare.

15α-glucosidase Activity of Nocardia seriolae Isolated From Seawater Fish in Japan
Yoshiko Shimahara*1, Yasuhiko Kawato2, Tohru Mekata2, Kei Yuasa2
12

Kamiura Field Station, Fisheries Technology Institute, FRA, Oita 879-2602, Japan
Nansei Field Station, Fisheries Technology Institute, FRA, Mie 516-0193, Japan*E-mail: shimahara@affrc.go.jp
Nocardiosis caused by a bacterial pathogen Nocardia seriolae has been confirmed in avariety of marine fishes, including yellowtail Seriola quinqueradiata, amberjack S. dumerili,-DSDQHVH flounder Paralichthys olivaceus, mackerel Scomber japonicus, bluefin tuna Thunnus
orientalis in Japan. N. seriolae is divided into two types based on the presence or absence of αglucosidase activity (α-glu), a carbohydrate-hydrolase, and strains isolated in Japan between
2000 and 2005 were α-glu negative except for some strains from amberjack (Shimahara et al.,2008). In this study, α-glu activity, DNA polymorphism, and pathogenicity of N. seriolae strains
isolated from 2006 to 2014 in Japan were determined. Of the 103 newly collected s trains
isolated from yellowtail, amberjack, and -DSDQHVH flounder, 26 strains (25%) were positive for αglu, which was a significant increase from the surveillance in 2000-2005 (6%). Many of the αglu positive strains were isolated from amberjack cultured in southern Kyushu, but also
confirmed in yellowtail and -DSDQHVH flounder raised in the same area. DNA polymorphism
analysis by pulsed-field gel electrophoresis (PFGE) showed that the α-glu positive and negative
strains were divided into 8 (type 1-8) and 9 pulsotypes (type 9-17), respectively. In experimental
infection, mortality of amberjack challenged with 3 α-glu positive strains were 25%, 88%, and100%, while those challenged with 3 α-glu negative strains were and 0%, 0% and 25%.
Although outbreaks of nocardiosis with α-glu positive strains were confirmed only in southern
Kyusyu, it is possible that α-glu positive strains, which shows high pathogenicity to amberjack,expand to many areas in Japan in future.
Annotated Bibliography of Key Work
Shimahara, Y., Nakamura, A., Nomoto, R., Itami, T., Yoshida, T. 2008. Genetic and phenotypiccomparison of Nocardia seriolae isolated from fish in Japan. J. Fish Diseases, 31:481-488.
The authors investigated phenotypic and genetic characterizations of fish pathogen
determinedby α-glucosidase
activity
and1970–2005.
biased sinusoidal
field gel andelectrophoresis(BSFGE)
Nocardia
seriolaeisolated in JapanfromThe phenotypic
genetic
characterizations
wereThere was no α-glucosidase activity in strains isolated from 2000–05 (n=50)
analysis, respectively.
with a few exceptions (n =3), while all strains isolated from 1970–90 (n=8) were positive. In
BSFGE analysis, all restriction patterns obtained from 50 strains isolated during 2000–05 wereunrelated to those obtained from eight strains isolated during 1970–90, with the exception of two
strains isolated during recent outbreaks. Based on the phenotypic and genetic characterizations,
recent outbreaks of nocardiosis in Japan are suggested to be epidemiologically unrelated to earlier
outbreaks in Japan.

16

Emergent Pathogens in Aquaculture: How Should We Identify Them?
Linda D. Rhodes
Northwest Fisheries Science Center, NOAA Fisheries, 2725 Montlake Boulevard East, Seattle,
WA 98112 USA
E-mail: linda.rhodes@noaa.gov
Identification of the causes of infectious disease has relied on demonstrating a relationshipthat exceeds simple correlation. The Henle-Koch's postulates have served as guidelines forestablishing causation, but over time, they have needed augmentation as awareness of etiologic
agents expanded to viruses, polymicrobial infections, and evolving technologies (Rivers, 1937,
Huebner, 1957, Evans, 1976, Fredricks and Relman, 1996).
There are several case studies demonstrating the complexities in establishing causation of disease
in aquatic organisms. One example is heart and skeletal muscle inflammation (HSMI) of Atlantic
salmon, a disease linked to a novel piscine orthoreovirus (PRV) through deep sequencing ofaffected tissues (Palacios et al.,, 2010). However, molecular genetic screening reveals the presenceof PRV sequences in animals exhibiting no signs of HSMI (Garseth et al.,, 2013, Purcell et al.,,2020). Another example is the difference in virulence in infectious salmon anemia virus (ISAV)
strains (McBeath et al.,, 2014), including possible strain evolution genetically and in pathogenicity(Christiansen et al.,, 2017, Markussen et al.,, 2008).
Broad spectrum screening using high throughput sequencing technologies further complicates
identification of true pathogens and can generate misconceptions about the prevalence of
pathogens (Mordecai et al.,, 2019). For aquatic animal health researchers and practitioners as well
as reviewers of scientific publications, it is important to establish clear criteria for pathogen
identification for both scientific accuracy and correct public understanding.
Annotated Bibliography of Key Works
Christiansen DH, McBeath AJA, Aamelfot M, Matejusova I, Fourrier M, White P, Petersen PE,
Falk K. 2017. First field evidence of the evolution from a non-virulent HPR0 to a virulent HPRdeleted infectious salmon anaemia virus. J Gen Virol 98: 595-606.
This publication reports on the first evidence that the avirulent HPR0 strain of ISAV could evolve
into a virulent phenotype under cultured fish conditions. Furthermore, the emergent subtype
possessed genetic changes that were consistent with virulence in other strains of ISAV. The work
provided the first support for HPR0 as a reservoir for virulent strains of ISAV.
Evans AS. 1976. Causation and disease: The Henle-Koch postulates revisited. Yale J Biol Med
49: 175-195.
This contains a nice historical timeline of the evolution of the Henle-Koch's postulates. It includes
an acknowledgment that new technologies will detect microorganisms that are not connected to adisease.

17

Fredricks DN, Relman DA. 1996. Sequence-based identification of microbial pathogens: A
reconsideration of Koch's postulates. Clin Microbiol Rev 9: 18-33.
An early but still highly relevant review of the role of sequence analysis in identifying microbial
pathogens. One of the authors, David Relman, is a leader in the human microbiome initiative in
the U.S., and he continues cutting edge research in this area.
Garseth AH, Fritsvold C, Opheim M, Skjerve E, Biering E. 2013. Piscine reovirus (PRV) in wild
Atlantic salmon, Salmo salar L., and sea-trout, Salmo trutta L., in Norway. J Fish Dis 36: 483493.
This publications reports on surveillance for PRV in wild-caught salmon, trout, and char in
Norway. It uncovered relatively high prevalences (13 - 55%) of PRV with nearly no evidence ofdisease or inflammation.
Huebner RJ. 1957. Criteria for etiologic association of prevalent viruses with prevalent diseases;
the virologist's dilemma. Ann N Y Acad Sci 67: 430-438.
In this publication, Huebner suggested that an epidemiological aspect could be added as a tool in
identifying and validating an etiologic agent.
Markussen T, Jonassen CM, Numanovic S, Braaen S, Hjortaas M, Nilsen H, Mjaaland S. 2008.
Evolutionary mechanisms involved in the virulence of infectious salmon anaemia virus (ISAV), a
piscine orthomyxovirus. Virology 374: 515-527.
This paper reports a detailed full genome analysis of the avirulent strain of ISAV, HPR0. It
provides genetic characterization of a virulence determinant by identifying a marker site that has
been pivotal in studying ISAV virulence.
McBeath AJ, Ho YM, Aamelfot M, Hall M, Christiansen DH, Markussen T, Falk K, Matejusova
I. 2014. Low virulent infectious salmon anaemia virus (ISAV) replicates and initiates the immune
response earlier than a highly virulent virus in Atlantic salmon gills. Vet Res 45: 83.
The publication reports on salmon immune responses to experimental exposure to strains of ISAVwith differential virulence. Low virulence strains replicated and disseminated more quickly,
stimulating a more rapid host response. In contrast, high virulence strains replicated lessefficiently, and the less effective host response ultimately allowed a higher viral load and greatermortality. The study provided a mechanistic explanation for differences in pathogenicity.
Mordecai GJ, Miller KM, Di Cicco E, Schulze AD, Kaukinen KH, Ming TJ, Li S, Tabata A, Teffer
A, Patterson DA et al. 2019. Endangered wild salmon infected by newly discovered viruses. Elife
8:347615.
This publications reports on a viral discovery screening of moribund and dead hatchery and wild
salmon. Many viral sequences were identified, including arenaviruses, nidoviruses, and 318

reoviruses. The publication implied these represented widespread sources of viral diseases, but inthe absence of evidence of disease being present in the sampled fish. The publication generated
considerable public concern about fish viruses and the role of aquaculture in amplifying andspreading fish viruses.
Palacios G, Lovoll M, Tengs T, Hornig M, Hutchison S, Hui J, Kongtorp RT, Savji N, Bussetti
AV, Solovyov A et al. 2010. Heart and skeletal muscle inflammation of farmed salmon isassociated with infection with a novel reovirus. PLoS One 5: e11487.
The first study to credibly inplicate PRV as a causative agent in HSMI, based on genetic
sequencing and bioinformatics.
Purcell MK, Powers RL, Taksdal T, McKenney D, Conway CM, Elliott DG, Polinski M, Garver
K, Winton J. 2020. Consequences of piscine orthoreovirus genotype 1 (PRV-1) infections inchinook salmon (Oncorhynchus tshawytscha), coho salmon (O. kisutch) and rainbow trout (O.
mykiss). J Fish Dis 43: 719-728.
This publication reports on experimental challenges of Pacific salmon with the PRV genotype thatcauses HSMI in Atlantic salmon. The outcomes demonstrated that although PRV-1 could establish
in Pacific salmon, there was no establishment of disease.
Rivers TM. 1937. Viruses and Koch's postulates. J Bacteriol 33: 1-12.
As president of the Society of American Bacteriologists, Thomas Rivers recognized that viral
etiology could not be demonstrated by strictly following Koch's postulates, especially when coinfections occurred or in carrier states. He proposed that a appropriate timing of the appearance ofthe putative agent and that a host response (i.e., antibodies) could be incorporated wheninvestigating causal relationships.

19

The Role of National Fisheries University and Studies of Fish Disease
Shinya Yasumoto
Department of Applied Aquabiology, National Fisheries University, Japan Fisheries Researchand Education Agency, Yamaguchi 759-6595, Japan
Email: yasumotos@fish-u.ac.jp
Two topics have been included in this presentation. The first topic is introduction of National
fisheries university (NFU) and second one is my research activities in NFU.
NFU has 80 years of history. Ministry of Agriculture, Forestry and Fisheries has jurisdiction
over NFU. Therefore, NFU has different policies (diploma, curriculum, and admissions policy)
from other universities which are under the jurisdiction of the Ministry of Education, Culture,
Sports, Science and Technology. NFU consists of four departments, advanced course, and graduate
school, and is a school that educates students to be the industry’s future leaders. NFU contributes
to the fisheries industry by producing many talents to the fisheries related company, government
office and academic.
In our laboratory, diseases of ornamental fish were studied mainly, and we are focusing on
koi carp (Nishikigoi; Cyprinus carpio) among them. Koi carp is internationally popular as an
ornamental fish, and the industry is unique. Product value is influenced by a number of factors,
including size, shape, color, and variety, and has recently been increasing. Many types of diseases,
including Koi herpesvirus (CyHV-3), have been reported in koi carp, and idiopathic diseases are
known among farmers and hobbyists.
Recently, we are studying about “Sinking disease” (swim bladder disorders) which has
become a large issue in the field of koi carp breeding. This disease was an idiopathic disease.
Diseased fish sink, become immobile, and stay at the bottom of a pond or tank with lethargy, buthave no external signs. Therefore, we herein examined the gross and histological features of
diseased fish from farms. At necropsy, their swim bladder was filled with liquid (swim bladder
fluid, SBF). Histopathologically, fish had hyperplastic connective tissue in the tunica intima of theswim bladder, and some were accompanied by inflammation. Furthermore, some species ofbacteria were isolated from SBF samples of diseased fish. To reproduce the disease, experimental
infections using the three major isolates (Streptococcus sp., Pseudomonas aeruginosa, andMycobacterium sp.) were also attempted. Immobility and the accumulation of SBF occurred in thelive bacteria-injected groups. Based on these results, we concluded that the major cause of sinking
disease is the accumulation of SBF due to bacterial invasion in the swim bladder and, as a result,a loss of buoyancy. Future studies are needed to elucidate the mechanisms underlying theaccumulation of SBF and develop measures to prevent and treat sinking disease in koi carp.
Annotated Bibliography of Key Words
Yasumoto, S. 2018. Remedy. In “Nishikigoi A to Z” (ed. by S. Mano). Shin Nippon Kyoiku Tosho,
Shimonoseki, pp 128-158.
This is the textbook of koi carp breeding. We introduced more than 30 kinds of diseases (noncommunicable, viral, bacterial, fungal, and parasitic disease) and explained those diagnostic andtreatment methods. This textbook is red by farmers and hobbyists and used in our lectures for NFU
students.
20

Yasumoto, S., Okada, M., Moroi, H., Shimizu, H., Kondo, M. 2021. A Preliminary Study on
Sinking Disease in Koi Carp. Fish. Pathol., 56(3):107-114.
Sinking disease has become a major issue in the field of koi carp breeding. Although many
diseased fish did not exhibit any significant changes in their appearance, some had ulcers anderosions on the abdomen and at the base of the fins due to physical scratching caused by being at
the bottom of a pond for a prolonged period of time. At necropsy, the swim bladder of the fish was
filled with fluid (swim bladder fluid, SBF) and very little gas. Histopathologically, fish had
hyperplastic connective tissue in the tunica intima of the swim bladder, and some wereaccompanied by inflammation. Furthermore, one to three species of bacteria were isolated fromsix out of fourteen SBF samples of diseased fish. To reproduce this disease, two experiments were
performed using the three major isolates (live or formalin-killed bacteria) from SBF as well as
SBF itself. Immobility and the accumulation of SBF occurred in the live bacteria-injected groups.
Based on these results, we concluded that the major cause of sinking disease is the accumulationof SBF due to bacterial invasion in the swim bladder and, as a result, a loss of buoyancy.

21

Aquaculture Medicine in the 21st Century: Training the Veterinary Community to Meetthe Needs of a Global Industry
Iske V Larkin1 and Ruth Francis-Floyd1,2*
1

Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of
Florida, Gainesville, FL 32610, USA
2
Program in Fisheries and Aquatic Sciences, School of Forest, Fisheries and Geomatic Sciences,
University of Florida, Gainesville FL 32603, USA*E-mail: rffloyd@ufl.edu
Globally, aquaculture is rapidly growing, diverse, and increasingly important as a source ofprotein and maintaining food security. In the United States, aquaculture was valued at $1.5 B in
2018, with the most valuable products produced being shellfish (oysters and clams) and Atlantic
salmon (NOAA Fisheries 2021). The loss of livestock by disease is a major threat to production,consequently there is a significant need for well-trained veterinarians and aquatic animal healthspecialists. We will describe how we have addressed this need by providing elective training for
veterinary students and graduate veterinarians.
Florida currently ranks 9th in aquaculture production by US states, with annual farm gate valueof approximately $72M in 2018 (Florida Department of Agriculture and Consumer Services,2020). Florida aquaculture is very diverse, with the major aquaculture product consisting of
hundreds of species and varieties of ornamental fish sold for the pet trade and valued at $28.7M in2018. Shellfish production is the major aquatic food product, valued at $15.5M in 2018. Other
species reared in Florida include food and game fish, alligators and other reptiles, and aquaticplants. The types of production systems used in Florida are also very diverse and include pond,
raceways, and recirculating systems as well as aquaponics and off shore “bottom” leases for clams.
This diversity creates an exceptional teaching opportunity for veterinarians and veterinary students
in aquaculture medicine. Further, Florida has famous theme parks and oceanaria that include large
collections of aquatic organisms, and a very engaged state fish and wildlife agency; the Florida
Fish and Wildlife Conservation Commission. These entities collectively create an exceptional
teaching opportunity for veterinary students, graduate veterinarians, and para-professionals
including veterinary technicians and biologists.
In the United States, there are currently 33 colleges of veterinary medicine that are accreditedby the American College of Veterinary Medicine. Of these, approximately 10 offer some elective
training to students in aquatic animal health. The University of Florida’s training programs areglobally recognized for excellence in this specialized area. Although some aspect of aquatic
medicine has always been offered to students at the University of Florida, specific training inaquaculture was initiated in the 1980s, and enhanced in the 1990s with the development of the UF
Tropical Aquaculture Laboratory (UF-TAL) that included a full-service veterinary diagnostic
laboratory. In 2000, through a state legislative appropriation for “Training in the care, treatment,
and rehabilitation of marine mammals” with an emphasis for species within the state of Florida,
especially the Florida manatee (Trichechus manatus), the existing program was enhanced toinclude learning opportunities with species beyond Florida’s aquaculture industries. Expanded
elective training for students included on-site clinical experiences at UF – TAL and with many of
our industry partners, including theme parks and government agencies.

22

The UF College of Veterinary Medicine (CVM) offers a unique 15-credit hour elective
certificate to veterinary students in Aquatic Animal Medicine. This is a popular program and has
an average of 8 students per year completing the training. This certificate is restricted to UF
veterinary students and includes two required courses (Sea Vet and Diseases of Warm Water Fish)and elective on-line course work, clinical externships, and research opportunities. Many students
who have earned the certificate have continued on to complete clinical residencies or graduate
programs and are emerging as leaders in the aquatic health field. For non-UF students and graduate
veterinarians, a new 12-credit hour certificate program entitled Care and Conservation of Aquatic
Animals has recently been approved.
Clinical training is also available but limited in scope. An annual clinical internship takes one
veterinarian each year and provides 12-months of training that includes fish and marine mammal
medicine. Advanced training is available through a 3-year residency program accredited by theAmerican College of Zoological Medicine (ACZM) but only one candidate is selected every three
years. Since 2004, 6 veterinarians have completed this program and a seventh is currently enrolled.
Five of our residents have been board certified by ACZM as specialists in aquatic medicine.
One of the most effective ways to provide post veterinary school training is through our
distance education program. The College of Veterinary Medicine and the School of Forest,
Fisheries and Geomatic Sciences collaborate and offer comprehensive on-line educational
programs for practicing veterinarians and working professionals. The development of strong
distance education programs has created significant learning opportunities for veterinarians andpara-professionals from around the globe. A fully on-line Master of Fisheries and Aquatic Sciences(MFAS) degree program is very popular with veterinarians seeking additional course work and an
academic credential in aquatic science. The MFAS degree requires 32 credits of course work anda technical paper reviewed by a supervisory committee and presented to the public. For
veterinarians who are not ready to commit to 32 hours of course work there is a certificate in
Aquaculture and Fish Health. This program consists of 4 graduate level courses. Required courses
are Aquaculture 1 and Diseases of Warm Water Fish, and then the student can select from a menu
of courses to complete the program. A veterinarian can complete the Aquaculture and Fish Healthcertificate and apply the academic credits to the MFAS degree if he/ she wishes to continue.
In 2017 the USDA provided funding for a national fellowship program to train practicing
veterinarians so that they could provide veterinary services to aquaculture farms in theircommunity. Eight aquaculture fellows have been trained and all are providing some level of
support to aquaculture industries. The fellows benefitted from academic and experiential training
in aquaculture production techniques, fish and shellfish disease diagnosis and health management,
and training to help them navigate the complex regulatory environment in the United States. All
fellows completed the Aquaculture and Fish Health certificate mentioned above as well as an onsite practicum offered at the UF-TAL in 2019. These veterinarians are willing and able to workwith aquaculture businesses and apply principles that have resulted in improved efficiency anddisease prevention in traditional livestock production to aquatic clientele. For maximum benefit,
aquaculture producers have to realize the benefits of establishing a veterinary-client-patient
relationship with well-trained veterinarians. Simply calling a veterinarian when they need a healthcertificate signed or a prescription issued will not be sufficient. Regular veterinary involvement indecision-making on aquatic farms will optimize production and profitability while minimizing theloss of livestock.
Annotated Bibliography of Key Works

23

Hartman, K., R.P.E. Yanong, C.A. Harms, G.A. Lewbart. 2006. The future of training for aquatic
veterinarians.
Journalof
Veterinary
Medical
Education
33(3):389-393.
https://doi.org//0.3138/jvme.33.3.389
The article provides an overview of employment opportunities and continuing education programsfor veterinarians in aquatic animal health.
DeHaven, W.R. and A.D. Scarfe. 2011. Professional education and aquatic animal health: a focus
on aquatic veterinarians and veterinary paraprofessionals. Proceedings OIE Global Conference on
Aquatic Animal Health, June 28030 2011, Panama, Pp 139-154.
This article provides a global perspective on the need for a “well educated and competentveterinary workforce” to meet the needs of expanding aquaculture production and the threat oflivestock loss through disease. It also describes several programs designed to provide supplemental
education to practicing veterinarians so they can develop the knowledge base required to do thiswork.
Sanders, J. 2018. Aquatic ambulatory practice. Vet Clin Exot Anim 21: 609-622.
https://doi.org/10.1016/j.cvex.2018.05.003
This article is a description of how a practicing veterinarian can adapt their business to provide
service to aquaculture and pet fish owning clients. The emphasis is on practice management andindividual animal care.

24Epitope Mapping of Anti-RSIV Monoclonal Antibody Using a Phage Display Library
Tomokazu Takano1*, Tomomasa Matsuyama1, Yasuhiko Kawato1, Takamitsu Sakai2, Jun
Kurita2, Yuta Matsuura1, Sachiko Terashima1, Kazuhiro Nakajima3, Chihaya Nakayasu1
1

Nansei Field Station, Fisheries Technology Institute, Japan Fisheries Research and Education
Agency, Mie 519-0193, Japan
2
Tamaki Field Station, Fisheries Technology Institute, Japan Fisheries Research and Education
Agency, Mie 519-0423, Japan
3
Hatsukaichi Field Station, Fisheries Technology Institute, Japan Fisheries Research and
Education Agency, Hiroshima 739-0452, Japan*Email: takanoto@fra.affrc.go.jp
Red sea bream iridoviral disease (RSIVD) spreads readily throughout the marine environment
because of its wide variety of host species. Use of a rapid diagnostic method, such as theimmunofluorescence antibody test (IFAT), helps to control the disease, so the anti-RSIV
monoclonal antibody M10 (MAb M10) has been developed. The IFAT test with MAb M10 fortissue smears from the diseased fish is introduced as a simple method in the OIE manual of
diagnostic tests for aquatic animals (CHAPTER 2.3.8. RED SEA BREAM IRIDOVIRALDISEASE). MAb M10 is available from our laboratory, which is part of the activities of the OIE
reference laboratory of the RSIVD. However, the antigens recognized by MAb M10 have not beencharacterized. Therefore, in order to define the specificity of IFAT, it is important to identify the
antigen and epitope of this antibody.
In the present study, we carried out epitope mapping using the phage display method to
identify the antigen of MAb M10. A phage display RSIV peptide library was constructed to cover
the entire genome of RSIV, then a total of 17 phage clones recognized by the antibody wereselected by biopanning. The selected clones harbored partial fragments of the laminin-type
epidermal growth factor-like domain (LEGFD) gene. N-terminal and C-terminal deletion peptides
were then prepared from the deduced amino acid sequence of the smallest fragment obtained fromthe biopanning to precisely determine the epitope. Finally, seven amino acids, EYDCPEY, locatedin the extracellular domain of the LEGFD protein were determined to be the epitope. Identicalresidues of the epitope were also identified from the LEGFD protein in other megalocytiviruses
including the infectious spleen and kidney necrosis virus (ISKNV) and turbot reddish body
iridovirus (TRBIV). MAb M10 is considered to be widely available for the diagnostics of these
megalocytivirus infections.
Annotated Bibliography of Key Works
Nakajima, K., Sorimachi M. 1995. Fish Pathology, 30(1):47-52.
A hybridoma clone secreting monoclonal antibody (MAb M10) recognizing an RSIV protein
was established.
Nakajima, K., Maeno, Y., Fukudome, M., Fukuda, Y., Tanaka, S., Matsuoka, S., Sorimachi, M.
1995. Fish Pathology, 30(2): 115-119.
IFAT with MAb M10 was shown to be a rapid and effective method for diagnosis of theiridovirus infection in cultured marine fishes.

25

Nakajima, K., Maeno, Y., Yokoyama, K., Kaji, C., Manabe S. 1998. Fish Pathology, 33(2):7378.
No cross-reactivity with other fish iridoviruses (EHNV, SFIV and GIV) was observed in the
IFAT and immunoprecipitation using MAb M10.
Takano, T., Matsuyama, T., Kawato, Y., Sakai, T., Kurita, J., Matsuura, Y., Terashima, S.,
Nakajima, K., Nakayasu C. 2020. Fish Pathology, 54(4):83-92.
Seven amino acids located in the extracellular domain of the LEGFD protein were determined tobe the epitope of MAb M10.

26

Managing the Threat of Emerging Diseases in Bivalve Mollusc Aquaculture
Ryan B. Carnegie
Virginia Institute of Marine Science, William & Mary, P.O. Box 1346, Gloucester Point,
Virginia 23062, U.S.A.
E-mail: carnegie@vims.edu
The history of diseases impacting, and sometimes devastating, fisheries and aquaculture formarine bivalve molluscs spans more than a century, and the full global geography of theseindustries. Yet even today, despite advances for example in biotechnology for pathogen detection,
we struggle to detect and manage emerging diseases, as exemplified most recently by the
emergence and spread over the last decade of the ostreid herpesvirus OsHV-1 microvariants in
Europe and other areas. Because of our recognized challenges, and lack of success, with emerging
pathogens, the shellfish health community has begun to focus attention on identifying andmitigating our greatest areas of weakness, through work at the regional level in the U.S.A., as wellas internationally through the International Council for the Exploration of the Sea (ICES) Working
Group on Pathology and Diseases of Marine Organisms (WGPDMO).
Initial work identified three areas of disease management that would appear to be strengths,
but which instead may represent structural flaws in our management paradigm. The use of
notifiable pathogen lists and advanced molecular diagnostics such as polymerase chain reaction
assays (PCR) are central to disease management, but, paradoxically, create blind spots with regard
to emerging diseases that are not listed or not targeted by the diagnostics (Carnegie et al. 2016).
Uncertainty about pathogen distributions and poorly studied aspects of pathogen biology cancreate regulatory paralysis that, while perhaps unnecessarily restrictive of reasonable commerce,
at least should maintain a high level of biosecurity, with commerce and thus pathogen transfers
limited; unless, paradoxically, this has the effect of reducing biosecurity by driving commerce tosurreptitious channels.
Subsequent work by the ICES WGPDMO, through a focused Workshop on Emerging
Mollusc Pathogens (WKEMOP, ICES 2019), identified thirty-six wide ranging priorities for
improvement in regional to national and international aquatic animal health management, in four
primary areas: improving communication and frameworks, essential infrastructure and expertise,
key research priorities, and the central role of husbandry. Highlights among the thirty-six priorities
were a continued relevance of foundational fields such as pathology, bacteriology and virology, as
opposed simply to molecular diagnostics; the critical need to maintain laboratory capacity,
especially at provincial governmental and academic institutions more deeply connected to local
environments and industries than major national laboratories would be; the great importance ofcommunication and information and data sharing at all levels, for example among labs andbetween regulators, the scientific community, and industry; and the need to better align funding
with the value of collecting long-term, continuous data for diseases and environments has value
beyond.
This presentation will synthesize work in this area and discuss as well an additional
contemporary challenge, our need to better understand and manage the strain-level diversity inaquatic pathogens, differences in virulence or pathogenicity potentially important contributors tooutbreak risks.

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Annotated Bibliography of Key Works
Carnegie, R.B., I. Arzul, and D. Bushek. 2016. Managing Marine Diseases in the Context ofRegional and International Commerce: Policy Issues and Emerging Concerns. PhilosophicalTransactions of the Royal Society B 371: 20150215.
http://doi:dx.doi.org/10.1098/rstb.2015.0215
This paper presents the paradox of the list, the paradox of advanced diagnostics, and the paradoxof uncertainty noted in the abstract above. It is a key publication highlighting the “blind spots” indisease management created by use of notifiable lists and advanced diagnostics in particular, andpresents unique perspective on the troubled response to OsHV-1.
ICES. 2019. Workshop on Emerging Mollusc Pathogens (WKEMOP). ICES Scientific Reports
1:57. 19 pp. http://doi.org/10.17895/ices.pub.5577
This paper highlights thirty-six priority areas for improvement in our detection and responses toemerging diseases. While mollusc-focused, the priorities are transcendent, equally relevant tofish and crustacean systems.
Gustafson, L.L., I. Arzul, C.A. Burge, R.B. Carnegie, J. Caceres-Martinez, L.H. Creekmore, B.
Dewey, R. Elston, C.S. Friedman, P. Hick, K. Hudson, C. Lupo, B. Rheault, K.A. Spiegel, and
R. Vásquez-Yeomans. 2021. Optimizing surveillance for early disease detection: Expert
guidance for Ostreid herpesvirus surveillance design and system sensitivity calculation.
Preventive Veterinary Medicine 194: 105419. http://doi.org/10.1016/j.prevetmed.2021.105419
This is an important new publication highlights a key area of potential focus for detection of
emerging pathogens, the passive surveillance represented by farm-level observations made every
day by aquaculturists.
Carnegie, R.B., S.E. Ford, R.K. Crockett, P.R. Kingsley-Smith, L.M. Bienlien, L.S.L. Safi, L.A.
Whitefleet-Smith, and E.M. Burreson. 2021. A rapid phenotype change in the pathogen
Perkinsus marinus was associated with a historically significant marine disease emergence in theeastern oyster. Scientific Reports 11: 12872. http://doi.org/10.1038/s41598-021-92379-6
This new paper provides entirely new perspective on the great intensification of oyster parasite
Perkinsus marinus in the 1980s, revealing it to be caused by emergence of a new, highly virulent
parasite phenotype. It highlights the importance of understanding sub-species or strain dynamics
of pathogens, which may be more important than we appreciate in driving disease dynamics infisheries and aquaculture.
Ben-Horin, T., C.A. Burge, D. Bushek, M.L. Groner, D.A. Proestou, L.I. Huey, G. Bidegain, and
R.B. Carnegie. 2018. Intensive oyster aquaculture can reduce disease impacts to sympatric wild
oysters. Aquaculture Environment Interactions 10: 557-567. http://doi.org/10.3354/aei00290
This recent paper presents counter-intuitive results from disease modeling, that aquaculture may
reduce, rather than exacerbate, disease impacts of certain pathogens. The results are relevant toour consideration of how aquaculture “fits” into ecosystems and potentially interacts withfisheries and restoration activities for wild natural populations of resource species.

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Advancing Epidemiological Tools for Marine Disease Management - Modeling Dispersionof OsHV-1μvar in Pacific Oyster Populations
Meghana P. Parikh and Gary H. Wikfors
Northeast Fisheries Science Center, NOAA Fisheries, Milford, CT 06460, USA*Email: meghana.parikh@noaa.gov
Ostreid herpesvirus-1 microvariant (OsHV-1 μvar), a significant pathogen of the Pacificoyster in Europe, Australia, and New Zealand, poses a major threat to the US oyster aquacultureindustry. Introduction of the virus into naive populations could leave growers vulnerable to awidespread epidemic accompanied by mass mortalities and major financial losses. For OsHV1μvar and other emerging pathogens, there is a great need for risk assessment to informprevention and management strategies. Classic statistical methods and models are regularly
employed for this purpose, and offer an opportunity to describe how specific factors influencedisease outcomes based on observational or experimental data. For marine diseases, this issometimes unable to be accomplished due to lack of sufficient data. Perhaps more importantly,
these methods may not always capture the complexity of marine disease dynamics, resulting in
uncertainty in their application to management practices. Mathematical simulation models offer
a solution to these challenges in understanding and managing aquatic animal diseases.
Significant advantages of this approach include the ability to investigate processes whereobservational data is limited, as well as the ability to incorporate uncertainty into the starting
conditions. There have been several studies detailing the construction of mathematical models
for marine disease, and these are typically particle tracking models which estimate an area’s riskof exposure based on the hydrodynamic connectivity of the region. There is great need to couple
these models with epidemiological models to better answer questions about disease risk,
potential outcomes, and implications of various management strategies. In this study, I willfurther discuss the need for advancement of these modeling tools for decision making, and I will
present our plans for such a model simulating spread of OsHV-1μvar in Pacific oyster
populations in the United States.
Annotated Bibliography of Key Works
Ben-Horin, T., Bidegain, G., de Leo, G., Groner, M. L., Hofmann, E., McCallum, H. & Powell,E. (2020). Modelling marine diseases. In D. C. Behringer, B. R. Silliman & K. D. Lafferty (Eds).
Marine Disease Ecology (pg. 223–255). Oxford University Press. DOI:
10.1093/oso/9780198821632.003.0012
In this review the authors discuss various modelling methods that have been used to describe
marine disease dynamics. They note that while the foundational principles apply to bothterrestrial and marine diseases, the physical ocean environment adds a layer of complexity that
requires an altered approach to disease transmission. Of particular interest is section 12.7 whichreferences many of the most well-known examples incorporating ocean hydrodynamics into theirmodeling methods.
ICES. 2019. Workshop on Emerging Mollusc Pathogens (WKEMOP). ICES Scientific Reports.
1:57. 19 pp. http://doi.org/10.17895/ices.pub.5577

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This report discusses the threat of emerging pathogens to the growth and sustainability ofshellfish aquaculture, and it identifies key priorities related to infrastructure, expertise, andresearch necessary for the management of these diseases. In particular, the report highlights thedevastation caused by OsHV-1μvar globally as well as the rapid assessment and preparation
required to manage the pathogen in North America, where most Pacific oyster populations are
currently unaffected.
Lupo, C., Dutta, B.L., Petton, S., Ezanno, P., Tourbiez, D., Travers, M.-A., Pernet, F., Bacher, C.
Spatial epidemiological modelling of infection by Vibrio aestuarianus shows that connectivity
and temperature control oyster mortality. Aquaculture Environment Interactions. Vol.12 (511527) 19 Nov 2020, https://doi.org/10.3354/aei00379
The aim of this study was to develop a spatial epidemiological model coupling hydrodynamic
connectivity with a traditional epidemiological compartment model to estimate disease spreadand resulting mortality caused by Vibrio aestuariansu in Pacific oysters. Sensitivity analyses
were conducted to evaluate the relative importance of each parameter to the model outcomes.
The authors propose that their methods can be used to evaluate the efficacy of proposedmanagement strategies. This model is one of the first to pair dispersion and epidemiologicalmodels, and it represents a significant advancement in marine modeling methods.
Peeler, EJ, Taylor, NG. The application of epidemiology in aquatic animal health -opportunities
and challenges. Veterinary Research vol. 42 (94). 11 Aug. 2011, https://doi.org/10.1186/12979716-42-94
This review discusses epidemiological approaches to disease management and applications toaquatic animal diseases. The paper details the short history of aquatic animal epidemiology,
method employed, and challenges to implementation. The author emphasizes that modern
epidemiological approaches are integral to addressing questions within complex aquatic diseasesystems, and they note particle tracking models as a valuable means of studying disease spread.
A lack of available data and systems for data collection is identified as a key constraint toadaptation of many epidemiological methods.

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Using a OneHealth Approach to Evaluate How Marine Pathogens and Parasites That Are
Infectious to Cultured Finfish, Shellfish, and Macroalgae Within the United States May Be
Affected by Climate Change.
Michelle Rub1*, Linda Rhodes1, Alexandra Arnold2,3,4, Kyle Carver2,3,4, Janet Whaley2, Seth
Theurerkauf2, Leo Goldsmith3, Julie Trtanj4 & Penny Swanson1
1

NOAA Fisheries Northwest Fisheries Science Center, 2725 Montlake Boulevard East, Seattle,
WA 98112 USA
2NOAA Fisheries Office of Aquaculture, 1315 East-West Highway, 12th Floor, Silver Spring,MD, 20910, USA
3
U.S. Global Change Research Program, 1800 G Street, NW, suite 9100 Washington, D.C.
20006 USA
4
NOAA Climate Program Office, 1315 East-West Highway, Suite 100, Silver Spring, MD,
20910, USA*E-mail: michelle.rub@noaa.gov
Climate change has led to an increase in the frequency of extreme events such as marine
heatwaves and harmful algal blooms (HABs). Such events can lead to catastrophic loss ofcultured and wild marine species. Due to their high visibility, these phenomena are well
documented and studied. However, there are more subtle effects of climate change that may
prove to be just as devastating to U.S. aquaculture in the future as these ‘extreme events’ aretoday. One such example includes the various impacts that marine pathogens and parasites canhave on cultured finfish, shellfish, and seaweeds.
Predicted climate related changes in ocean temperature, dissolved oxygen, salinity, pH,
ocean currents, estuarine flushing patterns, wind speed and direction, precipitation andfreshwater nutrient runoff (to name a few) will most certainly influence the virulence and spatial
reach of marine bacteria, viruses, and parasites. Novel pathogens and parasites may be‘introduced’ into a given area through expansion of their host species’ range or through othermechanisms. Changes in the physical environment of a cultured species can induce stress or alter
immune responsiveness, and this may influence their susceptibility to disease. These changes
could result in impacts to cultured animal health, reduced quality of cultured seafood products, or
impacts to human health.
In order to be better prepared for these climate related impacts, researchers from NOAA
Fisheries Office of Aquaculture, the NOAA Climate Program Office, NOAA Fisheries
Northwest Fisheries Science Center, and the US Global Change Research Program (USGCRP)
are working together using a OneHealth approach to identify current and emerging marine
pathogens and parasites that are important to cultured species and that are likely to be affected ineither their scope of influence/range or in their severity by climate change.
Information collected through this effort will be used to prioritize support for research, toidentify adaptive measures that can be taken to prepare for climate change related aquaculture
impacts, to inform aquaculture drug development and approval, and it will be used to inform thepermitting process and management of Aquaculture Opportunity Areas (AOAs). Our goal inpresenting our approach to is to learn how countries with a more developed marine aquacultureindustry maintain biosecurity on their farms. Additionally, we hope to learn about how others areincorporating climate change science into their aquaculture strategies and tactics.
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Annotated Bibliography of Key Works
Callaway R, Shinn AP, Grenfell SE, Bron JE and others. 2012. Review of climate change
impacts on marine aquaculture in the UK and Ireland. Aquat Conserv 22: 389−421.
This paper provides a comprehensive review of the various impacts that climate drivers such as
sea-level rise, storm damage, increased precipitation, and increasing ocean acidification may
have on marine aquaculture in the UK and Ireland. Key to this paper is an extensive section
describing the potential effects of climate change on the health of finfish and shellfish. In
particular, this paper covers climate induces changes in the virulence and range of parasite andmicrobial pathogens, changes in the host species, and changes in contact and interaction between
host species and pathogens.
Chiaramonte L, Munson D, Trushenski J. 2016. Climate change and considerations for fish
health and fish health professionals. Fisheries 41: 396−399.
This paper is short and succinct. The authors describe how a specific combination of pathogen,host, and environmental conditions is required in order for fish to succumb to disease. They
further describe how climate change might influence this relationship and warn that althoughisolated changes in the aquatic environment such as increased temperature can alone influence
fish health, fish health experts should prepare themselves for even more complex scenarios in the
future.
Marcogliese DJ. 2008. The impact of climate change on the parasites and infectious diseases ofaquatic animals. Rev Sci Tech 27: 467−484.
This paper presents a comprehensive look at how climate change is likely to impact the virulenceand spread of aquatic pathogens and parasites. The authors include predictions of environmental
changes and the biological effects associated with global warming from the perspective of thehost-pathogen-parasite relationship as well as illustrations to describe the mechanisms behindthese changes. The role of temperature in virulence is specifically discussed. Both marine andfreshwater systems are covered, and diseases of marine invertebrates are highlighted. The
authors also discuss how interactions between climate drivers and other stressors such aspollution, hypoxia, and habitat loss/fragmentation influence the disease process. Finally, there isdiscussion of the effects of climate change on human health in relation to diseases that aretransmitted through water.

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