Salmonellosis in a young Amazonian manatee (Trichechus inunguis) / Salmonelose em um filhote de peixe-boi-da-amazônia (Trichechus inunguis)

The objective of this study was to describe a case of salmonellosis in an Amazonian manatee (Trichechus inunguis). The animal was rescued from a tributary of the Capim River, in the municipality of Goianésia do Pará, Pará, Brazil, kept in a pool, and died approximately five months after being rescued. The main changes observed at necropsy were that the cecum and colon had serosal hyperemia, wall edema, thickened mucosa with ulcerated areas and covered by a thin layer of fibrin, and a yellowish liquid content. Histologically, there was moderate multifocal fibrinonecrotic typhlocolitis associated with intense bacterial colonization. Salmonella enterica subsp. enterica (rough) was isolated in cecum and colon fragments. This is the first report of salmonellosis in an Amazonian manatee.(AU)

O objetivo deste trabalho é descrever um caso de salmonelose em um peixe-boi-da-amazônia (Trichechus inunguis). O animal foi resgatado em um afluente do rio Capim, no município de Goianésia do Pará, Pará, Brasil, sendo mantido em uma piscina e morrendo aproximadamente cinco meses após o resgate. As principais alterações observadas na necropsia foram ceco e colón com hiperemia da serosa; edema da parede; mucosa espessada, com áreas ulceradas e revestida por uma fina camada de fibrina; e conteúdo líquido amarelado. Histologicamente havia tiflocolite fibrinonecrótica, multifocal, moderada, associada a intensa colonização bacteriana. Em fragmentos de ceco e cólon foi isolada Salmonella enterica subsp. enterica (rugosa). Este é o primeiro relato de salmonelose em peixe-boi-da-amazônia.(AU)

Using osteological measurements to estimate body length in Amazonian manatees / Utilização de medidas osteológicas para estimativa de comprimento corporal em peixe-boi da Amazônia

Body length is an important parameter in morphological, ecological and behavioral studies of a species and contributes to the understanding of the body condition of individuals. This parameter is essential for conservation and management strategies by informing studies evaluating growth rates, physical maturity and classification of individuals into age groups, promoting better accuracy for the biological parameters of the species. The aim of this study was to identify predictors of body length for Amazonian manatees (Trichechus inunguis) using metric characters of osteological materials. Eleven linear measurements of skull, jaw, scapulae and humeri were collected from 41 Amazonian manatee skeletons of different age classes (calf, juvenile and adult). Data were analyzed by simple linear regression. The condylobasal length was the best predictor of body length for the species (R = 0.943), however, all bones evaluated showed at least one measurement with the capacity to predict body size (R2 > 0.9). Results of this study are useful for inferring body length of Amazonian manatees using bones deposited in museums and biological collections, expanding the informative potential of these materials.(AU)

O comprimento corporal é um importante parâmetro em estudos morfológicos, ecológicos e comportamentais de uma espécie e contribui para o entendimento da condição corporal de um indivíduo. Este parâmetro é essencial para estratégias de conservação e manejo, informando estudos que avaliam taxas de crescimento, maturidade física e classificação dos indivíduos em grupos de idade, promovendo uma melhor precisão aos parâmetros biológicos de uma espécie. O objetivo desse trabalho foi identificar preditores de comprimento corporal para o peixe-boi da Amazônia (Trichechus inunguis) utilizando caracteres métricos de material osteológico. Onze medidas lineares do crânio, mandíbula, escápula e úmero foram coletadas de 41 esqueletos de peixe-boi da Amazônia de diferentes classes de idade (filhote, juvenil e adulto). Os dados foram analisados por regressão linear simples. O comprimento côndilobasal foi o melhor preditor de comprimento corporal para a espécie (R = 0,943), entretanto, todos os ossos avaliados mostraram pelo menos um caractere com capacidade de predizer o comprimento corporal (R2 > 0,9). Os resultados deste estudo são úteis para inferir o comprimento corporal de peixes-boi da Amazônia a partir de ossos depositados em museus e coleções biológicas, expandindo o potencial informativo destes materiais.(AU)

Doenças infecciosas e não infecciosas nos peixes-boi do Brasil / Infectious and non-infectious diseases in manatees in Brazil

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...

Piotórax em peixe-boi-marinho (Trichechus manatus) no Brasil / Pyothorax in Antillean manatee (Trichechus manatus) in Brazil

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...

Piotórax em peixe-boi-marinho (Trichechus manatus) no Brasil / Pyothorax in Antillean manatee (Trichechus manatus) in Brazil

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)

Bottlenecks in the migration routes of Amazonian manatees and the threat of hydroelectric dams / Gargalos na rota migratória do peixe-boi amazônico e a ameaça das barragens hidrelétricas

In a particular region within western Amazonia, Amazonian manatees (Trichechus inunguis) live in a floodplain environment that becomes inhospitable for them during the annual low-water season. To flee from it, they undergo a dangerous migration to a refuge while water levels are dropping fast. Our aim was to better understand the role of depth variation in this migratory process. We analyzed the sole tracking data on wild manatees (n=10 males), 30 years of Landsat images, a 14-year hydrograph and a 3-D bathymetric model. Migratory routes contained shallower segments, here called 'migratory bottlenecks', which dried out at the end of most lowering-water seasons, blocking the passage to the refuge. Manatees began migrating just in time to traverse the bottlenecks furthest away, suggesting they fine-tuned their departure so as to maximize time within the foraging home range without compromising safety. They apparently achieved this by estimating depth at the bottlenecks. Moreover, a bottleneck was created in >15 years, illustrating the environment's dynamism and the challenge this imposes upon manatees. Our results are probably generalizable to most of the species' range. We contend manatees possess an updatable cognitive map of their environment and are behaviorally plastic. Current dam-building plans, if implemented, would create more bottlenecks and make flooding less predictable, increasing manatee mortality from unsuccessful migrations. It would also partition the species into small populations, each prone to short-term extinction. The natural outcome would be the second species-level collapse. Economic growth should not come at the expense of the extinction of the iconic manatee.(AU)

Em uma região particular da Amazônia ocidental, peixes-boi amazônicos (Trichechus inunguis) vivem em um ambiente que se torna inóspito para eles durante a água-baixa anual. Para fugir dele, realizam uma migração perigosa para o refúgio enquanto o nível da água desce rapidamente. Nosso objetivo foi compreender melhor o papel da variação da profundidade neste processo migratório. Analisamos os únicos dados de rastreamento de peixes-boi selvagens (n=10 machos), 30 anos de imagens Landsat, 14 anos de hidrógrafa e um modelo batimétrico 3-D. As rotas migratórias possuíam trechos mais rasos, denominados gargalos migratórios, que secaram no final da maioria das vazantes, bloqueando o acesso ao refúgio. Os peixes-boi começaram a migração em tempo justo para atravessar os gargalos mais distantes, sugerindo que a sintonizaram para maximizar o período se alimentando sem comprometer a segurança. Para tal, parecem ter estimado a profundidade nos gargalos. Adicionalmente, um gargalo foi criado em 15 anos, ilustrando o dinamismo do ambiente e o desafio que isto impõe aos peixes-boi. Esses resultados provavelmente valem para boa parte da área de distribuição da espécie. Argumentamos que peixes-boi possuem um mapa cognitivo atualizável do ambiente e são comportamentalmente plásticos. Os planos de construção de barragens hidrelétricas, se concretizados, criariam mais gargalos e regimes de inundação menos previsíveis, dificultando a migração e consequentemente aumentando a mortalidade de peixes-boi. Também particionariam a espécie em populações pequenas, vulneráveis à extinção no curto-prazo. O desfecho seria o segundo colapso da espécie. O crescimento econômico não deve vir às custas da extinção do icônico peixe-boi.(AU)

Bottlenecks in the migration routes of Amazonian manatees and the threat of hydroelectric dams / Gargalos na rota migratória do peixe-boi amazônico e a ameaça das barragens hidrelétricas

In a particular region within western Amazonia, Amazonian manatees (Trichechus inunguis) live in a floodplain environment that becomes inhospitable for them during the annual low-water season. To flee from it, they undergo a dangerous migration to a refuge while water levels are dropping fast. Our aim was to better understand the role of depth variation in this migratory process. We analyzed the sole tracking data on wild manatees (n=10 males), 30 years of Landsat images, a 14-year hydrograph and a 3-D bathymetric model. Migratory routes contained shallower segments, here called 'migratory bottlenecks', which dried out at the end of most lowering-water seasons, blocking the passage to the refuge. Manatees began migrating just in time to traverse the bottlenecks furthest away, suggesting they fine-tuned their departure so as to maximize time within the foraging home range without compromising safety. They apparently achieved this by estimating depth at the bottlenecks. Moreover, a bottleneck was created in >15 years, illustrating the environment's dynamism and the challenge this imposes upon manatees. Our results are probably generalizable to most of the species' range. We contend manatees possess an updatable cognitive map of their environment and are behaviorally plastic. Current dam-building plans, if implemented, would create more bottlenecks and make flooding less predictable, increasing manatee mortality from unsuccessful migrations. It would also partition the species into small populations, each prone to short-term extinction. The natural outcome would be the second species-level collapse. Economic growth should not come at the expense of the extinction of the iconic manatee.

Em uma região particular da Amazônia ocidental, peixes-boi amazônicos (Trichechus inunguis) vivem em um ambiente que se torna inóspito para eles durante a água-baixa anual. Para fugir dele, realizam uma migração perigosa para o refúgio enquanto o nível da água desce rapidamente. Nosso objetivo foi compreender melhor o papel da variação da profundidade neste processo migratório. Analisamos os únicos dados de rastreamento de peixes-boi selvagens (n=10 machos), 30 anos de imagens Landsat, 14 anos de hidrógrafa e um modelo batimétrico 3-D. As rotas migratórias possuíam trechos mais rasos, denominados gargalos migratórios, que secaram no final da maioria das vazantes, bloqueando o acesso ao refúgio. Os peixes-boi começaram a migração em tempo justo para atravessar os gargalos mais distantes, sugerindo que a sintonizaram para maximizar o período se alimentando sem comprometer a segurança. Para tal, parecem ter estimado a profundidade nos gargalos. Adicionalmente, um gargalo foi criado em 15 anos, ilustrando o dinamismo do ambiente e o desafio que isto impõe aos peixes-boi. Esses resultados provavelmente valem para boa parte da área de distribuição da espécie. Argumentamos que peixes-boi possuem um mapa cognitivo atualizável do ambiente e são comportamentalmente plásticos. Os planos de construção de barragens hidrelétricas, se concretizados, criariam mais gargalos e regimes de inundação menos previsíveis, dificultando a migração e consequentemente aumentando a mortalidade de peixes-boi. Também particionariam a espécie em populações pequenas, vulneráveis à extinção no curto-prazo. O desfecho seria o segundo colapso da espécie. O crescimento econômico não deve vir às custas da extinção do icônico peixe-boi.

Changes in the blood parameters of the Amazonian manatee (Trichechus inunguis) after long-distance transportation / Alterações nos padrões sanguíneos do peixe-boi da Amazônia (Trichechus inunguis) após transporte de longa distância

In this study we report the hematological, biochemical and hormonal parameters in a juvenile male Amazonian manatee measured before transport, immediately after transport, and during adaptation to a new facility. The animal was transported from Manaus, Amazonas State, Brazil, to São Paulo, São Paulo State, Brazil, (2,733 km) within 6 hours. Among all blood parameters analyzed, we observed obvious neutrophilia, lymphopenia, and increases in the neutrophil/lymphocyte ratio and serum glucose and aspartate aminotransferase (AST) levels, but these parameters subsequently returned to normal. These results suggest that transport and changes in the environment are temporary stressful events for Amazonian manatees. We, therefore, recommend monitoring the hematological and biochemical parameters before and after translocation to minimize the effects of handling stressors in this species.

Neste estudo, são relatados os parâmetros hematológicos, bioquímicos e hormonais em um peixe-boi da Amazônia macho jovem, medidos antes do transporte, imediatamente depois, e durante a sua adaptação a um novo recinto. O animal foi transportado de Manaus, Estado do Amazonas, Brasil, para São Paulo, Estado de São Paulo, Brasil, (2.733 km) durante 6h de viagem. Entre os parâmetros sanguíneos analisados, foi observada clara neutrofilia, linfopenia e aumento na relação neutrófilo/linfócito e nos níveis de glicose e aspartato aminotransferase (AST), mas estes parâmetros posteriormente voltaram aos valores normais. Estes resultados sugerem que o transporte e as mudanças no ambiente são eventos estressantes para o peixe-boi da Amazônia. Desta forma, é recomendável o monitoramento dos parâmetros hematológicos e bioquímicos antes e após a translocação para minimizar os efeitos de agentes estressores nesta espécie.

Enfermidades em peixes-bois (Sirenia: Trichechidae) oriundos de encalhes na Amazônia Oriental

No Brasil são encontradas duas espécies de peixes-bois, uma ocorre na zona costeira das regiões Norte e Nordeste, denominada de peixe-boi-marinho (Trichechus manatus) e outra nos rios e estuários da Amazônia conhecido como peixe-boi-da-Amazônia (Trichechus inunguis). Essas espécies são consideradas criticamente ameaçadas por ações antropogênicas em todas as regiões onde elas ocorrem. O conhecimento sobre doenças que acometem peixes-bois vem crescendo e é constante. Este estudo tem como objetivo relatar enfermidades que levaram a óbito dois peixes-bois do gênero Trichechus oriundos de encalhes na Amazônia Oriental. Nos anos de 2014 e 2015, um exemplar de Trichechus spp. e um de Trichechus inunguis, peixes-boisr espectivamente, vieram a óbito. Fragmentos de diversos órgãos foram coletados em formalina a 10% e processados rotineiramente para histopatologia. As amostras foram incluídas em parafina, cortadas a 5 m de espessura e corados por hematoxilina e eosina (HE). Nos achados para Trichechus spp. foi observado opacidade das córneas, além de lesões irregulares nas regiões torácica e ventral sugerindo dermatite micótica. Microscopicamente, foi observada degeneração mixomatosa nas válvulas cardíacas, revelando endocardiose e congestão pulmonar. Nos achados de necropsia da espécie T. inunguis foi observado que no ceco e terço inicial do cólon a mucosa apresentava conteúdo líquido amarelo claro e erosões parcialmente recobertas por uma camada de material amarelado e elástico (pseudomembranas de fibrina). O exame histopatológico evidenciou intensa colonização bacteriana no estrato córneo, caracterizada pela presença de cocos e bacilos basofílicos entre as lâminas de queratina e sobre a mesma. Havia ainda áreas de necrose com desnudamento das criptas margeada por infiltrado inflamatório misto, contendo eosinófilos. Foi estabelecido diagnostico de Tiflocolite necrotizante multifocal moderada associada com intensa colonização bacteriana na mucosa. A bactéria Salmonella enterica subsp. enterica (rugosa) foi isolada do conteúdo do ceco. O relato de casos de enfermidades em peixes-bois é de suma importância para ampliar o conhecimento sobre a biologia desses animais. Além disso, auxilia na elaboração de novas estratégias de conservação na natureza e em centros de reabilitação. Portanto, mais pesquisas nesse contexto devem ser realizadas no Brasil e em particular na região amazônica, onde as duas espécies do genêro Trichechus ocorrem em simpatria.

In Brazil, two species of manatees are found, one in the coastal zone of the North and Northeast, called the manatee (Trichechus manatus) and the other in the Amazonian rivers and estuaries known as the Amazonian manatee (Trichechus inunguis). These species are considered critically endangered by anthropogenic actions in all regions where they occur. Knowledge about diseases that plague manatees has been growing steadily. This study aims to report diseases that led to the death of two manatees of the Trichechus genus from strandings in the Eastern Amazon. In the years 2014 and 2015, a copy of Trichechus spp. And one of Trichechus inunguis, fish steers specifically, came to death. Fragments of several organs were collected in 10% formalin and routinely processed for histopathology. Samples were embedded in paraffin, cut 5 m thick and stained with hematoxylin and eosin (HE). In the finds for Trichechus spp. Corneal opacity was observed, as well as irregular lesions in the thoracic and ventral regions suggesting fungal dermatitis. Microscopically, myxomatous degeneration was observed in the cardiac valves, revealing endocardiosis and pulmonary congestion. In the necropsy findings of the T. inunguis species, it was observed that in the cecum and early colon the mucosa presented light yellow liquid content and erosions partially covered by a layer of yellowish and elastic material (pseudomembranes of fibrin). The histopathological examination evidenced intense bacterial colonization in the stratum corneum, characterized by the presence of basophilic cocci and bacilli between the keratin laminae and on the same. There were also areas of necrosis with nudity of the crypts bordered by mixed inflammatory infiltrate containing eosinophils. A diagnosis of moderate multifocal necrotizing typhlocolitis associated with intense bacterial colonization of the mucosa was established. The bacterium Salmonella enterica subsp. Enterica (rugosa) was isolated from the contents of the cecum. The report of cases of diseases in manatees is of paramount importance to increase the knowledge about the biology of these animals. In addition, it assists in the development of new conservation strategies in nature and in rehabilitation centers. Therefore, more research in this context should be carried out in Brazil and in particular in the Amazon region, where the two species of the Trichechus genus occur in sympatry.

Single nucleotide polymorphisms from cytochrome b gene as a useful protocol in forensic genetics against the illegal hunting of manatees: Trichechus manatus, Trichechus inunguis, Trichechus senegalensis, and Dugong dugon (Eutheria: Sirenia)

The identification of mitochondrial DNA polymorphisms is one of the most efficient methods for species differentiation. Genotyping of molecular markers using PCR/RFLP is a reliable, sensitive and inexpensive method for the detection of species specific mutations. The major causes of decline in Sirenia populations are accidental and intentional catches, collisions with boats and habitat loss. The goal of the present study was to identify, in silico, nucleotide mutations in the cytochrome b gene that can be used for the future development of forensic tools capable of using small tissue fragments to discriminate manatee meat from domesticated species meat commonly used as food sources (bovine, ovine, caprine and swine). DNA sequence alignments revealed two polymorphic sites distinguishing the manatee species from domestic species. The present study reinforced the power of cytochrome polymorphisms as powerful markers for species identification, which may be particularly useful for identifying vulnerable/endangered species. The data provided herein also suggest such mtDNA markers as important conservation tools for combating predatory manatee hunting for illegal meat trade in the Americas

Single nucleotide polymorphisms from cytochrome b gene as a useful protocol in forensic genetics against the illegal hunting of manatees: Trichechus manatus, Trichechus inunguis, Trichechus senegalensis, and Dugong dugon (Eutheria: Sirenia)

The identification of mitochondrial DNA polymorphisms is one of the most efficient methods for species differentiation. Genotyping of molecular markers using PCR/RFLP is a reliable, sensitive and inexpensive method for the detection of species specific mutations. The major causes of decline in Sirenia populations are accidental and intentional catches, collisions with boats and habitat loss. The goal of the present study was to identify, in silico, nucleotide mutations in the cytochrome b gene that can be used for the future development of forensic tools capable of using small tissue fragments to discriminate manatee meat from domesticated species meat commonly used as food sources (bovine, ovine, caprine and swine). DNA sequence alignments revealed two polymorphic sites distinguishing the manatee species from domestic species. The present study reinforced the power of cytochrome polymorphisms as powerful markers for species identification, which may be particularly useful for identifying vulnerable/endangered species. The data provided herein also suggest such mtDNA markers as important conservation tools for combating predatory manatee hunting for illegal meat trade in the Americas

Single nucleotide polymorphisms from cytochrome b gene as a useful protocol in forensic genetics against the illegal hunting of manatees: Trichechus manatus, Trichechus inunguis, Trichechus senegalensis, and Dugong dugon (Eutheria: Sirenia)

The identification of mitochondrial DNA polymorphisms is one of the most efficient methods for species differentiation. Genotyping of molecular markers using PCR/RFLP is a reliable, sensitive and inexpensive method for the detection of species specific mutations. The major causes of decline in Sirenia populations are accidental and intentional catches, collisions with boats and habitat loss. The goal of the present study was to identify, in silico, nucleotide mutations in the cytochrome b gene that can be used for the future development of forensic tools capable of using small tissue fragments to discriminate manatee meat from domesticated species meat commonly used as food sources (bovine, ovine, caprine and swine). DNA sequence alignments revealed two polymorphic sites distinguishing the manatee species from domestic species. The present study reinforced the power of cytochrome polymorphisms as powerful markers for species identification, which may be particularly useful for identifying vulnerable/endangered species. The data provided herein also suggest such mtDNA markers as important conservation tools for combating predatory manatee hunting for illegal meat trade in the Americas

Morphology, morphometry and ultrastructure of the Amazonian manatee (Sirenia: Trichechidae) spermatozoa

This study describes the morphological, morphometric and ultrastructural characteristics of the Amazonian manatee Trichechus inunguis (Natterer, 1883) spermatozoon. The spermatozoa were obtained from a urine sample of an adult T. inunguis kept in captivity. The spermatozoa were analyzed by light and transmission electron microscopy. The head of Amazonian manatee spermatozoa had a flat oval shape and a well distinguishable midpiece. The mean dimensions of the spermatozoa were: head length, 7.49 ± 0.24 µm; head width, 3.53 ± 0.19 µm; head thickness, 1.61 ± 0.13 µm; midpiece length, 11.36 ± 0.34 µm; flagellum length, 40.91 ± 1.94 µm; total tail length, 52.16 ± 1.06 µm; total spermatozoon length, 60.08 ± 1.40 µm. The Amazonian manatee spermatozoa were similar in shape to other sirenian spermatozoa; however, presenting a different size. This study describes, for the first time, the morphometric and ultrastructural characteristics of the Amazonian manatee spermatozoa, and also demonstrates the possible use of spermatozoa retrieved from urine samples for biological studies.

Morphology, morphometry and ultrastructure of the Amazonian manatee (Sirenia: Trichechidae) spermatozoa

This study describes the morphological, morphometric and ultrastructural characteristics of the Amazonian manatee Trichechus inunguis (Natterer, 1883) spermatozoon. The spermatozoa were obtained from a urine sample of an adult T. inunguis kept in captivity. The spermatozoa were analyzed by light and transmission electron microscopy. The head of Amazonian manatee spermatozoa had a flat oval shape and a well distinguishable midpiece. The mean dimensions of the spermatozoa were: head length, 7.49 ± 0.24 µm; head width, 3.53 ± 0.19 µm; head thickness, 1.61 ± 0.13 µm; midpiece length, 11.36 ± 0.34 µm; flagellum length, 40.91 ± 1.94 µm; total tail length, 52.16 ± 1.06 µm; total spermatozoon length, 60.08 ± 1.40 µm. The Amazonian manatee spermatozoa were similar in shape to other sirenian spermatozoa; however, presenting a different size. This study describes, for the first time, the morphometric and ultrastructural characteristics of the Amazonian manatee spermatozoa, and also demonstrates the possible use of spermatozoa retrieved from urine samples for biological studies.

Morphology, morphometry and ultrastructure of the Amazonian manatee (Sirenia: Trichechidae) spermatozoa

This study describes the morphological, morphometric and ultrastructural characteristics of the Amazonian manatee Trichechus inunguis (Natterer, 1883) spermatozoon. The spermatozoa were obtained from a urine sample of an adult T. inunguis kept in captivity. The spermatozoa were analyzed by light and transmission electron microscopy. The head of Amazonian manatee spermatozoa had a flat oval shape and a well distinguishable midpiece. The mean dimensions of the spermatozoa were: head length, 7.49 ± 0.24 µm; head width, 3.53 ± 0.19 µm; head thickness, 1.61 ± 0.13 µm; midpiece length, 11.36 ± 0.34 µm; flagellum length, 40.91 ± 1.94 µm; total tail length, 52.16 ± 1.06 µm; total spermatozoon length, 60.08 ± 1.40 µm. The Amazonian manatee spermatozoa were similar in shape to other sirenian spermatozoa; however, presenting a different size. This study describes, for the first time, the morphometric and ultrastructural characteristics of the Amazonian manatee spermatozoa, and also demonstrates the possible use of spermatozoa retrieved from urine samples for biological studies.

Urinary parameters of Trichechus inunguis (Mammalia, Sirenia): reference values for the Amazonian Manatee

The Amazonian manatee, Trichechus inunguis (Natterer 1883) is endemic to the Amazon Basin and is currently considered a vulnerable species. In order to establish normality ranges of urinary parameters to help monitor the health of this species in captivity, chemical urinalyses were performed on twelve males and nine females of various age groups. Urine was collected once a month for twelve months in the tanks just after being drained, by placing stainless steel containers under the genital slit of females and applying abdominal massages to males in order to stimulate urination. Quantitative data of glucose, urea, creatinine, uric acid and amylase levels were obtained using colorimetric spectrophotometry. Dip strips were also useful for routine analyses, despite only providing qualitative results. Normal range to glucose levels, regardless of sex or age class, was 3.0 to 3.6 mg.dL-1, coinciding with qualitative values of glucose measured by dip strips. Statistical differences observed in some parameter levels suggest that some urine parameters analysed must take into consideration the sex and the age class of the animal studied, being these differences less remarkable in creatinine and amylase levels. To this last one, statistical difference was detected only in the calve's urine (7.0 to 11.5 mg.dL-1) compared to other age classes samples (4.1 to 5.3 mg.dL-1). The results presented here may be used as comparative data in future research on urinalysis in related species.

O peixe-boi da Amazônia, Trichechus inunguis (Natterer 1883) é endêmico da Bacia Amazônica e é considerado atualmente como espécie vulnerável. Com o objetivo de estabelecer valores de normalidade para os parâmetros urinários com vistas a ajudar no monitoramento da saúde desta espécie em cativeiro, foi realizada urinálise química em doze machos e nove fêmeas de várias classes etárias. A urina era coletada uma vez por mês, durante doze meses, nos tanques, logo após esvaziamento, colocando-se coletores de aço inoxidável sob o orifício genital das fêmeas e aplicando massagem abdominal nos machos para estimular a micção. Dados quantitativos dos níveis de glicose, ureia, creatinina, ácido úrico e amilase foram obtidos por meio de espectrofotometria colorimétrica. O uso de tiras reativas também foi útil para análises de rotina, apesar de somente fornecerem resultados qualitativos. A faixa de normalidade para os níveis de glicose, independente de classe etária e sexo foi de 3,0 a 3,6 mg.dL-1, coincidindo com os valores qualitativos identificados pelas tiras reativas. Diferenças estatísticas observadas nos níveis de alguns parâmetros analisados sugerem que estes devem levar em consideração o sexo e a classe etária do animal estudado. Estas diferenças foram menos marcantes nos níveis de creatinina e de amilase, sendo que para esta última foi detectada diferença estatística apenas nas urinas de filhotes lactentes (7,0 a 11,5 mg.dL-1), em comparação às amostras das demais classes etárias (4,1 a 5,3 mg.dL-1). Os resultados aqui apresentados podem ser usados como dados comparativos em futuras pesquisas sobre urinálise em espécies relacionadas.

AVALIAÇÃO HISTOPATOLÓGICA DE MAMÍFEROS MARINHOS ENCALHADOS NO LITORAL DO NORDESTE BRASILEIRO

O estudo anatomopatológico das carcaças dos mamíferos marinhos é uma ferramenta importante para conhecer a morfofisiologia natural, bem como as alterações morfológicas associadas com algumas doenças e, desta forma, evidenciar os possíveis impactos antropogênicos. O presente trabalho teve como intuito realizar estudos sobre histopatologia dos órgãos de cetáceos e sirênios. Para isso, foram utilizadas amostras pertencentes ao acervo biológico do Projeto Cetáceos da Costa Branca (PCCB), da Associação de Pesquisa e Preservação de Ecossistemas Aquáticos (AQUASIS) e do Centro de Mamíferos Aquáticos (CMA). Os fragmentos teciduais foram submetidos ao método convencional de preparação de lâminas histológicas. As peças passaram pelos processos de desidratação e diafanização, banho em parafina líquida, inclusão em bloco, e posterior corte em micrótomo rotativo. Os cortes, de quatro a cinco micra de espessura, foram colocados em lâminas lapidadas e submetidos ao método de coloração Hematoxilina-eosina (H.E.), sendo então montados utilizando-se lamínula e resina sintética. Considerando que os animais em estudo pertencem a diferentes espécies, faixas etárias e sexo, foi realizada uma análise descritiva geral dos aspectos patológicos encontrados em cada caso. De um total de 237 amostras pertencentes a 66 indivíduos da ordem Cetacea (n=28) e Sirenia (n=38), 34,17% (n=81) apresentaram alterações microscópicas, 54,85% (n=130) estavam normais e 10,97% (n=26) se encontravam autolisadas. Dentre as alterações microscópicas evidentes foi diagnosticado no sistema bucal 50% de estomatite (n=1), o coração apresentou 6,66% de fibrose intersticial (n=2), 10% de hemorragia (n=3) e 6,66% de miocardite (n=2). Os pulmões mostraram em 19% das análises edema pulmonar (n=11), 10% de congestão vascular (n=6), 13% de hemorragia (n=8), 3% de enfisema (n=2), 7% hemossiderose (n=4), 3% pneumonia intersticial (n=2), 2% de pneumonia eosinofílica (n=1), 2% de granuloma parasitário (n=1), 2% de cisto parasitário (n=1), 2% de traqueíte (n=1). No estômago foi visualizado em um sirênio áreas extensas de hemorragia (16,66%; n=1) e no intestino foi detectado hiperplasia nodular linfóide (7%; n=2), enterite (4%; n=1), congestão dos vasos (4%; n=1) e degeneração vacuolar (4%; n=1). Os fígados analisados mostraram 23% de degeneração vacuolar (n=5), 9% de necrose hepatocelular (n=2), 9% de depósito moderado de grânulos de melanina (n=2), 4,54% de hepatite crônica (n=1), 9% de fibrose periportal (n=2), 4,54% de abscesso (n=1), 4,54% de congestão (n=1), 4,54% de bilestase (n=1), e 4,54% de proliferação dos ductos biliares (n=1). Nos baços foi possível constatar 20% de rarefação de polpa branca (n=2), 10% de rarefação de polpa branca e vermelha (n=1), e 30% de deposição moderada de grânulos de melanina (n=3). No pâncreas foi observado uma rarefação das células centroacinosas em 50% (n=1) das amostras. Nos rins foi encontrado 14% de nefrose tubular tóxica (n=4), 7% de hemorragia multifocal (n=2), 14% de congestão vascular (n=4), 3,44% de presença de bactérias (n=1), 3,44% de glomerulonefrite proliferativa (n=1), 3,44% de microabscessos (n=1), 3,44% de edema severo (n=1), 3,44% de cistos renais (n=1) e 3,44% de nefrite intersticial crônica (n=1). Nos úteros avaliados foi observado 25% de calcificação distrófica focal (n=1) e 25% de metrite (n=1). Nos linfonodos foram observados alterações sugestivas de imunossupressão (25%; n=1), hemorragia (25%; n=1), e melanose (50%; n=2). Outros achados incluíram rarefação das células da adrenal (50%, n=1), poucos espermatozoides no lúmen (14,28%; n=2) ou presença apenas de espermatogônias nos túbulos seminíferos (21,42%; n=3), e 25% de hiperplasia focal da bexiga (n=1). Apesar das alterações presentes em diversos sistemas, as patologias pulmonares foram superiores as demais tanto em órgãos de cetáceos como em sirênios encalhados no litoral do nordeste brasileiro.

Pathological examination of marine mammals carcasses is an important tool to understand the natural morphophysiology as well as the morphological changes associated with some diseases and thus highlight the potential anthropogenic impacts. This work had as aim conduct studies on histopathology of the organs of the dead stranded cetaceans and Sirenia in the Brazilian northeast coast, or that died during the rehabilitation process in Specialized Centers of the Region. For this, samples were taken belonging to the biological collection of Projeto Cetáceos da Costa Branca (PCCB), Associação de Pesquisa e Preservação de Ecossistemas Aquáticos (AQUASIS) and from Centro de Mamíferos Aquáticos (CMA). The tissue samples were submitted to the conventional method of preparing histological slides. Whereas the studied animals belong to different species, age and sex, a general descriptive analysis of the pathological aspects found in each case was carried out. From a total of 237 samples belonging to 66 individuals of the Cetacea order (n = 28) and Sirenia (n = 38), 34,17% (n = 81) exhibited microscopic changes, 54,85% (n = 130) were normal and 10,97% (n = 26) were autolysed. Among the obvious microscopic changes was diagnosed 50% of stomatitis in the oral system (n = 1), the heart showed 6.66% of interstitial fibrosis (n = 2), 10% of haemorrhage (n = 3) and 6.66% myocarditis (n = 2). The lungs showed pulmonary edema in 19% of the analyzes (n = 11), 10% of vascular congestion% (n = 6), 13% of haemorrhage (n = 8), 3% of emphysema (n = 2), 7% full of hemosiderin (n = 4), 3% of interstitial pneumonia (n = 2), 2% of eosinophilic pneumonia (n = 1), 2% of parasitic granuloma (n = 1), 2% of parasitic cyst (n = 1), 2% of traqueíte (n = 1). In the stomach was visualized a hemorrhage (16.66%; n = 1) and in the gut was detected nodular lymphoid hyperplasia (7%; n = 2), enteritis (4%; n = 1), blood vessels congestion (4%; n = 1) and vacuolar degeneration (4%; n = 1). The livers analysis showed 23% of vacuolar degeneration (n = 5), 9% of hepatocellular necrosis (n = 2), 9% moderate deposit of melanin granules (n = 2), 4.54 % of chronic hepatitis (n = 1), 9% of periportal fibrosis (n = 2), 4.54% of abscess (n = 1), 4.54% of congestion (n = 1), 4.54 % of bilestase (n = 1), and 4.54% proliferation of the bile ducts (n = 1). It was possible to observe in the spleen 20% white pulp rarefaction (n = 2), 10% white and red pulp rarefaction (n = 1), and 30% moderate deposition of melanin granules (n = 3). In the pancreas was observed a 50% of centroacinosas cells rarefaction (n = 1). In the kidneys was found 14% of toxix tube nephrosis (n = 4), 7% of multifocal hemorrhaging (n = 2), 14% of vascular congestion (n = 4), 3.44% presence of bacteria (n = 1), 3.44% of proliferative glomerulonephritis (n = 1), 3.44% of micro abscesses (n = 1), 3.44% of severe edema (n = 1), 3.44% of renal cysts (n = 1) and 3.44% of chronic interstitial nephritis (n = 1). In the uteri evaluated were observed 25% of a focal dystrophic calcification (n = 1) and 25% of metritis (n = 1). In the lymph nodes were observed suggestive abnormalities of immunosuppression (25%; n = 1), bleeding (25%; n = 1) and melanose (50%; n = 2). Other findings include rarefaction of the adrenal cells (50%, n = 1), fewer sperm cells into the lumen (14,28%; n = 2) or presence of only spermatogonia in the seminiferous tubules (21,42%; n = 3), and 25% focal hyperplasia of the bladder (n = 1). Despite the changes present in many systems, lung diseases were higher in cetaceans bodies and sirenians stranded on the Brazilian northeast coast.

Urinary parameters of Trichechus inunguis (Mammalia, Sirenia): reference values for the Amazonian Manatee / Parâmetros urinários de Trichechus Inunguis (Mammalia, Sirenia): valores-referência para o peixe-boi da Amazônia

The Amazonian manatee, Trichechus inunguis (Natterer 1883) is endemic to the Amazon Basin and is currently considered a vulnerable species. In order to establish normality ranges of urinary parameters to help monitor the health of this species in captivity, chemical urinalyses were performed on twelve males and nine females of various age groups. Urine was collected once a month for twelve months in the tanks just after being drained, by placing stainless steel containers under the genital slit of females and applying abdominal massages to males in order to stimulate urination. Quantitative data of glucose, urea, creatinine, uric acid and amylase levels were obtained using colorimetric spectrophotometry. Dip strips were also useful for routine analyses, despite only providing qualitative results. Normal range to glucose levels, regardless of sex or age class, was 3.0 to 3.6 mg.dL-1, coinciding with qualitative values of glucose measured by dip strips. Statistical differences observed in some parameter levels suggest that some urine parameters analysed must take into consideration the sex and the age class of the animal studied, being these differences less remarkable in creatinine and amylase levels. To this last one, statistical difference was detected only in the calve's urine (7.0 to 11.5 mg.dL-1) compared to other age classes samples (4.1 to 5.3 mg.dL-1). The results presented here may be used as comparative data in future research on urinalysis in related species.(AU)

O peixe-boi da Amazônia, Trichechus inunguis (Natterer 1883) é endêmico da Bacia Amazônica e é considerado atualmente como espécie vulnerável. Com o objetivo de estabelecer valores de normalidade para os parâmetros urinários com vistas a ajudar no monitoramento da saúde desta espécie em cativeiro, foi realizada urinálise química em doze machos e nove fêmeas de várias classes etárias. A urina era coletada uma vez por mês, durante doze meses, nos tanques, logo após esvaziamento, colocando-se coletores de aço inoxidável sob o orifício genital das fêmeas e aplicando massagem abdominal nos machos para estimular a micção. Dados quantitativos dos níveis de glicose, ureia, creatinina, ácido úrico e amilase foram obtidos por meio de espectrofotometria colorimétrica. O uso de tiras reativas também foi útil para análises de rotina, apesar de somente fornecerem resultados qualitativos. A faixa de normalidade para os níveis de glicose, independente de classe etária e sexo foi de 3,0 a 3,6 mg.dL-1, coincidindo com os valores qualitativos identificados pelas tiras reativas. Diferenças estatísticas observadas nos níveis de alguns parâmetros analisados sugerem que estes devem levar em consideração o sexo e a classe etária do animal estudado. Estas diferenças foram menos marcantes nos níveis de creatinina e de amilase, sendo que para esta última foi detectada diferença estatística apenas nas urinas de filhotes lactentes (7,0 a 11,5 mg.dL-1), em comparação às amostras das demais classes etárias (4,1 a 5,3 mg.dL-1). Os resultados aqui apresentados podem ser usados como dados comparativos em futuras pesquisas sobre urinálise em espécies relacionadas.(AU)

Confirmação da relação entre peixes-bois, "hyraxes" e elefantes, por meio do estudo da proteína das lentes dos olhos ()

Summary The phylogenetic relationship between the members of the super-order Paenungulata, the manatees, elephants and hyraces were investigated using amino acid sequence analysis of the oc A crystallin of the eye lenses of these animals. The analysis confirms the monophyletic origin of the orders Hyracoidea, Sirenia and Proboscidea and places the edentates as the oldest offshoot from the main eutherian stem, followed by the Paenungulates as the next branch. The results suggest that the manatee may be closer to the hyrax than to the elephant due to the presence of 72-Leu in the hyrax and the manatee but not in the elephant. Morphological evidence would indicate a closer relationship between the elephant and the manatee which, if true, would require a back-substitution of 72 Leu Val in the elephant oc A chain or two parallel substitutions 72 Val Leu in the hyrax and manatee.

Resumo As relações filogenéticas entre os membros da super-ordem Paenungulata, os peixes-bois, elefantes e "hyraxes", foram investigadas, utilizando-se a seqüência de análises de aminoácidos da proteína oc A do cristalino da lente dos olhos destes animais. As análises confirmam a origem monofilética das ordens Hyracoidea, Sirenia e Proboscidea e colocam os edentatas como os mais antigos do ramo da linha principal dos euterianos, seguidos pelos Paenungulatas como ramo mais próximo. Os resultados sugerem que os peixes-bois podem estar mais próximos dos hyraxes que dos elefantes dada a presença dos 72-Leu nos hyraxes e peixes-bois, mas não nos elefantes. Evidências morfológicas indicariam uma relação mais próxima entre o elefante e peixe-boi, as quais, se verdadeira, exigirão uma retrossubstituição do 72-Leu Val na cadeia oc A dos elefantes ou 2 substituições paralelas na 72-Val Leu nos hyraxes e peixes-bois.

Confirmação da relação entre peixes-bois, "hyraxes" e elefantes, por meio do estudo da proteína das lentes dos olhos ()

Summary The phylogenetic relationship between the members of the super-order Paenungulata, the manatees, elephants and hyraces were investigated using amino acid sequence analysis of the oc A crystallin of the eye lenses of these animals. The analysis confirms the monophyletic origin of the orders Hyracoidea, Sirenia and Proboscidea and places the edentates as the oldest offshoot from the main eutherian stem, followed by the Paenungulates as the next branch. The results suggest that the manatee may be closer to the hyrax than to the elephant due to the presence of 72-Leu in the hyrax and the manatee but not in the elephant. Morphological evidence would indicate a closer relationship between the elephant and the manatee which, if true, would require a back-substitution of 72 Leu Val in the elephant oc A chain or two parallel substitutions 72 Val Leu in the hyrax and manatee.

Resumo As relações filogenéticas entre os membros da super-ordem Paenungulata, os peixes-bois, elefantes e "hyraxes", foram investigadas, utilizando-se a seqüência de análises de aminoácidos da proteína oc A do cristalino da lente dos olhos destes animais. As análises confirmam a origem monofilética das ordens Hyracoidea, Sirenia e Proboscidea e colocam os edentatas como os mais antigos do ramo da linha principal dos euterianos, seguidos pelos Paenungulatas como ramo mais próximo. Os resultados sugerem que os peixes-bois podem estar mais próximos dos hyraxes que dos elefantes dada a presença dos 72-Leu nos hyraxes e peixes-bois, mas não nos elefantes. Evidências morfológicas indicariam uma relação mais próxima entre o elefante e peixe-boi, as quais, se verdadeira, exigirão uma retrossubstituição do 72-Leu Val na cadeia oc A dos elefantes ou 2 substituições paralelas na 72-Val Leu nos hyraxes e peixes-bois.