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Brucella ceti infection is associated with a variety of disease outcomes in cetaceans globally. Multiple genotypes of B. ceti have been identified. This retrospective aimed to determine if specific lesions were associated with different B. ceti DNA sequence types (STs). Characterization of ST was performed on 163 samples from 88 free-ranging cetaceans, including common bottlenose dolphin Tursiops truncatus (T.t.; n = 73), common short-beaked dolphin Delphinus delphis (D.d.; n = 7), striped dolphin Stenella coeruleoalba (n = 3), Pacific white-sided dolphin Lagenorhynchus obliquidens (n = 2), sperm whale Physeter macrocephalus (n = 2), and harbour porpoise Phocoena phocoena (n = 1), that stranded along the coast of the US mainland and Hawaii. ST was determined using a previously described insertion sequence 711 quantitative PCR. Concordance with 9-locus multi-locus sequence typing was assessed in a subset of samples (n = 18). ST 26 was most commonly identified in adult dolphins along the US east coast with non-suppurative meningoencephalitis (p = 0.009). Animals infected with ST 27 were predominately perinates that were aborted or died shortly after birth with evidence of in utero pneumonia (p = 0.035). Reproductive tract inflammation and meningoencephalitis were also observed in adult T.t. and D.d. with ST 27, though low sample size limited interpretation. ST 23 infections can cause disease in cetacean families other than porpoises (Phocoenidae), including neurobrucellosis in D.d. In total, 11 animals were potentially infected with multiple STs. These data indicate differences in pathogenesis among B. ceti STs in free-ranging cetaceans, and infection with multiple STs is possible.

Microbiology records for 1127 cetaceans stranded on English and Welsh beaches and examined at the Institute of Zoology between 1990 and 2019 were reviewed to identify cases of Erysipelothrix rhusiopathiae, an uncommon but potentially fatal zoonotic pathogen. Once cases were identified, prevalence was calculated, corresponding postmortem reports were reviewed, common gross and histopathological findings were identified, and antibiotic susceptibilities were determined. Overall prevalence for E. rhusiopathiae was 0.62% (7/1127; 95% CI: 0.30-1.28%). It was isolated from 3 bottlenose dolphins Tursiops truncatus, 3 harbor porpoises Phocoena phocoena, and 1 short-beaked common dolphin Delphinus delphis, with a prevalence of 21.4% (3/14; 95% CI: 7.6-47.9%), 0.39% (3/779; 95% CI: 0.13-1.13%), and 0.47% (1/212; 95% CI: 0.08-2.62%) for each species, respectively. E. rhusiopathiae resulted in septicemia in all cases from which it was isolated. Gross necropsy findings included pulmonary edema (5/7), hemorrhage (5/7) and/or congestion of various organs (4/7), and serosanguineous effusion (3/7; pericardial: 3/7, pleural: 2/6, abdominal: 2/6). Congestion (5/5), bacterial emboli (4/5), and hemorrhage (4/5) were commonly observed on histopathology, and acute renal tubular injury (2/5) and pulmonary edema (2/5) were occasionally observed. Routine bacterial cultures were vital in identifying E. rhusiopathiae, since gross lesions were often subtle and nonspecific. The liver, kidney, and brain were key organs from which E. rhusiopathiae was consistently isolated. Antibiotic resistance was uncommon and was only observed for amikacin and trimethoprim sulfonamide. Penicillins were consistently effective, along with fluoroquinolones, macrolides, clindamycin, cephalexin, and oxytetracycline.

The physiological demands of pregnancy inevitably result in alterations in both biochemical and hematological parameters as fetal development occurs. The shifts observed in successful pregnancy in bottlenose dolphins Tursiops truncatus to support both fetal physiological needs and maternal basal requirements have been established according to each trimester. Detecting aberrations in blood-based biomarkers could help facilitate diagnosis of gestational abnormalities, improve our understanding of factors influencing reproductive outcomes and aid in prediction of reproductive failure. This study retrospectively analyzed 263 blood samples from 15 bottlenose dolphins in 21 failed pregnancies over 28 yr (1989-2017). Most samples remained within normal pregnancy reference ranges; however, significant shifts were observed between trimesters. Hematological alterations, compared to successful pregnancy reference ranges from previously published data, were consistent across failed pregnancies and included an increased prevalence of elevated 2nd and 3rd trimester neutrophils, elevated 2nd trimester monocytes and decreased 3rd trimester eosinophils. In addition, low hematocrit and low red blood cells were more prevalent in the 2nd trimester. Biochemical shifts included an increased prevalence of elevated creatine phosphokinase in the 3rd trimester outside of the normal reference ranges. Across failed pregnancies, calcium and iron were decreased in the 3rd trimester. Significantly decreased progesterone in the 3rd trimester was a negative prognostic indicator of pregnancy outcome with decreasing 3rd trimester progesterone associated with failed pregnancy. This study demonstrates the use of blood-based biomarkers as possible predictors of pregnancy outcome in bottlenose dolphins.

Brucellosis is a disease caused by the Gram-negative facultative intracellular bacterium Brucella spp. In terrestrial species, this zoonotic bacterium is a global public health risk, but there is also concern over the zoonotic potential of marine forms, such as B. ceti, which affects cetaceans. Due to the detection of B. ceti in samples from bottlenose dolphins Tursiops truncatus during the 2010-2014 Gulf of Mexico Unusual Mortality Event, a long-term study of the prevalence of Brucella in stranded bottlenose dolphins from South Carolina, USA, was conducted. From 2012 through 2017, 282 stranded bottlenose dolphins were tested for B. ceti via real-time PCR. Nearly 32% of the dolphins tested positive in at least one sample (brain, lung, blowhole swab). Very little information exists in the literature on the occurrence of Brucella spp. in marine mammals, though in terrestrial species, such as cattle and elk, higher prevalence is often reported in spring. Similar results were found in this study with the peak occurrence being between March and June, a known period of calving in South Carolina. Results from this study provide important insights into the occurrence of the marine bacterium B. ceti.

Between 2007 and 2013, before the 2013 cetacean morbillivirus outbreak, 26 fresh bottlenose dolphin carcasses were necropsied on the coast of Georgia, USA. Here, we present the pathological and microbiological findings associated with their most likely causes of death. The primary cause of death was determined in 25 individuals and included systemic bacterial infection (n = 7), verminous and bacterial bronchopneumonia (n = 5), drowning/entanglement (n = 5), disseminated histoplasmosis (n = 1), intestinal intussusception (n = 1), vegetative endocarditis (n = 1), meningitis (n = 1), necrotizing dermatitis (n = 1), disseminated angiomatosis (n = 1), emaciation (n = 1) and stingray spine trauma (n = 1). Histiocytic and eosinophilic bronchopneumonia associated with Halocerchus sp. infection was observed in 69% of the animals (18/26) and eosinophilic gastritis due to Anisakidae nematodes was found in 36% of the examined stomachs (8/22). Moderate to severe eosinophilic pancreatitis with fibrosis was observed in 4 animals infected with Brachycladiidae trematodes. Proliferative and ulcerative lymphoplasmacytic dermatitis was found in 5 animals and was considered to contribute to deteriorated health status in 2 calves. Pulmonary and lymph node angiomatosis were observed in 15 and 10 animals, respectively. In at least 2 animals, the concentration of polychlorinated biphenyls (PCBs) in the blubber exceeded 1500 µg g-1 of lipid. Bottlenose dolphins stranded on the Georgia coast have a wide range of inflammatory lesions associated with a variety of helminth, bacterial, and fungal pathogens. Some resident animals have also been exposed to high levels of PCB contamination, which could reduce host immunocompetence. Higher exposure to these or other pathogens could result in further decline in the health of resident and migrant dolphin populations in this region.

Photographs collected during a 23 yr photo-identification study in the Moray Firth were examined to assess the prevalence, type and severity of vertebral deformations present in bottlenose dolphin Tursiops truncatus calves. Fifteen cases of presenting spinal anomalies (scoliosis, kyphosis, lordosis and combinations thereof) of variable severity were identified in 7.4% of all known calves from the population. Thirteen of the 15 anomalies were either manifest from birth or acquired from an early age, as ascertained from longitudinal sightings histories of their mothers. Most afflicted calves died during early development or shortly after maternal separation. However, 3 survived to adulthood and persist in the population to date, in addition to 2 dependent infants whose fate remains to be established. At 15+ yr of age, the oldest surviving individual was remarkably one of the most severe cases identified, highlighting the ability of these delphinids for adaptation to such gross structural deformities. The aetiology of the observed conditions could be attributed to a range of causative factors that may have implications for the well-being and health of this North Sea coastal dolphin population, a topic which merits further investigation.

Pulmonary function testing was performed in 3 bottlenose dolphins Tursiops truncatus (1 female and 2 males) under managed care during a 2 yr period to assess whether these data provide diagnostic information about respiratory health. Pulmonary radiographs and standard clinical testing were used to evaluate the pulmonary health of each dolphin. The female dolphin (F1) had evidence of chronic pulmonary fibrosis, and 1 male (M2) developed pneumonia during the study. Pulmonary function data were collected from maximal respiratory efforts in water and from spontaneous breaths while beached. From these data, the flow-volume relationship, the flow measured between 25 and 75% of the expired vital capacity (mid forced expiratory flow, FEF25%-75%), and the percent of the vital capacity (VC) at the peak expiratory flow (%VCPEF), were evaluated and compared with the diagnostic assessment. For maximal respiratory manoeuvres in water, there were no differences in FEF25%-75% or %VCPEF, and the flow-volume relationship showed a consistent pattern for F1. Additionally, FEF25%-75% and %VCPEF decreased by 27 and 52%, respectively, and the flow-volume relationship showed clear flow limitations with emerging disease in M2. While spontaneously breathing on land, M2 also showed a 49% decrease in %VCPEF and changes in the flow-volume relationship, indicating flow limitations following the development of pneumonia. Based on these preliminary results, we suggest that pulmonary function testing should be given more attention as a non-invasive and possibly adjunctive diagnostic tool to evaluate lung health of dolphins under managed care and in the wild.

Erysipelas is an infection caused by Erysipelothrix rhusiopathiae that affects many different species around the world, including cetaceans. The acute septicemic form can rapidly cause death in bottlenose dolphins Tursiops truncatus. The ultimate goals of this long-term study were the development and identification of the most effective vaccination protocol against clinical erysipelas in T. truncatus using a commercially available swine vaccine, and to determine whether there is a need for a semi-annual vaccination versus an annual vaccination. The present study concentrated on the immunization of a dolphin population (7 wild-born and 22 captive-born individuals) with 2 swine vaccines, the European 'Eurovac Ery®' vaccine and the American 'ER Bac Plus®' vaccine, and immunological profile results over a 20-yr time period. The general protocol was a primo-vaccination (between 3 and 7 mo of age for calves) with or without a booster 1 mo post primo-vaccination and either annual or semi-annual vaccination thereafter. Sera were collected prior to vaccination, 2 wk post-vaccination and monthly. A dolphin-specific ELISA was developed to analyze the erysipelas-specific antibody response of vaccinated animals. The final ELISA results (n = 1362 samples from 29 animals at pre- and post-vaccination time) suggest that (1) there is a significant difference in antibody levels at the start of the vaccination between older and younger animals; (2) at least 3 vaccinations are necessary to obtain antibody levels above the levels at pre-vaccination; (3) thereafter, annual vaccinations seem sufficient to keep antibody levels above the levels at pre-vaccination; and (4) both vaccines induced similar responses. No case of erysipelas infection was observed in this population during the study.

Gastritis and gastric ulcers are well-recognized symptoms in cetaceans, and the genus Helicobacter is considered as the main cause. In this study, we examined the gastric fluid of captive common bottlenose dolphins Tursiops truncatus with gastric diseases in order to isolate the organisms responsible for diagnosis and treatment. Four Gram-negative, rod-shaped isolates (TSBT, TSH1, TSZ, and TSH3) with tightly coiled spirals with 2-4 turns and 2-6 bipolar, sheathed flagella, were obtained from gastric fluids of common bottlenose dolphins with gastric diseases. Phylogenetic analysis, based on 16S rRNA, atpA, and 60 kDa heat-shock protein (hsp60) genes, demonstrated that these isolates form a novel lineage within the genus Helicobacter. Analyses of 16S rRNA, atpA, and hsp60 gene sequences showed that isolate TSBT was most closely related to H. cetorum MIT99-5656T (98.5% similarity), H. pylori ATCC 43504T (76.7% similarity), and H. pylori ATCC 43504T (78.0% similarity), respectively. Type strains of Helicobacter showing resistance to 2% NaCl have not been reported previously; however, these novel isolates were resistant to 2% NaCl. Culture supernatant of some isolates induced intracellular vacuolization in mammalian cultured cells. These data, together with the different morphological and biochemical characteristics of the isolates, reveal that these isolates represent a novel species for which we propose the name Helicobacter delphinicola sp. nov. with type strain TSBT (= JCM 32789T = TSD-183T). Future studies will confirm whether H. delphinicola plays a role in lesion etiopathogenesis in cetaceans.

Dolphin tattoo lesions are superficial non-raised skin lesions caused by poxviruses. Their presentation can vary but typical lesions in bottlenose dolphins are circular to ovoid with concentric rings of black stippling. These lesions have at times been suggested as an indicator of overall dolphin health and welfare. This study explored the effect of water temperature on the extent of tattoo lesions in 25 dolphins, along with established hematological health parameters and food consumption. Study animals consisted of 9 males and 16 females with dolphin tattoo lesions ranging in age from 2 to 45 yr. A significant decrease (p < 0.01) in extent and appearance of tattoo lesions was documented following increase in water temperature from 21-24°C (70-75°F) to 25.5-26.5°C (78-80°F). Reduction in tattoo lesions could be noted as early as 2-3 wk following water temperature increase. Marked reduction to complete resolution of tattoo lesions was reproducibly seen 5-6 wk post temperature increase. Food consumption following temperature increase was variable: decrease in intake was noted in 48% of dolphins, increase in intake in 52%. Routine blood parameters (complete blood count, serum chemistry panel, fibrinogen, erythrocyte sedimentation rate) remained within normal limits. A significant increase (p < 0.05) in extent and appearance of tattoo lesions was documented in 3 dolphins as early as 4-5 wk following a decrease in water temperature from 26.1°C (79°F) to 21°C (70°F). Water temperature is a key environmental parameter affecting cetacean pox (‘tattoo’) lesions in bottlenose dolphins. The absence of changes in hematological parameters along with lack of correlation between extent of pox lesions and food intake indicates that dolphin tattoo lesions are not an appropriate indicator of overall health.

Lobomycosis and lobomycosis-like diseases (LLD) (also: paracoccidioidomycosis) are chronic cutaneous infections that affect Delphinidae in tropical and subtropical regions worldwide. In the Americas, these diseases have been relatively well-described, but gaps still exist in our understanding of their distribution across the continent. Here we report on LLD affecting inshore bottlenose dolphins Tursiops truncatus from the Caribbean waters of Belize and from the eastern tropical Pacific Ocean off the southwestern coast of Mexico. Photo-identification and catalog data gathered between 1992 and 2017 for 371 and 41 individuals, respectively from Belize and Mexico, were examined for the presence of LLD. In Belize, 5 free-ranging and 1 stranded dolphin were found positive in at least 3 communities with the highest prevalence in the south. In Guerrero, Mexico, 4 inshore bottlenose dolphins sighted in 2014-2017 were affected by LLD. These data highlight the need for histological and molecular studies to confirm the etiological agent. Additionally, we document a single case of LLD in an adult Atlantic spotted dolphin Stenella frontalis in southern Belize, the first report in this species. The role of environmental and anthropogenic factors in the occurrence, severity, and epidemiology of LLD in South and Central America requires further investigation.

A 5 yr old, 184 kg, and 262 cm total length female bottlenose dolphin Tursiops truncatus was found dead in a display after bloody discharge from the blowhole was observed 3 h prior to death. Pathological examination revealed fibrinous bronchopneumonia with prominent areas of necrosis (sequestra) and numerous Gram-negative bacilli within alveoli and in blood vessels of the lungs and liver and between muscle fibers. The cause of death was attributed to septicemia. Often, cases of fibrinous bronchopneumonia are characterized by bacteremia in the latter stages of infection, resulting in the death of the animal. Septicemia likely accounts for the ecchymoses and petechiae noted on the spleen, pancreas, forestomach, lungs, visceral peritoneum, and small intestine. Additional lesions included hemothorax, stable red frothy fluid in the trachea, and lymphoid depletion in the spleen and lymph nodes. Pure growth of Morganella morganii was isolated from the lungs, blood, liver, and blowhole mucosa. Sequencing of 16s rRNA of the isolated bacteria showed more than 99.6% identity with M. morganii strain FDAARGOS_172. To our knowledge, this is the first report of fatal fibrinonecrotizing bronchopneumonia associated with M. morganii infection in a cetacean.

Lobomycosis-like disease (LLD) is a chronic granulomatous skin disorder that affects Delphinidae worldwide. LLD has been observed in common bottlenose dolphins Tursiops truncatus from the Gulf of Guayaquil, Ecuador, since 1990. Although exogenous factors such as salinity and pollution may play a role in the pathogenesis of this disease in estuarine and coastal dolphin communities, we hypothesized that demography and social behaviour may also influence its epidemiology. To address this issue, the role of social behaviour in the distribution and prevalence of LLD was assessed through hierarchical cluster analysis and spatial distribution analysis in 7 dolphin communities inhabiting the inner estuary. Individuals with LLD lesions were observed in 5 of the 7 dolphin communities, with 13 of the 163 (8%) animals being positive, all adults. Among 8 dolphins of known sex, LLD affected mostly males (86%), who usually were found in pairs. Prevalence was low to moderate (5.1-13%) in dolphin communities where low-rank males had LLD. Conversely, it was high (44.4%, n = 9) in a small community where a high-rank male was infected. LLD affected both dolphins in 2 of the 4 male pairs for which large time series data were available, suggesting horizontal transmission due to contact. Thus, association with LLD-positive males seems to be an important risk factor for infections. Additionally, low-rank males had larger home ranges than high-rank males, indicating that low-status LLD-affected dolphins are likely responsible for the geographic dissemination of the disease in this population.

A captive 8 yr old male bottlenose dolphin Tursiops truncatus succumbed to septicemia with multisystemic inflammation including suppurative enteritis, encephalitis, and pneumonia with chronic pancreatitis. A pure culture of beta-hemolytic, catalase- and oxidase-negative, Gram-positive cocci was isolated from the hilar lymph nodes and pancreas. The isolate was identified by 16S rDNA sequencing as Streptococcus iniae. Histological examination of the digestive system revealed a mixed infection of both bacteria and fungus. Recognized as a pathogen in fish, dolphins, and humans, this is the first report of S. iniae in a dolphin in mainland China. As the number of managed animals in oceanariums is increasing, so is the frequency of contact with fish used as food for marine mammals and humans, highlighting the importance of education and appropriate personal protective protocols to minimize the risk of transmission. An understanding of marine mammal infectious disease organisms is essential to ensuring the health of marine mammals and humans coming into contact with such animals and their food. This study illustrates a systematic clinical, microbiological, and pathological investigation into a septicemic bottlenose dolphin infected with S. iniae. Our findings provide useful information for those involved in the diagnosis and control of infectious diseases in marine mammals and offer insight into an important zoonotic pathogen.

Erysipelothrix rhusiopathiae is the causative agent of erysipeloid in humans and of erysipelas in various animals, including bottlenose dolphins Tursiops truncatus, in which an infection has the potential to cause peracute septicemia and death. The purpose of this study was to evaluate the efficacy of using an off-label porcine (ER BAC PLUS®, Zoetis) E. rhusiopathiae bactrin in a bottlenose dolphin vaccination program by determining the anti-E. rhusiopathiae antibody levels in vaccinated dolphins over a 10 yr period. Serum samples (n = 88) were analyzed using a modified fluorescent microbead immunoassay from 54 dolphins, including 3 individuals with no history of vaccination and 51 dolphins with an average of 5 vaccinations, 3 of which had previously recovered from a natural E. rhusiopathiae infection. A mean 311-fold increase in the immunoglobulin G (IgG) antibody index was measured in a subsample of 10 dolphins 14 d after the first booster vaccination. Serum IgG antibody titers were influenced by number of vaccines received (r2 = 0.47, p < 0.05) but not by age, gender, history of natural infection, adverse vaccine reaction, vaccination interval or time since last vaccination. The commercial pig bacterin was deemed effective in generating humoral immunity against E. rhusiopathiae in dolphins. However, since the probability of an adverse reaction toward the vaccine was moderately correlated (p = 0.07, r2 = 0.1) with number of vaccines administered, more research is needed to determine the optimal vaccination interval.

Blowhole swabs are a simple and non-invasive method for collecting samples from cetaceans and can be used for screening large numbers of animals in the field. This study reports a real-time PCR assay for the detection of Brucella spp. using blowhole swab samples from bottlenose dolphins Tursiops truncatus stranded in the coastal region of Virginia, South Carolina and northern Florida, USA, between 2013 and 2015. We used real-time PCR results on lung samples from the same dolphins in order to estimate the relative sensitivity and specificity of real-time PCR of blowhole swabs. Brucella DNA was detected in lung tissue of 22% (18/81) and in blowhole swabs of 21% (17/81) of the sampled dolphins. The relative sensitivity and specificity of real-time PCR on blowhole swabs as compared to the real-time PCR on lung samples was 94% (17/18) and 100% (63/63), respectively. These results indicate that real-time PCR on blowhole swabs may be used as a non-invasive test for rapid detection of Brucella spp. in the respiratory tract of dolphins. To our knowledge, this is the first report on the use of blowhole swabs for detection of bacterial pathogens by real-time PCR in bottlenose dolphins.

Studies of skeletal pathology of marine mammals can contribute to conservation measures, yet few have focused on causative factors. Museums hold vast collections of skeletons relevant to this knowledge gap. Indo-Pacific bottlenose dolphin Tursiops aduncus carcasses (n = 162) were collected from Gulf St Vincent (GSV) and Spencer Gulf (SG), South Australia (SA), between 1988 and 2013, and post-mortem examinations were carried out. After preparing skeletons, their gross pathology was classified into 4 categories: lytic lesions, degenerative lesions, fractures and malformations. Comparisons were made between gulfs based on pathology type, relative age, period of collection and cause of death (anthropogenic vs. non-anthropogenic). Pathology prevalence (76%) was higher than reported in other studies but observed pathologies were similar. More than one pathology was observed in 88% of dolphins that had pathology. Vertebrae were often the site of pathology. Prevalence of lesions were: fractures (82%), lytic lesions (75%) including spondylo-osteomyelitis and osteomyelitis, degenerative lesions (57%) including spondylosis deformans, and malformations (17%). Prevalence of pathology increased with dolphin relative age, and GSV dolphins had more pathology than those in SG. In SG, anthropogenic cases had more pathology than non-anthropogenic cases, and dolphins collected after 2000 had more degenerative lesions than those collected before 2000. There were more malformations in SG dolphins than those from GSV, although this was not statistically significant. In one anomalous case, an individual was found with 5 detached transverse processes on 4 lumbar vertebrae. Heavy metal toxicity reported for T. aduncus from SG may be related to the malformations reported in this study.

We describe gross, histopathologic, ultrastructural, immunohistochemical, and microbiologic features of acute septicemia by Erysipelothrix rhusiopathiae in an Atlantic spotted dolphin Stenella frontalis and an Atlantic bottlenose dolphin Tursiops truncatus. Generalized lymphadenomegaly and widespread hemorrhages were the most consistent macroscopic findings. Tricavitary effusion and icterus were noted in one individual. Histologically, all organs examined showed numerous variably sized bacillary bacterial emboli (Gram-positive; Ziehl-Neelsen-negative), typically associated with systemic congestion, edema, hemorrhages, and fibrinocellular thrombi. These bacteria were frequently intravascular, either extracellular or intramonocytic/macrophagic, and to a lesser extent, free within the interstitium of parenchymal organs. In both cases, microbiological analysis yielded E. rhusiopathiae. A primary anti-E. rhusiopathiae antibody created in mice from one of the strains isolated allowed positive immunohistochemical detection. Electron microscopy and dual immunohistochemistry with lysozyme and MAC387 antibodies confirmed the intramacrophagic location of the bacilli. E. rhusiopathiae, a known multispecies and zoonotic agent, should be considered as a potential etiologic agent in septicemia cases in free-ranging individuals of these dolphin species.

Mucocutaneous lesions were biopsied from free-ranging Atlantic bottlenose dolphins Tursiops truncatus inhabiting the Indian River Lagoon (IRL), Florida, and estuarine waters of Charleston (CHS), South Carolina, USA, between 2003 and 2013. A total of 78 incisional biopsies from 58 dolphins (n=43 IRL, n=15 CHS) were examined. Thirteen dolphins had 2 lesions biopsied at the same examination, and 6 dolphins were re-examined and re-biopsied at time intervals varying from 1 to 8 yr. Biopsy sites included the skin (n=47), tongue (n=2), and genital mucosa (n=29). Pathologic diagnoses were: orogenital sessile papilloma (39.7%), cutaneous lobomycosis (16.7%), tattoo skin disease (TSD; 15.4%), nonspecific chronic to chronic-active dermatitis (15.4%), and epidermal hyperplasia (12.8%). Pathologic diagnoses from dolphins with 2 lesions were predominately orogenital sessile papillomas (n=9) with nonspecific chronic to chronic-active dermatitis (n=4), TSD (n=3), lobomycosis (n=1), and epidermal hyperplasia (n=1). Persistent pathologic diagnoses from the same dolphins re-examined and re-biopsied at different times included genital sessile papillomas (n=3), lobomycosis (n=2), and nonspecific dermatitis (n=2). This is the first study documenting the various types, combined prevalence, and progression of mucocutaneous lesions in dolphins from the southeastern USA. The data support other published findings describing the health patterns in dolphins from these geographic regions. Potential health impacts related to the observed suite of lesions are important for the IRL and CHS dolphin populations, since previous studies have indicated that both populations are affected by complex infectious diseases often associated with immunologic disturbances and anthropogenic contaminants.

Bottlenose dolphins Tursiops truncatus are the most common apex predators found in coastal and estuarine ecosystems along the southeastern coast of the USA, where these animals are exposed to multiple chemical pollutants and microbial agents. In this review, we summarize the results of investigations of environmental exposures evaluated in 360 free-ranging dolphins between 2003 and 2015. Bottlenose dolphins inhabiting the Indian River Lagoon, Florida (IRL, n = 246), and coastal waters of Charleston, South Carolina (CHS, n = 114), were captured, given comprehensive health examinations, and released as part of a multidisciplinary and multi-institutional study of individual and population health. High concentrations of persistent organic pollutants including legacy contaminants (DDT and other pesticides, polychlorinated biphenyl compounds) as well as 'emerging' contaminants (polybrominated diphenyl ethers, perfluorinated compounds) were detected in dolphins from CHS, with lower concentrations in the IRL. Conversely, the concentrations of mercury in the blood and skin of IRL dolphins were among the highest reported worldwide and approximately 5 times as high as those found in CHS dolphins. A high prevalence of resistance to antibiotics commonly used in humans and animals was detected in bacteria isolated from fecal, blowhole, and/or gastric samples at both sites, including methicillin-resistant Staphylococcus aureus (MRSA) at CHS. Collectively, these studies illustrate the importance of long-term surveillance of estuarine populations of bottlenose dolphins and reaffirm their important role as sentinels for marine ecosystems and public health.

From 2003 to 2015, 360 free-ranging Atlantic bottlenose dolphins Tursiops truncatus inhabiting the Indian River Lagoon (IRL, n = 246), Florida, and coastal waters of Charleston (CHS, n = 114), South Carolina, USA, were captured, given comprehensive health examinations, and released as part of a multidisciplinary and multi-institutional study of individual and population health. The aim of this review is to summarize the substantial health data generated by this study and to examine morbidity between capture sites and over time. The IRL and CHS dolphin populations are affected by complex infectious and neoplastic diseases often associated with immunologic disturbances. We found evidence of infection with cetacean morbillivirus, dolphin papilloma and herpes viruses, Chlamydiaceae, a novel uncultivated strain of Paracoccidioides brasiliensis (recently identified as the causal agent of dolphin lobomycosis/lacaziasis), and other pathogens. This is the first long-term study documenting the various types, progression, seroprevalence, and pathologic interrelationships of infectious diseases in dolphins from the southeastern USA. Additionally, the study has demonstrated that the bottlenose dolphin is a valuable sentinel animal that may reflect environmental health concerns and parallel emerging public health issues.

Lacaziosis (also known as lobomycosis) is a chronic dermal disease caused by the fungal agent Lacazia loboi, which affects both humans and dolphins. Photographic data have been used to identify lacaziosis-like disease (LLD) among dolphins in the waters of North and South America, and here we report LLD in bottlenose dolphins Tursiops truncatus off the coast of Costa Rica, the first reporting in Central American waters. During the periods of 1991 to 1992 and 2010 to 2011, 3 research teams conducted separate dolphin surveys in the Pacific tropical fiord Golfo Dulce, and each documented skin lesions in the resident population of bottlenose dolphins. Photo-ID records were used to identify LLD-affected bottlenose dolphins and to assess their lesions. Findings showed between 13.2 and 16.1% of the identified dolphins exhibited lesions grossly resembling lacaziosis. By combining efforts and cross-referencing photographic data, the teams explored the presence of LLD in Golfo Dulce over a time gap of approximately 20 yr. Our findings expand the geographical range of the disease and offer insight into its longevity within a given population of dolphins.

Nine striped dolphins Stenella coeruleoalba and 1 bottlenose dolphin Tursiops truncatus stranded along the Ligurian Sea coast of Italy were necropsied between February 2011 and April 2012. Macroscopic and histological findings were observed in the hearts of all animals and included saccular aneurysms of the pulmonary trunk (n = 3), cirsoid aneurysms (n = 1), right ventricular dilation (n = 1) associated with hypoplasia of the tricuspid chordae (n = 1), valvular fibrosis (n = 3), mitral leaflet thickening (n = 1), left ventricular hypertrophy (n = 1), lymphocytic myocarditis (n =1), and Lambl's excrescences (n = 4). To our best knowledge Lambl's excrescences, aneurysm of the pulmonary trunk, and cirsoid aneurysms have not previously been described in marine mammals, and some of these findings should be taken into account as possible causes of dolphin morbidity, mortality, and stranding.

Hepatitis E virus (HEV) infects several animal species that act as zoonotic reservoirs for viral transmission. Solid and liquid residues from infected animals could lead to HEV contamination of food and surface waters. Evidence of human HEV infection through ingestion of seafood (shellfish, mussels) has been reported. Dolphins generally feed on fish and squid but are able to adapt to an environment and consume whatever prey is available. Clinical signs of infected dolphins include lethargy, inappetence, behavioral aberrations and increased serum alanine aminotransferase (ALT). The dolphins examined in this study were maintained at the National Aquarium, Havana, Cuba. A total of 31 dolphins were evaluated for HEV markers. Sera were collected and screened for total immunoglobin (Ig) anti-HEV. Sera and liver homogenate were tested for HEV RNA by nested RT-PCR using primers targeting the open reading frame 1. Phylogenetic analysis was performed using partial nucleotide sequences at the amplified RNA-dependent RNA polymerase gene. Total anti-HEV Ig was detected in 32.2% (10 of 31), and 16.1% (5 of 31) of these dolphins were positive by both serology and HEV RNA testing. Nucleotide sequence analyses revealed that HEV strains identified in dolphins were genotype 3. This virus may represent an environmental contamination of food or wastewater as a source of HEV exposure and infection. Our findings provide evidence that HEV is associated with liver disorders in cetaceans and that it is advisable to screen for exposure of this virus in captive dolphins, particularly animals with elevated serum ALT or compromised liver function test results of undetermined cause.

An unusual mortality event (UME) involving primarily common bottlenose dolphins Tursiops truncatus of all size classes stranding along coastal Louisiana, Mississippi, and Alabama, USA, started in early 2010 and continued into 2014. During this northern Gulf of Mexico UME, a distinct cluster of perinatal dolphins (total body length

Marine-origin Brucella infections and serologic evidence of exposure have been doc- umented in multiple cetacean species. A dolphin-specific indirect enzyme-linked immunosorbent assay (ELISA) was developed to screen bottlenose dolphin sera for anti-Brucella antibodies. A total of 131 serum samples collected over a 2 to 18 yr period from 6 bottlenose dolphins Tursiops truncatus with confirmed Brucella infections were analyzed for the presence and magnitude of antibody titers against marine-origin Brucella to compare individual antibody responses to various disease manifestations. Additionally, an epidemiologic serologic survey of a managed population of 64 bottlenose dolphins was performed to evaluate for the presence of antibodies and to deter- mine whether there were any clinical pathology predictors for exposure or infection. The serologic results revealed that the dolphins with Brucella-associated abortions were seronegative for 7 to 18 yr until after the abortion and maintained positive titers for several years, with 2 of 3 animals returning to seronegative status. In contrast, the dolphins with Brucella-associated pulmonary or bone lesions maintained persistent positive titers for 2 to 18 yr. The population serosurvey revealed no significant differences in antibody levels among males and females, and dolphins between the ages of 17 and 25 yr were 6.8 times more likely to be Brucella antibody positive com- pared to those that were younger or older. Seropositive dolphins did not have significant inflam- mation compared to seronegative dolphins but were more likely to have higher levels of aspartate aminotransferase and gamma-glutamyl transpeptidase. Among 16 dolphins that tested seroposi- tive, 13 (81.3%) had previously been seropositive for at least 3 to 5 yr.

Lacaziosis (lobomycosis; Lacazia loboi) is a fungal skin disease that naturally occurs only in humans and dolphins. The first reported case of lacaziosis in a bottlenose dolphin Tursiops truncatus occurred in 1970 in Sarasota Bay, Florida, USA, and subsequent photo-ID monitoring of the Sarasota Bay dolphin population has revealed persistence of the disease. The objectives of this study were to estimate lacaziosis prevalence (P) in 2 bottlenose dolphin populations on the west coast of Florida (Sarasota Bay and Charlotte Harbor) and compare disease occurrence to other published estimates of lacaziosis in dolphin populations across the globe. Historic photographic records of dolphins captured and released for health assessment purposes (Sarasota Bay) and photo-ID studies (Charlotte Harbor) were screened for evidence of lesions consistent with lacaziosis. Health assessment data revealed a prevalence of lacaziosis in the Sarasota Bay bottlenose dolphin population between 2 and 3%, and analyses of photo-ID data provided a lacaziosis-like prevalence estimate of 2% for Charlotte Harbor dolphins. With the exception of lacaziosis prevalence estimates for dolphins inhabiting the Indian River Lagoon (P = 0.068; P = 0.12), no statistically significant differences were seen among Sarasota Bay, Charlotte Harbor, and other published estimates. Although lacaziosis is a rare disease among these dolphin populations, studies that assess disease burden among different populations can assist with the surveillance of this zoonotic pathogen.

Lacaziosis (lobomycosis) is a skin disease caused by Lacazia loboi, occurring naturally only in humans and dolphins. Attempts to culture the pathogen in vitro have been unsuccesstul, and inoculation studies of lacaziosis development in mice have provided only limited, short-term data on the progression and propagation of L. loboi. The present study used photographic data from long-term photo-identification and health assessment projects to model and quantify the progression of lacaziosis lesions in 3 common bottlenose dolphins Tursiops truncatus from Sarasota Bay, Florida, USA. Dorsal fin images throughout each animal's sighting history were examined for lesion growth, and the proportion of lesion coverage in each photograph was estimated using image analysis tools in Adobe Photoshop ® . The progression of lacaziosis lesions and lesion growth rates were modeled using a non-linear monomolecular growth model. As data on lacaziosis development and advancement are limited in humans and laboratory animals, dolphins with a long-term case history of the disease may serve as a good animal model to better understand lacaziosis progression. Furthermore, this study demonstrates the utility of long-term population monitoring data for tracking the progression of a poorly understood disease that is relevant to both dolphin and human health.

We report on the epidemiology of lobomycosis-like disease (LLD), a cutaneous disorder evoking lobomycosis, in 658 common bottlenose dolphins Tursiops truncatus from South America and 94 Indo-Pacific bottlenose dolphins T. aduncus from southern Africa. Photographs and stranding records of 387 inshore residents, 60 inshore non-residents and 305 specimens of undetermined origin (inshore and offshore) were examined for the presence of LLD lesions from 2004 to 2015. Seventeen residents, 3 non-residents and 1 inshore dolphin of unknown residence status were positive. LLD lesions appeared as single or multiple, light grey to whitish nodules and plaques that may ulcerate and increase in size over time. Among resident dolphins, prevalence varied significantly among 4 communities, being low in Posorja (2.35%, n = 85), Ecuador, and high in Salinas, Ecuador (16.7%, n = 18), and Laguna, Brazil (14.3%, n = 42). LLD prevalence increased in 36 T. truncatus from Laguna from 5.6% in 2007-2009 to 13.9% in 2013-2014, albeit not significantly. The disease has persisted for years in dolphins from Mayotte, Laguna, Salinas, the Sanquianga National Park and Bahia Malaga (Colombia) but vanished from the Tramandai Estuary and the Mampituba River (Brazil). The geographical range of LLD has expanded in Brazil, South Africa and Ecuador, in areas that have been regularly surveyed for 10 to 35 yr. Two of the 21 LLD-affected dolphins were found dead with extensive lesions in southern Brazil, and 2 others disappeared, and presumably died, in Ecuador. These observations stress the need for targeted epidemiological, histological and molecular studies of LLD in dolphins, especially in the Southern Hemisphere.

The number and prevalence of diseases is rapidly increasing in the marine ecosys- tem. Although there is an increase in the number of marine diseases observed world-wide, current understanding of the pathogens associated with marine mammals is limited. An important need exists to develop and apply platforms for rapid detection and characterization of pathogenic agents to assess, prevent and respond to disease outbreaks. In this study, a broad-spectrum molec- ular detection technology capable of detecting all sequenced microbial organisms, the Lawrence Livermore Microbial Detection Array, was used to assess the microbial agents that could be asso- ciated with wild Atlantic dolphins. Blowhole, gastric, and fecal samples from 8 bottlenose dolphins were collected in Charleston, SC, as part of the dolphin assessment effort. The array detected var- ious microbial agents from the dolphin samples. Clostridium perfringens was most prevalent in the samples surveyed using the microarray. This pathogen was also detected using microbiologi- cal culture techniques. Additionally, Campylobacter sp., Staphylococcus sp., Erwinia amylovora, Helicobacter pylori, and Frankia sp. were also detected in more than one dolphin using the micro- array, but not in culture. This study provides the first survey of pathogens associated with 3 tissue types in dolphins using a broad-spectrum microbial detection microarray and expands insight on the microbial community profile in dolphins.

Contamination of coastal waters can carry pathogens and contaminants that cause diseases in humans and wildlife, and these pathogens can be transported by water to areas where they are not indigenous. Marine mammals may be indicators of potential health effects from such pathogens and toxins. Here we isolated bacterial species of relevance to humans from wild bottlenose dolphins Tursiops truncatus and assayed isolated bacteria for antibiotic resistance. Samples were collected during capture-release dolphin health assessments at multiple coastal and estuarine sites along the US mid-Atlantic coast and the Gulf of Mexico. These samples were transported on ice and evaluated using commercial systems and aerobic culture techniques routinely employed in clinical laboratories. The most common bacteria identified were species belonging to the genus Vibrio, although Escherichia coli, Shewanella putrefaciens, and Pseudomonas fluorescens/putida were also common. Some of the bacterial species identified have been associated with human illness, including a strain of methicillin-resistant Staphylococcus aureus (MRSA) identified in 1 sample. Widespread antibiotic resistance was observed among all sites, although the percentage of resistant isolates varied across sites and across time. These data provide a baseline for future comparisons of the bacteria that colonize bottlenose dolphins in the southeastern USA.

Lung disease is common among wild and managed populations of bottlenose dol-phins Tursiops truncatus . The purpose of the study was to apply standardized techniques to theultrasound evaluation of dolphin lungs, and to identify normal and abnormal sonographic findingsassociated with pleuropulmonary diseases. During a 5 yr period (2005 to 2010), 498 non-cardiacthoracic ultrasound exams were performed on bottlenose dolphins at the Navy Marine MammalProgram in San Diego, California, USA. Exams were conducted as part of routine physical exams,diagnostic workups, and disease monitoring. In the majority of routine exams, no abnormal pleu-ral or pulmonary findings were detected with ultrasound. Abnormal findings were typicallydetected during non-routine exams to identify and track disease progression or resolution; there-fore, abnormal results are overrepresented in the study. In order of decreasing prevalence, abnor-mal sonographic findings included evidence of alveolar-interstitial syndrome, pleural effusion,pulmonary masses, and pulmonary consolidation. Of these findings, alveolar-interstitial syndromewas generally nonspecific as it represented several possible disease states. Pairing ultrasoundfindings with clinical signs was critical to determine relevance. Pleural effusion, pulmonarymasses, and pulmonary consolidation were relatively straightforward to diagnose and interpret.Further diagnostics were performed to obtain definitive diagnoses when appropriate, specificallyultrasound-guided thoracocentesis, fine needle aspirates, and lung biopsies, as well as radio -graphs and computed tomography (CT) exams. Occasionally, post mortem gross necropsy andhisto pathology data were available to provide confirmation of diagnoses. Thoracic ultrasound wasdetermined to be a valuable diagnostic tool for detecting pleural and pulmonary diseases in dolphins.KEY WORDS: Cetacean · Diagnostic imaging · Lung disease · Pulmonary

Pneumonia is one of the most common causes of morbidity in bottlenose dolphins Tursiops truncatus. To better understand associations of pneumonia with demographics, microbiology, pathology, and histopathology, a retrospective study on 42 dolphins from the US Navy Marine Mammal Program dolphin population was conducted (1980 to 2010). A total of 21 (50%) of the dolphins evaluated had pneumonia confirmed by histopathology. Bacterial and fungal pneumonia was present in 42.9 and 28.6% of cases (9 and 6 cases), respectively, with Staphylococcus aureus as the most common confirmed pathogen (4 cases, 19%). Other pathogens identified as the cause of pneumonia were Cryptococcus neoformans, Erysipelothrix rhusiopathiae, Histoplasma capsulatum, parainfluenza virus, Proteus species, Pseudomonas aeruginosa, and Streptococcus zooepidemicus. Neither sex nor age was a predictor of pneumonia. While many of the infections involved disseminated disease, lungs were consistently the most severely affected organs. The present study demonstrates the high susceptibility of dolphins to respiratory infections. Areas that warrant further investigation include eosinophilic pneumonia, chronic infections, co-infections, and metabolic or iron-storage diseases. There is a continuing need to improve the early diagnosis and effective treatment of pneumonia in dolphins.

An adult female bottlenose dolphin (Tursiops truncatus) stranded alive and subsequently died several minutes later on the Mediterranean coast of Spain on 14 July 2010. Clinical examination revealed foam through the blowhole and rales upon lung auscultation. On venipuncture, the blood was abnormally dense and dark. Hematological and biochemical abnormalities included dehydration, leukocytosis (48 600 leukocytes microl(-1)) characterized by neutrophilia (48 200 neutrophils microl(-1)), and elevated bilirubin (4.38 mg dl(-1)), alanine aminotransferase (382.3 U l(-1)), aspartate aminotransferase (1449.3 U l(-1)), lactate dehydrogenase (1631.3 U l(-1)), and creatine kinase (404.7 U l(-1)). The most relevant findings of the gross examination were rhomboid-shaped skin lesions, stable froth in the trachea, pulmonary congestion, abnormally thick and rough pleura with adhesions, edematous and congestive superficial cervical and tracheobronchial lymph nodes, red-tinged urine, and severe brain congestion. Histopathology of the kidney, lung, skin, and brain revealed multisystemic intravascular bacterial emboli. Samples of skin, brain, and lung were cultured on Columbia blood agar under both aerobic and anaerobic conditions, and pure and heavy bacterial cultures were obtained from skin and brain samples. The microorganism isolated was Gram-positive, catalase-negative, facultatively anaerobic, and rod-shaped. The isolates were identified as Erysipelothrix rhusiopathiae by the API Coryne biochemical system. Based on the gross and microscopic findings, a diagnosis of acute E. rhusiopathiae septicemia was made. To the best of our knowledge, this is the first report of E. rhusiopathiae septicemia in a free-ranging bottlenose dolphin.

Lobomycosis is a chronic dermal infection affecting humans and small cetaceans. In 1993, a study identified the presence of the etiologic agent of lobomycosis in a resident population of Tursiops truncatus (bottlenose dolphin) in Laguna, Brazil. Until now, no additional information relating to the persistence or prevalence of this pathogen in this population has been available. Numbering less than 60 animals, the residency of these dolphins in an impacted lagoon system has raised concerns about the health and viability of this small population. Using photo-identification data collected between September 2007 and September 2009, this study evaluated the occurrence of lobomycosis-like disease (LLD) throughout this population. Of 47 adult dolphins and 10 calves identified, 7 (12%) presented some form of epidermal lesion and 5 (9%) had evidence of LLD. The lesions were stable in all but 2 cases, in which a progressive development was recorded in a presumed adult female and her calf (referred to here as the LLD pair). During the first few months of observation, the lesion grew slowly and at a constant rate on the adult. However, in the fourteenth month, the growth rate increased rapidly and the first lesions appeared on the calf. Compared to the rest of the population, the LLD pair also presented a different spatial ranging pattern, suggesting a possible social or geographic factor. Current and previous records of LLD or lobomycosis indicate that the disease is endemic in this population. These findings highlight the importance of monitoring both the health of these cetaceans and the quality of their habitat.

This study represents the first systematic study of lacaziosis (lobomycosis) in bottlenose dolphins Tursiops truncatus in the Atlantic Ocean along the east-central coast of Florida, USA. Lacaziosis is a chronic infection of the skin caused by the fungus Lacazia loboi, which affects only dol- phins and humans. Previous studies have shown a high prevalence (6.8 to 12.0%) of lacaziosis in res- ident dolphins from the adjacent Indian River Lagoon Estuary (IRL), where the disease is endemic. We examined the prevalence of lacaziosis in this coastal area using photo-identification data col- lected between 2002 and 2008 to determine the prevalence of lacaziosis in coastal dolphins using photographic methodology shown to have high sensitivity and specificity in prior research. The prevalence of skin lesions compatible with lacaziosis estimated from photographic data was 2.1% (6/284), approximately 3 times lower than that described for the estuarine population using similar methods. To exclude potential bias introduced by differences in study duration and survey effort among areas, an 18 mo period when effort was most equal (January 2006 to June 2007) was chosen for statistical comparison. The prevalence of lacaziosis estimated from photographic data was signif- icantly lower (3.8%: n = 6/160) in the Atlantic Ocean compared to the IRL (12.0%: n = 20/167) (risk ratio = 3.19, 95% CI 1.32 to 7.75, p < 0.01 by chi-square analysis). The lower prevalence of lacaziosis in dolphins found in the Atlantic Ocean and the overall lack of movement of dolphins between these habitats suggests that environmental conditions within the estuary may favor viability of L. loboi, and/or that immune compromise in resident estuarine dolphins is a precursor to the disease.

Few cases of nephrolithiasis (renal calculi) have been reported in bottlenose dolphins Tursiops truncatus. A case-control study was conducted to compare ultrasonographic images and clinicopathologic serum and urine values among 14 dolphins with nephrolithiasis (mild cases: 1 to 19 nephroliths, n = 8; advanced cases:or = 20 nephroliths, n = 6) to 6 controls over an 18 mo period. Archived nephroliths collected postmortem from 7 additional bottlenose dolphins were characterized using quantitative analysis. All advanced cases had bilateral nephroliths, and 67% had visible collecting ducts. During the study, 2 of the advanced cases developed hydronephrosis, and 1 of these cases had ureteral obstruction due to a nephrolith. Compared to controls, cases (mild and advanced) were significantly more likely to have anemia (hematocrit [HCT]38%), high blood urea nitrogen (59 mg dl(-1)), high creatinine (1.9 mg dl(-1)), and low estimated glomerular filtration rate (150 ml min(-1) 2.78 m(-2)). Advanced-case urine samples were more likely to have erythrocytes, occult blood, and lower pH compared to mild cases and controls. Mean serum uric acid among all study groups was low (0.15 to 0.27 mg dl(-1)). Urinary uric acid concentrations were highest among mild cases (272 mg g(-1) creatinine), but advanced cases had levels lower than that of controls (40 and 127 mg g(-1) creatinine, respectively). All nephroliths were characterized as 100% ammonium acid urate. We conclude that nephrolithiasis is clinically relevant in dolphins and can decrease renal function and HCT. The presence of nephrolithiasis, presumably ammonium acid urate nephrolithiasis, in the face of low serum uric and relatively low urinary uric acid in advanced cases may indicate a metabolic syndrome similar to that reported in humans.

Parasitism of the respiratory system is a relatively common finding in stranded cetaceans; however, no systematic investigations regarding the severity, distribution, and clinical consequences of these infections in bottlenose dolphins Tursiops truncatus have been conducted pre- viously. The present study determined the prevalence of lungworm infections in dead stranded (n = 22) and live bottlenose dolphins (n = 44) from southwestern Florida, USA, during the period from 2003 to 2005. Dead stranded bottlenose dolphins were necropsied and lungs were examined visually, by palpation, and histologically for lesions consistent with verminous pneumonia. When present, nematodes were counted, measured, and identified to species based upon their morphology. Dolphin feces and blowhole swabs were collected and examined for nematode larvae. Lungworm prevalence was 77% in dead animals (n = 22). The lesions in most cases were mild, chronic, and not the primary cause of death. Only 13% of dead animals examined had patent infections, with larvae present in blowhole and fecal cytology, and only 18% of animals had intact worms present at necropsy, with a geometric mean intensity of infection of 22.6 worms animal -1 . Intact worms were identified as either Halocercus lagenorhynchi or Skrjabinalius cryptocephalus. The highest prevalence of active infec- tions was found in neonates and calves, including 1 stillborn calf. For free-ranging animals, all blow- hole swabs (n = 44) were negative, and fecal cytology (n = 22) showed a 3% prevalence of patent infection. Findings from the present study support the theory that bottlenose dolphins can be infected transplacentally by lungworms. The impact that such infections may have on neonatal survival is unknown; however, these infections could increase neonatal mortality.

Bacterial cultures of marine mammal samples often yield multiple genera and species, and it can be difficult to determine if a cultured bacterium is a primary pathogen or an incidental finding. To determine the relative risk of bacterial isolates among Atlantic bottlenose dolphins Tursiops truncatus at the United States Navy Marine Mammal Program (MMP), retrospective data on isolates cultured during June 1987 through June 2007 were organized into a novel, 5-tier risk categorization system limited to sole bacteria cultured from internal organ or fluid samples. Of 2586 bacterial isolates cultured, only 34 (1.3%) and 25 (1.0%) were sole isolates attributed to morbidity and mortality, respectively, and only 19 (0.7%) isolates were associated with mortalities without evidence of fungal or viral co-infections. Highest risk bacterial isolates were most likely to be identified in pleural fluid (33.3% of pleural fluid samples with bacterial isolates had only one genus), followed by renal (23.1%) and splenic (11.1%) tissue. Sole Staphylococcus aureus isolates were identified as the highest risk bacterial pathogens in the MMP dolphin population, accounting for 0.4 % of total bacterial isolates over a 20 yr period. In summary, isolation of sole bacterial isolates definitively associated with morbidity and mortality in marine mammals was uncommon in the MMP population. Our proposed risk categorization system may be useful in determining high risk pathogens among other marine mammal populations.