ESTABLISHING HEMATOLOGY AND SERUM CHEMISTRY REFERENCE INTERVALS FOR WILD HAWAIIAN MONK SEALS ( NEOMONACHUS SCHAUINSLANDI).

Hematology and serum chemistry reference intervals have been previously established for the endangered Hawaiian monk seal ( Neomonachus schauinslandi) as an imperative measure for health assessments. Monitoring the health of the wild population depends upon reference intervals that are context specific; hence we developed reference intervals from fresh samples, as opposed to frozen, from wild monk seals. This study builds on the number of parameters from previous efforts by using samples collected between 2004 and 2015 from wild monk seals. Blood samples were analyzed by a single veterinary diagnostic laboratory within 24 hr of collection from apparently healthy, wild seals during research activities. Reference intervals were determined based on the analytical steps outlined by the American Society for Veterinary Clinical Pathology. These comprehensive hematology and serum chemistry reference intervals enable more consistent and systematic interpretation of results, which will guide individual and population-level health assessment and decision-making research and recovery activities.

Survey for Placental Disease and Reproductive Pathogens in the Endangered Hawaiian Monk Seal ( Neomonachus schauinslandi).

There is considerable temporal and spatial variability in the reproductive rates of Hawaiian monk seals (HMS; Neomonachus schauinslandi). Poor reproductive performance limits the recovery of this endangered species; however, causal factors are not fully understood. There is serologic evidence that HMS are exposed to pathogens that can impact reproductive success, but the prevalence of placental infections in HMS has not been evaluated. Placental tissues ( n=50), including tissues from 25% of known HMS births, were opportunistically collected in 2011 from six Northwestern Hawaiian Islands and three main Hawaiian Islands. Reproductive histories of the sampled females were representative of the breeding population, as determined through comparisons in age of primiparity and mature reproductive rate. Placental tissues were examined histologically and screened by PCR for Coxiella burnetii, Brucella spp., Chlamydia spp., Leptospira spp., herpesviruses, and Toxoplasma gondii. There was no histologic evidence of placental pathology, and molecular analyses were negative. These negative results can be used to estimate pathogen prevalence in the nonsampled population. For an approximate population size of 1,300 HMS, we can estimate with 99% confidence that the prevalence of each pathogen tested is 9% or less. This is low relative to other pinnipeds and indicates that factors other than reproductive pathology, such as resource limitation, may drive variability in HMS reproductive rates. Further investigation into the cumulative impacts of resource limitation and other stressors on HMS reproduction is warranted.

Model recommendations meet management reality: implementation and evaluation of a network-informed vaccination effort for endangered Hawaiian monk seals.

Where disease threatens endangered wildlife populations, substantial resources are required for management actions such as vaccination. While network models provide a promising tool for identifying key spreaders and prioritizing efforts to maximize efficiency, population-scale vaccination remains rare, providing few opportunities to evaluate performance of model-informed strategies under realistic scenarios. Because the endangered Hawaiian monk seal could be heavily impacted by disease threats such as morbillivirus, we implemented a prophylactic vaccination programme. We used contact networks to prioritize vaccinating animals with high contact rates. We used dynamic network models to simulate morbillivirus outbreaks under real and idealized vaccination scenarios. We then evaluated the efficacy of model recommendations in this real-world vaccination project. We found that deviating from the model recommendations decreased the efficiency; requiring 44% more vaccinations to achieve a given decrease in outbreak size. However, we gained protection more quickly by vaccinating available animals rather than waiting to encounter priority seals. This work demonstrates the value of network models, but also makes trade-offs clear. If vaccines were limited but time was ample, vaccinating only priority animals would maximize herd protection. However, where time is the limiting factor, vaccinating additional lower-priority animals could more quickly protect the population.

MODELING A MORBILLIVIRUS OUTBREAK IN HAWAIIAN MONK SEALS (NEOMONACHUS SCHAUINSLANDI) TO AID IN THE DESIGN OF MITIGATION PROGRAMS.

We developed a stochastic susceptible-exposed-infectious-removed (SEIR) model to simulate a range of plausible morbillivirus outbreak scenarios in a randomly mixing population of 170 endangered Hawaiian monk seals (Neomonachus schauinslandi). We then modeled realistic vaccination and quarantine measures to determine the potential efficacy of such mitigation efforts. Morbillivirus outbreaks represent substantial risk to monk seals-91% of simulated baseline outbreaks grew (R0>1), and in one-third of the scenarios all, or nearly all, individuals were infected. Simulated vaccination efforts in response to an outbreak were not effective in substantially reducing infections, largely because of the prolonged interval between vaccination and immunity. Prophylactic vaccination, in contrast, could be an effective tool for preventing outbreaks. Herd immunity is practically achievable because of the small sizes of monk seal populations and the animals' accessibility on shore. Adding realistic spatial structure to the model, as informed by movement of seals tracked in the main Hawaiian Islands with the use of telemetry, greatly reduced the simulated impact of outbreaks (≤10 seals were infected in 62% of spatially structured simulations). Although response vaccination remained relatively ineffective, spatial segregation allowed herd immunity to be achieved through prophylactic vaccination with less effort. In a randomly mixing population of 170 seals, 86% would need to be vaccinated to achieve herd immunity in 95% of simulated outbreaks, compared to only approximately 60% in three spatially segregated subgroups with the same combined abundance. Simulations indicate that quarantining a modest number (up to 20) of ill seals has the potential to extinguish even fast-growing outbreaks rapidly. The efficacy of quarantine, however, is highly dependent upon rapid detection and response. We conclude that prophylactic vaccination combined with a quarantine program supported by vigilant surveillance and rapid, reliable diagnosis could greatly mitigate the threat of a morbillivirus outbreak in Hawaiian monk seals.

Protozoal-related mortalities in endangered Hawaiian monk seals Neomonachus schauinslandi.

Protozoal infections have been widely documented in marine mammals and may cause morbidity and mortality at levels that result in population level effects. The presence and potential impact on the recovery of endangered Hawaiian monk seals Neomonachus schauinslandi by protozoal pathogens was first identified in the carcass of a stranded adult male with disseminated toxoplasmosis and a captive monk seal with hepatitis. We report 7 additional cases and 2 suspect cases of protozoal-related mortality in Hawaiian monk seals between 2001 and 2015, including the first record of vertical transmission in this species. This study establishes case definitions for classification of protozoal infections in Hawaiian monk seals. Histopathology and immunohistochemistry were the primary diagnostic modalities used to define cases, given that these analyses establish a direct link between disease and pathogen presence. Findings were supported by serology and molecular data when available. Toxoplasma gondii was the predominant apicomplexan parasite identified and was associated with 100% of mortalities (n = 8) and 50% of suspect cases (n = 2). Incidental identification of sarcocysts in the skeletal muscle without tissue inflammation occurred in 4 seals, including one co-infected with T. gondii. In 2015, 2 cases of toxoplasmosis were identified ante-mortem and shared similar clinical findings, including hematological abnormalities and histopathology. Protozoal-related mortalities, specifically due to toxoplasmosis, are emerging as a threat to the recovery of this endangered pinniped and other native Hawaiian taxa. By establishing case definitions, this study provides a foundation for measuring the impact of these diseases on Hawaiian monk seals.

ESTIMATING CONTACT RATES OF HAWAIIAN MONK SEALS (NEOMONACHUS SCHAUINSLANDI) USING SOCIAL NETWORK ANALYSIS.

Understanding disease transmission dynamics, which are in part mediated by rates and patterns of social contact, is fundamental to predicting the likelihood, rate of spread, impacts, and mitigation of disease outbreaks in wildlife populations. Contact rates, which are important parameters required for epidemiologic models, are difficult to estimate. The endangered Hawaiian monk seal (Neomonachus schauinslandi) may be particularly vulnerable to morbillivirus outbreaks, due to its low abundance, lack of genetic diversity, and history of isolation from mammalian diseases. Morbillivirus epizootics have had devastating effects on other seal populations. We constructed social networks based on visual observations of individually identifiable monk seals associating onshore to estimate contact rates, assuming random mixing, and also to investigate contact patterns of different age and sex classes. Contact rates estimated from two island populations in 4 yr were remarkably similar, indicating any two individuals have about a one in 1,000 chance of making contact on any given day. Further, contact patterns within and among age and sex classes were statistically different from random. The methods we used could be broadly applied to empirically derive contact rates using association data. These rates are critical for epidemiologic modelling to simulate wildlife disease outbreaks and to inform science-based prevention and mitigation programs.