Genomic sequencing of a virus representing a novel type within the species Dyopipapillomavirus 1 in an Indian River Lagoon bottlenose dolphin.

Fecal samples collected from free-ranging Atlantic bottlenose dolphins (BDs) in the Indian River Lagoon of Florida were processed for viral discovery using a next-generation sequencing (NGS) approach. A 693-bp contig identified in the NGS data was nearly identical to the partial L1 gene sequence of a papillomavirus (PV) previously found in a penile papilloma in a killer whale (Orcinus orca). Based on this partial bottlenose dolphin papillomavirus (BDPV) sequence, a nested inverse PCR and primer-walking strategy was employed to generate the complete genome sequence. The full BDPV genome consisted of 7299 bp and displayed a typical PV genome organization. The BDPV E6 protein contained a PDZ-binding motif, which has been shown to be involved in carcinogenic transformation involving high-risk genital human PVs. Screening of 12 individual fecal samples using a specific endpoint PCR assay revealed that the feces from a single female BD displaying a genital papilloma was positive for the BDPV. Genetic analysis indicated that this BDPV (Tursiops truncatus papillomavirus 8; TtPV8) is a new type of Dyopipapillomavirus 1, previously sequenced from an isolate obtained from a penile papilloma in a harbor porpoise (Phocoena phocoena). Although only a partial L1 sequence has been determined for a PV detected in a killer whale genital papilloma, our finding of a nearly identical sequence in an Atlantic BD may indicate that members of this viral species are capable of host jumping. Future work is needed to determine if this virus is a high-risk PV that is capable of inducing carcinogenic transformation and whether it poses a significant health risk to wild delphinid populations.

Health and Environmental Risk Assessment Project for bottlenose dolphins Tursiops truncatus from the southeastern USA. I. Infectious diseases.

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.

Health and Environmental Risk Assessment Project for bottlenose dolphins Tursiops truncatus from the southeastern USA. II. Environmental aspects.

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.

Cutaneous Granulomas in Dolphins Caused by Novel Uncultivated Paracoccidioides brasiliensis.

Cutaneous granulomas in dolphins were believed to be caused by Lacazia loboi, which also causes a similar disease in humans. This hypothesis was recently challenged by reports that fungal DNA sequences from dolphins grouped this pathogen with Paracoccidioides brasiliensis. We conducted phylogenetic analysis of fungi from 6 bottlenose dolphins (Tursiops truncatus) with cutaneous granulomas and chains of yeast cells in infected tissues. Kex gene sequences of P. brasiliensis from dolphins showed 100% homology with sequences from cultivated P. brasiliensis, 73% with those of L. loboi, and 93% with those of P. lutzii. Parsimony analysis placed DNA sequences from dolphins within a cluster with human P. brasiliensis strains. This cluster was the sister taxon to P. lutzii and L. loboi. Our molecular data support previous findings and suggest that a novel uncultivated strain of P. brasiliensis restricted to cutaneous lesions in dolphins is probably the cause of lacaziosis/lobomycosis, herein referred to as paracoccidioidomycosis ceti.

Toward the identification, characterization and experimental culture of Lacazia loboi from Atlantic bottlenose dolphin (Tursiops truncatus).

Lobomycosis (lacaziosis) is a chronic, granulomatous, fungal infection of the skin and subcutaneous tissues of humans and dolphins. To date, the causative agent, the yeast-like organism Lacazia loboi, has not been grown in the laboratory, and there have been no recent reports describing attempts to culture the organism. As a result, studies on the efficacy of therapeutics and potential environmental reservoirs have not been conducted. Therefore, the objective of the current study was to utilize both classical and novel microbiological methods in order to stimulate growth of Lacazia cells collected from dolphin lesions. This included the experimental inoculation of novel media, cell culture, and the use of artificial skin matrices. Although unsuccessful, the methods and results of this study provide important insight into new approaches that could be utilized in future investigations of this elusive organism.

Mucocutaneous lesions in free-ranging Atlantic bottlenose dolphins Tursiops truncatus from the southeastern USA.

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.

Stress response of wild bottlenose dolphins (Tursiops truncatus) during capture-release health assessment studies.

There is a growing concern about the impacts of stress in marine mammals as they face a greater array of threats. The stress response of free-ranging dolphins (Tursiops truncatus) was examined by measuring their physiologic response to capture and handling. Samples were collected from 168 dolphins during capture-release health assessments 2003-2007 at two study sites: Charleston, SC (CHS) and the Indian River Lagoon, FL (IRL). Adrenocorticotropic hormone (ACTH), cortisol, aldosterone (ALD) and catecholamines (epinephrine (EPI), norepinephrine (NOR), dopamine (DA)), were measured in blood and cortisol in urine. Mean time to collect pre-examination samples after netting the animals was 22min; post-examination samples were taken prior to release (mean 1h 37min). EPI and DA concentrations decreased significantly with increased time to blood sampling. ACTH and cortisol levels increased from the initial capture event to the post-examination sample. EPI concentrations increased significantly with increasing time to the pre-examination sample and decreased significantly with time between the pre- and post-examination sample. Cortisol concentrations increased between the pre- and post-examination in CHS dolphins. Age- and sex-adjusted mean pre-examination values of catecholamines were significantly higher in CHS dolphins; ALD was higher in IRL dolphins. Significant differences related to age or sex included higher NOR concentrations in males; higher ALD and urine cortisol levels in juveniles than adults. Wild dolphins exhibited a typical mammalian response to acute stress of capture and restraint. Further studies that relate hormone levels to biological and health endpoints are warranted.

Clinicoimmunopathologic findings in Atlantic bottlenose dolphins Tursiops truncatus with positive Chlamydiaceae antibody titers.

Sera from free-ranging Atlantic bottlenose dolphins Tursiops truncatus inhabiting the Indian River Lagoon (IRL), Florida, and coastal waters of Charleston (CHS), South Carolina, USA, were tested for antibodies to Chlamydiaceae as part of a multidisciplinary study of individual and population health. A suite of clinicoimmunopathologic variables was evaluated in Chlamydiaceae-seropositive dolphins (n = 43) and seronegative healthy dolphins (n = 83). Fibrinogen, lactate dehydrogenase, amylase, and absolute numbers of neutrophils, lymphocytes, and basophils were significantly higher, and serum bicarbonate, total alpha globulin, and alpha-2 globulin were significantly lower in dolphins with positive Chlamydiaceae titers compared with seronegative healthy dolphins. Several differences in markers of innate and adaptive immunity were also found. Concanavalin A-induced T lymphocyte proliferation, lipopolysaccharide-induced B lymphocyte proliferation, and granulocytic phagocytosis were significantly lower, and absolute numbers of mature CD 21 B lymphocytes, natural killer cell activity and lysozyme concentration were significantly higher in dolphins with positive Chlamydiaceae antibody titers compared to seronegative healthy dolphins. Additionally, dolphins with positive Chlamydiaceae antibody titers had significant increases in ELISA antibody titers to Erysipelothrix rhusiopathiae. These data suggest that Chlamydiaceae infection may produce subclinical clinicoimmunopathologic perturbations that impact health. Any potential subclinical health impacts are important for the IRL and CHS dolphin populations, as past studies have indicated that both dolphin populations are affected by other complex infectious and neoplastic diseases, often associated with immunologic perturbations and anthropogenic contaminants.

Symptom frequency and exposure to a cyanobacteria bloom in Florida.

This investigation was undertaken to characterize health effects associated with a major bloom of blue-green algae due to the proliferation Microcystis aeruginosa that occurred in Florida in 2018. Cyanobacteria produce multiple toxins, including the potent hepatotoxic microcystins (MCs), that have been reported to cause illness in exposed persons worldwide. Widespread exposure to toxins released by blue-green algae during the 2018 bloom was shown by the presence of MCs in the nasal passages of 95 percent of the individuals studied previously in south Florida (Schaefer et al., 2020). The current analyses were conducted to determine whether self-reported symptoms were associated with activity patterns, direct contact with water, residential, recreational, and occupational exposure. The 125 persons who participated in the initial study reported an average of 4.94 (± 4.87) symptoms. Those reported most commonly included rhinorrhea, sneezing, headache, sore throat and dry cough. Respiratory symptoms were reported by 74%, ocular symptoms by 62%, and gastrointestinal symptoms by 35% of respondents. Residential and recreational exposures were associated with increased risks of respiratory, gastrointestinal, or ocular symptoms in univariate and adjusted multivariable analyses. Residential exposure was significantly associated with increased reporting of dry cough (p = 0.03), dyspnea (p < 0.01) and wheezy respirations (p = 0.04). Among persons reporting gastrointestinal symptoms, nausea (p = 0.02) and abdominal pain (p < 0.01) were significantly associated with residential exposure. Recreational exposure was significantly associated with sore throat and eye irritation. The findings add to the evidence that exposure to cyanobacteria at concentrations encountered during an algal bloom is associated with a diverse array of symptoms and that inhalation of aerosols constitutes an important exposure pathway.

Epidemiologic characterization of Colorado backyard bird flocks.

Backyard gallinaceous bird flocks may play an important role in the spread of infectious diseases within poultry populations as well as the transmission of zoonotic diseases to humans. An epidemiologic characterization was conducted of Colorado backyard flocks to gather information on general flock characteristics, human movement of birds, human-bird interaction, biosecurity practices, and flock health. Our results suggest that backyard poultry flocks in Colorado are small-sized flocks (68.6% of flocks had < 50 birds); consist primarily of layer chickens (85.49% of flocks), show chickens (32.18% of flocks), and waterfowl (34.07% of flocks); and are primarily owned for food (meat or egg) production for the family (86.44%) or as pet or hobby birds (42.27%). The backyard flock environment may promote bird-to-bird transmission as well as bird-to-human transmission of infectious disease. Birds are primarily housed with free access to the outside (96.85%), and many are moved from the home premises (46.06% within 1 yr). Human contact with backyard flocks is high, biosecurity practices are minimal, and bird health is negatively impacted by increased movement events. Increased knowledge of backyard bird characteristics and associated management practices can provide guidelines for the development of measures to decrease disease transmission between bird populations, decrease disease transmission from birds to humans, and increase the overall health of backyard birds.

Disseminated toxoplasmosis in Antillean manatees Trichechus manatus manatus from Puerto Rico.

Necropsies were conducted on 4 Antillean manatees Trichechus manatus manatus that were stranded in single events on the coastal beaches of Puerto Rico from August 2010 to August 2011. Three manatees were emaciated and the gastrointestinal tracts were devoid of digesta. Microscopically, all manatees had severe widespread inflammatory lesions of the gastrointestinal tract and heart with intralesional tachyzoites consistent with Toxoplasma gondii identified by histological, ultrastructural and immunohistochemical techniques. The gastrointestinal lesions included severe, multifocal to diffuse, chronic-active enteritis, colitis and/or gastritis often with associated ulceration, necrosis and hemorrhage. Enteric leiomyositis was severe and locally extensive in all cases and associated with the most frequently observed intralesional protozoans. Moderate to severe, multifocal, chronic to chronic-active, necrotizing myocarditis was also present in all cases. Additionally, less consistent inflammatory lesions occurred in the liver, lung and a mesenteric lymph node and were associated with fewer tachyzoites. Sera (n = 30) collected from free-ranging and captive Puerto Rican manatees and a rehabilitated/released Puerto Rican manatee from 2003 to 2012 were tested for antibodies for T. gondii. A positive T. gondii antibody titer was found in 2004 in 1 (3%) of the free-ranging cases tested. Disease caused by T. gondii is rare in manatees. This is the first report of toxoplasmosis in Antillean manatees from Puerto Rico. Additionally, these are the first reported cases of disseminated toxoplasmosis in any sirenian. The documentation of 4 cases of toxoplasmosis within one year and the extremely low seroprevalence to T. gondii suggest that toxoplasmosis may be an emerging disease in Antillean manatees from Puerto Rico.

Clinicopathologic findings from Atlantic bottlenose dolphins (Tursiops truncatus) with cytologic evidence of gastric inflammation.

As part of the Bottlenose Dolphin Health and Risk Assessment study, blood, gastric, fecal, and blowhole samples were collected from 114 Atlantic bottlenose dolphins (Tursiops truncatus) in the Indian River Lagoon, Florida (IRL), and from 73 dolphins from the estuarine waters around Charleston, South Carolina (CHS), to assess the presence and degree of cytologic evidence of gastric inflammation from 2003 to 2007. The prevalence of moderate and severe gastric inflammation was 9.6% in the IRL and 11.0% at CHS. A case-control study of 19 dolphins with cytologic evidence of gastric inflammation and 82 with normal cytology from the combined populations was conducted. Blood parameters evaluated included hematology, serum chemistry, serum protein electrophoresis, and stress hormones. Few differences of clinical or statistical significance were found between affected and unaffected dolphins. Serum norepinephrine and cortisol were significantly higher in cases compared to the controls, and aldosterone was marginally higher (P = 0.06) based on eight cases. None of the hematologic, serum chemistry, or serum electrophoresis results were significantly different. Gastric fluid pH was not significantly different between cases and controls. There were no clinically significant aerobic-anaerobic or fungal culture results from gastric contents; bacteria cultured from both groups were considered to represent normal flora. The prevalence of inflammation did not differ by gender. Historically, cytologic evidence of gastric inflammation has constituted a marker of systemic illness in dolphins; however, there was little evidence to indicate systemic illness among affected animals. The data obtained from this study provide a basis for further investigation and evaluation of gastric cytology in wild and managed bottlenose dolphins.

Menstrual cycle characteristics and reproductive hormone levels in women exposed to atrazine in drinking water.

Atrazine is the most commonly used herbicide in the U.S. and a wide-spread groundwater contaminant. Epidemiologic and laboratory evidence exists that atrazine disrupts reproductive health and hormone secretion. We examined the relationship between exposure to atrazine in drinking water and menstrual cycle function including reproductive hormone levels. Women 18-40 years old residing in agricultural communities where atrazine is used extensively (Illinois) and sparingly (Vermont) answered a questionnaire (n=102), maintained menstrual cycle diaries (n=67), and provided daily urine samples for analyses of luteinizing hormone (LH), and estradiol and progesterone metabolites (n=35). Markers of exposures included state of residence, atrazine and chlorotriazine concentrations in tap water, municipal water and urine, and estimated dose from water consumption. Women who lived in Illinois were more likely to report menstrual cycle length irregularity (odds ratio (OR)=4.69; 95% confidence interval (CI): 1.58-13.95) and more than 6 weeks between periods (OR=6.16; 95% CI: 1.29-29.38) than those who lived in Vermont. Consumption of >2 cups of unfiltered Illinois water daily was associated with increased risk of irregular periods (OR=5.73; 95% CI: 1.58-20.77). Estimated "dose" of atrazine and chlorotriazine from tap water was inversely related to mean mid-luteal estradiol metabolite. Atrazine "dose" from municipal concentrations was directly related to follicular phase length and inversely related to mean mid-luteal progesterone metabolite levels. We present preliminary evidence that atrazine exposure, at levels below the US EPA MCL, is associated with increased menstrual cycle irregularity, longer follicular phases, and decreased levels of menstrual cycle endocrine biomarkers of infertile ovulatory cycles.

Influences of biological variables and geographic location on circulating concentrations of thyroid hormones in wild bottlenose dolphins (Tursiops truncatus).

Thyroid hormones (TH) are key regulators of metabolism and development, yet our understanding of the variability in serum TH concentrations in free-ranging marine mammals is limited. Thus, we examined the interrelationships between TH and age, sex, reproductive status, geographic location, and ocean temperatures in wild bottlenose dolphins (Tursiops truncatus). Circulating concentrations of TH (total thyroxine (tT(4)), free T(4) (fT(4)), and total triiodothyronine (tT(3))) were determined in a total of 195 dolphins; 80 from the coastal waters of Charleston, South Carolina (CHS) and 115 from the Indian River Lagoon, Florida (IRL). Age had the most influence on circulating TH concentrations in dolphins at both sites with decreasing concentrations (p<0.0001) observed with increasing age for all TH. No significant differences were found between males and non-reproductive females. Geographic location significantly influenced tT(4) and tT(3) concentrations; CHS dolphins had higher concentrations than IRL animals. These TH differences between CHS and IRL dolphins may be attributed to the colder year-round water temperature that CHS dolphins inhabit compared to IRL dolphins and could constitute an adaptive response to their colder environment. Results from this study highlight the importance of establishing reference values for dolphins in different geographic locations to support valid comparisons. This initial assessment provides a foundation of how biological and environmental variables could affect circulating TH in dolphins, which will help to elucidate the impacts of disease, pollution, and climate change on the thyroid hormone system of aquatic mammals.

Animal sentinels for environmental and public health.

Studies of the effects of environmental exposures on domestic and wild animals can corroborate or inform epidemiologic studies in humans. Animals may be sensitive indicators of environmental hazards and provide an early warning system for public health intervention, as exemplified by the iconic canary in the coal mine. This article illustrates the application of animal sentinel research to elucidate the effects of exposure to traditional and emerging contaminants on human health. Focusing on environmental issues at the forefront of current public health research, the article describes exposures to community air pollution, environmental tobacco smoke, and pesticides and associations with cancer, reproductive outcomes, and infectious diseases. Finally, it covers the role of marine mammals in monitoring the health of the oceans and humans.

Clinicoimmunopathologic findings in Atlantic bottlenose dolphins Tursiops truncatus with positive cetacean morbillivirus antibody titers.

Sera from free-ranging Atlantic bottlenose dolphins Tursiops truncatus inhabiting the Indian River Lagoon (IRL), Florida were tested for antibodies to cetacean morbilliviruses from 2003 to 2007 as part of a multidisciplinary study of individual and population health. A suite of clinicoimmunopathologic variables were evaluated in morbillivirus-seropositive dolphins (n = 14) and seronegative healthy dolphins (n = 49). Several important differences were found. Serum alkaline phosphatase, creatine phosphokinase, chloride, albumin and albumin/globulin ratios were significantly lower in seropositive dolphins. Innate immunity appeared to be upregulated with significant increases in lysozyme concentration and marginally significant increases in monocytic phagocytosis. Adaptive immunity was also impacted in dolphins with positive morbillivirus antibody titers. Mitogen-induced T lymphocyte proliferation responses were significantly reduced in dolphins with positive morbillivirus antibody titers, and marginally significant decreases were found for absolute numbers of CD4+ lymphocytes. The findings suggest impairment of cell-mediated adaptive immunity, similar to the immunologic pattern reported with acute morbillivirus infection in other species. In contrast, dolphins with positive morbillivirus antibody titers appeared to have at least a partially upregulated humoral immune response with significantly higher levels of gamma globulins than healthy dolphins, which may represent an antibody response to morbillivirus infection or other pathogens. These data suggest that subclinical dolphin morbillivirus infection in IRL dolphins may produce clinicoimmunopathologic perturbations that impact overall health.

Associations between mercury and hepatic, renal, endocrine, and hematological parameters in Atlantic bottlenose dolphins (Tursiops truncatus) along the eastern coast of Florida and South Carolina.

We evaluated associations between total mercury (Hg) concentrations in blood and skin and endocrine, hepatic, renal, and hematological parameters in free-ranging bottlenose dolphins (Tursiops truncatus). Dolphins in Indian River Lagoon, FL had higher concentrations of Hg in blood (0.67 µg/l wet wt) and skin (7.24 µg/g dry wt) compared with those from Charleston Harbor, SC (0.15 µg/l wet wt, 1.68 µg/g dry wt). An inverse relationship was observed between blood and skin Hg concentrations and total thyroxine, triiodothyronine, absolute numbers of lymphocytes, eosinophils, and platelets. Adrenocorticotropic hormone (ACTH), blood urea nitrogen, and gamma-glutamyl transferase increased with increasing concentrations of Hg in blood and skin; lactate dehydrogenase and neutrophils increased with concentrations in skin only. Hemoglobin and mean corpuscular hemoglobin increased with increasing concentrations of Hg in blood. Selenium was negatively associated with free T4, progesterone, and absolute numbers of monocytes, and positively correlated with absolute numbers of eosinophils and lymphocytes, and mean corpuscular volume. The results suggest the potential for a deleterious effect of Hg in highly exposed dolphins.

Lacaziosis in bottlenose dolphins Tursiops truncatus along the coastal Atlantic Ocean, Florida, USA.

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 dolphins and humans. Previous studies have shown a high prevalence (6.8 to 12.0%) of lacaziosis in resident 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 collected 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 significantly 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.

Exposure to microcystin among coastal residents during a cyanobacteria bloom in Florida.

Florida has experienced multiple cyanobacteria blooms in recent years the most severe of which occurred in 2016 and 2018. Several toxins are produced by proliferating cyanobacteria, including the hepatotoxin microcystin (MC). Harmful algal blooms (HABs) caused by cyanobacteria have the potential to impact public health. However, to date there have been limited attempts to quantify exposure in human populations. This study investigated potential exposure to the cyanobacterial toxin, MC by measuring concentrations in swabs of the nasal mucosa. In addition, the relationships between nasal concentrations of MC, environmental concentrations and activity patterns were assessed. Participants (n = 125) were recruited in 2018 during a cyanobacterial bloom of Microcystis aeruginosa and completed a questionnaire which included location, type, and duration of recreational or occupational contact with impacted waterways within the last 10 days. Water samples were collected concurrently. A sterile swab was used to collect a sample from the nasal mucosa. Concentrations of MC were measured by ELISA. Of the 121 participants who provided nasal swabs, 115 (95.0 %) had concentrations of MC above the limit of detection with a mean concentration of 0.61 + 0.75 ppb. There were significant differences (p < 0.01) in mean MC concentration between individuals with direct contact with impacted waters (0.77 + 0.88 ppb) compared to those with no recent contact (0.37 + 0.49 ppb). Higher concentrations were observed among occupationally exposed individuals. Nasal concentrations of MC varied significantly over time and location of exposure to the bloom, concordant with concentrations in water samples. The results suggest that inhalation of aerosols may be an important pathway for exposure to MC. Nasal MC concentrations were generally highest during periods when concentrations in the surrounding waters peaked. Further research is needed to characterize the public health implications of exposure to cyanobacterial blooms.

Seasonal distributions and other risk factors for Giardia duodenalis and Cryptosporidium spp. infections in dogs and cats in Chiang Mai, Thailand.

The objectives of this study were to explore risk factors associated with Giardia and Cryptosporidium infections in dogs and cats in Chiang Mai, Thailand, to describe the seasonal distributions of Giardia and Cryptosporidium prevalence, and to determine the potential for zoonotic transmission through genetic characterization of isolates. Fecal samples from 301 dogs and 66 cats were collected between August 2009 and February 2010. The presence of Giardia cysts and Cryptosporidium oocysts was determined using zinc sulfate centrifugal flotation and immunofluorescent assay (IFA). Genotype/species were determined by DNA sequence analyses of PCR products from Giardia glutamate dehydrogenase (gdh), beta-giardin (bg), and triosephosphateisomerase (tpi) and Cryptosporidium heat shock protein 70KDa (hsp70) and small subunit-rRNA (SSU-rRNA) genes. Information related to specific risk factors was collected from owners of each animal using a questionnaire. The risk factor data were analyzed for associations with Giardia and Cryptosporidium infections using logistic regression. The overall estimated prevalence of Giardia and Cryptosporidium in dogs was 25.2% and 7.6%, respectively and in cats, 27.3% and 12.1%, respectively. The estimated prevalence of Giardia infection in dogs in the rainy season (31.7%) was significantly higher than in the drier, winter season (17.2%) (p < 0.01). The estimated prevalence of Cryptosporidium infection in dogs and of Giardia and Cryptosporidium infections in cats was not associated with season (p > 0.05). Multivariable analysis indicated that Giardia cysts were more likely to be detected in fecal samples of dogs that resided in high-density environments, drank untreated water, were shedding Cryptosporidium oocysts, were having acute diarrhea or a history of chronic diarrhea, and were collected in the rainy season. All 19 Giardia PCR positive samples typed as G. duodenalis canine adapted genotypes (assemblages C or D). In cats, of six Giardia PCR positive samples, five typed as dog assemblages and one typed as assemblage AI. Of ten dogs with Cryptosporidium PCR positive samples, eight typed as C. canis, one as C. parvum (a zoonotic species) and one had both C. canis and C. parvum. Of three Cryptosporidium PCR positive samples in cats, one typed as C. felis and two typed as C. parvum. The presence of zoonotic G. duodenalis assemblage AI in a cat, and C. parvum in feces of dogs and cats suggests a potential role for a reservoir for zoonotic transmission. Whether or not these presences were from exposure to other animal or human hosts or environment are needed to be confirmed.