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Abstract Knowledge of infectious diseases in wildlife provides important information for preventing potential outbreaks of zoonotic diseases. Adiaspiromycosis is a neglected human disease caused by dimorphic Onygenales fungi. The disease is produced by the inflammatory response against growing adiaspores, leading to granulomatous pneumonia. In humans, adiaspiromycosis is relevant in immunosuppressed patients. In animals, it is associated with pneumonia in fossorial species. Given the potential role of armadillos in the epidemiology of adiaspiromycosis, in this study, we sought to investigate the occurrence and pathological features of adiaspiromycosis in roadkilled armadillos. In total, 54 armadillo carcasses were suitable for postmortem pathologic examinations between February 2017 and 2020. Adiaspores, associated with granulomatous lesions, were observed in ten six-banded (Euphractus sexcinctus) and two southern naked-tailed armadillos (Cabassous unicinctus). A previously uncharacterized Onygenales species was molecularly identified in two E. sexcinctus. In summary, herein we report 12 cases of pulmonary adiaspiromycosis (PA) in two species of free-living armadillos in Brazil. Both, the morphology of the fungus, as well as the histopathological findings (granulomatous inflammatory response to adiaspores) are consistent with PA; however, as the molecular identification differs from the reported species, the potential impact of this fungus for human PA is unknown, and we cannot rule out its impact on public health.

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Polyomaviruses (PyVs) are small double-stranded DNA viruses able to infect species across all vertebrate taxa. In cetaceans, PyVs have been reported only in short-beaked common dolphin (Delphinus delphis), common bottlenose dolphin (Tursiops truncatus) and killer whale (Orcinus orca). Herein, we surveyed PyV in 119 cetaceans (29 mysticetes and 90 odontocetes) stranded along the Brazilian coast, from 2002 to 2022, comprising 18 species. DNA extracted from the lungs was tested using a nested PCR targeting the major capsid protein gene of PyV. Polyomavirus was detected in lung samples of 1.7% (2/119) cetaceans: two juvenile female Guiana dolphins (Sotalia guianensis) stranded in Rio de Janeiro (Rio de Janeiro state) and Guriri (Espírito Santo state), in 2018. Both retrieved sequences were identical and presented 93.3% amino acid identity with Zetapolyomavirus delphini, suggesting a novel species. On histopathology, one of the PyV-positive individuals presented basophilic intranuclear inclusion bodies morphologically consistent with polyomavirus in the lungs. Other available tissues from both cases were PyV-PCR-negative; however, both individuals tested positive for Guiana dolphin morbillivirus. To our knowledge, this is the first report of PyV infection in cetaceans of the Southern Hemisphere and the first description of a co-infection with morbillivirus.

Mycoplasma spp. are wall-less bacteria able to infect mammals and are classified as hemotropic (hemoplasma) and nonhemotropic. In aquatic mammals, hemoplasma have been reported in California sea lions (Zalophus californianus) and river dolphins (Inia spp.). We investigated Mycoplasma spp. in blood samples of West Indian manatees (Trichechus manatus), pinnipeds (5 species), and marine cetaceans (18 species) that stranded or were undergoing rehabilitation in Brazil during 2002–2022. We detected Mycoplasma in blood of 18/130 (14.8%) cetaceans and 3/18 (16.6%) pinnipeds. All tested manatees were PCR-negative for Mycoplasma. Our findings indicate that >2 different hemoplasma species are circulating in cetaceans. The sequences from pinnipeds were similar to previously described sequences. We also detected a nonhemotropic Mycoplasma in 2 Franciscana dolphins (Pontoporia blainvillei) that might be associated with microscopic lesions. Because certain hemoplasmas can cause disease and death in immunosuppressed mammals, the bacteria could have conservation implications for already endangered aquatic mammals.

Opossums are considered resistant to rabies. Nonhematophagous bats are reservoirs of rabies in urban areas of South America. We analyzed bats and opossums tested for rabies during 2021 in a highly urbanized city in Brazil to understand spillover in an urban setting. Wildlife surveillance is necessary to prevent rabies in humans and domestic animals.

A one-year-old female miniature goat was presented to an emergency service after calving a dead goatling. Physical and ultrasonographic examination revealed the presence of a viable fetus; therefore, the goat was submitted to an emergency cesarean section. In the postoperative period, the animal had septic peritonitis caused by Enterococcus faecium and Enterococcus casseliflavus. Both bacterial strains showed contrasting antimicrobial resistance profiles. Laparohysterectomy and abdominal cavity lavage were performed, but, once the animal had adhesions and necrotic lesions in abdominal organs, euthanasia was executed. A post-mortem examination revealed fibrino-necrotic septic peritonitis secondary to uterine rupture. To the authors’ knowledge, this is the first detailed report of polymicrobial septic peritonitis in a miniature goat and the first report of septic peritonitis caused by E. faecium and E. casseliflavus.

The giant anteater (Myrmecophaga tridactyla) is a vulnerable species in South America and is considered endangered or near extinction in Central America. Therefore, studies describing the reproductive characteristics of this species are pivotal for its conservation. Thus, this study aimed to provide a morphological description of the female reproductive tissues of this species. We collected tissue samples from six female giant anteaters and performed gross, morphological, and histochemical analyses. Five adult subjects and one juvenile were included in the study. In the ovary, classifications were made according to the follicle and oocyte sizes: primordial, primary, secondary, early antral, or antral. Typical follicles with a single oocyte surrounded by a simple or stratified layer of cubic epithelium, atretic follicles, corpora lutea, corpora albicans, and ovarian cysts were also observed. No ovarian lesions were observed. By contrast, endometritis, metritis, mucometra, and endometrial cysts were identified in the uterus. Uterine alterations in these subjects were frequent and could affect reproduction.

Abstract River dolphins are a highly threatened polyphyletic group comprised of four odontocete families: Iniidae, Pontoporiidae, Lipotidae, and Platanistidae, the first two endemic to South America. To address the knowledge gap regarding infectious agents in this cetacean group, we surveyed the presence of herpesviruses by PCR in skin and/or blood samples of live-captured Amazon (Inia geoffrensis, n = 25) and Bolivian (Inia boliviensis, n = 22) river dolphins of the Amazon basin and in selected tissue samples of franciscanas (Pontoporia blainvillei, n = 27) stranded or bycaught in southeastern Brazil. Additionally, available franciscana tissue samples were examined by histopathology. Herpesvirus DNA was amplified in 13 Bolivian river dolphins (59.1%, 95% CI 38.5–79.6%) and 14 franciscanas (51.9%, 95% CI 33.0–70.7%). All Amazon river dolphins were herpesvirus-negative. Two different herpesviruses were found in Bolivian river dolphins: a previously known gammaherpesvirus detected in blood and/or skin samples of all positive individuals and a novel alphaherpesvirus in the skin of one animal. A new gammaherpesvirus was found in several franciscana samples—the first herpesvirus recorded in Pontoporiidae. Intranuclear inclusion bodies consistent with herpesvirus were observed in the lymph node of one franciscana. The high divergence among the obtained herpesviruses and those previously described can be explained by viral-host coevolution, and by the fact that these populations are fairly isolated.

Antimicrobial resistance is an ancient natural phenomenon increasingly pressured by anthropogenic activities. Escherichia coli has been used as markers of environmental contamination and human-related activity. Seabirds may be bioindicators of clinically relevant bacterial pathogens and their antimicrobial resistance genes, including extended-spectrum-beta-lactamase (ESBL) and/or plasmid-encoded AmpC (pAmpC), in anthropized and remote areas. We evaluated cloacal swabs of 20 wild magnificent frigatebirds (Fregata magnificens) of the Alcatrazes Archipelago, the biggest breeding colony of magnificent frigatebirds in the southern Atlantic and a natural protected area with no history of human occupation, located in the anthropized southeastern Brazilian coast. We characterized a highly virulent multidrug-resistant ST648 (O153:H9) pandemic clone, harboring blaCTX–M–2, blaCMY–2, qnrB, tetB, sul1, sul2, aadA1, aac(3)-VIa and mdfA, and virulence genes characteristic of avian pathogenic (APEC) (hlyF, iroN, iss, iutA, and ompT) and other extraintestinal E. coli (ExPEC) (chuA, kpsMII, and papC). To our knowledge, this is the first report of ST648 E. coli co-producing ESBL and pAmpC in wild birds inhabiting insular environments. We suggest this potentially zoonotic and pathogenic lineage was likely acquired through indirect anthropogenic contamination of the marine environment, ingestion of contaminated seafood, or by intra and/or interspecific contact. Our findings reinforce the role of wild birds as anthropization sentinels in insular environments and the importance of wildlife surveillance studies on pathogens of critical priority classified by the World Health Organization.

After the sudden death of captive marmosets in São Paulo, Brazil, we conducted a histologic and microbiologic study. We found hyperacute septicemia caused by hypermucoviscous sequence type 86 K2 Klebsiella pneumoniae. We implemented prophylactic antimicrobial therapy, selected dedicated staff for marmoset interactions, and sanitized the animals’ fruit to successfully control this outbreak.

Abstract Background Penguin interaction with gillnets has been extensively reported in the Atlantic and Pacific Oceans, and is considered a major conservation threat. Among penguin species, Magellanic penguins (Spheniscus magellanicus) are currently considered of great concern, particularly in Brazil, where they are highly susceptible to gillnet bycatch. Nevertheless, information about drowning-associated microscopic findings in penguins is limited. Results We describe the anatomopathological findings of 20 Magellanic penguins that drowned after getting entangled in a drift gillnet while wintering along the Brazilian shelf and washed ashore still enmeshed in Santa Catarina, Brazil. All 20 birds (19 juveniles and 1 adult; 18 females and 2 males) were in good body condition. Major gross findings were abrasion, bruising, and local erythema and edema of the wings, multiorgan congestion, jugular vein engorgement, pulmonary edema and hemorrhage, splenomegaly and hepatomegaly, fluid in the trachea, serous bloody fluid in the lungs, gastrointestinal parasites (nematodes, cestodes and trematodes), and debris in the stomach. The most common histopathological findings were cerebral and pulmonary congestion, pulmonary edema, splenic histiocytosis, lymphoid splenic hyperplasia, acute splenitis, extramedullary hepatic hematopoiesis, and parasitic enteritis. Although unspecific, the observed multiorgan congestion and pulmonary edema are consistent with previous reports of drowning in birds and may be indicative of this process. Conclusions Drowning may be a challenging diagnosis (e.g., carcass decomposition, predation), but must be considered as a differential in all beach-cast seabird postmortem examinations. To the authors’ knowledge this is the largest anatomopathological study based on microscopic examination in drowned penguins.

Antimicrobial resistance genes (ARGs) are environmental pollutants and anthropization indicators. We evaluated human interference in the marine ecosystem through the ocurrence and quantification (real-time PCRs) of 21 plasmid-mediated ARGs in enema samples of 25 wild seabirds, upon admission into rehabilitation: kelp gull (Larus dominicanus, n = 14) and Magellanic penguin (Spheniscus magellanicus, n = 11). Overall, higher resistance values were observed in kelp gulls (non-migratory coastal synanthropic) in comparison with Magellanic penguins (migratory pelagic non-synanthropic). There were significant differences between species (respectively, kelp gull and Magellanic penguin): ARGs occurrence (blaTEM [p = 0.032]; tetM [p = 0.015]; tetA [p = 0.003]; and sulII [p = 0.007]), mean number of ARGs per sample (p = 0.031), ARGs mean load percentage (aadA [p = 0.045], tetA [p = 0.031], tetM [p = 0.016], blaTEM [p = 0.032], sulII [p = 0.008]), percentage of genes conferring resistance to an antimicrobial class (betalactams [p = 0.036] and sulfonamides [p = 0.033]), mean number of genes conferring resistance to one or more antimicrobial classes (p = 0.024]), percentage of multiresistant microbiomes (p = 0.032), and clustering (p = 0.006). These differences are likely due to these species' contrasting biology and ecology - key factors in the epidemiology of ARGs in seabirds. Additionally, this is the first report of mecA in seabirds in the Americas. Further studies are necessary to clarify the occurrence and diversity of ARGs in seabirds, and their role as potential sources of infection and dispersal within the One Health chain of ARGs.

Knowledge of reproductive health in wild southern tamanduas (Tamandua tetradactyla; Mammalia: Myrmecophagidae) is fragmentary. During necropsies of roadkill xenarthran species in Brazil, a case of ovarian filariasis in an adult female southern tamandua was observed. Macroscopically, both ovaries were irregularly enlarged and had numerous smooth protuberances. Histologically, the affected ovarian parenchyma presented adult nematodes (including females with microfilaria) surrounded by pleocellular inflammatory infiltrates. The morphological characteristics of the nematodes were consistent with the superfamily Filarioidea (order Spirurida). The adjacent ovarian parenchyma had developing and atretic follicles at different stages of maturation. Filarial nematodes were not observed in other tissues. The cause of death of this tamandua was fatal acute polytrauma as a consequence of the motor vehicle collision. This case adds to a prior report of ovarian filariasis in two southern tamanduas in Nicaragua and Guatemala, dating back almost 100 years, and suggests filarial infections could potentially have an impact on reproductive success in southern tamanduas and possibly other xenarthrans. Several xenarthran species are under different levels of threat and knowledge of their basic reproductive health is crucial for conservation programs.

Osteitis deformans (Paget’s disease) is a chronic bone disorder characterized by excessive osteoclast-mediated bone resorption followed by new bone formation. The present paper reports this condition in an 18-year-old captive golden lancehead (Bothrops insularis) from Brazil. This patient initially exhibited anorexia and swelling in the middle third of the spine associated with locomotor disability. For diagnosis, radiography, ultrasound, computed tomography, cytology, and microbiological culture were performed. Diagnostic imaging showed bone changes, vertebral fusion, and bone proliferation. Cytology revealed blood cells how toxic heterophiles, reactive monocytes, young red blood cells, and polychromasia compatible with an infectious process. A bacterial culture identified an ampicillin-susceptible strain of Enterococcus faecalis. Antibiotic treatment was promptly started, but the snake died 25 days later. Histopathologically, the bone tissue showed a generalized thickening of the vertebral trabeculae. For the first time, the presence of E. faecalis associated with the development of osteitis deformans in snakes was presented.

Abstract Avian malaria is one of the most important diseases of captive penguins. We employed morphometric techniques to evaluate hepatic hemosiderosis in rehabilitating wild Magellanic penguins (Spheniscus magellanicus) that were negative (n = 9) or naturally infected by different subgenera of Plasmodium spp. (n = 24), according with: Plasmodium subgenera (Haemamoeba, Huffia, Other lineages, and Unidentified lineages), severity of Plasmodium histopathological lesions, and concurrent diseases, age class (juvenile or adult plumage), sex (male, female or not determined), body score (emaciated, thin, good, excellent, not available), molt, presence or absence of oil contamination upon admission, iron supplementation, and rehabilitation center. The percentage of the area occupied by hemosiderin was called ‘Index of Hepatic Hemosiderosis (IHH)’. Plasmodium-positive females presented significantly higher IHH values (17.53 ± 12.95%) than males (7.20 ± 4.25%; p = 0.041). We observed higher levels of congestion (p = 0.0182) and pneumonia (p = 0.0250) severity between Unidentified lineages vs. Huffia. We believe that the hepatic hemosiderosis observed in this study was multifactorial, the result of pathological processes caused by malaria, molting, hemoglobin and myoglobin catabolism during migration, anemia, concomitant diseases, and iron supplementation, all possibly potentiated by decreased liver mass. Further studies are needed to clarify the mechanisms of these hypotheses.

Abstract Background Tuberculosis is widely known as a progressive disease that affects endothermic animals, leading to death and/or economical losses, while mycobacterial infections in amphibians are commonly due to nontuberculous mycobacteria. To the authors’ knowledge, this report describes the first case of bovine tuberculosis in a poikilothermic animal. Case presentation An adult female captive American bullfrog (Lithobates catesbeianus Shaw, 1802) died in a Brazilian aquarium. Multiple granulomas with acid-fast bacilli were observed in several organs. Identification of Mycobacterium bovis was accomplished by culture and PCR methods. The other animals from the same enclosure were euthanized, but no evidence of mycobacterial infection was observed. Conclusions The American bullfrog was introduced in several countries around the world as an alternative husbandry, and its production is purposed for zoological and aquarium collections, biomedical research, education, human consumption and pet market. The present report warns about an episode of bovine tuberculosis in an amphibian, therefore further studies are necessary to define this frog species’ role in the epidemiology of M. bovis.

In January 2017, a yellow fever outbreak occurred in Espirito Santo, Brazil, where human immunization coverage is low. Histologic, immunohistologic, and PCR examinations were performed for 22 deceased nonhuman New World primates; typical yellow fever features were found in 21. Diagnosis in nonhuman primates prompted early public health response.

Abstract Background Feline leukemia virus (FeLV) is an exogenous gammaretrovirus of domestic cats (Felis catus) and some wild felids. The outcomes of FeLV infection in domestic cats vary according to host susceptibility, virus strain, and infectious challenge dose. Jaguarundis (Puma yagouaroundi) are small wild felids from South and Central America. We previously reported on FeLV infections in jaguarundis. We hypothesized here that the outcomes of FeLV infection in P. yagouaroundi mimic those observed in domestic cats. The aim of this study was to investigate the population of jaguarundis at Fundação Parque Zoológico de São Paulo for natural FeLV infection and resulting outcomes. Methods We investigated the jaguarundis using serological and molecular methods and monitored them for FeLV-related diseases for 5 years. We retrieved relevant biological and clinical information for the entire population of 23 jaguarundis held at zoo. Post-mortem findings from necropsies were recorded and histopathological and immunohistopathological analyses were performed. Sequencing and phylogenetic analyses were performed for FeLV-positive samples. For sample prevalence, 95% confidence intervals (CI) were calculated. Fisher’s exact test was used to compare frequencies between infected and uninfected animals. P-values

Abstract Background The study of neoplasia in wildlife species contributes to the understanding of cancer biology, management practices, and comparative pathology. Higher frequencies of neoplasms among captive non-domestic felids have been reported most commonly in aging individuals. However, testicular tumours have rarely been reported. This report describes a metastatic testicular sex cord–stromal tumour leading to fatal haemorrhage and thrombosis in a captive African lion (Panthera leo). Case presentation During necropsy of a 16-year-old male African lion, the left testicle and spermatic cord were found to be intra-abdominal (cryptorchid), semi-hard and grossly enlarged with multiple pale-yellow masses. Encapsulated haemorrhage was present in the retroperitoneum around the kidneys. Neoplastic thrombosis was found at the renal veins opening into the caudal vena cava. Metastases were observed in the lungs and mediastinal lymph nodes. Histology revealed a poorly differentiated pleomorphic neoplasm comprised of round to polygonal cells and scattered spindle cells with eosinophilic cytoplasm. An immunohistochemistry panel of inhibin-α, Ki-67, human placental alkaline phosphatase, cytokeratin AE1/AE3, cKit, vimentin and S100 was conducted. Positive cytoplasmic immunolabeling was obtained for vimentin and S100. Conclusions The gross, microscopic and immunohistochemical findings of the neoplasm were compatible with a poorly differentiated pleomorphic sex cord–stromal tumour. Cause of death was hypovolemic shock from extensive retroperitoneal haemorrhage and neoplastic thrombosis may have contributed to the fatal outcome. To our knowledge, this is the first report of sex cord–stromal tumour in non-domestic felids.

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, TGFβ, 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.

Non-human primates (NHPs) have been shown to be infected by parasites of the genus Plasmodium, the etiological agent of malaria in humans, creating potential risks of zoonotic transmission. Plasmodium brasilianum, a parasite species similar to P. malariae of humans, have been described in NHPs from Central and South America, including Brazil. The merozoite surface protein 1 (MSP1), besides being a malaria vaccine candidate, is highly immunogenic. Due to such properties, we tested this protein for the diagnosis of parasite infection. We used recombinant proteins of P. malariae MSP1, as well as of P. falciparum and P. vivax, for the detection of antibodies anti-MSP1 of these parasite species, in the sera of NHPs collected in different regions of Brazil. About 40% of the NHP sera were confirmed as reactive to the proteins of one or more parasite species. A relatively higher number of reactive sera was found in animals from the Atlantic Forest than those from the Amazon region, possibly reflecting the former more intense parasite circulation among NHPs due to their proximity to humans at a higher populational density. The presence of Plasmodium positive NHPs in the surveyed areas, being therefore potential parasite reservoirs, needs to be considered in any malaria surveillance program.

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.

Blood parasites are potential threats to the health of penguins and to their conservation and management. Little penguins Eudyptula minor are native to Australia and New Zealand, and are susceptible to piroplasmids (Babesia), hemosporidians (Haemoproteus, Leucocytozoon, Plasmodium) and kinetoplastids (Trypanosoma). We studied a total of 263 wild little penguins at 20 sites along the Australian southeastern coast, in addition to 16 captive-bred little penguins. Babesia sp. was identified in seven wild little penguins, with positive individuals recorded in New South Wales, Victoria and Tasmania. True prevalence was estimated between 3.4% and 4.5%. Only round forms of the parasite were observed, and gene sequencing confirmed the identity of the parasite and demonstrated it is closely related to Babesia poelea from boobies (Sula spp.) and B. uriae from murres (Uria aalge). None of the Babesia-positive penguins presented signs of disease, confirming earlier suggestions that chronic infections by these parasites are not substantially problematic to otherwise healthy little penguins. We searched also for kinetoplastids, and despite targeted sampling of little penguins near the location where Trypanosoma eudyptulae was originally reported, this parasite was not detected.

This study describes the pathologic findings of 24 humpback whales (Megaptera novaeangliae) found stranded along the Brazilian coast from 2004 to 2016. Eighteen (75%) animals evaluated were found stranded alive. From these, 13 died naturally on shore and five were euthanized. Six died at sea and were washed ashore. Of the 24, 19 (79.2%) were calves, four (16.7%) were juveniles, and one (4.2%) was an adult. The most probable cause of stranding and/or death (CSD) was determined in 23/24 (95.8%) individuals. In calves, CSD included neonatal respiratory distress (13/19; 68.4%), infectious disease (septicemia, omphaloarteritis and urachocystitis; 3/19; 15.8%), trauma of unknown origin (2/19; 10.5%), and vehicular trauma (vessel strike; 1/19; 5.3%). In juveniles and adult individuals, CSD was: emaciation (2/5; 40%), sunlight-thermal burn shock (1/5; 20%); and discospondylitis (1/5; 20%). In one juvenile, the CSD was undetermined (1/5; 20%). This study integrates novel findings and published case reports to delineate the pathology of a South-western Atlantic population of humpback whales. This foundation will aid in the assessment of the population health and establish a baseline for development of conservation policies.

Babesia spp. are tick-borne parasites, and 16 avian-infecting species have been described to date, including one species (Babesia peircei) that infects penguins. Considering the results of a recent study reporting Babesia sp. in penguins on Deception Island, South Shetland Islands, we re-examined the samples obtained in a previous investigation on the occurrence of blood parasites in adult Adélie (Pygoscelis adeliae), chinstrap (Pygoscelis antarcticus) and gentoo penguins (Pygoscelis papua) on King George and Elephant islands, South Shetland Islands. Notwithstanding a comprehensive re-examination of the blood smears, Babesia sp. was not detected. When we employed two nested PCR tests targeting the 18S rRNA gene of Babesia, a considerable proportion of the samples produced positive results; however, gene sequencing revealed these were due to cross-amplification of non-target organisms. We therefore did not detect Babesia sp. infection in penguins on King George and Elephant islands. Additional studies will be valuable to clarify the distribution and epidemiology of tick-borne pathogens in sub-Antarctic and Antarctic seabirds.

This retrospective study describes the biological and epidemiological aspects, gross and microscopical findings, and most likely causes of death (CD) in two species of Neotropical deer in Brazil. The animals were collected between 1995 and 2015 and represented 75 marsh deer (MD) and 136 brown brocket deer (BBD). Summarized, pneumonia was diagnosed microscopically in 48 MD and 52 BBD; 76 deer suffered trauma, involving dog attack (14 BBD) and vehicle-collision (14 BBD). Pulmonary edema (50 MD; 55 BBD) and congestion (57 MD; 78 BBD) were the most common findings for both species. Additionally, we diagnosed ruminal and myocardial mycosis in MD and BBD, respectively; ovarian dysgerminoma and pancreatic trematodiasis in BBD; and lesions suggestive of malignant catarrhal fever and orbiviral hemorrhagic disease in both species. The main CD in MD was: respiratory (41/75), alimentary, nutritional, trauma and euthanasia (3/75 each). Correspondingly, in BBD were: trauma (34/131), respiratory (30/131) and euthanasia (9/131). Respiratory disease was often defined by pulmonary edema and pneumonia. We provide evidence that respiratory disease, mainly pneumonia, is a critical pathological process in these Neotropical deer species. Although no etiological agents were identified, there is evidence of bacterial and viral involvement. Our results show trauma, mainly anthropogenic, as a common ailment in BBD. We propose to prioritize respiratory disease in future research focused on South American deer health aspects. We believe anthropogenic trauma may be a primary threat for populations of BBD.

Recent studies have discussed the importance of snakes, going beyond the context of Public Health (snakebites). Promising research demonstrates the utility of snake venom in several fields, including oncology, diagnosis, anticoagulant therapy, and pain treatment. Nevertheless, there are still gaps in the basic knowledge on these animals, specifically regarding the histological characterization of the gastrointestinal tract, which ultimately hinder comparative histology and pathology studies. In this context the present study intends to contribute with the advance of current knowledge on snake histology by describing and analyzing histological samples of the gastrointestinal tracts of Bothrops jararaca and Crotalus durissus. Samples were collected from 12 individuals (six from each species), up to 6 months of age and equally distributed between males and females. Histological slides were prepared from the organs collected and were stained with hematoxylin and eosin, periodic acid-Schiff and alcian blue. The slides were photographed with a high-resolution camera to create a portfolio representative of the histology of the gastrointestinal systems of these animals.

Amphibians are potentially reliable and efficient bioindicators. Existing anuran white blood cell morphology studies are limited, with only a few morphometric studies available. We employed morphometric techniques to characterize leukocytes of selected Neotropical anurans from Brazil and compare our findings with the exotic American Bullfrog (Lithobates catesbeianus), genus Ranidae. We compared blood smears of 28 specimens from six different genera (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus, and Proceratophrys) with samples from L. catesbeianus. Leukocyte average diameter was calculated by an image analysis software. One-way analyses of variance and Bonferroni tests were used on statistical analyses. Lymphocytes, neutrophils, eosinophils, and basophils were significantly smaller than the reference ranges reported for other amphibian genera, including Lithobathes, whereas monocyte diameters did not differ significantly between genera. This is the first study to evaluate leukocyte morphometrics of Brazilian anuran species. Our findings suggest that geographical separation could possibly influence leukocyte morphometry.

Anisakiasis and Pseudoterranovosis are human diseases caused by the ingestion of live Anisakidae larvae in raw, undercooked or lightly marinated fish. Larvae were collected from one salted cod sold for human consumption in a Sao Paulo market in 2013. One section of one brownish larva was used for molecular analyses. The partial COX2 gene sequence from the larva had a nucleotide identity of 99.8 % with Pseudoterranova azarasi, which belongs to the Pseudoterranova decipiens species complex. The risk of allergy when consuming dead larvae in salted fish is not well known and should be considered.

Different herpesviruses have been associated with respiratory and enteric disease and mortality among seabirds and waterfowl. In 2011, a respiratory disease outbreak affected 58.3% (98/168) of the Magellanic penguins undergoing rehabilitation due to an oil spill off the southern Brazilian coast. Etiology was attributed to a novel herpesvirus identified by histopathology, immunohistochemistry, electron microscopy and molecular studies with partial DNA sequencing. Since migration, rehabilitation and translocation may facilitate the spread of pathogens between populations and trigger the onset of clinical disease in animals with latent infections, investigation of herpesvirus occurrence in asymptomatic seabirds was performed. Samples from free-ranging seabirds were collected in Argentinian Patagonia (Magellanic penguins) and the Abrolhos Archipelago in Brazil (Brown boobies, Masked boobies, Red-billed tropicbirds, White-tailed tropicbirds and South American tern). Furthermore, asymptomatic seabirds housed at the facility where the outbreak occurred were also sampled. In total, 354 samples from eight seabird species were analyzed by PCR for herpesvirus. Four different sequences of herpesviruses were identified, one in Yellow-nosed Albatross, one in Boobies and Tropicbirds and two in Magellanic penguins. Magellanic penguin herpesvirus 1 was identified during the penguin outbreak at the rehabilitation facility in Brazil, while Magellanic penguin herpesvirus 2 was recovered from free-ranging penguins at four reproduction sites in Argentina. Phylogenic analysis of the herpesviruses sequences tentatively identified suggested that the one found in Suliformes and the one associated with the outbreak are related to sequences of viruses that have previously caused seabird die-offs. These findings reinforce the necessity for seabird disease surveillance programs overall, and particularly highlight the importance of quarantine, good hygiene, stress management and pre-release health exams in seabirds undergoing rehabilitation.

The sound producing apparatus of the dwarf sperm whale (Kogia sima) presents a complex anatomic structure composed of melon, spermaceti, phonic lips, vocal cap, case, papillae, spermaceti chamber and other airspaces, as well as facial muscles involved in sound production. The spermaceti chamber rests on the caudal portion of the premaxilla, with part of its mucosa covered with spherical/oval-shaped structures (approximately 1 to 2 mm in diameter), compatible with vesicles (previously referred to as “papillae”). Macroscopical examination revealed whitish, firm, widely and irregularly distributed vesicular mucosa on the premaxillary portion of the spermaceti chamber of a K. sima specimen stranded on the coast of Santos (southeastern Brazilian coast). Upon microscopic examination, walls of connective tissue with abundant type I collagen forming vesicles with an internal space or cavity filled with a small amount of eosinophilic substance compatible with mucoproteic fluid were observed. The base of such vesicles presented glands within the connective tissue, probably responsible for fluid production. This study describes the histology of the mucosa of the spermaceti chamber of a K. sima specimen and characterizes the glands associated with fluid production.

The capture and quarantine of non-human primates could be necessary in some circumstances including those required under environmental permit. Mortality is undesirable for wildlife management programs and could be related to opportunistic pathogens, for example, deaths due to intestinal protozoa infection outbreaks as described here. Parasitological, necroscopic, microbiological, and molecular tests were used in the diagnosis of severe necrotic enteritis leading to death of three female and two male black howler monkeys (Alouatta caraya) captured and held in quarantine prior to translocation. Parasitological tests showed the presence of cysts of Entamoeba histolytica/dispar (5/5), Entamoeba coli (5/5) and Giardia duodenalis (1/5). Necroscopic assessment revealed areas of severe multifocal necrosis in the intestinal mucosa and submucosa. Histopathological examination revealed the presence of structures morphologically compatible to Entamoeba spp. in all individuals examined. Furthermore, G. duodenalis (1/5) was demonstrated by the nested PCR technique. During temporary captivity of non-human primates in management programs, proper handling protocols, including fast or immediate destination, are suggested in order to mitigate the negative effects of stress and decrease the risk of infections.

The Indirect Fluorescence Assay (IFA) and the indirect ELISA were comparatively used to detect IgG and IgM antibodies for Toxoplasma gondii in experimentally and naturally infected primates. In the experimentally infected group, antibodies of diagnostic value were detected at day 9 post-infection (PI) with the IFA (IgG and IgM) and with IgG-ELISA. IgM-ELISA detected antibodies for T. gondii starting at day 3 PI until the end of the experiment (102 days PI). Of the 209 naturally infected sera tested, from many zoos of State of Sao Paulo, 64.59 and 67.94% were positive in the IgG-IFA test and IgG-ELISA respectively. IgM-ELISA test detected seropositivity in 52.63% of the sera although IgM-IFA test detected it in only in 0.96% of the samples. The differential toxoplasmosis diagnosis was accomplished with Neospora caninum by IFA, observing 61 (29.2%) seropositive animals for this parasite and 149 (70.8%) negative. Sixty animals were positive for both T. gondii and N. caninum. Pneumonia, splenomegaly, and intestinal ulcers were macroscopically observed. Unremarkable interstitial pneumonia, enteritis, colitis, splenitis, and glomerulitis were microscopically observed. The immunohistochemical stain could not detect the presence of T. gondii in the tissues of the animals infected experimentally.Detectou-se anticorpos das classes IgG e IgM anti-Toxoplasma gondii em primatas experimentalmente e naturalmente infectados, utilizando-se como técnicas comparativas a RIFI e o ELISA-teste. No grupo dos primatas experimentalmente infectados, anticorpos de valor diagnóstico foram detectados a partir do 9º dia de infecção tanto na RIFI (IgG e IgM) como no ELISA-IgG. O ELISA IgM detectou anticorpos a partir do 3º dia de infecção até o final do experimento (102 dias pós-infecção). Dos 209 soros dos primatas naturalmente infectados, de diversos zoológicos do Estado de São Paulo, 64,59 e 67,94% mostraram-se positivos na RIFI-IgG e no ELISA-IgG, respectivamente. O ELISA-IgM detectou soropositividade em 52,63% dos soros ao passo que a RIFI-IgM detectou apenas 0,96%. Foi realizado também diagnóstico diferencial para Neospora caninum, através da RIFI, observando-se 61 (29,2%) animais soropositivos para este parasita e 149 (70,8%) animais negativos. Sessenta animais foram positivos para T. gondii e N. caninum. Pneumonia, esplenomegalia e úlceras intestinais foram observadas macroscopicamente. Pneumonia intersticial, enterite, colite, esplenite e glomerulite foram os achados microscópicos. A imunoistoquímica não revelou a presença do T. gondii nos tecidos dos animais experimentalmente infectados.

Heartworm disease is caused by the intravascular nematode Dirofilaria immitis, a pathogen of public health importance usually associated to domestic dogs and cats, and to a lesser extend to other mammal species. The oncilla (Leopardus tigrinus) is a threatened neotropic felid species that naturally occurs in Brazil. Here, we report the encounter of adult and larval stages of heartworms in a female specimen of L. tigrinus, probable of free-ranging origin, from Ubatuba, São Paulo, Brazil, which died showing clinical signals compatible with heartworm disease. This was the first reported case of D. immitis infection and associated disease in L. tigrinus, also suggesting that the oncilla acted as a definitive host for this parasite. The present findings confirmed D. immitis as a pathogenic agent for this felid species, thus supporting the recommendation for the inclusion of diagnostic testing for this pathogen in routine health screening procedures for captive and free-ranging oncillas in Brazil, especially in those localities where climate conditions support the occurrence of the parasite. Potential reservoirs as oncillas are established beyond the reach of veterinary care, thus representing a continuing risk for domestic animals and humans acquiring heartworm infection. We encourage further serologic and molecular studies aiming to establish D. immitis prevalences in L. tigrinus and other wild carnivores in the region of Ubatuba, as well as ecological and veterinary studies to access the role of this pathogen for the survival of this threatened felid species.A doença do verme do coração é causada pelo nematódeo intravascular Dirofilaria immitis, um patógeno de importância em Saúde Pública geralmente associado a cães e gatos domésticos e, em menor extensão, a outras espécies de mamíferos. O gato-do-mato-pequeno (Leopardus tigrinus) é uma espécie ameaçada de felídeo neotropical que ocorre naturalmente no Brasil. Aqui relatamos o encontro de estágios adultos e larvais de vermes do coração em uma fêmea de L. tigrinus, provavelmente de vida livre e originária de Ubatuba, São Paulo, Brasil, que veio a óbito demonstrando sinais clínicos compatíveis desta doença. Este é o primeiro caso relatado de infecção e doença associada com D. immitis em L. tigrinus, sugerindo também que o gato-do-mato-pequeno agiu como hospedeiro definitivo para este parasita. Estes achados confirmaram que D. immitis é um agente patogênico para esta espécie de felino, sugerindo a recomendação para a inclusão de testes diagnósticos para este patógeno em procedimentos rotineiros de avaliação da saúde para gatos-do-mato-pequenos mantidos em cativeiro e de vida livre no Brasil, especialmente em localidades onde as condições climáticas permitam a ocorrência do parasita. Os gatos-do-mato-pequenos são potenciais reservatórios da D. immitis, mas vivem fora do alcance veterinário, representando desta forma um risco contínuo para animais domésticos e humanos. Encorajamos a realização de outros estudos epidemiológicos a fim de estabelecer as prevalências de infecção por D. immitis em L. tigrinus e outros carnívoros selvagens na região de Ubatuba, assim como estudos ecológicos e médicos veterinários para se acessar o papel deste patógeno para a sobrevivência desta espécie ameaçada de felídeo.

Seabirds may be responsible for the spread of pathogenic/resistant organisms over great distances, playing a relevant role within the context of the One World, One Health concept. Diarrheagenic E. coli strains, known as STEC (shiga toxin-producing E. coli), and the extraintestinal pathogenic E. coli (ExPEC and the subpathotype APEC), are among the E. coli pathotypes with zoonotic potential associated with the birds. In order to identify health threats carried by frigates and to evaluate the anthropic influence on the southern coast of Brazil, the aim of this work was to characterize E. coli isolated from free-ranging frigates in relation to virulence genotypes, serotypes, phylogenetic groups and antimicrobial resistance. Cloacal and choanal swabs were sampled from 38 Fregata magnificens from two oceanic islands and one rescue center. Forty-three E. coli strains were recovered from 33 out of the 38 birds (86.8%); 88.4% of strains showed some of the virulence genes (VGs) searched, 48.8% contained three or more VGs. None of the strains presented VGs related to EPEC/STEC. Some of the isolates showed virulence genotypes, phylogenetic groups and serotypes of classical human ExPEC or APEC (O2:H7, O1:H6, ONT:H7, O25:H4). Regarding antimicrobial susceptibility, 62.8% showed resistance, and 11.6% (5/43) were multidrug-resistant. The E. coli present in the intestines of the frigates may reflect the environmental human impact on southeast coast of Brazil; they may also represent an unexplored threat for seabird species, especially considering the overlap of pathogenic potential and antimicrobial resistance present in these strains.

Avian malaria is a mosquito-borne disease caused by Plasmodium spp. Avian plasmodia are recognized conservation-threatening pathogens due to their potential to cause severe epizootics when introduced to bird populations with which they did not co-evolve. Penguins are considered particularly susceptible, as outbreaks in captive populations will often lead to high morbidity and rapid mortality. We used a multidisciplinary approach to investigate an outbreak of avian malaria in 28 Magellanic penguins (Spheniscus magellanicus) at a rehabilitation center during summer 2009 in Florianópolis, Brazil. Hemosporidian infections were identified by microscopic and molecular characterization in 64% (18/28) of the penguins, including Plasmodium (Haemamoeba) tejerai, Plasmodium (Huffia) elongatum, a Plasmodium (Haemamoeba) sp. lineage closely related to Plasmodium cathemerium, and a Haemoproteus (Parahaemoproteus) sp. lineage closely related to Haemoproteus syrnii. P. tejerai played a predominant role in the studied outbreak and was identified in 72% (13/18) of the hemosporidian-infected penguins, and in 89% (8/9) of the penguins that died, suggesting that this is a highly pathogenic parasite for penguins; a detailed description of tissue meronts and lesions is provided. Mixed infections were identified in three penguins, and involved P. elongatum and either P. tejerai or P. (Haemamoeba) sp. that were compatible with P. tejerai but could not be confirmed. In total, 32% (9/28) penguins died over the course of 16 days despite oral treatment with chloroquine followed by sulfadiazine-trimethoprim. Hemosporidian infections were considered likely to have occurred during rehabilitation, probably from mosquitoes infected while feeding on local native birds, whereas penguin-mosquito-penguin transmission may have played a role in later stages of the outbreak. Considering the seasonality of the infection, rehabilitation centers would benefit from narrowing their efforts to prevent avian malaria outbreaks to the penguins that are maintained throughout summer.

O presente trabalho teve por objetivo investigar a modulação exercida pela temperatura ambiental sobre a cinética celular inflamatória experimentalmente induzida em Rana catesbeiana, rã-touro gigante. Para tanto, 120 espécimes pós-metamórficos foram mantidos a 6ºC e 24ºC e tratados pela transfixação do tecido muscular da coxa por fio de sutura ou injeção intramuscular de carragenina. Os resultados obtidos através de avaliações qualitativa e quantitativa do foco lesional mostraram que a baixa temperatura ambiental modula significativamente a evolução do processo inflamatório. Animais mantidos a 6ºC, em ambos os modelos, apresentaram números de células inflamatórias significativamente menores que os verificados a 24ºC, independentemente do tempo de avaliação. Por outro lado, algum fator pertencente aos mecanismos de defesa do hospedeiro não deve ter sido bloqueado pela temperatura, visto que a área de reação à injúria mostrou-se equivalente na maioria dos tempos pesquisados.

É relatado um caso de toxoplasmose ocorrido em sagüi-de-bigode (Saguinus imperator). O animal foi encontrado morto sem que tenham sido observados sinais clínicos. Achados de necropsia e histopatológicos incluíram pneumonia intersticial moderada, hepatite necrótica multifocal e linfadenite necrosante multifocal a coalescente. A técnica imunoistoquímica (strepto-avidina-biotina peroxidase) realizada em pulmão, fígado, linfonodos, baço, coração, intestino e tecido adiposo foi fortemente positiva para Toxoplasma gondii.

Abstract New World Nonhuman Primates are highly susceptible to clinical toxoplasmosis. Serum samples from 126 recently captured Leontopithecus chrysomelas, from an exotic and invasive population, were tested for Toxoplasma gondii antibodies by the modified agglutination test (MAT, cut-off 1:25); all were seronegative. The MAT is highly specific and is not species-specific. This is the first report of T. gondii survey in this tamarin in the wild. This result is consistent with prior reports that showed the high susceptibility of the species to infection by T. gondii usually with high mortality rates.

Abstract Toxoplasmosis is a parasitic disease caused by the protozoan Toxoplasma gondii. In cetaceans, T. gondii infection is a significant cause of morbidity and mortality. Despite the worldwide range and broad cetacean host record of T. gondii infection, there is limited information on toxoplasmosis in cetaceans from the Southern hemisphere. We investigated the occurrence of T. gondii by histopathology and immunohistochemistry in tissue samples of 185 animals comprising 20 different cetacean species from Brazil. Three out of 185 (1.6%) animals presented T. gondii-associated lesions: a captive killer whale Orcinus orca, a free-ranging common bottlenose dolphin Tursiops truncatus and a free-ranging Guiana dolphin Sotalia guianensis. The main lesions observed in these animals were necrotizing hepatitis, adrenalitis and lymphadenitis associated with protozoal cysts or extracellular tachyzoites presenting immunolabeling with anti-T. gondii antibodies. This study widens the spectrum of species and the geographic range of this agent in Brazil, and provides the first reports of T. gondii infection in a captive killer whale and in a free-ranging common bottlenose dolphin in South America.

Abstract A juvenile subantarctic fur seal (Arctocephalus tropicalis) found dead in Santa Catarina state, southern Brazil, presented with disseminated verminous pneumonia due to Parafilaroides sp. A concomitant infection with two different gammaherpesviruses was identified by PCR in different tissues; one of them possibly a novel species (tentatively named Otariid herpesvirus 7). Sarcocystis sp. DNA was identified molecularly in skeletal muscle samples with intrasarcoplasmic bradyzoites and no apparent tissue response. All analyzed samples (mandibular, laryngeal, tracheal, and mesenteric lymph nodes, and lung) were PCR-negative for Brucella spp. The most likely cause of death was severe pulmonary parafilaroidiasis. The pathogenic role of the gammaherpesviruses in several of the tissues was not evident. This study describes the pathogenicity of Parafilaroides sp. in a subantarctic fur seal, widens the host range of herpesvirus in pinnipeds, and reports the first molecular identification of Sarcocystis sp. in this species.