Use of point of care device in Otostrongylus circumlitus infected Northern Seal (Mirounga angustirostris) and ( vitulina) Denver Coleman1, Cara Field2, Dan Fletcher3, and Karl Jandrey1 1School of Veterinary Medicine, University of , Davis, California, USA 2The Marine Center, Sausalito, California, USA 3Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, USA

MATERIALS AND METHODS RESULTS CONT.

Study Population Results Continued • Measurements from coagulation curves were compared: clot time (CT), clot • The control population was composed of juvenile NES and HS undergoing formation time (CFT), α angle (Alpha), maximum clot firmness (MCF), lysis index at 30 rehabilitation at between May–August 2017. & 60 minutes (Li30 & Li60, respectively, amplitude 10 (A10) and amplitude 20 (A20) • The study population were undergoing parallel rehabilitation. Animals were for healthy and O. circumlitus infected NES and HS (Table 1). presumably diagnosed with O. circumlitus arteritis based on clinical criteria • There were differences between NES and HS with each measurement, as well developed by The Marine Mammal Center or necropsy. Samples were collected at as differences between healthy and O. circumlitus affected individuals within a time of O. circumlitus diagnosis, during therapy and post-recovery. species.

Sample Collection Table 1. Median coagulation parameters from (A) healthy and O. circumlitus affected HS and from (B) • healthy and O. circumlitus affected NES. Image 1. Juvenile (Mirounga angustirostris) and Harbor Seal (Phoca Samples were collected from the extradural intravertebral sinus using a 20g 1.5” vitulina) undergoing rehabilitation at The Marine Mammal Center between May – August 2017. needle and 3cc syringe. All samples were run within 2 minutes of collection. A B Viscoelastic Machine • Coagulation curves were performed using an Entegrion Portable Coagulation Monitor BACKGROUND (PCM; Entegrion Incorporated, Durham, NC, 27703, USA). All assays run for 60 minutes following manufacturer specifications. • Otostrongylus circumlitus is a common nematode found in the trachea, bronchi and bronchioles of Harbor Seals (HS; Phoca vitulina) and Northern Elephant Seals (NES; Limitations Mirounga angustirostris) (Gulland et al., 1997). Infected individuals typically may be • There is no ante-mortem test for O. circumlitus, therefore we assume animals are subclinical, but many animals have moderate to severe disease including obstructive infected based on physical exam findings and progression of clinical signs during bronchitis and briochiolitis as well as pneumonia and pulmonary abscesses secondary rehabilitation. We assume the control population represent the wild population and to concurrent bacteria infections (Gulland et al., 1997). are absent or subclinical of O. circumlitus infection. The point of care device is still in SUMMARY AND ONGOING WORK • Both species are afflicted by pulmonary O. circumlitus infections, but infections in testing phase and has not been tested against a reference standards. NES may result in disseminated intravascular coagulation (DIC) associated with larval migration (Gulland et al., 1996). Clinical signs include anorexia, emaciation, Summary depression, respiratory distress, bronchospasm, expulsion of blood-tinged mucus and RESULTS • A viscoelastic point of care device is valuable in the diagnosis of a coagulation defect in these species nasal discharge (Measures, 2001) and may contribute to the development and monitoring of therapies targeted at Otostrongylus circumlitus infections. • No ante-mortem test for prepatent O. circumlitus infections exists for NES because Results Ongoing Work the arteritis develops during larval migration, prior to maturation and reproduction. • A total of 25 samples were collected (NES n = 16; HS n = 9). Samples were collected • We continue to collect more of these data to establish a reference interval in healthy populations for Diagnosis and treatment are based solely on physical exam, hematological and during admission, treatment and release. Samples were collected for each group, the pilot study to compare against affected animals. We plan to carry over research into the biochemistry findings. healthy NES (n = 10), HS (n = 6), and O. circumlitus affected NES (n = 6), HS (n = 3). following year to better support diagnostics and treatment protocols (Kaye et al., 2016). • We developed a pilot study to test a point of care device for comparing coagulability • O. circumlitus individuals show hypocoagulability and hyperfibrinolysis compared to • We plan to investigate the consistency and accuracy compared to traditional thromboelastography in between healthy and O. circumlitus infected individuals. We sought to establish healthy controls for both NES and HS (Fig. 1). both species. reference ranges for NES and HS as well as elucidate signs associated with infection. 60 A 40 REFERENCES AND ACKOWLEDGEMENTS 20 0 Acknowledgements 0 600 1200 1800 2400 3000 3600 • We would like to thank the staff and volunteers at The Marine Mammal Center for allowing us to -20 participate in rehabilitation and collection of blood samples for this pilot study. -40 • UC Davis STAR (Student Training in Advanced Research) Program, UC Davis Veterinary Scientist Training Program Firmness (mm) -60 Time (s) References B 60 1. Gulland FMD, Beckmen K, Burek K, Lowenstine L, Werner L, Spraker T, Dailey M, and Harris E. 1997. 40 Nematode (Otostrongylus circumlitus) infestation of Northern Elephant Seals (Mirounga angustirostris) stranded along the Central California Coast. Marine Mammal Science 13(3): 446-459. 20 2. Gulland FMD, Wener L, O’Neill S, Lowenstine J, Trupkiewitz J, Smith D, Royal B, and Strubel I. 1996. 0 Baseline coagulation assay values for Northern Elephant Seals (Mirounga angustirostris) and Disseminated Intravascular Coagulation in this species. Journal of Wildlife Disease (3): 536-540. -20 0 600 1200 1800 2400 3000 3600 3. Kaye S, Johnson s, Arnold RD, Nie B, Davis JT, Gulland FMD, Abou-Madi N, and Fletcher DJ. 2016. Adult O. circumlitus -40 Pharmacokinetic study of oral ε-aminocaproic acid in Northern Elephant Seal (Mirounga Firmness (mm) Time (s) angustirostris). Journal of Zoo and Wildlife Medicine 47(2): 438-446. Image 2. Lung from a fatal case of O. circumlitus infection in a NES. Lungs are diffusely congested -60 4. Measures, LN. 2001. Lungworms of marine . In Parasitic Diseases of Wild Mammals 2nd and swollen with large areas of consolidation. Adult O. circumlitus parasites are visible on gross Figure 1. Coagulation curves from (A) healthy (blue) and O. circumlitus infected (red) NES edition. Eds W.M. Samuel, M.J. Pybus, A.A. Kocan. Ames, Iowa State University Press. pg 279-300. examination of pulmonary parenchyma. Courtesy Dr. Padraig Duignan at The Marine Mammal Center. and from (B) healthy (blue) and O. circumlitus affected (red) HS.