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A shock trial of a US Navy Destroyer, the USS Winston S. Churchill, was conducted offshore of northern Florida in May and June 2001. The shock trial consisted of three underwater detonation tests, spaced approximately one week apart. Environmental mitigation to minimise the impact of the shock trial on marine mammals and sea turtles was based on a Safety Range of 3.7km (2 n.miles) radius around the detonation site, and a Buffer Zone of an additional 1.85km (1 n.mile) radius beyond the Safety Range. Mitigation included site selection surveys, pre-detonation aerial, vessel and bio-acoustic monitoring, and post-detonation aerial and vessel monitoring. Six species of odontocete and two species of sea turtle were identified during mitigation monitoring, as well as several sightings that could not be identified by species. Site selection aerial surveys were implemented to select a test site with the lowest abundance of marine mammals and sea turtles. Nearly 300 animals were seen during site selection surveys. Pre-detonation aerial and vessel monitoring was implemented to sight any marine mammal and sea turtle within the Safety Range on designated test days, and track the animals until they could be verified to be outside the Safety Range. Approximately 1,200 marine mammals and 32 sea turtles were sighted during pre-detonation monitoring. Pre-detonation bio-acoustic monitoring was implemented to detect large cetaceans within the Safety Range and Buffer Zone; the only calls heard were from dolphins that could not be localised. Post-detonation monitoring was implemented to determine the effectiveness of mitigation procedures. No injured or dead marine mammals or turtles were detected during approximately 185 hours of post-detonation aerial and vessel visual monitoring. Post-detonation monitoring resulted in observations of 767 marine mammals and 42 sea turtles. With only two exceptions, the same marine mammal and sea turtle species were observed prior to, during and after the shock trial test time period. Factors leading to the success of this environmental mitigation effort are summarised, and recommendations for improvements to mitigation efforts of this size and scope are suggested. These recommendations include the use of a simultaneous second aircraft for improved coverage during pre-detonation surveys, increased post-detonation aerial monitoring, equitable survey data for all test sites under consideration during planning stages, and reassessment of bio-acoustic monitoring need and purpose.

A juvenile male Gervais’ beaked whale (Mesoplodon europaeus) was found dead in Barnstable Harbor, Cape Cod Bay, Massachusetts, USA. It apparently died from a septicaemia, possibly induced by a puncture wound of unknown origin. The previous most northerly stranding for this species in the western North Atlantic ocean was from New York State, USA.

Cryptococcus gattii is a fungal pathogen that primarily affects the respiratory and nervous systems of humans and other animals. C. gattii emerged in temperate North America in 1999 as a multispecies outbreak of cryptococcosis in British Columbia (Canada) and Washington State and Oregon (USA), affecting humans, domestic animals, and wildlife. Here we describe the C. gattii epizootic in odontocetes. Cases of C. gattii were identified in 42 odontocetes in Washington and British Columbia between 1997 and 2016. Species affected included harbor porpoises Phocoena phocoena (n = 26), Dall’s porpoises Phocoenoides dalli (n = 14), and Pacific white-sided dolphins Lagenorhynchus obliquidens (n = 2). The probable index case was identified in an adult male Dall’s porpoise in 1997, 2 yr prior to the initial terrestrial outbreak. The spatiotemporal extent of the C. gattii epizootic was defined, and cases in odontocetes were found to be clustered around terrestrial C. gattii hotspots. Case-control analyses with stranded, uninfected odontocetes revealed that risk factors for infection were species (Dall’s porpoises), age class (adult animals), and season (winter). This study suggests that mycoses are an emerging source of mortality for odontocetes, and that outbreaks may be associated with anthropogenic environmental disturbance.

E. coli was isolated from the Salish Sea (Puget Sound) ecosystem, including samples of marine and fresh water, and wildlife dependent on this environment. E. coli isolates were assessed for phenotypic and genotypic resistance to antibiotics. A total of 305 E. coli isolates was characterized from samples collected from: marine water obtained in four quadrants of the Salish Sea, select locations near beaches, fresh water from streams near marine beaches, and fecal samples from harbor porpoises (Phocoena phocoena), harbor seals (Phoca vitulina), river otters (Lontra canadensis), and English sole (Parophrys vetulus). Isolates were evaluated using antimicrobial susceptibility typing, whole-genome sequencing, fumC, and multilocus sequence typing. Resistance and virulence genes were identified from sequence data. Of the 305 isolates from Salish Sea samples, 20 (6.6%) of the E. coli were intermediate, and 31 (10.2%) were resistant to ≥1 class of antibiotics, with 26.9% of nonsusceptible (resistant and intermediate resistant) E. coli isolates from marine mammals and 70% from river otters. The proportion of nonsusceptible isolates from animals was significantly higher than samples taken from marine water (p &lt, 0.0001). A total of 196 unique STs was identified including 37 extraintestinal pathogenic E. coli (ExPEC)-associated STs [ST10, ST38, ST58, ST69, ST73, ST117, ST131, and ST405]. The study suggests that animals may be potential sentinels for antibiotic-resistant and ExPEC E. coli in the Salish Sea ecosystem.

Journalists play a crucial role in the dissemination of health-related information. In developing countries, such as Nepal, the media are integral in shaping the national agenda and informing the public of important health issues. With an increasing need for a collaborative effort to attain optimal health for people, animals, and the environment, the One Health approach was used to characterize health reporting in Nepal. A comprehensive survey was administered to health journalists regarding their public, animal, and environmental health reporting habits. Seventy-one journalists completed the survey across three study sites. Many journalists indicated a history of reporting across all three sectors but did not routinely focus on health reporting in general. The majority of journalists perceived the quality and overall coverage of health-related topics increased over the last five years. However, few journalists reported receiving specialized training in any health sector. Although the overall quality of health reporting in the Nepali media showed improvements, many journalists acknowledged a lack of understanding of common health topics and a desire to learn more skills related to accurate health reporting. One Health provides a conceptual framework for understanding and promoting health communication through mass media to benefit humans, animals, and ecosystems.

Primary fungal diseases in marine mammals are rare. Mucormycosis, a disease caused by fungi of the order Mucorales, has been documented in few cetaceans and pinnipeds. In 2012, the first case of mucormycosis in the Pacific Northwest was documented in a dead stranded harbor porpoise (Phocoena phocoena) in Washington state. Since then, mucormycosis has been detected in a total of 21 marine mammals; fifteen harbor porpoises, five harbor seals (Phoca vitulina), and one southern resident killer whale (Orcinus orca). Infected animals were predominately found in the inland waters of Washington and British Columbia, and one harbor seal was recovered in northern Oregon. Fungal hyphae were detected histologically in a variety of tissues, including brain, lung, spleen, pancreas, kidneys, muscle, lymph nodes, and skin. Three fungal species were identified from seven cases by PCR screening or fungal culture; Rhizomucor pusillus (four cases), Lichtheimia corymbifera (two cases), and Cunninghamella bertholletiae. Underlying conditions such as emaciation, current or recent pregnancy, multisystemic parasitism, protozoal infection, and herpesvirus were found in several affected animals. Reasons for the appearance and subsequent increase of these fungal infections in marine mammals are unknown. The emergence of this disease as a source of marine mammal mortality in the Pacific Northwest is of particular concern for endangered southern resident killer whales that spend time in this region. Current population-level stressors such as insufficient prey, high levels of contaminants, and noise pollution, could predispose them to these fatal infections.

We evaluated harbor porpoise (Phocoena phocoena (Linnaeus, 1758)) strandings in the Salish Sea to determine calving seasonality (1980–2015). A total of 443 strandings were analyzed, of which 134 were calves and 53 were neonates. Stranded calves were reported every month, but peaked in July, August, and September. Based on fetal size and an estimated fetal growth rate of 80 mm/month, mean (±SD) conception date (and range) was back-calculated to 11 October ± 30 days (16 August – 31 December) and was later than in most other studies. Using mean (±SD) length at birth (80 ± 5.8 cm), gestation was estimated to be approximately 10.8 months. Estimated birthing period was 16 July – 27 November, with a mean (±SD) birth date of 10 September (±30.7 days) and a birth length of 80.0 cm. Estimated pregnancy rate (0.28–0.29) is lower than reported in other areas and is likely an underestimate due to missed early embryos, poor postmortem condition of a large proportion of the stranded adult females, and potential biases related to the animals that strand and are available. This study of harbor porpoise reproduction and calving in the Salish Sea is the first assessment of calving seasonality for this species in the northeast Pacific Ocean.

Necropsy of a female adult pregnant harbor porpoise Phocoena phocoena revealed a multicentric plasmacytoma. The plasmacytoma infiltrated the cranial lung lobes, mediastinal lymph nodes and the spleen. Diagnosis was based on gross, histopathologic and immunohistochemical studies. Histopathology revealed a diffuse proliferation of atypical pleomorphic neoplastic round cells with plasmacytic features. Positive immunohistochemistry with anti-CD79a and anti-CD20 antibody markers and anti-multiple myeloma oncogene 1 (MUM-1) for plasmacytoma confirmed this neoplasm to be of B-cell origin. This is the first recorded case of a plasmacytoma in a harbor porpoise. Routine viral screening was negative via standard PCR for herpesvirus and reverse transcriptase PCR for morbillivirus. Retroviral screening was not performed.

Tags have been used to examine migration routes and habitat use of large whales for >40 yr, however, evaluation of tag wound healing has largely been short-term, anecdotal or generalized. This study developed methods for systematic photographic assessment of long-term external consequences of tag placement, to determine potential differences in wound healing between species and tag types and thus advise future tagging efforts to possibly minimize undesirable side effects. Tag site appearance and healing characteristics were evaluated by two reviewers and a time series evaluated by five veterinarians from photographs during 995 postdeployment encounters with 34 gray and 63 blue whales tagged in the North Pacific. Blue whale resightings were less frequent, but spanned a longer time period due to earlier tag deployments than the more frequent gray whale follow-up observations. Swelling occurred in 74% of reencountered gray whales, with the highest frequency 6 mo postdeployment. Swellings were common in blue whales with early tag designs but rare with current models. Depressions occurred in 82% of gray and 71% of blue whales. This study demonstrates the value of follow-up studies of tagged animals and systematic scoring of photographs to quantitatively compare tag response.

Aging is associated with significant bone loss and increased fracture risk, which has been attributed to a diminished response to anabolic mechanical loading. In adults, skeletal progenitors proliferate and differentiate into bone-forming osteoblasts in response to increasing mechanical stimuli, though the effects of aging on this response are not well-understood. Here we show that both adult and aged mice exhibit load-induced periosteal bone formation, though the response is significantly attenuated with age. We also show that the acute response of adult bone to loading involves expansion of Sca-1+Prrx1+ and Sca-1-Prrx1+ cells in the periosteum. On the endosteal surface, loading enhances proliferation of both these cell populations, though the response is delayed by 2 days relative to the periosteal surface. In contrast to the periosteum and endosteum, the marrow does not exhibit increased proliferation of Sca-1+Prrx1+ cells, but only of Sca-1-Prrx1+ cells, underscoring fundamental differences in how the stem cell niche in distinct bone envelopes respond to mechanical stimuli. Notably, the proliferative response to loading is absent in aged bone even though there are similar baseline numbers of Prrx1 + cells in the periosteum and endosteum, suggesting that the proliferative capacity of progenitors is attenuated with age, and proliferation of the Sca-1+Prrx1+ population is critical for load-induced periosteal bone formation. These findings provide a basis for the development of novel therapeutics targeting these cell populations to enhance osteogenesis for overcoming age-related bone loss. © 2019 The Authors. JBMR Plus published by Wiley Periodicals, Inc. on behalf of American Society for Bone and Mineral Research.

Background Mental health issues can reach epidemic proportions in developed countries after natural disasters, but research is needed to better understand the impact on children and adolescents in developing nations. Methods A cross-sectional study was performed to examine the relationship between earthquake exposures and depression, PTSD, and resilience among children and adolescents in Phulpingdanda village in Nepal, 1 year after the 2015 earthquakes, using the Depression Self-Rating Scale for Children, Child PTSD Symptom Scale, and the Child and Youth Resilience Measure, respectively. To quantify exposure, a basic demographic and household questionnaire, including an earthquake exposure assessment tool for children and adolescents, was created. Results Of the 62 respondents interviewed, 3.23% and 4.84% displayed symptomatology of depression and PTSD. A large number of respondents interviewed scored high for resiliency (80.65%). All 62 respondents were displaced from their household and witnessed severe damage of both their homes and village. The number of earthquake exposures had a strong, positive correlation with PTSD symptomatology. Conclusions Although the number of respondents who showed signs of depression and PTSD symptomatology was lower than anticipated, resilience scores were considerably higher. Future research should explore which protective factors may contribute to high resiliency in Nepali children and adolescents. Electronic supplementary material The online version of this article (10.1186/s13034-018-0257-9) contains supplementary material, which is available to authorized users.

In 2015, two strong earthquakes, as well as continuous, high magnitude aftershocks, struck Nepal. Phulpingdanda village was greatly impacted due to its lack of infrastructure and environmental remoteness. Adults from sampled households were surveyed 1-year later to examine the association between earthquake exposures and indicators of depression, post-traumatic stress disorder (PTSD), and resilience. Results showed 33% of surveyed residents screened positive for depression, 9% screened positive for severe PTSD, and 46% displayed moderate to high resilience. Additionally, participants experienced resource loss (100%), damaged home and goods (99%), and exposure to the grotesque (82%). Traumatic earthquake experiences related to personal harm were associated with symptoms of depression and PTSD and resource loss was associated with depressive symptoms. Earthquake experiences associated with less damage to home and goods, but greater exposure to the grotesque were associated with increased resilience. This research adds to our knowledge of the relationship between traumatic exposures and indicators of psychological distress and resilience following a disaster.

Brucella ceti and Brucella pinnipedialis have been documented as occurring in marine mammals, and B. ceti has been identified in 3 naturally acquired human cases. Seroconversion and infection patterns in Pacific Northwest harbor seals ( Phoca vitulina richardii) and North Atlantic hooded seals ( Cystophora cristata) indicate post-weaning exposure through prey consumption or lungworm infection, suggesting fish and possibly invertebrates play an epizootiologic role in marine Brucella transmission and possible foodborne risk to humans. We determined if real-time quantitative PCR (qPCR) assays can detect marine Brucella DNA in fish DNA. Insertion sequence (IS) 711 gene and sequence type (ST)27 primer–probe sets were used to detect Brucella associated with marine mammals and human zoonotic infections, respectively. First, DNA extracts from paired-species fish (containing 2 species) samples were tested and determined to be Brucella DNA negative using both IS 711 and ST27 primer–probe sets. A representative paired-species fish DNA sample was spiked with decreasing concentrations of B. pinnipedialis DNA to verify Brucella detection by the IS 711 primer–probe within fish DNA. A standard curve, developed using isolated DNA from B. pinnipedialis, determined the limit of detection. Finally, the IS 711 primer–probe was used to test Atlantic cod ( Gadus morhua) DNA extracts experimentally infected with the B. pinnipedialis hooded seal strain. In culture-positive cod tissue, the IS 711 limit of detection was ~1 genome copy of Brucella. Agreement between culture and PCR results for the 9 positive and 9 negative cod tissues was 100%. Although a larger sample set is required for validation, our study shows that qPCR can detect marine Brucella in fish.

B-cell lymphoma, a common morphologic variant of non-Hodgkin lymphoma, has been associated with persistent pollutants in humans, but this association is not well-characterized in top-level predators sharing marine resources with humans. We characterized and compared blubber contaminants and hormones of a pregnant harbor porpoise (Phocoena phocoena) with B-cell lymphoma, with those in two presumed healthy fishery by-caught porpoises with no lymphoma: a pregnant adult and female juvenile. Common historic use compounds, including polychlorinated biphenyls, polybrominated diphenyl ethers, and pesticides, were evaluated in blubber samples from three porpoises. In addition, blubber cortisol and progesterone levels (ng/g) were determined in all three animals. Total pollutant concentrations were highest in the juvenile porpoise, followed by the lymphoma porpoise and the nonlymphoma adult. Blubber cortisol concentrations were 191% greater in the pregnant with lymphoma porpoise compared with the pregnant no lymphoma porpoise, and 89% greater in the juvenile female compared with the pregnant no lymphoma porpoise. Although both adults were pregnant, progesterone levels were substantially greater (90%) in the healthy compared with the lymphoma adult. Health monitoring of top-level marine predators, such as porpoise, provides a sentinel measure of contaminants that serve as indicators of potential environmental exposure to humans.

We collected blood from 18 beluga whales (Delphinapterus leucas), live-captured in Bristol Bay, Alaska, USA, in May and September 2008, to establish baseline hematologic and serum chemistry values and to determine whether there were significant differences in hematologic values by sex, season, size/age, or time during the capture period. Whole blood was collected within an average of 19 min (range=11-30 min) after the net was set for capture, and for eight animals, blood collection was repeated in a later season after between 80-100 min; all blood was processed within 12 hr. Mean hematocrit, chloride, creatinine, total protein, albumin, and alkaline phosphatase were significantly lower in May than they were in September, whereas mean corpuscular hemoglobin concentration, monocytes, phosphorous, magnesium, blood urea nitrogen, alanine aminotransferase, aspartate aminotransferase, γ-glutamyltranspeptidase, and creatinine kinase were significantly higher. Mean total protein, white blood cell count, neutrophils, and lymphocytes were significantly higher early in the capture period than they were later. No significant differences in blood analyte values were noted between males and females. Using overall body length as a proxy for age, larger (older) belugas had lower white blood cell, lymphocyte, and eosinophil counts as well as lower sodium, potassium, and calcium levels but higher creatinine levels than smaller belugas. These data provide values for hematology and serum chemistry for comparisons with other wild belugas.

This is the publisher’s final pdf. The published article is copyrighted by John Wiley & Sons, Inc. and can be found at: http://onlinelibrary.wiley.com/journal/10.1111/%28ISSN%291748-7692. To the best of our knowledge, one or more authors of this paper were federal employees when contributing to this work.

Leptospirosis is one of the most widespread bacterial zoonoses in the world and affects most mammalian species. Although leptospirosis is well documented and characterized in terrestrial species, less information is available regarding the distribution and impact of leptospirosis in marine mammals. Additionally, the role of animal migrations on the geographical spread of leptospirosis has not been reported. Periodic epizootic outbreaks of acute leptospirosis among California sea lions (Zalophus californianus) have been reported since 1971. In this study, we collected samples from California sea lions stranded along the Pacific coast of North America during the most recent epidemic in 2004, and maintained leptospirosis surveillance of the California sea lion population along the California coast through 2007. Several isolates of Leptospira interrogans serovar Pomona were obtained from kidney and urine samples collected during this study, a finding consistent with serological evidence that California sea lions are persistently exposed to this leptospiral serovar. Combined, these data support a model whereby California sea lions are maintenance hosts for L. interrogans serovar Pomona, yet periodically undergo outbreaks of acute infection. During the 2004 outbreak, the incidence of new leptospirosis cases among California sea lions coincided with the seasonal movement of male sea lions from rookeries along the coast of central and southern California north as far as British Columbia. These data show that seasonal animal movement contributes to the distribution of leptospirosis across a large geographical region.

Leptospirosis, caused by the spirochete Leptospira , is a geographically widespread disease that affects a broad range of mammals, including marine mammals. Among pinniped populations, periodic epizootics of leptospirosis are responsible for significant die-offs. Along the west coast of North America, the most recent leptospirosis epizootic occurred in 2004, during which samples were collected from cases ranging from California to British Columbia. The primary objective of this study was to use this well-defined sample set to determine the feasibility of using PCR techniques to diagnose Leptospira infection among pinniped populations in comparison with diagnostic methodologies commonly used for marine mammals. Successful amplification was achieved from a variety of samples, including freshly collected urine, urine stored at −80°C for less than 6 months, and kidney (freshly collected, frozen, and decomposed), as well as feces- and urine-contaminated sand collected in the vicinity of a live-stranded animal. Pathological examination of tissue collected from Leptospira -infected animals revealed the presence of leptospiral antigen in the kidneys. The use of species-specific primer pairs revealed a pattern of host specificity for Leptospira interrogans in sea lions and Leptospira kirschneri in elephant seals. These studies indicate PCR is a sensitive and specific diagnostic tool for the detection of Leptospira infection in pinnipeds and reveal a potential source for epizootic, enzootic, and zoonotic spread of leptospirosis in a marine environment.

Ship strikes of large whales cause mortalities worldwide, but there is uncertainty regarding the frequency and species involved. We examined 130 records (from 1980–2006) of large whale strandings in Washington State. Nineteen strandings (seven species) had evidence of ship-strikes. Fin whales (Balaenoptera physalus) had the highest incidence of ante-mortem ship strike (five of seven, with the remaining two possibly post-mortem) and all but one occurring since 2002. Six grey whales (Eschrichtius robustus) suffered ‘possible ship strike’ injuries, likely the result of their large numbers in the area, rather than high levels of ship strikes. Only one possible ship-struck humpback whale was recorded, despite concentrations of humpbacks feeding within shipping lanes in this region. This study shows dramatic differences in occurrences of ship-struck large whales by species, which we believe results from a combination of species' vulnerability to ship strikes, and how likely a struck whale is to be caught up on the bow of a ship and brought to waters where it can be examined.

In 2006, a marked increase in harbor porpoise Phocoena phocoena strandings were reported in the Pacific Northwest of the USA, resulting in the declaration of an unusual mortality event (UME) for Washington and Oregon to facilitate investigation into potential causes. The UME was in place during all of 2006 and 2007, and a total of 114 porpoises stranded during this period. Responders examined 95 porpoises; of these, detailed necropsies were conducted on 75 animals. Here we review the findings related to this event and how these compared to the years immedi- ately before and after the UME. Relatively equal numbers among sexes and age classes were rep- resented, and mortalities were attributed to a variety of specific causes, most of which were cate- gorized as trauma or infectious disease. Continued monitoring of strandings during 4 yr following the UME showed no decrease in occurrence. The lack of a single major cause of mortality or evi- dence of a significant change or event, combined with high levels of strandings over several post- UME years, demonstrated that this was not an actual mortality event but was likely the result of a combination of factors, including: (1) a growing population of harbor porpoises; (2) expansion of harbor porpoises into previously sparsely populated areas in Washington's inland waters; and (3) a more well established stranding network that resulted in better reporting and response. This finding would not have been possible without the integrated response and investigation under- taken by the stranding network.

A large number of vaginal calculi were observed in a juvenile harbor porpoise (Phocoena phocoena) stranded on Whidbey Island, Washington. Vaginal calculi have been reported in other species, but not in harbor porpoises. Histologic examination of the urinary tract revealed mucosal hyperplasia most likely attributable to the calculi. The calculi were numerous (.30), composed completely of struvite (magnesium ammonium phosphate), and on culture yielded Enterococcus spp., a bacterium not usually associated with struvite urolith formation in domestic animals. The only other lesion of note was severe hepatic lipidosis, and its relationship to the development of the vaginal calculi is unknown.

Blood analytes are critical for evaluating the general health of cetacean populations, so it is important to understand the intrinsic variability of hematology and serum chemistry values. Previous studies have reported data for follow-up periods of several years in managed and wild populations, but studies over long periods of time (20 yr) have not been reported. The study objective was to identify the influences of partitioning characteristics on hematology and serum chemistry analytes of apparently healthy managed beluga (Delphinapterus leucas). Blood values from 31 managed belugas, at three facilities, collected over 22 yr, were assessed for seasonal variation and aging trends, and evaluated for biologic variation among and within individuals. Linear mixed effects models assessed the relationship between the analytes and sex, age, season, facility location, ambient air temperature, and photoperiod. Sex differences in analytes and associations with increasing age were observed. Seasonal variation was observed for hemoglobin, hematocrit, mean corpuscular volume, monocytes, alkaline phosphatase, total bilirubin, cholesterol, and triglycerides. Facilities were associated with larger effects on analyte values compared to other covariates, whereas age, sex, and ambient temperature had smaller effects compared to facility and season. Present findings provide important baseline information for future health monitoring efforts. Interpretation of blood analytes and animal health in managed and wild populations over time is aided by having available typical levels for the species and reference intervals for the degree to which individual animals vary from the species average and from their own baseline levels during long-term monitoring.

Fecal pathogens are transported from a variety of sources in multi-use ecosystems such as upper Cook Inlet (CI), Alaska, which includes the state's urban center and is highly utilized by humans and animals. This study used a novel water quality testing approach to evaluate the presence and host sources of potential fecal pathogens in surface waters and sediments from aquatic ecosystems in upper CI. Matched water and sediment samples, along with effluent from a municipal wastewater treatment facility, were screened for Salmonella spp., Vibrio spp., Cryptosporidium spp., Giardia spp., and noroviruses. Additionally, Bacteroidales spp. for microbial source tracking, and the fecal indicator bacteria Enterococcus spp. as well as fecal coliforms were evaluated. Overall, Giardia and Vibrio were the most frequently detected potential pathogens, followed by Cryptosporidium and norovirus, while Salmonella was not detected. Sample month, matrix type, and recent precipitation were found to be significant environmental factors for protozoa or host-associated Bacteroidales marker detection, whereas location and water temperature were not. The relative contribution of host-associated markers to total fecal marker concentration was estimated using a Monte Carlo method, with the greatest relative contribution to the Bacteroidales marker concentration coming from human sources, while the remainder of the universal fecal host source signal was uncharacterized by available host-associated assays, consistent with wildlife fecal sources. These findings show how fecal indicator and pathogen monitoring, along with identifying contributing host sources, can provide evidence of coastal pathogen pollution and guidance as to whether to target human and/or animal sources for management.

Leptospirosis has been reported in California sea lions (Zalophus californianus) since 1970; however, the source of infection and mode of transmission remain unknown. To elucidate these features, demographic and environmental risk factors for leptospirosis were evaluated. California sea lion stranding records from northern California for 2004 were used to identify cases of leptospirosis (n = 316) and controls (n = 143). Demographic characteristics (age class, sex) and environmental factors, representing surrogates for exposure to dogs, cattle, rainfall, and freshwater sources, were compared between cases and controls with the use of a geographic information system (GIS) and logistic regression. Multivariate analyses revealed that summer and autumn seasons, juvenile age class, male sex, high dog-park density, and close proximity to dog parks were significantly associated with leptospirosis in sea lions, whereas county farmland cattle density, rainfall levels 30 days prior to stranding, human density, and proximity to freshwater sources were not associated. Thus, dogs and dog parks, or factors associated with them, might be further investigated to assess their relationship to leptospirosis in sea lions.

The use of spatial epidemiology and geographical information systems (GIS) facilitates the incorporation of spatial relationships into epidemiological investigations of marine mammal diseases and conservation medicine. Spatial epidemiology is the study of the spatial variation in disease risk or incidence and explicitly addresses spatial structures and functions that factor into disease. The GIS consists of input, management, analysis, and presentation of spatial disease data and can act as an integrative tool so that a range of varied data sources can be combined to describe different environmental aspects of wild animals and their diseases. The use of modern spatial analyses and GIS is becoming well developed in the field of marine mammal ecology and biology, but has just recently started to gain more use in disease research. The use of GIS methodology and spatial analysis in nondisease marine mammal studies is briefly discussed, while examples of the specific uses of these tools in mapping, surveillance and monitoring, disease cluster detection, identification of environmental predictors of disease in wildlife populations, risk assessment, and modeling of diseases, is presented. Marine mammal disease investigations present challenges, such as less consistent access to animals for sampling, fewer baseline data on diseases in wild populations, and less robust epidemiologic study designs, but several recommendations for future research are suggested. Since location is an integral part of investigating disease, spatial epidemiology and GIS should be incorporated as a data management and analysis tool in the study of marine mammal diseases and conservation medicine.

To the Editor: We report maternal–fetal transmission of Cryptococcus gattii and death in a wild porpoise. Cryptococcus neoformans and C. gattii are 2 environmental, encapsulated yeasts that cause invasive, potentially life-threatening infections in humans and animals (1). C. neoformans causes disease in immunocompromised hosts, and C. gattii is also pathogenic in immunocompetent hosts (2). Since 1999, cryptococcosis caused by C. gattii has appeared on southern Vancouver Island (British Columbia, Canada) and nearby surrounding areas (2,3). Spread beyond Vancouver Island has been documented along the Pacific Northwest Coast, but the mechanism remains undetermined (4). A pregnant, dead, stranded, harbor porpoise (Phocoena phocoena) was reported on February 22, 2007, on western Whidbey Island, in Puget Sound, Washington State (48.2833°N, 122.7283°W). The carcass was iced and necropsy was performed on February 24. Sampled tissues from the adult and fetus were divided: half fixed in 10% formalin for histopathologic analysis, and half frozen for ancillary studies. For histologic analysis, tissues were embedded in paraffin, sectioned to 3–5 μm, and stained with hematoxylin and eosin. Selected sections were stained with mucicarmine. The adult porpoise (length 177 cm, weight ≈57.7 kg) was in poor condition (reduced blubber layer). Both lungs were exposed, extensively scavenged, firm, and nodular; a sectioned surface exuded clear to slightly opaque gelatinous to mucinous discharge. Mediastinal lymph nodes were grossly enlarged, multinodular, and firm with large numbers of yeasts visible by microscopy (online Figure A1 panel A, www.cdc.gov/EID/content/17/2/304-appF.htm). The first stomach chamber contained two 3.5 cm × 2.5 cm raised, centrally umbilicated ulcers and several embedded anisakid nematodes. The uterus was gravid in the right horn with a mid-term fetus. No other gross lesions were identified. Microscopically, the lung lesions correlated with granulomatous to pyogranulomatous infiltrates, often with a myriad of yeasts. The male fetus (length 30 cm, weight 2.4 kg), was examined separately at a different facility than the dam. It appeared grossly normal externally and was at a gestation of ≈5–6 months. Mediastinal lymph nodes had mild granulomatous inflammation and contained numerous yeasts morphologically consistent with Cryptococcus spp. (Figure A1, panel B). The lymph nodes were partially replaced with intracellular and extracellular mulilobulated yeast aggregates (length 8–20 μm) with pale eosinophilic central regions and a thin refractile wall peripherally bound by a 5-μm nonstaining capsule. Around the periphery of these aggregates, there were small numbers of macrophages and lymphocytes and fewer neutrophils. Specific staining showed a prominent mucicarminophilic capsule consistent with Cryptococcus spp. Yeasts were found in the amniotic fluid and interspersed within the chorioallantoic villi and submucosal vasculature of the placenta. Mild multifocal nonsuppurative myocarditis was detected. However, no yeasts were seen in inflamed areas. There were no overt lesions in the remaining organs. Maternal and fetal tissues were cultured for fungi, and diagnosis was based on Gram stain (budding yeast-like cells), India ink stain (positive for encapsulated cells), hydrolysis of urea (positive), and final confirmation by using API 20C Aux V3.0 (bioMerieux, Marcy l’Etoile, France). Canavanine-glycine-bromthymol blue agar was used to differentiate between C. gattii and C. neoformans (5). Molecular typing by restriction fragment length polymorphism was used to definitively speciate and subtype C. gattii (6). Fungal culture showed heavy growth of Cryptococcus spp. from the dam (lungs, mediastinal lymph nodes, and placenta) and fetus (mediastinal lymph nodes). Genotyping of primary isolates identified VGIIa C. gattii in both animals. Test results for enteric pathogens, intestinal nematodes, morbillivirus, and Brucella spp. were negative. Fetal infection was most likely hematogenous, disseminated from a primary maternal pulmonary source to the uterus and subsequently to placental vasculature and internal fetal tissues. Infection by aspiration or ingestion of contaminated amniotic fluid was also possible. Although close evaluation of the lung did not show any discernible yeasts, the organism may have been present in an area other than that sectioned. During 1998–2007, ≈450 harbor and Dall’s porpoises (Phocoenoides dalli) and Pacific white-sided dolphins (Lagenorhynchus obliquidens) along the Pacific Northwest Coast were recovered and subjected to necropsy. Disseminated cryptococcosis caused by C. gattii since 2000 was diagnosed in 15 harbor porpoises, 10 Dall’s porpoises, 2 adult Pacific white-sided dolphins, and 3 unrecorded species (10 females, 15 males, and 5 unknown sex; 24 adults, 4 juveniles, 1 fetus, and 1 undocumented age; S. Raverty, unpub. data). Wild porpoises in the Pacific Northwest Region, being near shore inhabitants in waters surrounding Vancouver Island, may come into contact with air containing C. gattii at the air–water interface or ingest seawater containing yeasts while feeding (7). Their proximity to a habitat containing Coastal Douglas fir (Pseudotsuga menziesii) and Western hemlock (Tsuga heterophylla) may play a role in the epidemiology of C. gattii because these trees have been associated with cases of C. gattii (8). Cryptococcal infection during pregnancy has been reported in humans and horses (9,10). This fetal case of cryptococcosis may have major human and animal health implications. Further studies should be undertaken to assess possible fetal involvement, identify infections in pregnant females, and provide information on risk reduction and improving diagnosis and treatment.