CHARACTERIZATION OF THE FUNGAL MICROBIOTA IN THE NOSTRILS AND RECTUM OF AMAZONIAN MANATEES (TRICHECHUS INUNGUIS) FROM A REHABILITATION PROGRAM IN BRAZIL.

The present study characterized the filamentous and yeast-like fungal microbiota of the nasal cavity and rectum of Amazonian manatees (Trichechus inunguis) undergoing rehabilitation at the Laboratory of Aquatic Mammals, National Institute of Amazonian Research, Manaus, Amazonas, and determined the antifungal susceptibility of these organisms. Nasal and rectal swabs were collected from 22 calves and three juveniles. The samples were seeded in Sabouraud agar supplemented with chloramphenicol 10%, incubated at 26°C, and observed daily for up to 7 d. The growth of different filamentous and yeast-like fungi was observed among the two anatomical sites. Filamentous fungi were categorized by macro- and microscopic characteristics of the colonies. Representatives of each group were selected for molecular identification based on the internal transcribed spacer region. Yeast identification was performed using MALDI-TOF MS and molecular analyses. Thirteen genera of filamentous fungi and six genera of yeasts were isolated and identified. The dominant filamentous species were Fusarium spp., Aspergillus spp., and Cochliobolus lunatus in the nostril samples and Aspergillus melleus in the rectal samples. Candida was the dominant genus among the identified yeasts at both anatomical sites. In the antifungal susceptibility test, 28 isolates showed resistance to fluconazole (78%), itraconazole (39%), and nystatin (42%). The knowledge of fungal microbiota composition of Amazonian manatees provides information that assists in monitoring the health status of individuals maintained in captivity, as these organisms can behave either as opportunists or as primary pathogens. Moreover, the composition and resistance of these organisms may vary among different rehabilitation institutions or different time frames of search, reinforcing the importance of constant in loco surveillance of these microorganisms. This study provides new perspectives on the fungal diversity in the microbiota of manatees and supports future studies concerning the clinical and epidemiological aspects and the impacts of these agents on the health of Amazonian manatees undergoing rehabilitation.

Hemotropic Mycoplasma spp. in Aquatic Mammals, Amazon Basin, Brazil.

Hemotropic Mycoplasma spp. (hemoplasmas) are uncultivable bacteria that infect mammals, including humans. We detected a potentially novel hemoplasma species in blood samples from wild river dolphins in the Amazon River Basin, Brazil. Further investigation could determine pathogenicity and zoonotic potential of the detected hemoplasma.

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.

Health assessment of juvenile green turtles in southern São Paulo State, Brazil: a hematologic approach.

We assessed the health status and hematologic and blood chemistry values of 48 juvenile green turtles (Chelonia mydas) on the southeastern coast of Brazil (25°S 48°W). We investigated the impact of size and weight, nutritional condition, and presence of epibionts and fibropapilloma tumors. Nutritional status was good for 39 animals; these animals had higher serum albumin, cholesterol, and phosphorus concentrations than 9 animals with a fair nutritional score. Reference values for hematology and biochemistry were calculated for 39 individuals without fibropapillomas (FPs). Turtles with epibionts (n = 33) had lower hemoglobin (Hb) than turtles without epibionts (n = 15; t = -2.09, p = 0.04), and the area occupied by epibionts was positively correlated with the white blood cell count (r = 0.37, p = 0.03). FP turtles had significantly lower hematocrit (Hct), Hb, and red blood cell (RBC) counts than non-FP turtles; serum albumin and cholesterol were higher in non-FP than FP turtles. A negative correlation between curved carapace length (CCL) and Hct (r = -0.51, p = 0.0002) and RBC (r = -0.47, p = 0.0007), and between serum cholesterol, sodium, and uric acid and CCL (r = -0.53, p = 0.0001; r = -0.38, p = 0.007; r = -0.35, p = 0.014, respectively) were identified. The health of turtles appears to deteriorate as they get larger, which manifests in more FPs, decreased body condition, and systemic physiologic changes consistent with chronic disease including lower Hct, RBC counts, serum cholesterol, sodium, and uric acid.

Hematologic profile of Amazon river dolphins Inia geoffrensis and its variation during acute capture stress.

Hematological values are of primary importance when investigating the health and physiological status of populations as they reflect the biological equilibrium of aquatic ecosystems. The objectives of this study are to produce baseline values for hematological parameters of the Amazon River dolphin (Inia geoffrensis), as well as to investigate significant variations according to sex, age, reproductive status and stress level. One-hundred-and-ten dolphins from Mamirauá Sustainable Development Reserve (3°3'S, 64°51'W), Central Amazon, Brazil, were live captured and sampled in November 2004 and 2005. Further, the means, standard deviations, minimum and maximum values and reference values (90% CI) were calculated. Correlations were performed to assess the relationships among blood values and cardiac rate (CR), respiratory frequency (RF), handling time and level of stress. No significant differences were found between sexes. Also, no differences occurred among pregnant and non-pregnant females, pregnant females and adult males or non-pregnant females and adult males. Calves had a higher white blood cell (WBC) count, and the neutrophil and lymphocyte absolute counts were significantly higher in calves than adults. The level of stress determined by empirical observation positively correlated with the WBC, neutrophil, lymphocyte and monocyte absolute counts and CR and RF. It was found that less stressed animals tend to present lower platelet counts and lower CR. The handling time of the dolphins was positively correlated with hematocrit (Hct), red blood cells (RBC) and Hb level. The hematological and physiological parameters varied according to time of handling and proved to be a good bioindicator of acute stress in Amazon River dolphins. The data provided here can complement long-term monitoring and identify the early warning indicators of health problems at the population level.