Sores of boreal moose reveal a previously unknown genetic lineage of parasitic nematode within the genus Onchocerca.

Long-standing reports of open sores on the hind legs of moose (Alces alces) have been recorded in Alaska (as well as Canada, Europe, and Michigan), eliciting concerns about causes and infection. We used histological and genomic methods to investigate the sores from 20 adult moose on the Kenai Peninsula, Alaska. We paired this with thermal imagery and molt scoring of adult moose to further describe sore formation and understand its timing. Severe, ulcerative and eosinophilic dermatitis was found in all moose with sores present, and microfilariae within intraepidermal pustules were additionally found in four samples. Genetic analysis of sores from moose revealed a previously unknown genetic lineage of Onchocerca. Adult moose molt and lose their barrier of protection against flies in June and July during peak fly activity, leaving them vulnerable and allowing the development of sores. In summary, our results indicate that the cause for the sores on the hindleg of moose is a previously unknown genetic lineage of Onchocerca, probably transmitted by black flies, in timing with the molt cycle of adult moose. These sores leave moose exposed to pathogens, making them vulnerable, and challenging their health and fitness.

CLINICO-PATHOLOGIC FINDINGS AND PATHOGEN SCREENING IN FUR SEALS (ARCTOCEPHALUS AUSTRALIS AND ARCTOCEPHALUS TROPICALIS) STRANDED IN SOUTHEASTERN BRAZIL, 2018.

Among pinnipeds, four Otariidae species (eared seals) have been reported as occasional or frequent vagrants along the coast of Brazil, mainly in the southern region. These animals usually arrive debilitated during winter and are directed to rehabilitation. Nevertheless, available information on sanitary aspects of stranded pinnipeds in Brazil is limited. Increased fur seal strandings (n=23) were recorded during the 2018 winter season in southeast Brazil (Iguape, Ilha Comprida, and Ilha do Cardoso, Sao Paulo State) compared to 2017 (n=2). Of these 23 fur seals, two were found dead and were in a good postmortem condition, and four died during rehabilitation and were subsequently necropsied. The remaining fur seals were not analyzed due to advanced decomposition (9/23) or successful rehabilitation (8/23). Herein, we report the antemortem hematology (n=4) and postmortem pathologic, parasitologic, and molecular analysis results as well as the most likely cause of stranding and/ or death (CSD) in five free-ranging juvenile South American fur seals (Arctocephalus australis) and one free-ranging juvenile subantarctic fur seal (Arctocephalus tropicalis). All animals were males, and all but one had poor body condition. Pathologic examinations revealed a variety of lesions, predominantly hemodynamic disturbances, endoparasitism, and inflammatory disease processes of suspected infectious nature. Molecular analyses detected gammaherpesvirus infections in two South American seals and one subantarctic fur seal, Sarcocystis sp. in one subantarctic fur seal, and Neospora spp. in two South American fur seals. All seals were PCR-negative for morbillivirus, flavivirus, and Toxoplasma gondii. The most likely CSDs were: starvation (2), aspiration pneumonia (1), asphyxia (1), predator attack (1), and presumed systemic infectious disease (1). These findings expand the geographic range of various pathogens of pinnipeds and may be of value to first responders, clinicians, and diagnosticians.

Crassicaudiasis in three geographically and chronologically distant Cuvier's beaked whales (Ziphius cavirostris) stranded off Brazil.

The Cuvier's beaked whale (CBW; Ziphius cavirostris) is a cosmopolitan marine mammal found in deep tropical and temperate waters of all oceans. CBW strandings have been recorded sporadically in Brazil; however, there is lack of information available regarding their causes of stranding and/or death. Herein, we report the epidemiologic, pathologic, morphologic parasitologic features and molecular identification of arterial and renal crassicaudiasis by Crassicauda sp. in three geographically and chronologically distant CBW stranded off Brazil. CBW-1 was an adult male stranded dead in Rio Grande do Sul State. CBW-2 was an adult female that stranded alive in Sergipe State and died shortly after. CBW-3 was and adult male that stranded dead in Santa Catarina State. The most relevant pathologic findings in these three CBW were severe, chronic proliferative mesenteric and caudal aortic endarteritis and chronic granulomatous and fibrosing interstitial nephritis with renicular atrophy and loss, and numerous intralesional Crassicauda sp. nematodes. Furthermore, CBW-1 had concomitant gram-negative bacterial pneumonia and pulmonary and hepatic thromboembolism. Morphologic analysis of renal adult nematodes identified Crassicauda sp. in the three CBW. Molecular analyses targeting the 18S and ITS-2 ribosomal loci of renal nematodes in CBW-2 and CBW-3 identified C. anthonyi. It is believed that severe arterial and renal crassicaudiasis likely resulted or contributed significantly to morbidity and death of these animals. These results expand the known geographical range of occurrence of crassicaudiasis in CBW. Specifically, the present study provides the first accounts of arterial and renal crassicaudiasis in CBW off the southern hemisphere, specifically in CBW off Brazil, and to the authors' knowledge, it is the first record of C. anthonyi in the southern Atlantic Ocean.

Retrospective Survey of Amphibian Pathology Cases at Texas A&M University System (2016-2020).

Dramatic declines in amphibians worldwide highlight the need for a better understanding of diseases affecting these species. To delineate the health issues of amphibians submitted to Texas A&M University System (2016-2020), the databases were queried on the basis of defined selection criteria. A total of 502 anurans (157 frogs [44 species] and 345 toads [10 species]) and 30 caudatans (23 salamanders [6 species] and 7 newts [4 species]) were reviewed. A most likely cause of death or major pathological finding (CD-MPF) leading to euthanasia was identified in 295 (55%) Anura cases and 15 (50%) Caudata cases. Of the 532 records reviewed, anurans included 492 captive, seven free-ranging and three undetermined specimens. All caudatans were captive. The most common CD-MPF in anurans was infectious/inflammatory (228/295; 77%), involving mycobacteriosis (73/228; 32%), chlamydiosis (44/228; 19%) and mycosis (32/228; 14%). Neoplasia was less common (28/295; 9%). Infectious/inflammatory lesions (14/15; 93%) were the main CD-MPFs in caudatans. Infectious diseases are a significant threat to captive amphibians in Texas and these results may aid personnel involved in amphibian conservation programmes, veterinarians and diagnosticians.

Cetacean morbillivirus in Humpback whales' exhaled breath.

The humpback whale (HW; Megaptera novaeangliae) population that seasonally resides along the Brazilian coast concentrates in the Abrolhos Bank (Bahia and Espírito Santo states) for breeding during austral winter and spring. Cetacean morbillivirus (CeMV, Paramyxoviridae family) is currently one of the most significant biological threats to cetaceans worldwide with high infection and mortality rates. CeMV is pleiotropic yet it has special tropism for the respiratory, lymphoid and nervous system and is primarily transmitted by the aerogenous route. A new lineage of CeMV, the Guiana dolphin morbillivirus (GDMV), is known to affect cetaceans off Brazil. GDMV was first detected in a Guiana dolphin (Sotalia guianensis) stranded in the Abrolhos Bank region, in 2010. In addition to pathologic examinations on stranded HW, pathogen survey of free-ranging HW may provide valuable insight into the epidemiology of diseases. We hypothesized that HW in the Brazilian breeding ground could be exposed to CeMV. Thus, in the present study, we investigated the presence of CeMV in exhaled breath condensates (EBC) of HW in the Abrolhos Bank. Overall, 73 samples of EBC from 48 groups of HW were collected during the breeding seasons of 2011 (n = 16) and 2012 (n = 57). One sample failed to have the reference gene amplified and was excluded from the study. CeMV was detected by a RT-qPCR method in 2 EBC samples, representing 2 whale groups. Phylogenetic analysis of partial morbillivirus phosphoprotein gene showed 100% homology to GDMV. Our results show that HW in Brazil are infected by CeMV with a relative prevalence of 4.3% (2/47) and demonstrate the suitability of using EBC and RT-qPCR as a non-invasive tool for CeMV survey in free-ranging whales. This pioneer study provides scientific basis for non-invasive CeMV monitoring of HW, suggests HW may play a role in the dynamics of CeMV and raises concern for potential conservation implications for this species.

Characterization of canine epidermal organoid cultures by immunohistochemical analysis and quantitative PCR.

BACKGROUND: Keratinocyte organoids can be used as a tool to evaluate epidermal structure, function and dysfunction. OBJECTIVES: To optimize the canine keratinocyte organoid system and produce organoids that are structurally equivalent to in vivo canine epidermis, in order to enable studies that focus on epidermal diseases and diseases resulting from an impaired epidermal barrier. ANIMALS: Skin biopsies were obtained from five recently euthanized dogs of different breeds with no skin abnormalities. METHODS AND MATERIALS: Cells derived from microdissected interfollicular epidermis were seeded in basement membrane extract and epidermal organoids were grown under different media conditions. Organoids were characterized to assess cell morphology and architecture in haematoxylin and eosin-stained slides and expression of selected epidermal markers (keratin 5, keratin 10, loricrin and filaggrin) by immunohistochemical analysis and quantitative reverse transcription PCR. RESULTS: The selected epidermal markers were expressed in the same epidermal layers in the organoids cultured in expansion medium and differentiation medium as in normal interfollicular epidermis, yet restriction to the distinct layers was best achieved with expansion medium. Comparison of the mRNA expression levels of these markers revealed that relative expression is similar in organoids cultured in expansion medium and normal canine epidermis, while it differs in organoids cultured in differentiation medium. CONCLUSION AND CLINICAL IMPORTANCE: Organoids cultured in expansion medium have an equivalent structure to the interfollicular epidermis and express key marker proteins in similar proportions. Epidermal organoids are therefore a promising in vitro model to study epidermal structure, function and dysfunction.

Cetacean Morbillivirus Infection in a Killer Whale (Orcinus orca) from Brazil.

We provide pathological, immunohistochemical and molecular evidence of cetacean morbillivirus (CeMV) infection in a live-stranded adult female killer whale (Orcinus orca), which stranded alive in Espírito Santo State, Brazil, in 2014. Although attempts were made to release the animal, it stranded again and died. The main pathological findings were severe pulmonary oedema, pleural petechiation, multifocal, lymphoplasmacytic meningoencephalitis and leptomeningomyelitis with perivascular cuffing and gliosis, chronic lymphocytic bronchointerstitial pneumonia and multicentric lymph node and splenic lymphoid depletion. Other pathological findings were associated with the 'live-stranding stress response'. Immunohistochemical analysis revealed multifocal morbilliviral antigen in neurons and astrocytes, and in pneumocytes, histiocytes and leukocytes in the lung. CeMV was detected by a novel reverse transcriptase polymerase chain reaction method in the brain and kidney. Phylogenetic analysis of part of the morbillivirus phosphoprotein gene indicates that the virus is similar to the Guiana dolphin (Sotalia guianensis) morbillivirus strain, known to affect cetaceans along the coast of Brazil. To the authors' knowledge, this is the first report of morbillivirus disease in killer whales.

Outbreak of Paranannizziopsis australasiensis Infection in Captive African Bush Vipers (Atheris squamigera).

We report the epidemiological, clinical and pathological features of an outbreak of Paranannizziopsis australasiensis (order Onygenales) in captive African bush vipers (Atheris squamigera) (ABVs) that died suddenly. The snakes had multifocal, raised, white-grey to dark brown discoloured cutaneous patches. Microscopically, all had integumentary lesions characterized by multifocal to coalescent necroheterophilic epidermitis with superficial and intraepidermal fungal elements and bacteria. Concurrent epidermal hyperplasia, hyperkeratosis and intracellular and intercellular oedema, often leading to vesiculation, and fasciitis/superficial myositis, were consistent findings in all snakes, while ulceration (9/11) and dysecdysis (5/11) varied. A panfungal polymerase chain reaction targeting the internal transcribed spacer-2 region, and gene sequencing, confirmed P. australasiensis infection in three cases. This is the first report of P. australasiensis in the USA and the first record of paranannizziopsis infection in African bush vipers. P. australasiensis should be considered in the differential diagnosis of dermatomycosis in snakes and represents a potential threat to reptile conservation programmes.

The Pathology of Cetacean Morbillivirus Infection and Comorbidities in Guiana Dolphins During an Unusual Mortality Event (Brazil, 2017-2018).

Cetacean morbillivirus (CeMV; Paramyxoviridae) is the most significant pathogen of cetaceans worldwide. The novel "multi-host" Guiana dolphin (Sotalia guianensis; GD)-CeMV strain is reported in South American waters and infects Guiana dolphins and southern right whales (Eubalaena australis). This study aimed to describe the pathologic findings, GD-CeMV viral antigen distribution and detection by RT-PCR (reverse transcriptase polymerase chain reaction), and infectious comorbidities in 29 Guiana dolphins that succumbed during an unusual mass-mortality event in Rio de Janeiro state, Brazil, between November 2017 and March 2018. The main gross findings were lack of ingesta, pulmonary edema, ascites, icterus, hepatic lipidosis, multicentric lymphadenomegaly, as well as pneumonia, polyserositis, and multiorgan vasculitis caused by Halocercus brasiliensis. Microscopically, the primary lesions were bronchointerstitial pneumonia and multicentric lymphoid depletion. The severity and extent of the lesions paralleled the distribution and intensity of morbilliviral antigen. For the first time in cetaceans, morbilliviral antigen was detected in salivary gland, optic nerve, heart, diaphragm, parietal and visceral epithelium of glomeruli, vulva, and thyroid gland. Viral antigen within circulating leukocytes suggested this as a mechanism of dissemination within the host. Comorbidities included disseminated toxoplasmosis, mycosis, ciliated protozoosis, and bacterial disease including brucellosis. These results provide strong evidence for GD-CeMV as the main cause of this unusual mass-mortality event.

A novel real-time PCR to detect Cetacean morbillivirus in Atlantic cetaceans.

Cetacean morbillivirus (CeMV, family Paramyxoviridae) is a re-emergent pathogen associated with severe epizootic outbreaks causing high mortality among cetaceans worldwide. Recently, CeMV caused an unusual mortality event of Guiana dolphins (Sotalia guianensis) in Brazil. Partial sequence of the viral phosphoprotein (P) gene showed that the Guiana dolphin morbillivirus (GDMV) might represent a new lineage of CeMV. This study aimed to develop a molecular technique to detect the most common CeMV strains known to circulate in the Atlantic Ocean: GDMV, Dolphin morbillivirus (DMV) and Pilot-whale morbillivirus (PWMV). A sensible real-time reverse transcription polymerase chain reaction (RT-qPCR) method based on intercalating dye, targeting the P gene was described. This assay successfully detected GDMV, PWMV and DMV from field samples. Its performance was compared to a RT-qPCR method that specifically detects GDMV. Both assays had high sensibility and excellent intra- and inter-assay reproducibility. A total of 109 field samples from 32 Guiana dolphins were screened for CeMV by conventional RT-PCR in parallel with the RT-qPCR assay. The detection rate increased from 32% to 60% by use of the novel RT-qPCR. The RT-qPCR assay described herein allows rapid and sensitive detection of Atlantic CeMV strains, and is potentially suitable for screening of CeMV globally.

Transcriptome Profiling and Differential Gene Expression in Canine Microdissected Anagen and Telogen Hair Follicles and Interfollicular Epidermis.

The transcriptome profile and differential gene expression in telogen and late anagen microdissected hair follicles and the interfollicular epidermis of healthy dogs was investigated by using RNAseq. The genes with the highest expression levels in each group were identified and genes known from studies in other species to be associated with structure and function of hair follicles and epidermis were evaluated. Transcriptome profiling revealed that late anagen follicles expressed mainly keratins and telogen follicles expressed GSN and KRT15. The interfollicular epidermis expressed predominately genes encoding for proteins associated with differentiation. All sample groups express genes encoding for proteins involved in cellular growth and signal transduction. The expression pattern of skin-associated genes in dogs is similar to humans. Differences in expression compared to mice and humans include BMP2 expression mainly in telogen and high KRT17 expression in the interfollicular epidermis of dogs. Our data provide the basis for the investigation of the structure and function of canine skin or skin disease and support the use of dogs as a model for human cutaneous disease by assigning gene expression to specific tissue states.

Pathologic findings and causes of death in southern right whales Eubalaena australis, Brazil.

Southern right whales Eubalaena australis (SRWs) migrate to southern Brazil for breeding and calving from June through November. Overall, there is scarce knowledge on health status and pathologic conditions in SRWs. We report the pathologic and molecular investigation results of 8 SRWs that were necropsied between 2010 and 2017 within a breeding and calving ground in Santa Catarina state, Brazil. The animals were of various ages (7 newborns/calves, 1 adult) and sex (3 females, 5 males). Five whales stranded dead; 3 stranded alive and died shortly after (n = 2) or were euthanized (n = 1). The causes of stranding and/or death were neonatal respiratory distress syndrome with meconium aspiration (n = 3) with concomitant congenital hepatopathy in one of them; trauma of unknown origin (n = 3), infectious renal and lung disease with presumed sepsis (n = 1), and euthanasia (n = 1). Three animals were PCR-positive for cetacean morbillivirus; one of them also had morbilliviral antigen in kidney via immunohistochemical analysis. These results, integrating novel findings and a published report, contribute to the pathology knowledge of this species.

Comparative Immunopathology of Cetacean morbillivirus Infection in Free-Ranging Dolphins From Western Mediterranean, Northeast-Central, and Southwestern Atlantic.

Cetacean morbillivirus (CeMV; Paramyxoviridae) causes epizootic and interepizootic fatalities in odontocetes and mysticetes worldwide. Studies suggest there is different species-specific susceptibility to CeMV infection, with striped dolphins (Stenella coeruleoalba), bottlenose dolphins (Tursiops truncatus), and Guiana dolphins (Sotalia guianensis) ranking among the most susceptible cetacean hosts. The pathogenesis of CeMV infection is not fully resolved. Since no previous studies have evaluated the organ-specific immunopathogenetic features of CeMV infection in tissues from infected dolphins, this study was aimed at characterizing and comparing immunophenotypic profiles of local immune responses in lymphoid organs (lymph nodes, spleen), lung and CNS in CeMV-molecularly (RT-PCR)-positive cetaceans from Western Mediterranean, Northeast-Central, and Southwestern Atlantic. Immunohistochemical (IHC) analyses targeted molecules of immunologic interest: caspase 3, CD3, CD20, CD57, CD68, FoxP3, MHCII, Iba1, IFNγ, IgG, IL4, IL10, lysozyme, TGFß, and PAX5. We detected consistent CeMV-associated inflammatory response patterns. Within CNS, inflammation was dominated by CD3+ (T cells), and CD20+ and PAX5+ (B cells) lymphocytes, accompanied by fewer Iba1+, CD68+, and lysozyme+ histiocytes, mainly in striped dolphins and bottlenose dolphins. Multicentric lymphoid depletion was characterized by reduced numbers of T cells and B cells, more pronounced in Guiana dolphins. Striped dolphins and bottlenose dolphins often had hyperplastic (regenerative) phenomena involving the aforementioned cell populations, particularly chronically infected animals. In the lung, there was mild to moderate increase in T cells, B cells, and histiocytes. Additionally, there was a generalized increased expression of caspase 3 in lymphoid, lung, and CNS tissues. Apoptosis, therefore, is believed to play a major role in generalized lymphoid depletion and likely overt immunosuppression during CeMV infection. No differences were detected regarding cytokine immunoreactivity in lymph nodes, spleen, and lung from infected and non-infected dolphins by semiquantitative analysis; however, there was striking immunoreactivity for IFNγ in the CNS of infected dolphins. These novel results set the basis for tissue-specific immunophenotypic responses during CeMV infection in three highly susceptible delphinid species. They also suggest a complex interplay between viral and host's immune factors, thereby contributing to gain valuable insights into similarities, and differences of CeMV infection's immunopathogenesis in relation to body tissues, CeMV strains, and cetacean hosts.

Molecular, serological, pathological, immunohistochemical and microbiological investigation of Brucella spp. in marine mammals of Brazil reveals new cetacean hosts.

Brucella-exposure and infection is increasingly recognized in marine mammals worldwide. To better understand the epidemiology and health impacts of Brucella spp. in marine mammals of Brazil, molecular (conventional PCR and/or real-time PCR), serological (Rose Bengal Test [RBT], Competitive [c]ELISA, Serum Agglutination Test [SAT]), pathological, immunohistochemical (IHC) and/or microbiological investigations were conducted in samples of 129 stranded or by-caught marine mammals (orders Cetartiodactyla [n = 124], Carnivora [n = 4] and Sirenia [n = 1]). Previous serological tests performed on available sera of 27 of the 129 animals (26 cetaceans and one manatee), indicated 10 seropositive cetaceans. Conventional PCR and/or real-time PCR performed in cases with available organs (n = 119) and/or blood or swabs (n = 10) revealed 4/129 (3.1%) Brucella-infected cetaceans (one of them with positive serology; the remaining three with no available sera). Pathological, IHC and/or microbiological analyses conducted in PCR/real-time PCR and/or seropositive cases (n = 13) revealed Brucella-type lesions, including meningitis/meningoencephalitis, pneumonia, necrotizing hepatitis, pericarditis and osteoarthritis in some of those animals, and positive IHC was found in all of them (excepting two live-stranded animals without available organs). Brucella spp. culture attempts were unsuccessful. Our results demonstrated exposure, asymptomatic, acute and chronic Brucella sp. infection in several cetacean species in the Brazilian coast, highlighting the role of this pathogen in stranding and/or death, particularly in Clymene dolphin (Stenella clymene) and short-finned pilot whale (Globicephala macrorhynchus) off Ceará State. Novel hosts susceptible to Brucella included the franciscana (Pontoporia blainvillei), the Guiana dolphin (Sotalia guianensis) and the spinner dolphin (Stenella longirostris). Additionally, three coinfection cases involving Brucella spp. and cetacean morbillivirus, Edwarsiella tarda and Proteus mirabilis were detected. To the best of our knowledge, this is the first long-term and large-scale survey of Brucella spp. in marine mammals of South America, widening the spectrum of susceptible hosts and geographical distribution range of this agent with zoonotic potential.

Comparative histopathologic and viral immunohistochemical studies on CeMV infection among Western Mediterranean, Northeast-Central, and Southwestern Atlantic cetaceans.

Cetacean morbillivirus (CeMV) is a major natural cause of morbidity and mortality in cetaceans worldwide and results in epidemic and endemic fatalities. The pathogenesis of CeMV has not been fully elucidated, and questions remain regarding tissue tropism and the mechanisms of immunosuppression. We compared the histopathologic and viral immunohistochemical features in molecularly confirmed CeMV-infected Guiana dolphins (Sotalia guianensis) from the Southwestern Atlantic (Brazil) and striped dolphins (Stenella coeruleoalba) and bottlenose dolphins (Tursiops truncatus) from the Northeast-Central Atlantic (Canary Islands, Spain) and the Western Mediterranean Sea (Italy). Major emphasis was placed on the central nervous system (CNS), including neuroanatomical distribution of lesions, and the lymphoid system and lung were also examined. Eleven Guiana dolphins, 13 striped dolphins, and 3 bottlenose dolphins were selected by defined criteria. CeMV infections showed a remarkable neurotropism in striped dolphins and bottlenose dolphins, while this was a rare feature in CeMV-infected Guiana dolphins. Neuroanatomical distribution of lesions in dolphins stranded in the Canary Islands revealed a consistent involvement of the cerebrum, thalamus, and cerebellum, followed by caudal brainstem and spinal cord. In most cases, Guiana dolphins had more severe lung lesions. The lymphoid system was involved in all three species, with consistent lymphoid depletion. Multinucleate giant cells/syncytia and characteristic viral inclusion bodies were variably observed in these organs. Overall, there was widespread lymphohistiocytic, epithelial, and neuronal/neuroglial viral antigen immunolabeling with some individual, host species, and CeMV strain differences. Preexisting and opportunistic infections were common, particularly endoparasitism, followed by bacterial, fungal, and viral infections. These results contribute to understanding CeMV infections in susceptible cetacean hosts in relation to factors such as CeMV strains and geographic locations, thereby establishing the basis for future neuro- and immunopathological comparative investigations.

Novel herpesviruses in riverine and marine cetaceans from South America.

Herpesvirus (HV) infections in cetaceans are frequently associated with skin and mucosal lesions. Although HV infections have been reported worldwide, their occurrence in southern Atlantic marine mammals is still poorly understood. We tested skin, oral and genital mucosal beta-actin PCR-positive samples from 109 free-ranging Brazilian cetaceans using a universal herpesvirus DNA polymerase PCR. Herpesvirus-positive skin samples from a Guiana dolphin (Sotalia guianensis), a dwarf sperm whale (Kogia sima), a Bolivian river dolphin (Inia boliviensis), and a lingual sample from an Atlantic spotted dolphin (Stenella frontalis) were histologically evaluated. Additional tissue samples from these animals were also PCR-positive for HV, including a novel sequence obtained from the dwarf sperm whale's stomach and mesenteric lymph node. Four novel HV species were detected in the Guiana dolphin (one), the dwarf sperm whale (two) and the Bolivian river dolphin (one). The cutaneous lesions (marked, focally extensive, chronic proliferative dermatitis) of the Guiana dolphin and the Bolivian river dolphin were similar to previous HV reports in cetaceans, despite the absence of intranuclear inclusion bodies. This is the largest HV survey in South American cetaceans and the first detection of HV infection in riverine dolphins worldwide.

Cetacean morbillivirus in Southern Right Whales, Brazil.

Cetacean morbillivirus (CeMV) has caused repeated epizootics and interepizootic fatalities in a variety of cetacean species worldwide. Recently, a novel CeMV strain (GD-CeMV) was linked to a mass die-off of Guiana dolphins (Sotalia guianensis) in Brazil. Southern right whales (SRWs; Eubalaena australis) migrate to the southern Brazilian coast during austral winter and spring (June through November) for breeding and calving. Because unexplained high calf mortality rates have recurrently been documented in SRWs, we hypothesized they could be infected with CeMV. We developed a novel real-time RT-PCR method based on SYBR® GREEN for detection of CeMV and identified the virus in three out of five stranded SRWs from Santa Catarina state, Brazil. The partial sequences of the morbillivirus phosphoprotein gene suggest that the virus is similar to the GD-CeMV strain. Our results indicate CeMV can infect SRWs and should be considered in the differential aetiologic diagnosis of infectious diseases in this species. It also raises concern for potential conservation implications for this species in its main coastal breeding area off Southern Brazil.

Survey of Brucella spp. and Leptospira spp. antibodies in cetaceans and manatees of the Amazon basin and Atlantic Ocean, Brazil.

Aquatic mammals can act as sentinels of emerging and resurging pathogens in the environment. Brucella spp. and Leptospira spp. are 2 zoonotic pathogens relevant to aquatic mammals, and their detection can be used to assess pathogen exposure. In this study, serum from 84 individuals-63 cetaceans (families Iniidae, n = 37; Delphinidae, n = 22; and Kogiidae, n = 4) and 21 West Indian manatees Trichechus manatus-was tested by the Rose Bengal Test (RBT) and a commercial competitive enzyme-linked immunosorbent assay (c-ELISA) for detecting Brucella spp. antibodies, and the microscopic agglutination test (MAT) for screening Leptospira spp. exposure. Overall, 4.8% (3/63) of cetaceans were positive by RBT and 15.9% (10/63) by c-ELISA for Brucella spp. Serum from 8 c-ELISA positive cetaceans (with available serum) was further tested via serum agglutination test (SAT) and 1 individual was positive. c-ELISA was more sensitive than RBT. Exposure to Brucella spp. was found in 5 cetacean species: Clymene dolphin Stenella clymene, short-finned pilot whale Globicephala macrorhynchus, pygmy killer whale Feresa attenuata, melon-headed whale Peponocephala electra and Atlantic bottlenose dolphin Tursiops truncatus in the Atlantic Ocean, Brazil, expanding the range of known Brucella seropositive aquatic hosts. No evidence of Brucella spp. exposure was found in Iniidae and Kogiidae odontocetes and manatees. Antibodies against Leptospira spp. were not detected in cetaceans and sirenians by MAT. These results contribute to the evaluation of different Brucella spp. serological methods in cetaceans and manatees and highlight the epidemiology of zoonotic pathogens in aquatic mammals of the southwestern Atlantic Ocean and the Amazon basin.

Guiana Dolphin Unusual Mortality Event and Link to Cetacean Morbillivirus, Brazil.

During November-December 2017, a mass die-off of Guiana dolphins (Sotalia guianensis) began in Rio de Janeiro, Brazil. Molecular and pathologic investigations on 20 animals indicated that cetacean morbillivirus played a major role. Our findings increase the knowledge on health and disease aspects of this endangered species.

Pulmonary and systemic fungal infections in an Atlantic spotted dolphin and a Bryde's whale, Brazil.

We report the gross and microscopic findings and molecular identification of 2 cases of hyphate fungal infection in cetaceans from Brazil. The first case involved an adult male Atlantic spotted dolphin Stenella frontalis with localized pulmonary disease characterized by pyogranulomatous and necrotizing bronchopneumonia with intralesional hyphae. The second case involved an adult male Bryde's whale Balaenoptera edeni with orchitis, periorchitis, mesenteric lymphadenitis and pyogranulomatous bronchopneumonia with intralesional hyphae. PCR analysis from the dolphin's lung yielded Aspergillus fumigatus, and the fungus from the whale's mesenteric lymph node showed the greatest identity to Nanniziopsis obscura and Stagonosporopsis cucurbitacearum These cases represent the first reports of pulmonary aspergillosis by A. fumigatus in an Atlantic spotted dolphin and systemic mycosis by a possibly novel Onygenales in marine mammals.