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Anthropogenic actions, especially inadequate waste disposal, cause permanent effects on aquatic fauna, resulting in a significant loss in their population. In this scenario, in situ and ex situ conservation strategies have been developed for these species. Among these strategies is the formation of somatic cell and tissue banks derived from skin collection that act complementarily to other biotechnologies. These banks contain all the information for genomic, genetic, and proteomic analyses. They are useful in the assessment of the toxicity of pollutants on the physiology of the species and regenerative and reproductive biotechnologies. The formation of these cryobanks involves different steps, including cryopreservation, with the optimization of all steps occurring in a species-specific manner. There is a diversity of studies on aquatic mammals; however, a low quantity compared to the number of studies on land mammals, with more than 80% of species still unexplored. This is mainly due to the difficulty of execution and asepsis in collecting skin from aquatic mammals and the in vitro culture, which seems to require more particularities for it to be successful. Therefore, this review aims to address the current scenario and the steps involved in the conservation of somatic cells and tissues derived from aquatic mammal skin, as well as results that have been achieved in recent years and the prospects.

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Morphological study of the tongue is an interesting way of understanding evolutionary processes associated with feeding habits. Therefore, the aim of the present study was to describe the tongue morphology of the Antillean manatee and to understand possible morphological relationships with its way of capturing food. Macroscopic dissections and light and scanning electron microscopy analyses of seven manatee tongues were performed. The tongue in Antillean manatees is a muscular and robust organ, divided into apex, body, and root. It is firmly adhered to the floor of the oral cavity. Lingual papillae were distributed over the entire tongue surface. They were identified as filiform papillae concentrated in the apex. Fungiform papillae were present on the apex and lateral regions. Foliate papillae were located on the dorsolateral portion of the root. Lentiform papillae were located across the dorsal tongue surface. The mucosa was lined by a keratinized stratified squamous epithelium presenting compound tubuloacinar glands and taste buds in the foliate papillae. The tongue of the Antillean manatee is similar to other Sirenia species, both of which share a completely herbivorous diet.

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Morphological studies of the oropharyngeal cavity of chelonians have become an interesting tool in the understanding of evolutionary processes associated with feeding habits in aquatic animals and the transition from aquatic to terrestrial forms. In this context, the aim of the present study was to describe the oropharyngeal cavity floor morphology of hawksbill sea turtle (Eretmochelys imbricata) hatchlings. Ten dead hatchlings of undefined sex were obtained from nests hatched on the coast of the state of Rio Grande do Norte, Brazil. The heads of each specimen were fixed, dissected, and analyzed at the macroscopic and microscopic levels. The oropharyngeal cavity floor of the hawksbill sea turtle hatchlings is formed by the tongue, pharynx, floor muscles, and hyolingual skeleton, delimited in the rostral and lateral directions by a keratinized beak, called the rhamphotheca, and in the caudal region at the limit between the pharynx and the esophagus. The tongue muscles and the muscles that support the floor of the oral cavity comprise the following: m. hypoglossohyoideus, m. hypoglossoglossus, m. hyoglossus, m. genioglossus, m. constrictor laryngis, m. geniohyoideus pars lateralis, and m. intermandibularis. The oropharyngeal cavity floor mucosa is formed by keratinized stratified squamous epithelium and the lamina propria is formed by loose connective tissue. The floor mucosa is devoid of taste buds. We believe that the basic oropharyngeal cavity floor characteristics in hawksbill sea turtle hatchlings may comprise indications that these animals are plesiomorphic and that semiaquatic and terrestrial turtles may have undergone adaptations to feed out of water.

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Morphological studies concerning the digestive system can further information on animal diets, thus aiding in the understanding of feeding behavior. Given the scarcity of information on sea turtle digestive system morphology, the aim of the present study was to describe the digestive tube (DT) morphology of Eretmochelys imbricata hatchlings to further understand the diet of these individuals in the wild. DT samples from 10 stillborn turtles (undefined sex) were analyzed at the macro and microscopic levels. The esophagus, stomach, small intestine (SI), and large intestine (LI) are described. Histologically, the DT is formed by four tunics, the mucosa, submucosa, muscular, and adventitia or serosa. The esophagus is lined by keratinized stratified squamous epithelium, while the remainder of the DT is lined by a simple columnar epithelium. The esophagus mucosa is marked by conical, pointed papillae. The stomach comprises three regions, the cardiac, fundic, and pyloric and is covered by neutral mucous granular cells. The intestinal mucosa presents absorptive cells with microvilli, neutral and acidic goblet cells, and mucosa-associated lymphoid tissue. The SI is significantly longer than the LI (p value = 0.006841). These morphological findings are strong indications of adaptations to a carnivorous diet in this hawksbill turtle age group.

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Among the bacterial infections that impair the health status of marine mammals, those caused by Brucella spp. are the most reported worldwide. Brucella infections in marine mammals can result in acute or chronic disease and are associated with variable clinical outcomes, depending on the organ involved during the infectious process, infection route, host immunity, and strain pathogenicity. Asymptomatic infections may also occur. The current study expands the investigation of Brucella infection in northeast Brazil by analyzing 19 dead, stranded cetaceans and 52 Antillean manatees Trichechus manatus manatus. The manatees included 8 dead, captive manatees and 44 live specimens, of which 10 were analyzed only after reintroduction into the wild as part of a rehabilitation program, 9 were analyzed both while in captivity or semi-captivity and after reintroduction, 20 were sampled only in captivity or semi-captivity, and 5 were free-living manatees. Serological tests were used to screen for antibodies against smooth Brucella spp. Whole blood, swabs, and tissue samples were screened for Brucella spp. DNA by PCR. Samples with positive PCR results were cultured for Brucella spp. isolation. All manatees yielded negative results in serological and molecular tests. Brucella spp. DNA was detected in the kidney of one adult Guiana dolphin Sotalia guianensis exhibiting necrosis in the liver. No growth of Brucella spp. was observed via microbiological culturing. This study is the first report of Brucella spp. DNA detection in cetaceans in the state of Pernambuco, and it highlights the importance of conducting systematic monitoring for the presence of Brucella infection in marine mammals along the Brazilian coast, especially in the northeast region, where several cases have been reported.

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Amazonian (Trichechus inunguis) and West Indian (Trichechus manatus) manatees are aquatic mammals vulnerable to extinction found in the Amazon basin and the coastal western Atlantic. Toll-like receptors (TLR) play a key role in recognizing pathogen-associated molecular patterns using leucine-rich repeats (LRRs). We described the diversity of TLR4 and TLR8 genes in these two species of manatee. Amazonian manatee showed seven SNPs in TLR4 and the eight in TLR8, while West Indian manatee shared four and six of those SNPs, respectively. In our analysis, TLR4 showed one non-conservative amino acid replacement substitution in LRR7 and LRR8, on the other hand, TLR8 was less variable and showed only conserved amino acid substitutions. Selection analysis showed that only one TLR4 site was subjected to positive selection and none in TLR8. TLR4 in manatees did not show any evidence of convergent evolution compared to species of the cetacean lineage. Differences in TLR4 and TLR8 polymorphism may be related to distinct selection by pathogens, population reduction of West Indian manatees, or an expected consequence of population expansion in Amazonian manatees. Future studies combining pathogen association and TLR polymorphism may clarify possible roles of these genes and be used for conservation purposes of manatee species.

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Reports of intussusception in sea turtles are generally linked to marine debris ingestion; therefore, only a few cases of the disease are associated with parasitic infestations. The objective of this study was to describe the necropsy findings of the first reported case of colocolic intussusception in a green sea turtle Chelonia mydas, associated with the helminth Octangium sp. A juvenile female green sea turtle, which was registered and rescued by the team from the Cetaceans Project of Costa Branca, State University of Rio Grande do Norte, was examined. The animal died 1 d after being treated and was immediately submitted for necropsy. Our findings indicated that parasitic infestation by Octangium sp. in the green sea turtle caused intussusception and consequently led to the animal's death. Early diagnosis and surgical correction are fundamental for a good prognosis and, consequently, for successful rehabilitation of marine species.

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A ocorrência de prolapso peniano em repteis é observada tanto em quelônios terrestres, quanto em squamatas, porém há a escassez de relatos em tartarugas marinhas documentados até o momento. Desta forma, objetivou-se relatar o processo de reabilitação de uma tartaruga-verde (Chelonia mydas) com prolapso peniano, de modo a nortear procedimentos clínicos e terapêuticos que possam propiciar maior sucesso na reabilitação destes animais. Uma tartaruga-verde (C. mydas), adulto, macho, foi resgatada no dia 02 de maio de 2020 na praia de Muriú, Ceará Mirim, Rio Grande do Norte, Brasil. O espécime encontrava-se ativo, responsivo, com bom escore corpóreo, hidratação moderada, presença de prolapso peniano com mucosa associada hiperêmica e parte necrosada, além disso, encontrava-se com a nadadeira anterior esquerda amputada. O animal recebeu suporte terapêutico à base de analgésicos, antibióticos, anti-inflamatórios, polivitamínicos e fluidoterapia. Foram realizados procedimentos cirúrgicos para remoção total do úmero e redução mecânica do prolapso peniano. Após 50 dias de reabilitação o animal teve alta médica e foi reintroduzido de volta ao mar. Conclui-se que o protocolo utilizado, tanto na reabilitação, quanto no tratamento conservativo do prolapso peniano mostrou-se eficaz na redução do edema, evitando assim a recidiva do prolapso.

The occurrence of penile prolapse in reptiles has been observed both in terrestrial turtles and squamates. However, there are few reports of penile prolapse in sea turtles published to date. This study aimed at reporting the rehabilitation process of a green sea turtle (Chelonia mydas) with penile prolapse, in order to guide clinical and therapeutic measures that can provide greater success in the rehabilitation of these animals. An adult male green sea turtle (C. mydas) was rescued on May 2, 2020 at Muriú beach, Ceará Mirim, Rio Grande do Norte, Brazil. The specimen was active and responsive, with a good body score and moderate hydration, and showed the presence of penile prolapse with associated hyperemic mucosa and necrosis. It also had the left anterior fin amputated. The animal received therapeutic support based on antibiotics, analgesics, anti-inflammatories, multivitamins, and fluid therapy. The surgical procedure for the complete removal of the humerus and mechanical reduction of penile prolapse was performed. After 50 days of rehabilitation, the animal was discharged and was reintroduced back to the sea. It is concluded that the protocol used was effective in reducing edema, thus preventing the recurrence of prolapse, both in rehabilitation and in the conservative treatment of penile prolapse

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A ocorrência de prolapso peniano em repteis é observada tanto em quelônios terrestres, quanto em squamatas, porém há a escassez de relatos em tartarugas marinhas documentados até o momento. Desta forma, objetivou-se relatar o processo de reabilitação de uma tartaruga-verde (Chelonia mydas) com prolapso peniano, de modo a nortear procedimentos clínicos e terapêuticos que possam propiciar maior sucesso na reabilitação destes animais. Uma tartaruga-verde (C. mydas), adulto, macho, foi resgatada no dia 02 de maio de 2020 na praia de Muriú, Ceará Mirim, Rio Grande do Norte, Brasil. O espécime encontrava-se ativo, responsivo, com bom escore corpóreo, hidratação moderada, presença de prolapso peniano com mucosa associada hiperêmica e parte necrosada, além disso, encontrava-se com a nadadeira anterior esquerda amputada. O animal recebeu suporte terapêutico à base de analgésicos, antibióticos, anti-inflamatórios, polivitamínicos e fluidoterapia. Foram realizados procedimentos cirúrgicos para remoção total do úmero e redução mecânica do prolapso peniano. Após 50 dias de reabilitação o animal teve alta médica e foi reintroduzido de volta ao mar. Conclui-se que o protocolo utilizado, tanto na reabilitação, quanto no tratamento conservativo do prolapso peniano mostrou-se eficaz na redução do edema, evitando assim a recidiva do prolapso.(AU)

The occurrence of penile prolapse in reptiles has been observed both in terrestrial turtles and squamates. However, there are few reports of penile prolapse in sea turtles published to date. This study aimed at reporting the rehabilitation process of a green sea turtle (Chelonia mydas) with penile prolapse, in order to guide clinical and therapeutic measures that can provide greater success in the rehabilitation of these animals. An adult male green sea turtle (C. mydas) was rescued on May 2, 2020 at Muriú beach, Ceará Mirim, Rio Grande do Norte, Brazil. The specimen was active and responsive, with a good body score and moderate hydration, and showed the presence of penile prolapse with associated hyperemic mucosa and necrosis. It also had the left anterior fin amputated. The animal received therapeutic support based on antibiotics, analgesics, anti-inflammatories, multivitamins, and fluid therapy. The surgical procedure for the complete removal of the humerus and mechanical reduction of penile prolapse was performed. After 50 days of rehabilitation, the animal was discharged and was reintroduced back to the sea. It is concluded that the protocol used was effective in reducing edema, thus preventing the recurrence of prolapse, both in rehabilitation and in the conservative treatment of penile prolapse(AU)

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Background: Manatees are the most endangered aquatic mammals in Brazil. The current conservation scenario, together with their biological characteristics, raises concern with the future of this species. Pyothorax, also known as septic pleural effusion or pleural empyema, is characterized by the accumulation of a septic purulent exudate within the pleural space. Although this infection often has a multifactorial etiology, it is most commonly associated with respiratory tract disorders and trauma. Here, we report a case of pyothorax in a Antillean manatee (Trichechus manatus) held in captivity for acclimatization in Brazil. Case: A young, male Antillean manatee, aged 4 years and 11 months, measuring 227 cm in total length, and weighing 258 kg was held in captivity for acclimatization (natural environment) in Porto de Pedras in the State of Alagoas, Brazil. The animal died in February 2013 and was sent for necropsy at the CMA/ICMBio laboratory. The study was developed with authorisation from the Ethics Committee on Animal Use (License number 020/2009) and under the license SISBIO/ ICMBio number 20685-1. Externally, the carcass of the animal showed swelling and bulging of the right antimere and purulent secretion from the right nostril. After external examination, a window was opened in the layers of skin, fat, and muscles from the level of the anus to the navel. Subsequently, the abdominal cavity was opened for inspecting the internal organs. The subcutaneous muscles had a slightly friable, swollen, and pale texture, in addition to petechial hemorrhage, suffusions, and marked edema of the subcutaneous tissue. The right hemidiaphragm was elevated, filling the abdominal and thoracic spaces. Depending on the elevation, all organs were moved to...(AU)

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Background: Antillean manatee (Trichechus manatus) and Amazonian manatee (Trichechus inunguis) are the two species of manatees that occur in Brazil, and both are endangered. Studies on manatee disease in Brazil were previously based mainly on Florida manatees. In recent years, these studies have increased in Brazil. Thus, the objective was to carry out a survey of the publications of infectious and non-infectious diseases in the two species of manatees that occur in the country, to assist in the identification of threats to the species and in the health assessment of manatees in the country. Review: Regarding infectious and contagious diseases, parasites have affected both species of manatees in Brazil, such as Pulmonicola cochleotrema in wild T. manatus, Toxoplasma gondii in both species, and Cryptosporidium spp. also in both species, this being an important agent of zoonotic character. Giardia sp., another zoonotic parasite, has also been reported in both species of manatees in Brazil. Mycotic lesions caused by B. hawaiiensis have been described in calves of T. manatus in captivity. Meanwhile, Candida sp., Trichosporon sp., Rhodotorula sp., and Cryptococcus sp. have been identified in T. inunguis. Studies on viruses in manatees are still not widespread compared to other species; however, it has been verified that rotavirus is present in T. manatus in captivity. In studies carried out to identify bacteria in T. manatus, the following bacteria were isolated: Arizona sp., Bacillus sp., Corynebacterium equi, Corynebacterium pseudodiphteriticum, Corinebacterium sp., Edwasiella tarda, Enterobacter aerogenes, Enterobacter agglomerans, Enterobacter sakasakii, Escherichia coli, Leptospira spp., Proteus sp., Proteus vulgaris, Providencia rettgeri...

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Background: Manatees are the most endangered aquatic mammals in Brazil. The current conservation scenario, together with their biological characteristics, raises concern with the future of this species. Pyothorax, also known as septic pleural effusion or pleural empyema, is characterized by the accumulation of a septic purulent exudate within the pleural space. Although this infection often has a multifactorial etiology, it is most commonly associated with respiratory tract disorders and trauma. Here, we report a case of pyothorax in a Antillean manatee (Trichechus manatus) held in captivity for acclimatization in Brazil. Case: A young, male Antillean manatee, aged 4 years and 11 months, measuring 227 cm in total length, and weighing 258 kg was held in captivity for acclimatization (natural environment) in Porto de Pedras in the State of Alagoas, Brazil. The animal died in February 2013 and was sent for necropsy at the CMA/ICMBio laboratory. The study was developed with authorisation from the Ethics Committee on Animal Use (License number 020/2009) and under the license SISBIO/ ICMBio number 20685-1. Externally, the carcass of the animal showed swelling and bulging of the right antimere and purulent secretion from the right nostril. After external examination, a window was opened in the layers of skin, fat, and muscles from the level of the anus to the navel. Subsequently, the abdominal cavity was opened for inspecting the internal organs. The subcutaneous muscles had a slightly friable, swollen, and pale texture, in addition to petechial hemorrhage, suffusions, and marked edema of the subcutaneous tissue. The right hemidiaphragm was elevated, filling the abdominal and thoracic spaces. Depending on the elevation, all organs were moved to...

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The aortic arrangement is an important structure associated with the maintenance of homeostasis. Based on this information, this study was conducted to describe the collateral arteries of the aortic arch of Antillean manatee and define the standard model for the species. Three specimens, an adult male, adult female and a male neonate, all strandings on the coast of the state of Rio Grande do Norte, Brazil, were used. The study was performed in the Laboratory of Morphophysiology of Vertebrates of the Federal University of Rio Grande do Norte, where in situ photographs were taken to demonstrate their topography. Subsequently, their hearts were removed and fixed in 10% formaldehyde and after 72 hr were dissected and analysed, obtaining schematic drawings and photographs of the vascular arrangement. The aortic arch was represented by three collateral arteries identified as the brachiocephalic trunk, left common carotid artery and left subclavian artery. This arrangement was similar to that found for other sirenians, and yet, for other mammals like hooded seal, murine, margarita island capuchin, black-handed tamarin, Mongolian gerbil and human. The morphological similarity presented in this study with different species of mammals, including humans, may contribute valuable information from an evolutionary point of view.

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This work aimed at evaluating the antifungal susceptibility and production of virulence factors by Candida spp. isolated from sirenians in Brazil. The isolates (n = 105) were recovered from the natural cavities of Amazonian and West Indian manatees and were tested for the susceptibility to amphotericin B, itraconazole, and fluconazole and for the production of phospholipases, proteases, and biofilm. The minimum inhibitory concentrations (MICs) for amphotericin B ranged from 0.03 to 1 µg/mL, and no resistant isolates were detected. Itraconazole and fluconazole MICs ranged from 0.03 to 16 µg/mL and from 0.125 to 64 µg/mL, respectively, and 35.2% (37/105) of the isolates were resistant to at least one of these azole drugs. Concerning the production of virulence factors, phospholipase activity was observed in 67.6% (71/105) of the isolates, while protease activity and biofilm production were detected in 50.5% (53/105) and 32.4% (34/105) of the isolates, respectively. Since the natural cavities of manatees are colonized by resistant and virulent strains of Candida spp., these animals can act as sources of resistance and virulence genes for the environment, conspecifics and other animal species, demonstrating the potential environmental impacts associated with their release back into their natural habitat.

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The Antillean manatee inhabits coastal regions of North and Northeastern Brazil and currently is considered an endangered species in the country. Aiming to gather information for the development of public policies focusing on the conservation of manatees, the National Center for Research and Conservation of Aquatic Mammals of the Chico Mendes Institute for Biodiversity has been rescuing, rehabilitating and releasing these mammals since the 1980s. Over the last 36 years, 40 manatees were released by the CMA/ICMBio and four of them were rescued again due to debris ingestion. Two of these manatees died and the other two were taken back into captivity for a new rehabilitation process. The four mammals had confirmed diagnosis of plastic debris ingestion. These findings demonstrate that the environment where the manatees live after being released had a significant amount of garbage which may hinder the success of the species conservation in Brazil.

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The aim of this study was to characterize the yeast microbiota of natural cavities of manatees kept in captivity in Brazil. Sterile swabs from the oral cavity, nostrils, genital opening, and rectum of 50 Trichechus inunguis and 26 Trichechus manatus were collected. The samples were plated on Sabouraud agar with chloramphenicol and incubated at 25 °C for 5 days. The yeasts isolated were phenotypically identified by biochemical and micromorphological tests. Overall, 141 strains were isolated, of which 112 were from T. inunguis (Candida albicans, Candida parapsilosis sensu stricto, Candida orthopsilosis, Candida metapsilosis, Candida guilliermondii, Candida pelliculosa, Candida tropicalis, Candida glabrata, Candida famata, Candida krusei, Candida norvegensis, Candida ciferri, Trichosporon sp., Rhodotorula sp., Cryptococcus laurentii) and 29 were from T. manatus (C. albicans, C. tropicalis, C. famata, C. guilliermondii, C. krusei, Rhodotorula sp., Rhodotorula mucilaginosa, Rhodotorula minuta, Trichosporon sp.). This was the first systematic study to investigate the importance of yeasts as components of the microbiota of sirenians, demonstrating the presence of potentially pathogenic species, which highlights the importance of maintaining adequate artificial conditions for the health of captive manatees.

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The aim of this work was to analyze whether there are differences between the development pattern of Antillean manatee (Trichechus manatus) calves born in captivity and those rescued and kept under rehabilitation. Biometrics data were collected from 1990 to 2010 from 38 calves, 29 of which still had the remnants of the umbilical cord and had been rescued from the Brazilian northeastern coastline (Group I), and nine individuals that were born in captivity and remained with their mothers (Group II). Among the measures obtained through biometry, the total length and weight of the animal were recorded. Given that the breastfeeding of calves occurs approximately until the age of 2 yr, data obtained until the 24th month of life of each individual were evaluated. An average increase in weight of 53.50 +/- 38.54 kg (mean +/- standard deviation [SD]) was detected in Group I and a gain of 106.87 +/- 47.21 kg (mean +/- SD) in Group II. From months 13 to 24, no significant difference in the weight increment was observed. A similar pattern occurred with regard to the increase in the overall length during the first year, where animals from Group I grew 34.81 +/- 17.94 cm (mean +/- SD) and from Group II grew 83.83 +/- 28.21 cm, a statistically significant difference. The growth was not significantly different from 13 to 24 mo. The results found in this study identified the need for a review of the nutritional diet offered to orphaned calves rescued and kept in captivity. The results also support the need for a better adequacy of facilities for these animals as a way to encourage the management strategies adopted for manatee calves maintained in captivity.

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