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You may use the information and images contained in this document for non-commercial, personal, or educational purposes only, provided that you (1) do not modify such information and (2) include proper citation. If material is used for other purposes, you must obtain written permission from the author(s) to use the copyrighted material prior to its use.

DOCUMENTATION OF FIBROPAPILLOMATOSIS IN GREEN TURTLES (CHELONIA MYDAS) CAPTURED FROM TEXAS INSHORE WATERS Tasha L. Metz 1,2, Mandi L. Gordon 1, George Guillen 1 and Joel D. Anderson 3 1 Environmental Institute of Houston, University of Houston Clear Lake 2 Department of Marine Biology, Texas A&M University at Galveston 3 Perry R. Bass Fisheries Research Station, Texas Parks and Wildlife, Palacios, TX

GREEN TURTLES IN TEXAS Green turtles utilize Texas waters primarily as juvenile foraging habitat Green turtle fishery and turtle canneries present in Texas during the late 1800s, but fishery had collapsed by the turn of the 20 th century (Hildebrand 1982)

1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011 2012 CPUE (# turtles/ km-hr) GREEN TURTLES IN TEXAS Exponential increase in green turtle CPUE (turtles/ km-hr) from entanglement netting surveys since 1990 (Metz and Landry, 2013) 12 10 8 y = 0.2604e0.1314x R2 = 0.7239 p = 0.001 6 4 2 0

GREEN TURTLES IN TEXAS Greatest genetic contribution from Florida and Mexico rookeries based on mixed stock analyses, although confidence intervals for these estimates was large (Anderson and Shaver 2013)

FIBROPAPILLOMATOSIS Etiology of the disease not completely understood, but a virus in the Herpes family (with geographic variants) has been isolated from tumors Additional contributing factors have been identified with the occurrence of FP: Environmental habitat degradation related to elevated nutrient levels and ingestion of species that have tumor promoting compounds (i.e. cyanobacteria Lyngbya majuscula in Australia and Hawaii) Environmental stressors, including cold stunning events and pollution Occurrence of a parasite vector (i.e. marine leeches in the genus Ozobranchus) First documented in Texas in 2010 2 stranded turtles and 11 of 33 turtles captured by entanglement netting displayed tumors (Tristan et al. 2010)

STUDY OBJECTIVES The impact of FP infection on the health and continued growth of Texas green turtle population was assessed via directed capture operations designed to: 1) determine the geographic extent as well as within-assemblage rate of FP infection in Texas green turtle population 2) assess size composition of FP-affected population constituents 3) potentially identify the herpesvirus variants associated with FP in Texas green sea turtle assemblages in an effort to document the possible geographic source(s) of this outbreak

Study Areas TEXAS Aransas Bay Complex Lower Laguna Madre Lavaca- Matagorda Bay Gulf of Mexico Port Mansfield Gulf of Mexico MEXICO Port Isabel Mexiquita Flats/ Brazos- Santiago Pass

Catch-Per-Unit-Effort (CPUE) = # turtles/km-hour Entanglement Nets: 91.4 m long; 2.9-3.6 m deep 17.8 cm bar mesh 2-4 nets deployed Net check every 20 minutes Tumor and skin biopsies were collected using a 3-5 mm punch (Work and Balazs 1999; Hirama and Ehrhart 2007)

Lavaca- Matagorda Bay 0 Greens Aransas Bay Complex 14 Greens 8 entangled 6 cast-netted at jetty 33 entangled Lower Laguna Madre 27 Greens 25 entangled 2 cast-netted at jetty 8 cast-netted at jetty

Mean SCL (cm) Mean SCL (cm) % Length Frequency GREEN TURTLE SIZE 100% 90% 80% 70% 60% 50% 40% 30% 20% 10% 0% Overall Mean SCL = 38.2 cm (range = 25.2 65.0 cm, n = 41) 20.0-29.9 30.0-39.9 40.0-49.9 50.0-59.9 60.0-69.9 SCL size classes (cm) No significant difference in mean SCL between study areas for turtles captured at either seagrass habitat or jetties Green turtles captured at jetties were significantly smaller than those captured at seagrass habitats (t 36 = 7.14, p < 0.001) 70 Entanglement Netting: Seagrass Habitat 70 Cast Netting: Jetty Habitat 60 60 50 n = 8 n = 25 50 40 40 n = 6 n = 2 30 30 20 20 10 10 0 ABC LLM 0 ABC LLM

% with FP GREEN TURTLES WITH FP 11 of 41 turtles had FP tumors (26.8%); all from the LLM 11 of 33 green turtles had tumors (33.3%) in 2010 (Tristan et al. 2010) Length Frequency of Turtles with FP 80% 70% 60% 50% 40% 30% n = 10 n = 3 20% 10% 0% n = 19 n = 8 n = 2 20.0-29.9 30.0-39.9 40.0-49.9 50.0-59.9 60.0-69.9 SCL Size Classes (cm) Size of turtles with FP was consistent with those afflicted in other Atlantic locations (Foley et al. 2005; Hirama and Ehrhart 2007)

TUMOR SCORES # of Tumors in each size class A B C D Tumor Turtle ID (<1 cm) (>1-4 cm) (>4-10 cm) (>10 cm) Score PI14-7-2w 19 7 0 0 2 PI14-7-3r 5 2 0 0 1 PI14-7-9w 1 1 0 0 1 PI14-7-10w 5 0 0 0 1 PI14-7-11w 1 1 0 0 1 PI14-7-12r 58 74 0 0 3 PM14-8-1w 17 19 1 0 3 PM14-8-2w 7 4 0 0 2 PM14-8-5w 3 5 0 0 1 PM14-8-7w 18 35 3 0 3 PM14-8-12w 6 14 0 0 2 TUMOR SCORES FOR TEXAS GREEN TURTLES (2014) TS 0 TS 1 TS 2 TS 3 % of turtles 73.2 12.2 7.3 7.3 Majority of turtles observed with FP only mildly afflicted

GREEN TURTLES WITH FP FP herpesvirus variant B, (Ene et al. 2005) was found in all tumor samples and unaffected skin samples from the 11 turtles with tumors Variant B is prevalent in the eastcentral FL and Indian River Lagoon green turtle foraging population 14 of 41 green turtles had marine leeches (Ozobranchus spp); 6 of these had both leeches and tumors Typically located on the flippers and axillary/inguinal regions of the turtle Some leeches were found attached to tumors Marine Leeches

SUMMARY AND CONCLUSIONS Size of green turtles with FP in Texas was consistent with those afflicted in other locations with juveniles (40-60 cm SCL) most affected Based on the results of this study, Texas green turtles have an FP occurrence of ~ 30% and are mostly mildly afflicted Presence of FP herpesvirus variant B in Texas green turtles suggest a connection with east-central FL and Indian River Lagoon green turtles or a dispersion of this variant to more distant regions Genetic link between Texas greens and east-central FL nesting populations (Anderson and Shaver 2013) Raises new questions about the spread of this virus and the overall epidemiology of this disease Need additional samples and analyses

SUMMARY AND CONCLUSIONS On-going research is assessing other contributing factors, including: The role of marine leeches as a vector of virus transfer Presence of tumor promoting algal species in the environment Nutrient levels and environmental stressors, such as cold stunning events

ACKNOWLEDGEMENTS Texas State Aquarium Wildlife Care, Conservation and Research Fund (Leslie Peart) Texas Parks and Wildlife Department (Joel Anderson) Sea Turtle, Inc. (Jeff George and Kat Lillie) EIH Staff - Patti Koch, Jenny Oakley, James Yokley 2014 Turtle Crew: Brian Alleman Rachel George Sherah Loe Jim DelBene Rachel Byrne Suzanne Tenison Adriana Leiva (TPWD)