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.

Persistent long-term habitat use by Florida manatees at Fort Pierce, Florida from 1997 to 2020.

To survive cold winters, Florida manatees (Trichechus manatus latirostris) depend on artificial (i.e., power plants) and natural warm water sources such as springs and passive thermal basins. Passive thermal basins can provide critical habitat for manatees for short or extended periods of time. The Henry D. King Powerplant in Fort Pierce, Florida discharged warm water into Moore's Creek until it went offline in 1995. However, it is unknown to what degree manatees continue to occupy this area and how environmental factors influence their occurrence in the creek. To explore this, we examined the habitat use of Florida manatees in Moore's Creek after the shutdown from November 1997 to March 2020 from daily counts of manatees. In addition, we correlated local environmental data (ambient air, temperature, salinity) to assess if Moore's Creek had properties indicative of a passive thermal basin. Results indicated there was not an increase or decrease in habitat use over twenty years in the Creek. The consistent use of Moore's Creek over the study period suggests that this habitat possesses thermal and freshwater resources to support manatee occurrence long-term. These findings provide robust support for the importance of this habitat and passive thermal basins for Florida manatees.

Metal and metalloid maternal transfer in a newborn West Indian manatee (Trichechus manatus) two years after the northeastern oil spill disaster of 2019 in Brazil.

A large-scale oil spill along the northeastern Brazilian coast in 2019-2020 severely impacted primary manatee habitats in Brazil. This study aimed to assess metal and metalloid contamination in a dead manatee calf found following this oil spill. Several elements were detected, including chromium, iron, nickel, lead, and vanadium, which are crude oil components, and thallium, a component of dispersants used to degrade and dissipate crude oil. The presence of these contaminants in the manatee calf is indicative of maternal exposure, metabolism, and newborn transference via placenta and lactation. This is the first report of mother-calf metal transfer in manatees, highlighting the species' vulnerability and potential long-term population dynamics effects, and the first report for several metals and metalloids in these animals, laying the foundation for future research efforts. These findings highlight the urgent need for continued environmental monitoring, ecotoxicological assessments, and conservation initiatives.

Agonistic interactions initiated by adult bottlenose dolphins on Antillean manatee calves in the Caribbean Sea.

The dynamics and drivers of inter-species interactions in the wild are poorly understood, particularly those involving social animal species. Inter-species interactions between cetaceans and sirenians have rarely been documented and investigated. Here, we report 10 cases of interaction initiated by adult bottlenose dolphins (Tursiops truncatus) towards Antillean manatee (Trichechus manatus manatus). Interactions were documented through behavioral observations in the wild (n = 7) and from the examination of orphaned calves (i.e., tooth rake marks on their body; n = 4) that entered a rehabilitation facility, one individual both observed interacting with dolphins and found stranded with bite marks. Bottlenose dolphins were observed interacting with orphan manatee calves and with mother-calf pairs, exhibiting agonistic behavior (n = 2), affiliative or neutral behaviors (n = 1), but the behavioral contexts of these interactions remain unclear in most cases (n = 7). Information on stranded individuals was collected from four calves (of 13 examined calves) recovered in poor condition with bottlenose dolphin tooth rakes and bite wounds on their bodies, one of which died. Injury from bite wounds varied in extent and severity, ranging from superficial scratches leaving rake marks to deep lacerations. Our findings suggest the regular occurrence of agonistic behaviors initiated by adult bottlenose dolphins and directed toward manatee calves. However, the drivers of these interactions remain unknown and need to be further investigated.

Gross and microanatomic description of the dental pads of the Florida manatee, Trichechus manatus latirostris, and their importance to mastication.

OBJECTIVE: Florida manatee feeding ecology is critical to species survival, but the role of dental pads in feeding has received limited attention. This study characterized the gross and microscopic anatomy of the manatee's dorsal and ventral dental pad in relation to these structures' importance in mastication, which furthers our understanding of manatee feeding and health. DESIGN: Whole heads from 6 animals (4 male and 2 female) of varying sizes were examined grossly. Sections (5 µm) from throughout the dorsal and ventral dental pads were stained with Hematoxylin and Eosin to document microanatomy. The thickness of the epithelium and stratum corneum were measured. RESULTS: The ventral dental pad epidermal (1129-3391 µm) and stratum corneum (331-1848 µm) thickness increased with increased body size. The dorsal dental pad epidermal (690-1988 µm) and stratum corneum (121-974 µm) thickness varied relative to size. The dental pad anatomy, including the thickened stratum corneum, indicates an importance similar to molars in grinding and physically breaking up plant material. Extensive appendages including filiform-like papillae and well-developed rete were observed and likely provide physical support for mastication. CONCLUSION: While the sample size limits specific conclusions based on sex or age, it provides a good overview of the anatomy of the dental pads. The manatee is the only mammal known to have a ventral dental pad and the well-developed grinding surfaces demonstrates a crucial role in mastication for these structures. These dental pads should be evaluated during health checks and necropsies and considered in future research on manatee's feeding mechanisms.

First molecular identification of the trematode Pulmonicola cochleotrema (Platyhelminthes: Opisthotrematidae) in West Indian manatees (Trichechus manatus, Sirenia: Trichechidae) from Puerto Rico and Florida.

West Indian manatees (Trichechus manatus) harbor a variety of endoparasites, including the nasal trematode Pulmonicola cochleotrema, which infects the respiratory tract, especially the nasal passages. Previous studies have described and identified this digenean using morphological data only. This study presents the first molecular identification of P. cochleotrema in West Indian manatees from Puerto Rico and Florida. Samples of the trematode were collected from seven manatees found stranded dead at both locations. The small subunit ribosomal DNA (18S rDNA) was amplified from each sample using universal primers for different regions of the gene, resulting in a consensus sequence of 1871 base pairs. The phylogenetic reconstruction was carried out using DNA sequences of other species of digenean parasites from other hosts, including a trematode of the same taxonomic family from another sirenian species. Specimens collected from both locations show the same molecular identity using SSU rDNA sequence data. The identity of P. cochleotrema was confirmed using the Basic Local Alignment Search Tool from the National Center for Biotechnology Information database, yielding a high similarity of 98.8 % with Opisthotrema dujonis and 98.2 % with Lankatrema mannarense located in the same clade in our analysis. The latter two digeneans belong to the Opisthotrematidae as does P. cochleotrema and previous studies reported them infecting the Eustachian tubes, esophagus, and digestive tract in dugongs (Dugong dugon). These findings evidence that the nasal trematode of manatees in Florida and Puerto Rico and the dugong, all inhabiting and feeding in marine environments, will have a marine mollusk as an intermediate host, probably a gastropod. The question remains, which species of nasal trematode are harbored by lotic-dwelling manatees in other parts of their distribution like South America.

Abdominal ultrasound in amazonian manatee (Trichechus inunguis) (Natterer, 1883) / Ultrassonografia abdominal em peixe boi amazônico (Trichechus inunguis (Natterer, 1883)

Morphophysiological species researches are fundamental, and diagnostic imaging is an excellent technique, already used in wild animals, with great application, not invasive and provide real-time information of each body. Amazonian manatees are on the list of endangered animals classified in the vulnerable category and knowledge of the normal pattern of ultrasound anatomy of organs and tissues is important for the maintenance and well-being of captive specimens contributing to reintroduction actions. The objective of the study was to standardize the examination technique and describe the ultrasound findings of the liver, gallbladder, stomach, urinary bladder and the subcutaneous tissue of the abdominal region in Trichechus inunguis, in order to contribute with the anatomical and sonographic knowledge and assist in the diagnosis and prognosis diseases. The study used 18 animals to describe the normal sonographic anatomy in the abdominal cavity of the Amazonian manatee. During abdominal scan, it was possible to visualize the features of the liver, gallbladder, stomach, urinary bladder obtained satisfactory results in this study. Therefore, other structures were not primarily identified by the reduced time, lots of fat and gases in intestines of animals.

Pesquisas morfofisiológicas em espécies selvagens são fundamentais, e o diagnóstico por imagem é uma excelente técnica, já usada e com grande aplicação, não invasiva e que fornece informações em tempo real de cada órgão. Peixes-boi-amazônico encontram-se na lista de animais ameaçados de extinção classificados na categoria vulnerável e o conhecimento do padrão normal da anatomia ultrassonográfica de órgãos e tecidos é importante para a manutenção e bem-estar de espécimes em cativeiro contribuindo para ações de reintrodução. O objetivo deste estudo foi padronizar a técnica de exame e descrever os achados ultrassonográficos do fígado, vesícula biliar, estômago, vesícula urinária e o tecido subcutâneo da região abdominal em Trichechus inunguis, de modo a contribuir com o conhecimento anátomo-sonográfico e auxiliar no diagnóstico e prognóstico de doenças. O estudo utilizou 18 animais para descrever a anatomia ultrassonográfica normal na cavidade abdominal de peixe-boi amazônico. Durante a varredura abdominal foi possível visualizar as características dos órgãos obtendo resultados satisfatórios neste estudo, concluindo ser uma técnica eficiente para avaliação de determinados órgãos abdominais em peixe-boi amazônico. Entretanto, outras estruturas não foram identificadas principalmente pelo tempo reduzido, muita gordura e gases nos intestinos dos animais.

Enfermedades Autoinmunes, tratamiento con Trichuris suis y otros helmintos / Autoimmune diseases, treatment with Trichuris suis and some other helminths

Objetivos: La "Hipótesis de la Higiene" postula sobre los efectos inmunomoduladores inducidos por agentes infecciosos en los seres humanos. El objetivo principal de este trabajo es indagar sobre las evidencias de esa hipótesis y sobre sus aplicaciones en el campo del tratamiento de las enfermedades autoinmunes, haciendo especial hincapié tanto en los mecanismos de acción en los que se basan estas aplicaciones como en los resultados reales obtenidos. Además analizará la posible evolución de estas terapias, especialmente a la sombra de las controversias éticas que surgen de la aplicación de estos tratamientos en relación a si es lícito causar una infección para curar otra patología, como es el caso que nos ocupa. Resultados: Actualmente existen resultados prometedores de ensayos clínicos sobre terapias helmínticas aplicadas al tratamiento de enfermedades autoinmunes como son la enfermedad de Crohn y la colitis ulcerosa, resumidos en el apartado de resultados de este artículo. Igualmente prometedora es la gran variedad de ensayos clínicos que actualmente están en curso sobre la aplicación de la terapia helmíntica al tratamiento de diversas patologías en las que está involucrado el sistema inmunológico, como son: asma, rinitis alérgica, artritis reumatoide, esclerosis múltiple, diabetes Mellitus tipo I, encefalomielitis autoinmune, obesidad, autismo, etc que han llevado a identificar cuáles son los parásitos indicados en el tratamiento de este amplio espectro de enfermedades. Sin embargo es necesario indicar que no todos los helmintos son inmunorreguladores y, por lo tanto útiles en el tratamiento de estas enfermedades y que, los que lo son, no son útiles en el tratamiento de todas las enfermedades de origen inmunológico sino que presentan una marcada especificidad. Es más, la utilidad de éstos presenta una variabilidad importante, no sólo dependiente de la enfermedad de origen inmunológico a tratar, sino también de las condiciones del paciente. Es por ello que actualmente no existen terapias helmínticas aprobadas por las principales agencias del medicamento; aún quedan muchos aspectos por desvelar, lo que hace que la hipótesis de la higiene no haya pasado de ser una hipótesis. Sin embargo lo prometedor de estas terapias ha traído consigo la autorización de algunas de ellas como "productos en fase de investigación clínica". Conclusiones: Es necesario seguir investigando sobre las terapias helmínticas. Además es especialmente prometedor el empleo de productos solubles de parásitos en el tratamiento de estas enfermedades en lugar de la infección helmíntica e incluso, el diseño de análogos sintéticos que sean más efectivos y presenten menos riesgos. La utilización de este último tipo de productos traerá consigo un aumento en el número de ensayos clínicos, justificados por las facilidades de patente de éstos frente a la utilización directa de estados de los parásitos. Sin embargo todavía es necesaria más investigación para conocer las biomoléculas presentes en los parásitos que son responsables de su efecto terapéutico

Objectives: Hygiene hypothesis postulates about immunomodulatory effects induced by some infectious agents in human beings. The main objective of this review is to find out what the evidence of this hypothesis is and its applications in the field of autoimmune diseases, paying especial attention not only in the action mechanisms in which they are based upon but also in the real outcome obtained. Moreover, the likely evolution of these therapies will be analyzed, especially in what is related to ethical disputes arising from these treatment applications regarding whether the causing infection is fair in order to heal another pathology, as the case may be. Outcome: Promising outcomes currently exist in regards to helminth therapy applied to treat autoimmune diseases such as Crohn's disease and ulcerative colitis, summarised in this review. Equally promising is the high amount of outstanding clinical trials for the application of helminthic therapy to a wide range of different pathologies in which the immune system is involved, such as: asthma, allergical rhinitis, rheumatoid arthritis, type I diabetes mellitus, autoimmune encephalomyelitis, obesity, autism, etc, which have identified useful parasites for the treatment of such a wide spectrum of diseases. However it is needed to highlight that helminths are not always immunoregulators and therefore useful in all autoimmune disease treatments. Those which are immunoregulators, are not useful in any autoimmune disease treatment because they are highly specific. Moreover, its usefulness shows high variability, which means it is not only dependent on disease but also dependent upon patient conditions. It is because of this that helminthic therapies are not currently approved by major medical agencies; still many outstanding aspects need to be unveiled, because hygiene hypothesis has not overcome the hypothesis status. However these promising therapies have gained the status of "investigational products". Conclusions: Additional investigation is needed. Especially promising are parasitic soluble products employed in autoimmune disease treatments in place of helminthic infection and even, less risky and more effective new analogue synthetic drug designs. This latter kind of product utilisation will increase the number of clinical trials due to patent ease of this product versus direct utilisation of parasitic phases. However more investigation is still needed to know which parasitic biomolecules are responsible for the therapeutic effect