Gornik KR, Pirie CG, Marrion RM, Ocial JN, Innis CJ. 2016. Ophthalmic variables in rehabilitated juvenile Kemp’s ridley sea turtles (Lepidochelys kempii). J Am Vet Met Assoc 248: 673-680.

OBJECTIVE To determine central corneal thickness (total corneal thickness [TCT], epithelial thickness [ET], and stromal thickness [ST]), anterior chamber depth (ACD), and intraocular pressure (IOP) in Kemp's ridley sea turtles (Lepidochelys kempii).

DESIGN Prospective cross-sectional study.

ANIMALS 25 healthy rehabilitated juvenile Kemp's ridley sea turtles.

PROCEDURES Body weight and straight-line standard carapace length (SCL) were recorded. All turtles underwent a complete anterior segment ophthalmic examination. Central TCT, ET, ST, and ACD were determined by use of a spectral-domain optical coherence tomography device. Intraocular pressure was determined with a rebound tonometer; the horse setting was used to measure IOP in all 25 turtles, and the undefined setting was also used to measure IOP in 20 turtles. For each variable, 3 measurements were obtained bilaterally. The mean was calculated for each eye and used for analysis purposes.

RESULTS The mean ± SD body weight and SCL were 3.85 ± 1.05 kg (8.47 ± 2.31 lb) and 29 ± 3 cm, respectively. The mean ± SD TCT, ET, ST, and ACD were 288 ± 23 μm, 100 ± 6 μm, 190 ± 19 μm, and 581 ± 128 μm, respectively. Mean ± SD IOP was 6.5 ± 1.0 mm Hg when measured with the horse setting and 3.8 ± 1.1 mm Hg when measured with the undefined setting.

CONCLUSIONS AND CLINICAL RELEVANCE

Results provided preliminary reference ranges for objective assessment of ophthalmic variables in healthy juvenile Kemp's ridley sea turtles.

- Key Points:o Cold-stunned turtles often have multiple physiologic derangements including

metabolic and respiratory acidosis, electrolyte imbalances, dehydration, and impaired renal function, and many have concurrent lesions of varying severity in the resp, GI, integumentary, skeletal, and urinary systems.

o Occasionally develop ocular disease. Chelonians – keratopathies, uveitis, cataracts subsequent to exposure to

cold. Sea turtles – corneal fibropapillomatosis (alphaherpesvirus).

Focal, necrotizing lesions with infiltration of heterophils and bacteria in cornea, sclera, conjunctiva, eyelids.

o Purpose: Determine central corneal thickness (TCT, ET, ST, ACD) and IOP (tonometer) in juvenile Kemp’s ridley sea turtles.

o 12/25 had iridescent cells within cornea/anterior lens capsule, normal variation. Reported in corneas of several fish and amphibians.

Located either throughout corneal stroma, as a separate layer between stroma and Descemets membrane, or within either the DM or corneal endothelium.

Teleost fish – iridescent cells form a layer between anterior and posterior corneal stroma (aka autochthonous layer).

Proposed functions – reduction of intraocular glare to increase visual range underwater, birefringence, filtration or polarization of color, camouflage, and enhancement or suppression of reflections.

o 8/25 turtles had minor paraxial corneal scarring, consistent with Florida keratopathy.

FL keratopathy – commonly identified in cornea of dogs and cats in tropical and subtropical climates, described also in horses and birds.

Multifocal, round-to-oval, gray-to-white, well-delineated opacities with uniform density in the anterior cornea.

In Kemp’s, possibly due to migration north from Gulf of Mexico, although lesions originate from corneal epithelium (unlike dogs, throughout the corneal stroma).

o Histo would be required to confirm whether or not the corneal lesions were FL keratopathy in these turtles (lesions would contain vacuoles and rod-like structures).

o TCT thinner than dogs, cats, horses; thicker than Hermann’s tortoises, Greek tortoises, red-footed tortoises, serrated hingeback tortoises.

A-mode US - TCT of RES considerably thinner than Kemp’s. Histo - TCT juvenile leatherbacks and greens substantially thinner, adult

leatherbacks and greens similar vs juvenile Kemp’s.o ACD widely variable in this study.

Aquatic chelonians have evolved ability to use iris sphincter muscle to alter anterior profile of lens to dramatically change curvature and refractive ability of the lens for sight in water (compensation for hyperopia).

Potentially could account for variation within and between turtles.o IOP on horse setting consistently higher than IOP on undefined setting.

IOPs in Kemp’s substantially lower than IOPs previously reported for several land tortoise and turtle spp.

Other studies used applanation tonometry. Only studies in Hermann’s tortoises and RES used rebound

tonometry. Direct measurement of IOP by manometry (gold standard) would

be required to determine which method of tonometry and which calibration setting is most accurate.

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Innis CJ, McGowan JP, Burgess EA. 2019. Cold-stunned loggerhead sea turtles (Caretta caretta): initial vs. convalescent physiologic status and physiologic findings associated with death. J Herp Med Surg 29: 105-112.

Abstract: This study evaluated the physiologic status of 155 loggerhead sea turtles (Caretta caretta) that stranded on Cape Cod, Massachusetts, between 2008 and 2016 after exposure to naturally cold temperatures. One hundred thirty-five turtles (87%) survived to be released into the wild, whereas 20 turtles (13%) died during the first week of hospitalization. Comparisons of the initial data for turtles that died vs. those that survived indicated that turtles that died had significantly higher blood glucose, potassium, lactate, and partial pressure of carbon dioxide as well as significantly lower pH, partial pressure of oxygen (pO2), heart rate, and respiratory rate. Convalescent data for 80 turtles were acquired 10.5 days (median) after admission (interquartile range: 6–17 days; range: 5–66 days). Convalescent turtles had significantly higher body temperature, blood urea nitrogen (BUN), sodium, chloride, pH, pO2, and ionized calcium (iCa), as well as significantly lower glucose, lactate, and bicarbonate. BUN, pH, and iCa were positively correlated with the number of days in the hospital, whereas glucose and lactate were negatively correlated. Results of this study indicate that the majority of cold-stunned loggerhead sea turtles had a favorable prognosis with medical management. More severely affected turtles showed a variety of physiologic derangements and had a worse prognosis.

- Introduction:o Goal of study – evaluation of initial and convalescent physiologic status of

loggerhead sea turtles that survived cold-stunning as well as initial status of turtles that did not survive.

o Hypothesized initial status of survivors would be significantly different than that of turtles that died and that convalescent data for survivors would demonstrate significant changes from their initial values.

- M+M:o Retrospective, loggerheads NEAq 2008-2016.

Data – body temp, weight, straight carapace length, HR, RR, BUN, iCa, iMg, Cl, Cr, Glu, K, lac, Na, pCO2, pO2, pH, bicarbonate.

For survivors, convalescent data defined as last available data acquired at least 5 days after admission to the hospital.

Turtles medically assessed, gradually warmed. Treated for physiologic derangements and pathologic conditions.

Surviving turtles introduced into rehabilitation pools, health status serially evaluated.

Majority transferred to secondary facilities for completion of rehabilitation before release.

- Results and Discussion:o 155 turtles met inclusion criteria.o 135 (87%) survived to be released.o Turtles that died – significantly higher glucose, K, lactate; significantly lower pH,

pO2, HR, RR.

o Convalescent turtles had significantly higher body temp, BUN, Na, Cl, pH, pO2, iCa; significantly lower Glu, lactate, bicarb.

o BUN and iCa positively correlated with number of days in hospital.o Glucose and lactate negatively correlated with number of days in hospital.o Aside from relatively low BUN and hyperglycemia, average initial data for

survivors consistent with previously published data for healthy conspecifics.o Prognosis for cold-stunned loggerheads good overall (87%).o Turtles that died showed more severe physiologic derangements.

Severe respiratory acidosis, hypoxia, hyperkalemia, hypermagnesemia. More severe derangements than other studies in response to anesthesia,

pound net capture, oil exposure, debilitation, general stranding, slightly less severe than trawl net capture and forced submergence.

o Initial vs convalescent data showed improvement over time. pH, pO2, BUN, most electrolytes, glucose, lactate changed over time and

converged on values typical of healthy individuals. Many convalescent turtles were still under medical management and may

not yet have been eating. Convalescent data may not be representative of final physiologic status

prior to release.o Glucose highly variable, consistent with previous reports in Kemp’s and greens.

In this study, higher in turtles that died. Cautious insulin therapy may be considered for loggerheads with

persistent hyperglycemia. Increased glucose has been correlated to increased corticosterone

during transport of Kemp’s.o Low BUN on intake be due to anorexia, although clinical observations indicate

BUN increased during convalescence before resumption of feeding. Reduced hepatic function due to low body temperature or illness also

possible.o Due to time period of this study, different analyzer used vs previous study.

Author suggests analyzer results can be considered valid regardless of whether or not they are in numerical agreement with another gold-standard analyzer if reflective of patient status clinically and outcome.

Journal of the American Veterinary Medical Association. 250(8): 909-917, 2017.                    Effects of alfaxalone administered intravenously to healthy yearling loggerhead sea turtles (Caretta caretta) at three different dosesB. E. Phillips, L. P. Posner, G. A. Lewbart, E. F. Christiansen and C. A. Harms

Phillips PE, Posner LP, Lewbart GA, Christiansen EF, Harms CA. 2017. Effects of alfaxalone administered intravenously to healthy yearling loggerhead sea turtles (Caretta caretta) at three different doses. J Am Vet Med Assoc 250: 909-917.

OBJECTIVE: To compare physiologic and anesthetic effects of alfaxalone administered IV to yearling loggerhead sea turtles (Caretta caretta) at 3 different doses. DESIGN: Randomized crossover study. ANIMALS: 9 healthy yearling loggerhead sea turtles. PROCEDURES: Animals received each of 3 doses of alfaxalone (3 mg/kg [1.4 mg/lb], 5 mg/kg [2.3 mg/lb], or 10 mg/kg [4.5 mg/lb]) administered IV in randomly assigned order, with a minimum 7-day washout period between

doses. Endotracheal intubation was attempted following anesthetic induction, and heart rate, sedation depth, cloacal temperature, and respirations were monitored. Times to first effect, induction, first voluntary muscle movement, first respiration, and recovery were recorded. Venous blood gas analysis was performed at 0 and 30 minutes. Assisted ventilation was performed if apnea persisted 30 minutes following induction. RESULTS: Median anesthetic induction time for all 3 doses was 2 minutes. Endotracheal intubation was accomplished in all turtles following induction. Heart rate significantly increased after the 3- and 5-mg/kg doses were administered. Median intervals from alfaxalone administration to first spontaneous respiration were 16, 22, and 54 minutes for the 3-, 5-, and 10-mg/kg doses, respectively, and median intervals to recovery were 28, 46, and 90 minutes, respectively. Assisted ventilation was required for 1 turtle after receiving the 5-mg/kg dose and for 5 turtles after receiving the 10-mg/kg dose. The 10-mg/kg dose resulted in respiratory acidosis and marked hypoxemia at 30 minutes. CONCLUSIONS AND CLINICAL RELEVANCE: IV alfaxalone administration to loggerhead sea turtles resulted in a rapid anesthetic induction and dose-dependent duration of sedation. Assisted ventilation is recommended if the 10 mg/kg dose is administered. Introduction/Background:

Alfaxalone has been evaluated in reptiles as an anesthetic agent and used clinically as an anesthetic induction agent

o Dose-dependent sedation was observed in green iguanas (Iguana iguana) and Horsfield tortoises (Agrionemys horsfieldii) following IM administration

o In red-eared sliders (Trachemys scripta elegans), IM alfaxalone administration resulted in rapid anesthetic induction, increased loss of muscle tone at increased doses, and longer interval from administration to anesthetic recovery when given at lower temperatures

o In juvenile estuarine crocodiles (Crocodylus porosus) and Australian freshwater crocodiles (Crocodylus johnstoni), IV alfaxalone administration at 4 temperatures resulted in variable duration of sedation, hypersensitivity to stimulation during anesthetic recovery, and apnea 1 to 2 hours following recovery

 Discussion/key points: n=9 yearling loggerheads in random crossover for IV alfaxalone at 3, 5, and 10 mg/kg Endotracheal intubation was successfully achieved in all turtles at all doses between 6 and 7

minutes after alfaxalone administration Cloacal temperatures decreased significantly from baseline to the 30-minute assessment point for

all doses and from baseline to the 60-minute assessment point for the 10-mg/kg dose

 Take home message: IV alfaxalone administration at each of the 3 doses evaluated would be suitable as an induction

agent in this species and may be followed by endotracheal intubation and inhalation anesthesia (3,5 mg/kg recommended…10mg/kg can be used if maintained with IPPV)

INHALANT ANESTHETIC RECOVERY FOLLOWING INTRAMUSCULAR EPINEPHRINE IN THE LOGGERHEAD SEA TURTLE (CARETTA CARETTA)Balko JA, Gatson BJ, Cohen EB, Griffith EH, Harms CA, Bailey KM.Journal of Zoo and Wildlife Medicine. 2018 Sep;49(3):680-8.

Balko JA, Gatson BJ, Cohen EB, Griffith EH, Harms CA, Bailey KM. 2018. Inhalant anesthetic recovery following intramuscular epinephrine in the loggerhead sea turtle (Caretta caretta). J Zoo Wildl Med 49: 680-688.

Abstract: Prolonged anesthetic recovery time is a common complication of chelonian inhalant anesthesia and may be exacerbated by right-to-left intracardiac shunting of blood. Epinephrine may decrease intracardiac shunting, which may shorten anesthetic recovery time. The study objective was to assess inhalant anesthetic recovery time following intramuscular epinephrine compared with saline in the loggerhead sea turtle (Caretta caretta). With the use of a prospective, randomized, blinded, crossover

design with a 1-wk washout period, six turtles were anesthetized with intravenous (IV) alfaxalone 3 mg/kg, orotracheally intubated, manually ventilated with 3.5% isoflurane inhalant in 100% oxygen for 90 min, and administered either intramuscular (IM) epinephrine 0.1 mg/kg or IM saline 0.1 ml/kg. Isoflurane administration was immediately discontinued and turtles were manually ventilated with room air until extubation. Physiologic variables, sedation scores, end-tidal carbon dioxide (ETCO2) and isoflurane (ETISO) concentrations, time to first movement, and time to extubation were recorded and two-time-point venous blood gas analyses performed. Data were compared with the use of paired t-tests and repeated-measures analyses of variance (ANOVA) (P < 0.05). No morbidity, mortality, or adverse events occurred. ETCO2 and ETISO did not significantly change over time during the isoflurane delivery period (P=0.990). Mean time to first movement was significantly faster following epinephrine (69.24 6 12.28 min) compared with saline (87.71 6 27.05 min, P = 0.047). Although differences were not statistically significant (P = 0.133), time to extubation was at least 30 min faster (31–123 min) in 4/6 turtles following epinephrine compared with saline. Intramuscular epinephrine significantly reduces time to first movement during isoflurane anesthetic recovery in loggerhead sea turtles. Intro

Turtle have one ventricle with incomplete septum When PO2 is low, increased parasympathetic tone constricts the pulmonary arterial sphincter.

o Rise in pulmonary vascular resistance decreases pulmonary circulationo Promotes right-to-left shunting of oxygen-poor blood to systemic circulation

Under anesthesia, this decreases respiratory elimination of anesthetic gas Epi reduces shunt by increasing sympathetic tone.  Speeds recovery in snapping turtles and gators

Results/Discussion Time to first movement was faster following epinephrine compared with saline. Time to extubation was faster in 4/6 turtles following epinephrine (not statically significant).

o Lack of significance could be due to low sample size. Supports use of IM epinephrine in loggerheads to reduce isoflurane recovery time. No negative side effects End-tidal and venous blood gasses throughout the anesthetic event did not support a significant

change in right-to-left intracardiac shunting.o Unexpected.  Mechanism epi improves recover may not be by reversing shuntingo Or limitations of measurement of gasses or gas dilution

Take home: intramuscular epinephrine significantly decreased time to first movement in isoflurane-anesthetized loggerhead sea turtles. Also faster time to extubation noted in 4/6 turtles. No adverse effects. Intramuscular epinephrine may be a beneficial adjunct to isoflurane anesthetic recovery in loggerhead sea turtles.  Study this in combination with the snapping turtle and alligator epinephrine recovery papers.

J Wildl Dis 51: 509-512Pharmacokinetic behavior of meloxicam in loggerhead sea turtles (Caretta caretta) after intramuscular and intravenous administrationLai OR, Di Bello A, Soloperto S, Freggi D, Marzano G, Cavaliere L, Crescenzo G

Data on reptile analgesia are scarce for nonsteroidal anti-inflammatory drugs (NSAIDs) and opioids and almost completely lacking in sea turtles, even though emergencies requiring correct pain management are very frequent in their rehabilitative medicine; therefore, dosage regimens extrapolated from other species involve the risk of clinical failure and damage to the animals. We describe the pharmacokinetic behavior of meloxicam in the loggerhead sea turtle (Caretta caretta). We chose meloxicam because of its selective anti-cyclooxygenase-2 activity and lesser adverse side effects. No data are available on the capacity of turtles to tolerate NSAIDs, so we chose a dose of 0.1 mg/kg of meloxicam. Plasma concentrations of meloxicam were unexpectedly low both for intravenous (IV; maximum concentration [Cmax] = 0.04±0.02

µg/mL) and intramuscular (IM; Cmax = 0.07±0.09 µg/mL) administration. A double-peak phenomenon occurred after both IV (time for second peak concentration Tmax2 = 10.33±10.89 h) and IM (Tmax2 = 1.17±0.75 h). The second peak after IM injection was premature, so some difficulty and delay in absorption appears to be an appropriate explanation. Furthermore, the area under the curve, and therefore systemic bioavailability (F = 31.82±28.24%), after both IV (0.30±0.29) and IM (0.10±0.03) injection appeared particularly limited. Terminal elimination slope and mean residence time indicated fast elimination after IM dosing; as a consequence, plasma concentrations dropped below analytic limits in 8 h. Considering that IM is the favored route of administration of drugs in rescue centers, it is unlikely that meloxicam at 0.1 mg/kg is an appropriate choice, particularly in long-term pain management protocols.

Introduction: Meloxicam- cyclooxygenase-2 selective NSAID Extensive hepatic metabolism, then excreted in urine and feces Most reptiles are uricotelic- only few thousand nephrons (compared to millions in mammals), so

potential for nephrotoxic effects, as well as reptiles eat infrequently and more likely to be given on empty stomach.

Materials & Methods: 8 clinically healthy loggerhead sea turts; randomized crossover design with 15 day wash-out

o Each animal received 0.1mg/kg either IV or IMResults/Discussion

No local or systemic reactions Plasma concentrations were low Double peak occurred after both routes Potential difficulty in absorption from the IM site could explain presence of early second peak

Take Home: Meloxicam at 0.1mg/kg is not a good choice

Case Descriptions of Fibropapillomatosis in Rehabilitating Loggerhead Sea Turtles Caretta Caretta in the Southeastern USA

Annie Page-Karjian, Terry M Norton, Craig Harms, Doug Mader, Larry H Herbst, Nancy Stedman, Nicole L Gottdenker

Dis Aquat Organ, 115 (3), 185-91 2015 Aug 20

Taxa: Reptilia → Testudines → Cryptodira (suborder) → Chelonioidea (superfamily) → Cheloniidae → Carrettinae (subfamily)Abstract: Fibropapillomatosis (FP) is a debilitating neoplastic disease that affects all species of hard-shelled sea turtles, including loggerhead turtles Caretta caretta. FP can represent an important clinical concern in rehabilitating turtles, since managing these infectious lesions often requires special husbandry provisions including quarantine, and FP may affect clinical progression, extend rehabilitation duration, and complicate prognoses. Here we describe cases of rehabilitating loggerhead turtles with FP (designated FP+). Medical records of FP+ loggerhead cases from 3 sea turtle rehabilitation facilities in the southeastern USA were reviewed. Between 2001 and 2014, FP was observed in 8 of 818 rehabilitating loggerhead turtles (0.98% overall prevalence in admitted patients). FP+ loggerhead size classes represented were large juvenile (straight carapace length, SCL: 58.1-80 cm; n=7) and adult (SCL>87 cm; n=1). Three turtles presented with FP, and 5 developed tumors during rehabilitation within a range of 45 to 319 d. Sites of new tumor growth included the eyes, sites of trauma, neck, and glottis. FP+ turtles were scored as mildly (3/8), moderately (4/8), or heavily (1/8) afflicted. The mean total time in rehabilitation was 476±355 d (SD) (range: 52-1159 d). Six turtles were released without visible evidence of FP, 1 turtle was released with mild FP, and 1 turtle with internal FP was euthanized. Clinical decision-making for FP+ loggerhead patients can be aided by such information as time to tumor development, anatomic locations to monitor for new tumor growth, husbandry considerations, diagnostic and treatment options, and comparisons to FP in rehabilitating green turtles Chelonia mydas.

Background:- Fibropapillomatosis (FP): transmissible neoplastic disease of hard-shelled sea turtles

- Etiologic agent: chelonid fibropapilloma-associated herpesvirus (CFPHV)- Clinical signs: tumors on eyes, skin, and internal organs- Clinical pathology: anemia, lymphocytopenia, neutrophilia, monocytosis,

hypoproteinemia, hyperglobulinemia- FP is occasionally seen in loggerheads in the southeastern US

Methods: Retrospective analysis of rehabilitated loggerhead sea turtles (n=818) in the southeastern US to describe case characteristics of FP+ turtles

Key Points:- Only 7 loggerhead sea turtles (0.98%) had or developed FP lesions, which were

confirmed via histology- Most FP+ turtles were large juveniles, similar to green sea turtles- Variety of presenting complaints (floating, anthropogenic, etc.)- Most cases were mild to moderate FP lesions- Most common tumor locations: ocular > flipper/shoulder = neck = tail base > glottis =

head fracture lesion = plastron = inguinal region (1 each)- High ocular predilection is also seen in green sea turtles- Dorsoanterior body is most common region for tumor occurrence

- CT was better for diagnosing internal tumors than radiographs- Some turtles developed FP following admission to rehabilitation facility- Most turtles were released without evidence of FP because regrowth following excision

was rare- One female turtle was euthanized due to severe FP (>50% of skin affected) with FP

tumors throughout most internal organs- Green sea turtles are typically more severely affected and have poorer survival rates

(28%)

Conclusions: Fibropapillomatosis in rehabilitated loggerhead sea turtles is rare, presents with mild to moderate clinical signs, and is rarely fatal, unlike the severe disease seen in green sea turtles.

INTRAVENOUS LIPID EMULSION TREATMENT REDUCES SYMPTOMS OF BREVETOXICOSIS IN TURTLES (TRACHEMYS SCRIPTA).Cocilova CC, Flewelling LJ, Granholm AA, Manire CA, Milton SL.

J Zoo Wildl Med. 2019 Mar 1;50(1):33-44.

Taxa: Testudines → Cryptodira (suborder) → Testudinoidea (subfamily) → Emydidae Abstract: Harmful algal blooms (HABs) occur when excess nutrients allow dinoflagellates to reproduce in large numbers. Marine animals are affected by blooms when algal toxins are ingested or inhaled. In the Gulf of Mexico, near annual blooms of Karenia brevis release a suite of compounds (brevetoxins) that cause sea turtle morbidity and mortality. The primary treatment at rehabilitation facilities for brevetoxin-exposed sea turtles is supportive care, and it has been difficult to design alternative treatment strategies without an understanding of the effects of brevetoxins in turtles in vivo. Previous studies using the freshwater turtle as a model species showed that brevetoxin-3 impacts the nervous and muscular systems, and is detoxified and eliminated primarily through the liver, bile, and feces. In this study, freshwater turtles (Trachemys scripta) were exposed to brevetoxin (PbTx-3) intratracheally at doses causing clear systemic effects, and treatment strategies aimed at reducing the postexposure neurological and muscular deficits were tested. Brevetoxin-exposed T. scripta displayed the same behaviors as animals admitted to rehabilitation centers for toxin exposure, ranging from muscle twitching and incoordination to paralysis and unresponsiveness. Two treatment regimes were tested: cholestyramine, a bile acid sequestrant; and an intravenous lipid emulsion treatment (Intralipid) that provides an expanded circulating lipid volume. Cholestyramine was administered orally 1 hr and 6 hr post PbTx-3 exposure, but this regime failed to increase toxin clearance. Animals treated with Intralipid (100 mg/kg) 30 min after PbTx-3 exposure had greatly reduced symptoms of brevetoxicosis within the first 2 hr compared with animals that did not receive the treatment, and appeared fully recovered within 24 hr compared with toxin-exposed control animals that did not receive Intralipid. The results strongly suggest that Intralipid treatment for lipophilic toxins such as PbTx-3 has the potential to reduce morbidity and mortality in HAB-exposed sea turtles.Background:

- Harmful algal blooms (HABs): Overabundance of dinoflagellates produce toxins- Red tide = HAB caused by Karenia brevis, which produces brevetoxins- Brevetoxins binds to voltage-gated sodium channels, causing muscle twitching, ataxia,

paralysis, an coma- Brevetoxins readily cross mucous membranes and can be ingested or inhaled from

ocean aerosols- Brevetoxins are excreted by the liver bile and feces

- Loggerheads exposed to brevetoxins have increased oxidative stress- Once exposure stops, brevetoxins are cleared from tissues in 24-48 hours but can remain

in feces for longer- Dehydration via furosemide may help brevetoxicosis turtles- Cholestyramine, an anion exchange resin that binds bile salts, can be used to prevent bile

reabsorption- Intralipid, a soybean oil/egg yolk protein, may draw toxins out of fatty membranes of

cells

Objective: Evaluate the efficacy of cholestyramine and/or Intralipid to reduce the clinical signs of sliders (n=46) inoculated with brevetoxins.Key Points:

- Turtles inoculated with brevetoxins showed clinicals signs at 5 min postexposure then signs decreased in the following 24 hours post exposure

- Clinical signs: head bobbing, muscle twitching, ataxia, circling, and paralysis- Brevetoxin was distributed through all tissues by 1 hour- Cholestyramine did not reduce clinical signs or reduce bile toxin concentrations- Intralipid-treated turtles had more toxin in the bile/feces after 24 hours than other

treatments- Turtles that received Intralipid 30 minute after exposure had improved signs within 2

hours and no clinical signs at 6 hours- 100 mg/kg was better than 50 mg/kg

- Turtles that received Intralipid 30 minutes before exposure has almost no clinical signs- 1 hr post toxin exposure, 100 mg/kg intralipid had lower toxin levels in

kidney/liver/brain/heart- When treated with intralipid before toxin administration, lower tissue concentrations

overall

Conclusion: Intralipid IV treatment after brevetoxin exposure reduces clinical signs and increases toxin tissue clearance resulting in lower tissue toxin concentrations overall, and works best at the higher 100 mg/kg dose in freshwater turtles.

OZOBRANCHUS MARGOI INFECTIONS IN LOGGERHEAD TURTLES (CARETTA CARETTA) IN GREECE AND POTENTIAL TREATMENT OPTIONSSterioti A, Doxa CC, Grigoriou P, Vardanis G, Cascarano MC, Katharios P. Journal of Exotic Pet Medicine. 2017 Jul 1;26(3):196-9.Abstract: Two injured loggerhead turtles, Caretta caretta, were presented to the public aquarium in Crete, infested by the parasitic marine leech, Ozobranchus margoi. This report describes these 2 loggerhead turtles and parasitic marine leeches along with the results of various treatment trials. Iodine ointment was the most efficient treatment against all life stages of O. margoi, taking into account animal welfare, ease of application, and efficacy. Monitoring the occurrence of parasitic leeches is important, as apart from the direct clinical affects to the host animal, they have been reported as vectors of threatening viral diseases.

- Case report of two turtles with Ozobranchus margoi infection.- Genus Ozobranchus consists of 9 species, 2 are permanent parasites of sea turtles:

Ozobranchus branchiatus and Ozobranchus margoi.- One turtle covered with > 1,000 leeches- Treatment trial: leeches were placed in different culture plates with different treatments

to see which most effectively killed the parasites.

Results

- The most prominent morphologic structure was the gills: 5 pairs of lateral branching appendages

- Leech body divided into two main parts: anterior (trachelosome) and posterior (urosome).

- 2 suckers: a small ventral anterior sucker and a larger posterior sucker.- Eggs were brownish and attached in patches on the plastron.- 100% leech mortality with all treatments

o Treatments with iodine, hydrogen peroxide, and ethanol were the most effectiveo Iodine most environmentally safe and practical

Application of a thin film of iodine ointment killed leeches in 20 min Effective on all life stage, no adults or eggs.

COCCIDIAL INFECTION OF THE ADRENAL GLANDS OF LEATHERBACK SEA TURTLES (DERMOCHELYS CORIACEA)

Ferguson SD, Wellehan Jr JF, Frasca Jr S, Innis CJ, Harris HS, Miller M, Weber ES, Walden HS, Greiner EC, Merigo C, Stacy BA.Journal of wildlife diseases. 2016 Oct;52(4):874-82.

Abstract: Histologic lesions incidental to the cause of death were observed in the adrenal glands of 17 subadult and adult leatherback sea turtles (Dermochelys coriacea) found dead or moribund on or near shore in North America. Round bodies, 250–300 lm in diameter composed of an outer capsule and large multinucleated cells surrounding a central mass of acellular material were distributed throughout the affected glands. Protozoal etiology was suspected based on some resemblance to coccidia; however, features diagnostic for coccidial infection were lacking in all but one case, which had a focal area of adrenalitis containing zoites. A novel eucoccidian partial 18S rRNA genetic sequence was consistently detected in adrenal glands with lesions. With the use of quantitative PCR, a specific area of the V4 region of the coccidian 18S gene was quantified in affected adrenal glands and correlated significantly with density of the histologic lesions. A second distinct, but closely related, 18S sequence was also amplified from the adrenal gland of one turtle and from a fecal sample containing unsporulated coccidian oocysts. The two 18S sequences identified from leatherback sea turtles form a clade within the family Eimeriidae.

Further investigation is required to understand better the morphology of the life stages, life cycle, and potential effects of this coccidian parasite on adrenal function.

Introduction/Methods Sea turtle parasites: most significant coccidian is Caryospora cheloniae in green sea turtles

Associated w/ epizootics and mortalities in captive and wild populations  Other coccidian: Eimeria caretta: found in a loggerhead; significance unknown. 

Evaluated adrenal lesions found in multiple leatherback sea turtle adrenal glands and used PCR to identify coccidian DNA within the lesions

Results/Discussion Adrenal lesions, consistent with protozoal infection, observed microscopically in 17 subadults

and adults, but none in juveniles. All adrenals were unremarkable grossly. Organisms: round, 250+ micron, prominent outer capsule, central material in core.  3 turtles had a few individual suspected protozoal lesions in other tissues: small intestine

submucosa, connective tissue of dermis, phallus, tongue PCR amplified several segments of the 18S gene that confirmed this parasite to belonging to the

family Eimeriidae One turtle had coccidian oocyst found in a fecal sample Authors assume that capsule and multinucleate giant cells component are affected host cells

altered by infection, as lesions lacked any intact parasitic stages  Other animals with adrenal coccidians reported:

Virginia opossum (w/ Besnoittia darling) Golden hamsters (experimental infxn w/ Besnoittia jellisoni)

Takeaways Report of a coccidian infection of the adrenal glands in leatherback sea turtles found dead or

moribund Suggests that the species may be novel, but further work is needed to confirm this

COCCIDIOSIS IN GREEN TURTLES (CHELONIA MYDAS) IN AUSTRALIA: PATHOGENESIS, SPATIAL AND TEMPORAL DISTRIBUTION, AND CLIMATE-RELATED DETERMINANTS OF DISEASE OUTBREAKS.Ban de Gouvea Pedroso S, Phalen DN, Terkildsen M, Blyde D, March DT, Gordon AN, Chapman PA, Mills PC, Owen H, Gillett A, Lloyd HB. Journal of Wildlife DiseasesAbstract: An epizootic of coccidiosis in free-ranging green turtles (Chelonia mydas) occurred in Australia in 1991 and the parasites were thought to be Caryospora cheloniae. Recurring outbreaks over an increased geographic range followed. We used medical records and temporal and spatial data of turtles diagnosed with coccidiosis between 1991 and 2014 to characterize the disease and factors associated with outbreaks. Most affected animals were subadults or older. Neurologic signs with intralesional cerebral coccidia were observed. Coccidia associated with inflammation and necrosis were predominantly found in the intestine, brain, kidney, and thyroid. Cases occurred in the spring and summer. Three major outbreaks (1991, 2002, and 2014) were concentrated in Port Stephens, New South Wales (NSW) and Moreton Bay, Queensland (QLD), but cases occurred as far south as Sydney, NSW. Coccidiosis cases were more likely during, or 1 mo prior to, El Nino–like events. Molecular characterization of the 18S rRNA locus of coccidia from tissues of 10 green turtles collected in 2002 and 2004 in Port Stevens and Sydney imply

that they were Schellackia-like organisms. Two sequences were identified. The genotype 3 sequence was most common (in eight of 10 turtles), with 98.8% similarity to the 18S sequence of Schellackia orientalis. The Genotype 4 sequence was less common (in two of 10 turtles) with 99.7% similarity to the 18S sequence of the most common genotype (Genotype 1) detected in turtles from the 2014 Moreton Bay outbreak. Our study will help with the identification and management of future outbreaks and provide tools for identification of additional disease patterns in green turtles.

Intro- Caryospora cheloniae was fist described as parasite of green turtles in Cayman Islands.

o Coccidia located in hindgut associated with hemorrhagic enteritiso Never molecularly identified.

- 1991: epizootic of coccidiosis in Moretone Bay, Queensland, Auso Lesions throughout the intestine, also extraintestinal lesions containing organisms,

particularly in the braino Oocyte morphology not identical to Cayman Island outbreako Genetics: novel coccidian (Genotype 1) relate to Schellackia spp.o Second coccidian (Genotype 2) assoc with lesion in kidney and thyroid.

Parasites from Leatherbacks (Ferguson et al., JWD 2016) related - Coccidiosis in green turtle in Aus: mostly subadults in El Nino years

Results- Review of cases of coccidiosis in green turtles in Aus. from 1991-2014. 63 cases

identified.- Most (54) died. About half were depressed/moribund. Almost half floating on

presentation. - Neuro signs – circling, head tilt were common. Almost all with neuro signs died/euth.

Most had histo evidence of cerebral parasites.- Lesions: all portions of the GI tract usually assoc with parasites

o White foci in the brain gross, Most had histo brain lesionso Less frequent, white foci in kidney and thyroid. Inflammation on histo.

- Coccidia cases assoc. with El Nino events.- Molecular: two genotypes (3 and 4), grouped with Schellackia spp.

Discussion- Coccidiosis should be suspected in green turtles stranded Sept-Feb, peaking in October,- Australian eastern coast. Between Sydney and Fraser Island. Moreton Bay.- Clinical signs, neuro and GI. Body condition not useful for diagnosis.- Prognosis for clinically ill turtles is grave - Suspect when enteritis or gastritis is grossly present.

o Diagnosed via fecal, GI mucosal scrape, histo, PCRo Although absence of coccidia in these lesions does not exclude infection

- Brain lesions more severe, with more organisms present than previously described.- Age (subadult) suggests exposer following recruitment to coastal feeding grounds.

Baseline plasma thromboelastography in Kemp’s ridley (Lepidochelys kempii), green (Chelonia mydas) and loggerhead (Caretta caretta) sea turtles and its use to diagnose coagulopathies in

cold-stunned Kemp’s ridley and green sea turtles

Barratclough A, Tuxbury K, Hanel R, Stacy NI, Ruterbories L, Christiansen E, Harms CA

J. of Zoo and Wildlife Medicine, 50(1):62-68 (2019)

Abstract: Cold-stunning in sea turtles is a frequent natural cause of mortality and is defined as a hypothermic state due to exposure to water temperatures <12C. Derangements of biochemistry and hematology data by cold stunning have been well documented, although the effects on coagulation have not yet been investigated. The objectives of this study were to characterize the hemostatic state of non–cold-stunned sea turtles and to compare cold-stunned sea turtles at admission and after successful rehabilitation via a sea turtle–specific thromboelastography (TEG) protocol. TEG enables evaluation of the entire coagulation process, and the methodology has recently been established in sea turtles. Initially, 30 wild and apparently healthy sea turtles were sampled as controls: loggerhead sea turtles (Caretta caretta), n¼17; Kemp’s ridley sea turtles (Lepidochelys kempii), n¼8; and green turtles (Chelonia mydas), n¼5. In addition, paired TEG samples were performed on 32 Ch. mydas and 14 L. kempii at admission and prerelease after successful rehabilitation from cold stunning. Statistically significant differences in reaction time, kinetics, angle, and maximum amplitude parameters in L. kempii and Ch. mydas species demonstrated that the time taken for blood clot formation was prolonged and the strength of the clot formed was reduced by cold stunning. These findings indicate that cold stunning may cause disorders in hemostasis that can contribute to the severity of the condition. Early diagnosis of coagulopathies in the clinical assessment of a cold-stunned sea turtle may influence the treatment approach and clinical outcome of the case.

Summary:Intro:

Cold stunning o hypothermic state due to exposure to cold water temperatures <12Co Kemp’s ridley sea turtles, green turtles, and loggerhead sea turtleso juveniles are most affected – failure to migrate to warmer water in late fallo chronic cold stun - > 2week in cold water, usually in higher latitudeso acute cold stun – <1-2weeks, short time frame, usually in lower latitudes o clinical observations consistent with coagulopathies have been described in cold-

stunned turtles (hyphema, DIC) Clotting cascade

o Reptiles lack intrinsic system and factors XI and XII Thromboelastography (TEG)

o only coagulation test to provide global evaluation of the hemostatic processo four main parameters

reaction time (R) clot formation time (K) clot formation rate [angle(a)] maximum amplitude (MA) - indicates clot strength

Goals of studyo determine baseline TEG values in three species of sea turtles o characterize hemostasis of cold-stunned turtles at admission & after rehab

M + M: turtles of 3 species (>1kg and PCV >24%) were sampled on admission and after rehab for

TEG

Results and discussion: significantly slower clot formation rate in cold-stunned in green and KR turtles MA significantly lower at the time of cold stunning in green and KR turtles Acute and chronic cold stun greens were not statistically different

o However, chronic cases showed longer time to reach maximum clot strength and acute cases achieved less overall blood clot strength

TEG between admission and prerelease demonstrated significantly reduced clot formation rate and MA in cold-stunned animals compared with the time of release

Greens had significant differences in R and K - time to initiate clot formation also affected by cold stunning

o May be adaptive - maintain microcirculation by inhibiting micro thrombi effects of thrombocytopenia due to hypothermia on clotting not evaluated as TEG

performed on plasma rather than whole blood there were individuals with abnormal TEG values as severe as the non-survivors that did

survive – serial TEG’s may be more useful for prognosis

Advancing transfusion medicine in sea turtles: optimization of a cross-matching protocol

Donnelly, Kyle A., Norton, Terry M., Zirkelbach, Bette, and Stacy, Nicole I.

Journal of Zoo and Wildlife Medicine, 50(2) : 315-321

Abstract: Whole blood transfusions are an essential treatment modality during rehabilitation of stranded sea turtles, however, standardized protocols for transfusions are not available in reptile medicine. The objective of this study was to optimize a cross-matching protocol for sea turtle blood transfusions. Fresh venous blood samples from 15 turtles (n ¼ 14 green turtles, Chelonia mydas [Cm]; n ¼ 1 loggerhead sea turtle, Caretta caretta [Cc]) were tested using a temperature-appropriate (i.e., reflecting body temperature), time-sensitive protocol in 26 reactions using two procedures for cross-match evaluation at 30 and 60 min at ambient air and water bath temperature. There were no significant differences between both protocols at 30- and 60-min incubation times or between microscopic evaluations at 2 or 5 min. The major cross-match identified 7/22 incompatible Cm–Cm reactions as observed by microscopic erythrocyte agglutination. Minor cross-matches resulted in 6/22 incompatible Cm–Cm reactions. About half of all Cm–Cm reactions (12/22) were compatible by major and minor cross-match. All Cc– Cm reactions (4/4) were incompatible. A higher than expected proportion of incompatible Cm cross-matching reactions suggests preexisting antibodies to nonself erythrocyte antigens in this species, or other factors promoting erythrocyte aggregation or agglutination. Preliminary data across Cm and Cc suggests cross-species incompatibility. These results indicate that sea turtles may react to donor erythrocytes even at the first transfusion. Concurrent major and minor cross-matching using the proposed protocol at 30-min incubation at room temperature should be considered a necessary and effective way to test for patient and donor incompatibilities in sea turtles.

Summary:Intro:

● slide agglutination technique – historically only to test for compatibility in reptiles

● no described blood types in the non-avian reptiles

● major cross-match – test for reaction between donor RBC and recipient antibodies

● minor cross-match - test for reaction between donor circulating antibodies and recipient RBC

● positive cross-match reactions – hemolysis, macroscopic and microscopic RBC agglutination

M+M:

● Blood collected from 15 turtles (14 greens, 1 loggerhead)

●

Results:

● major crossmatch identified 7/22 incompatible Cm–Cm reactions

● minor cross–matches different from major cross-match incompatibilities resulted in 6/22 Cm–Cm reactions

● half of all Cm–Cm reactions (12/22) were compatible by major and minor cross-match

● all cross-species reactions were incompatible with at least one major or one minor crossmatch

● Hemolysis noted in Cm–Cc reactions but not the Cc–Cm reactions

● agglutination scores tended to be higher in the longer incubation times

●

Discussion:

● protocol using a 30-min incubation with microscopic evaluations at 2 or 5 min is sufficient to detect cross-match compatibilities in sea turtles

● all cross-species (Cc–Cm) reactions were incompatible

● 30-min incubation protocol is sufficient

● ambient temperature used to reflect sea turtle body temperature

● sea turtles may react to donor RBC, even at the first transfusiono ~half of all Cm–Cm cross-matches had incompatible resulto Cm may have preexisting antibodies to non-self RBC’s or other factors promoting

RBC aggregation or agglutination

● transfusion reactions have been observed in sea turtleso cross-matching - inexpensive and time efficient method to minimize adverse

reactions