Breathing Air and Living Underwater: Molecular Evolution of Genes Related to Antioxidant Response in Cetaceans and Pinnipeds.

Cetaceans and pinnipeds are lineages of mammals that have independently returned to the aquatic environment, acquiring varying degrees of dependence on it while sharing adaptations for underwater living. Here, we focused on one critical adaptation from both groups, their ability to withstand the ischemia and reperfusion experienced during apnea diving, which can lead to the production of reactive oxygen species (ROS) and subsequent oxidative damage. Previous studies have shown that cetaceans and pinnipeds possess efficient antioxidant enzymes that protect against ROS. In this study, we investigated the molecular evolution of key antioxidant enzyme genes (CAT, GPX3, GSR, PRDX1, PRDX3, and SOD1) and the ROS-producing gene XDH, in cetaceans and pinnipeds lineages. We used the ratio of non-synonymous (dN) to synonymous (dS) substitutions as a measure to identify signatures of adaptive molecular evolution in these genes within and between the two lineages. Additionally, we performed protein modeling and variant impact analyzes to assess the functional consequences of observed mutations. Our findings revealed distinct selective regimes between aquatic and terrestrial mammals in five of the examined genes, including divergences within cetacean and pinniped lineages, between ancestral and recent lineages and between crowns groups. We identified specific sites under positive selection unique to Cetacea and Pinnipedia, with one site showing evidence of convergent evolution in species known for their long and deep-diving capacities. Notably, many sites under adaptive selection exhibited radical changes in amino acid properties, with some being damaging mutations in human variations, but with no apparent detrimental impacts on aquatic mammals. In conclusion, our study provides insights into the adaptive changes that have occurred in the antioxidant systems of aquatic mammals throughout their evolutionary history. We observed both distinctive features within each group of Cetacea and Pinnipedia and instances of convergence. These findings highlight the dynamic nature of the antioxidant system in response to challenges of the aquatic environment and provide a foundation for further investigations into the molecular mechanisms underlying these adaptations.

Establishment and Characterization of a Primary Fibroblast Cell Culture from the Amazonian Manatee (Trichechus inunguis).

The vulnerable status of the Amazon manatee, Trichechus inunguis, indicates the need to seek measures to guarantee its conservation. In this context, the cultivation of cells in vitro is a strategy that should at least guarantee the preservation of their genetic material. Thus, we established for the first time a primary culture of Amazonian manatee fibroblasts (TINsf) from a skin biopsy of a young male. Karyotypic analysis of the 3rd, 7th, and 12th passages confirmed the taxonomic identity of the species T. inunguis (2n = 56/NF = 92) and indicated that this culture presents genomic stability. Gene and protein expression of vimentin at the 13th passage show the predominant presence of fibroblasts in TINsf. To test the cell line's responsiveness to materials and demonstrate a possible application of this culture, it was exposed to andiroba seed oil (ASO), and its viability and proliferative capacity were evaluated. ASO demonstrated toxic effects at the highest concentrations and longest exposure times tested, reproducing results observed in human cultures, indicating the applicability of TINsf in toxicological and biotechnological studies. After cryopreservation, the TINsf line maintained its proliferative potential, indicating the establishment of a new culture available for future studies.

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

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

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...

Ontogeny of the skull of Sotalia guianensis (Cetartiodactyla: Delphinidae) on the Northern coast of Brazil

This study aimed to investigate the ontogenetic development of the skull in the Guiana dolphin (Sotalia guianensis Van Bénéden, 1864 ) through the comparison of 29 morphological non-metrical characters together with the estimated age of 54 specimens derived from four areas on the northern coast of Brazil. The state of each character was associated with the age of each specimen to assess the formation of bone structures, and the degree of fusion of the sutures in each state of the character studied. Nineteen qualitative morphological characters presenting state ontogenetic variations have been analyzed statistically. The analysis depicted that the characters associated with the parietal-frontal suture, parietosupraoccipital, parietoexoccipital, supraoccipital-exoccipital and basioccipital-pterygoid varied according with the age. The results suggest that the physical maturity of the cranium was more premature in the specimens from the Northern Coast, suggesting a geographical adjustment of the species, or a biological response to an intense pressure of by-catch in fisheries over decades on the populations.

O objetivo do estudo foi investigar o desenvolvimento ontogenético craniano do boto-cinza (Sotalia guianensis Van Bénéden, 1864), comparando 29 caracteres morfológicos não métricos aliados às estimativas de idade de 54 espécimes provenientes de quatro áreas da Costa Norte do Brasil. Cada estado de caráter foi associado à idade de cada espécime para avaliar a formação de estruturas ósseas e o grau de fusionamento das suturas em cada estado de caráter analisado. Foram analisados estatisticamente 19 caracteres morfológicos qualitativos que apresentaram variação nos estados de caráter. A análise revelou que os caracteres que diferiram de acordo com a idade foram aqueles ligados à sutura parietal-frontal, parietal-supraoccipital, parietal-exoccipital, supraoccipital-exoccipital e basioccipital-pterigóide. Os resultados indicaram que a maturidade física do crânio foi mais precoce nos espécimes da Costa Norte, sugerindo uma estruturação geográfica da espécie ou uma resposta biológica a uma intensa pressão de captura acidental em redes de pesca sobre as populações.

Ontogeny of the skull of Sotalia guianensis (Cetartiodactyla: Delphinidae) on the Northern coast of Brazil

ABSTRACT This study aimed to investigate the ontogenetic development of the skull in the Guiana dolphin (Sotalia guianensis Van Bénéden, 1864 ) through the comparison of 29 morphological non-metrical characters together with the estimated age of 54 specimens derived from four areas on the northern coast of Brazil. The state of each character was associated with the age of each specimen to assess the formation of bone structures, and the degree of fusion of the sutures in each state of the character studied. Nineteen qualitative morphological characters presenting state ontogenetic variations have been analyzed statistically. The analysis depicted that the characters associated with the parietal-frontal suture, parietosupraoccipital, parietoexoccipital, supraoccipital-exoccipital and basioccipital-pterygoid varied according with the age. The results suggest that the physical maturity of the cranium was more premature in the specimens from the Northern Coast, suggesting a geographical adjustment of the species, or a biological response to an intense pressure of by-catch in fisheries over decades on the populations.

RESUMO O objetivo do estudo foi investigar o desenvolvimento ontogenético craniano do boto-cinza (Sotalia guianensis Van Bénéden, 1864), comparando 29 caracteres morfológicos não métricos aliados às estimativas de idade de 54 espécimes provenientes de quatro áreas da Costa Norte do Brasil. Cada estado de caráter foi associado à idade de cada espécime para avaliar a formação de estruturas ósseas e o grau de fusionamento das suturas em cada estado de caráter analisado. Foram analisados estatisticamente 19 caracteres morfológicos qualitativos que apresentaram variação nos estados de caráter. A análise revelou que os caracteres que diferiram de acordo com a idade foram aqueles ligados à sutura parietal-frontal, parietal-supraoccipital, parietal-exoccipital, supraoccipital-exoccipital e basioccipital-pterigóide. Os resultados indicaram que a maturidade física do crânio foi mais precoce nos espécimes da Costa Norte, sugerindo uma estruturação geográfica da espécie ou uma resposta biológica a uma intensa pressão de captura acidental em redes de pesca sobre as populações.

Ontogeny of the skull of Sotalia guianensis (Cetartiodactyla: Delphinidae) on the Northern coast of Brazil

This study aimed to investigate the ontogenetic development of the skull in the Guiana dolphin (Sotalia guianensis Van Bénéden, 1864 ) through the comparison of 29 morphological non-metrical characters together with the estimated age of 54 specimens derived from four areas on the northern coast of Brazil. The state of each character was associated with the age of each specimen to assess the formation of bone structures, and the degree of fusion of the sutures in each state of the character studied. Nineteen qualitative morphological characters presenting state ontogenetic variations have been analyzed statistically. The analysis depicted that the characters associated with the parietal-frontal suture, parietosupraoccipital, parietoexoccipital, supraoccipital-exoccipital and basioccipital-pterygoid varied according with the age. The results suggest that the physical maturity of the cranium was more premature in the specimens from the Northern Coast, suggesting a geographical adjustment of the species, or a biological response to an intense pressure of by-catch in fisheries over decades on the populations.(AU)

O objetivo do estudo foi investigar o desenvolvimento ontogenético craniano do boto-cinza (Sotalia guianensis Van Bénéden, 1864), comparando 29 caracteres morfológicos não métricos aliados às estimativas de idade de 54 espécimes provenientes de quatro áreas da Costa Norte do Brasil. Cada estado de caráter foi associado à idade de cada espécime para avaliar a formação de estruturas ósseas e o grau de fusionamento das suturas em cada estado de caráter analisado. Foram analisados estatisticamente 19 caracteres morfológicos qualitativos que apresentaram variação nos estados de caráter. A análise revelou que os caracteres que diferiram de acordo com a idade foram aqueles ligados à sutura parietal-frontal, parietal-supraoccipital, parietal-exoccipital, supraoccipital-exoccipital e basioccipital-pterigóide. Os resultados indicaram que a maturidade física do crânio foi mais precoce nos espécimes da Costa Norte, sugerindo uma estruturação geográfica da espécie ou uma resposta biológica a uma intensa pressão de captura acidental em redes de pesca sobre as populações.(AU)

Survey of Brucella spp. and Leptospira spp. antibodies in cetaceans and manatees of the Amazon basin and Atlantic Ocean, Brazil.

Aquatic mammals can act as sentinels of emerging and resurging pathogens in the environment. Brucella spp. and Leptospira spp. are 2 zoonotic pathogens relevant to aquatic mammals, and their detection can be used to assess pathogen exposure. In this study, serum from 84 individuals-63 cetaceans (families Iniidae, n = 37; Delphinidae, n = 22; and Kogiidae, n = 4) and 21 West Indian manatees Trichechus manatus-was tested by the Rose Bengal Test (RBT) and a commercial competitive enzyme-linked immunosorbent assay (c-ELISA) for detecting Brucella spp. antibodies, and the microscopic agglutination test (MAT) for screening Leptospira spp. exposure. Overall, 4.8% (3/63) of cetaceans were positive by RBT and 15.9% (10/63) by c-ELISA for Brucella spp. Serum from 8 c-ELISA positive cetaceans (with available serum) was further tested via serum agglutination test (SAT) and 1 individual was positive. c-ELISA was more sensitive than RBT. Exposure to Brucella spp. was found in 5 cetacean species: Clymene dolphin Stenella clymene, short-finned pilot whale Globicephala macrorhynchus, pygmy killer whale Feresa attenuata, melon-headed whale Peponocephala electra and Atlantic bottlenose dolphin Tursiops truncatus in the Atlantic Ocean, Brazil, expanding the range of known Brucella seropositive aquatic hosts. No evidence of Brucella spp. exposure was found in Iniidae and Kogiidae odontocetes and manatees. Antibodies against Leptospira spp. were not detected in cetaceans and sirenians by MAT. These results contribute to the evaluation of different Brucella spp. serological methods in cetaceans and manatees and highlight the epidemiology of zoonotic pathogens in aquatic mammals of the southwestern Atlantic Ocean and the Amazon basin.

Cryptosporidium spp. and Giardia sp. in aquatic mammals in northern and northeastern Brazil.

Cryptosporidium and Giardia are protozoans that can infect humans and wild and domestic animals. Due to the growing importance of diseases caused by protozoan parasites in aquatic species, we aimed to evaluate the frequency of infection by Cryptosporidium spp. and Giardia sp. in aquatic and marine mammals in the northern and northeastern regions of Brazil. We collected 553 fecal samples from 15 species of wild-ranging and captive aquatic mammals in northern and northeastern Brazil. All samples were analyzed by the Kinyoun technique for identification of Cryptosporidium spp. oocysts. Giardia sp. cysts were identified by means of the centrifugal-flotation technique in zinc sulfate solution. Subsequently, all samples were submitted for direct immunofluorescence testing. The overall frequency of infection was 15.55% (86/553) for Cryptosporidium spp. and 9.04% (50/553) for Giardia sp. The presence of Cryptosporidium spp. was detected in samples from 5 species: neotropical river otter Lontra longicaudis (15.28%), giant otter Pteronura brasiliensis (41.66%), Guiana dolphin Sotalia guianensis (9.67%), Amazonian manatee Trichechus inunguis (16.03%), and Antillean manatee T. manatus (13.79%). Giardia sp. was identified in L. longicaudis (9.23%), P. brasiliensis (29.16%), pygmy sperm whale Kogia breviceps (100%), dwarf sperm whale K. sima (25%), S. guianensis (9.67%), T. inunguis (3.81%), and T. manatus (10.34%). This is the first report of Cryptosporidium spp. in L. longicaudis, P. brasiliensis, and S. guianensis, while the occurrence of Giardia sp., in addition to the 2 otter species, was also identified in manatees, thus extending the number of hosts susceptible to these parasitic agents.

Morphology of the eyeball from the Humpback whale (Megaptera novaeangliae).

Aquatic mammals underwent morphological and physiological adaptations due to the transition from terrestrial to aquatic environment. One of the morphological changes regards their vision since cetaceans' eyes are able to withstand mechanical, chemical, osmotic, and optical water conditions. Due to insufficient information about these animals, especially regarding their sense organs, this study aimed to describe the morphology of the Humpback whale (Megaptera novaeangliae) eyeball. Three newborn females, stranded dead on the coast of Sergipe and Bahia, Brazil, were used. Samples were fixed in a 10% formalin solution, dissected, photographed, collected, and evaluated through light and electron microscopy techniques. The Humpback whale sclera was thick and had an irregular surface with mechanoreceptors in its lamina propria. Lens was dense, transparent, and ellipsoidal, consisting of three layers, and the vascularized choroid contains melanocytes, mechanoreceptors, and a fibrous tapetum lucidum. The Humpback whale eyeball is similar to other cetaceans and suggests an adaptation to diving and migration, contributing to the perception of differences in temperature, pressure, and lighting.

Enfermidades e impactos antrópicos em cetáceos no Brasil / Diseases and human impacts in cetaceans in Brazil / Enfermedades e impactos antrópicos en cetáceos de Brasil

Pouco se conhece sobre as enfermidades e a influência dos impactos antrópicos na saúde dos cetáceos que habitam a costa brasileira. Com o objetivo de avaliar o conhecimento atual sobre as enfermidades em cetáceos no Brasil, esta revisão reúne e acrescenta informações sobre doenças infecciosas e parasitárias, entre outras. Há registro de lesões semelhantes às causadas por poxvírus e papilomavírus, e foram detectados anticorpos para Morbillivirus sp. Bactérias como Aeromonas sobria, Pseudomonas aeruginosa e Pasteurella pneumotropica, o fungo Lacazia loboi, os protozoários Giardia sp e Toxoplasma gondii e diversos metazoários foram associados a lesões. Impactos antrópicos também influenciam nas taxas de mortalidade desses animais. O estado de conservação das carcaças e a ausência de patologistas capacitados nas equipes de pesquisa dificultam a avaliação do estado de saúde dos cetáceos.(AU)

Little is known about the diseases and how human impact influences the health of cetaceans that inhabit the Brazilian coast. This review gathers and adds information about infectious and parasitic diseases, among others, in order to evaluate the current knowledge about the major diseases that affect cetaceans in Brazil. There are reports of lesions similar to the ones caused by pox- and papillomavirus, and antibodies against Morbillivirus sp have been detected. Bacteria such as Aeromonas sobria, Pseudomonas aeruginosa and Pasteurella pneumotropica, the fungus Lacazia loboi, the protozoa Giardia sp and Toxoplasma gondii, as well as several metazoans have been associated with lesions. The anthropic impact also influences cetacean mortality rate. The state of decomposition and lack of trained pathologists in research teams make it difficult to assess the health status of these animals.(AU)

Es poco lo que se conoce sobre las enfermedades y la influencia de los impactos antrópicos en la salud de los cetáceos que habitan la costa brasilera. Con el objetivo de analizar el conocimiento actual sobre las enfermedades de los cetáceos en Brasil, la presente revisión reúne y adiciona informaciones de varias enfermedades, entre ellas aquellas de origen infeccioso y las parasitarias. Existen registros de lesiones parecidas a las ocasionadas por poxvirus y papilomavirus, y también fueron detectados anticuerpos contra Morbillivirus sp. Ciertas bacterias como Aeromonas sobria, Pseudomona aeruginosa y Pasteurella pneumotropica, el hongo Lacazia loboi, protozoarios como Giardia sp y Toxoplasma gondii, y diversos metoazoarios, han sido asociados a lesiones en cetáceos. Los impactos antrópicos también influyen en las tasas de mortalidad de estos animales. El mal estado de conservación de los cadáveres y la ausencia de patólogos capacitados en los equipos de investigación, dificultan la evaluación del estado de salud de los cetáceos.(AU)

Enfermidades e impactos antrópicos em cetáceos no Brasil / Diseases and human impacts in cetaceans in Brazil / Enfermedades e impactos antrópicos en cetáceos de Brasil

Pouco se conhece sobre as enfermidades e a influência dos impactos antrópicos na saúde dos cetáceos que habitam a costa brasileira. Com o objetivo de avaliar o conhecimento atual sobre as enfermidades em cetáceos no Brasil, esta revisão reúne e acrescenta informações sobre doenças infecciosas e parasitárias, entre outras. Há registro de lesões semelhantes às causadas por poxvírus e papilomavírus, e foram detectados anticorpos para Morbillivirus sp. Bactérias como Aeromonas sobria, Pseudomonas aeruginosa e Pasteurella pneumotropica, o fungo Lacazia loboi, os protozoários Giardia sp e Toxoplasma gondii e diversos metazoários foram associados a lesões. Impactos antrópicos também influenciam nas taxas de mortalidade desses animais. O estado de conservação das carcaças e a ausência de patologistas capacitados nas equipes de pesquisa dificultam a avaliação do estado de saúde dos cetáceos.

Little is known about the diseases and how human impact influences the health of cetaceans that inhabit the Brazilian coast. This review gathers and adds information about infectious and parasitic diseases, among others, in order to evaluate the current knowledge about the major diseases that affect cetaceans in Brazil. There are reports of lesions similar to the ones caused by pox- and papillomavirus, and antibodies against Morbillivirus sp have been detected. Bacteria such as Aeromonas sobria, Pseudomonas aeruginosa and Pasteurella pneumotropica, the fungus Lacazia loboi, the protozoa Giardia sp and Toxoplasma gondii, as well as several metazoans have been associated with lesions. The anthropic impact also influences cetacean mortality rate. The state of decomposition and lack of trained pathologists in research teams make it difficult to assess the health status of these animals.

Es poco lo que se conoce sobre las enfermedades y la influencia de los impactos antrópicos en la salud de los cetáceos que habitan la costa brasilera. Con el objetivo de analizar el conocimiento actual sobre las enfermedades de los cetáceos en Brasil, la presente revisión reúne y adiciona informaciones de varias enfermedades, entre ellas aquellas de origen infeccioso y las parasitarias. Existen registros de lesiones parecidas a las ocasionadas por poxvirus y papilomavirus, y también fueron detectados anticuerpos contra Morbillivirus sp. Ciertas bacterias como Aeromonas sobria, Pseudomona aeruginosa y Pasteurella pneumotropica, el hongo Lacazia loboi, protozoarios como Giardia sp y Toxoplasma gondii, y diversos metoazoarios, han sido asociados a lesiones en cetáceos. Los impactos antrópicos también influyen en las tasas de mortalidad de estos animales. El mal estado de conservación de los cadáveres y la ausencia de patólogos capacitados en los equipos de investigación, dificultan la evaluación del estado de salud de los cetáceos.

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.

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.

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.

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)

Ocorrência de infecção Cryptosporidium spp. em peixe-boi marinho (Trichechus manatus) / Occurrence of Cryptosporidium spp. infection in antillean manatee (Trichechus manatus)

A criptosporidiose constitui-se como uma zoonose que pode afetar o homem e uma ampla variedade de animais domésticos e silvestres, principalmente indivíduos imunodeficientes. O objetivo desse trabalho foi registrar a ocorrência de infecção por Cryptosporidium em peixe-boi marinho. Após ser constatada a mudança de comportamento de um peixe-boi marinho mantido nos oceanários do Centro Mamíferos Aquáticos, ICMBio - FMA, animal foi submetido à exame clínico e, posteriormente, à coleta de amostra fecal. As amostras fecais foram analisadas pela técnica de Kinyoun, teste de imunofluorescência direta e pelo corante 4'.6'-Diamidino-2-Phenilindole (DAPI). No exame clínico, o animal apresentou sinais de desconforto abdominal. Os resultados obtidos nas análises de microscopia de luz e fluorescente revelaram a presença de oocistos de Cryptosporidium nas fezes desse peixe-boi.

Cryptosporidiosis is a zoonosis which can affect man and a wide range of domestic and wild animals, mainly immunodeficient individuals. The objective of this paper was reported the occurrence of a Cryptosporidium infection in Antillean manatee. After an unusual behavior of an Antillean manatee kept in captivity at the Centro Mamíferos Aquáticos, ICMBio - FMA, clinical examination and posterior fecal sampling was performed. Fecal samples were examined by the Kinyoun technique, Direct Immunofluorescence Test and also examined by 4'.6'-Diamidino-2-Phenylindole (DAPI) staining. At the clinical examination, the animal showed signs of abdominal pain. The results obtained by light and fluorescence microscopy analysis showed the presence of Cryptosporidium spp. oocyst in feces of this manatee.

Ocorrência de infecção Cryptosporidium spp. em peixe-boi marinho (Trichechus manatus)

Cryptosporidiosis is a zoonosis which can affect man and a wide range of domestic and wild animals, mainly immunodeficient individuals. The objective of this paper was reported the occurrence of a Cryptosporidium infection in Antillean manatee. After an unusual behavior of an Antillean manatee kept in captivity at the Centro Mamíferos Aquáticos, ICMBio - FMA, clinical examination and posterior fecal sampling was performed. Fecal samples were examined by the Kinyoun technique, Direct Immunofluorescence Test and also examined by 4'.6'-Diamidino-2-Phenylindole (DAPI) staining. At the clinical examination, the animal showed signs of abdominal pain. The results obtained by light and fluorescence microscopy analysis showed the presence of Cryptosporidium spp. oocyst in feces of this manatee.

A criptosporidiose constitui-se como uma zoonose que pode afetar o homem e uma ampla variedade de animais domésticos e silvestres, principalmente indivíduos imunodeficientes. O objetivo desse trabalho foi registrar a ocorrência de infecção por Cryptosporidium em peixe-boi marinho. Após ser constatada a mudança de comportamento de um peixe-boi marinho mantido nos oceanários do Centro Mamíferos Aquáticos, ICMBio - FMA, animal foi submetido à exame clínico e, posteriormente, à coleta de amostra fecal. As amostras fecais foram analisadas pela técnica de Kinyoun, teste de imunofluorescência direta e pelo corante 4'.6'-Diamidino-2-Phenilindole (DAPI). No exame clínico, o animal apresentou sinais de desconforto abdominal. Os resultados obtidos nas análises de microscopia de luz e fluorescente revelaram a presença de oocistos de Cryptosporidium nas fezes desse peixe-boi.

Embarcações motorizadas: uma ameaça aos peixes-boi marinhos (Trichechus manatus) no Brasil / Motorboat: a threat to Antillean manatees (Trichechus manatus) in Brazil

O aumento do tráfego de embarcações motorizadas em áreas utilizadas para alimentação, descanso e reprodução pelos peixes-boi marinhos tem não só afugentado estes animais, mas resultado em potenciais casos de colisões. Este trabalho tem como objetivo relatar casos de interações entre embarcações motorizadas e peixes-boi marinhos (Trichechus manatus) no litoral Nordeste do Brasil. As interações ocorreram nos Estados do Ceará, Paraíba e Sergipe com três espécimes, sendo um destes com quatro registros. Foram observados transtornos clínicos como edema da região frontal, proptose ocular, emagrecimento e múltiplos cortes ao longo do corpo. Sendo assim, o aumento do tráfego de embarcações, e a falta de normativas e de fiscalização que assegurem a efetiva proteção de determinadas áreas, representam um risco direto para a conservação dos peixes-boi marinhos no litoral Nordeste do Brasil.

The increasing traffic of motorboats on feeding, resting and reproduction areas used by manatees causing serious problems for these animals, so scaring them away and leading to potential collisions. The purpose of this study was to describe cases interaction between boat and Antillean manatees (Trichechus manatus) living in the Northeastern coast of Brazil. These interactions took place in the State of Ceará, Paraíba and Sergipe, involving three specimens, one of them with four records. Clinical alterations were observed, such as edema in the frontal part of the body, ocular proptosis, weight loss and multiple cuts all over the body. Therefore, the increasing traffic of motorboats, allied to the lack of normative rules and effective law enforcement assuring the protection of specific areas, represent a direct risk to the conservation of Antillean manatees living in the Northeastern coast of Brazil.

Cytogenetic study of the giant otter Pteronura brasiliensis Zimmermann 1780 (Carnivora, Mustelidae, Lutrinae)

The giant otter, Pteronura brasiliensis Zimmermann 1780 (Carnivora, Mustelidae, Lutrinae), was widely distributed in South America but stable populations are now only found in the Pantanal and Amazon regions and the species is classified as endangered. There is only one recognized species of giant otter, although two subspecies of doubtful value have also been cited in the literature. We present the first karyotype of four captive P. brasiliensis specimens, all of which posses 2n = 38 chromosomes as 14M+8SM+6ST+8A and one pair of sexual chromosomes. An heteromorphic secondary constriction, associated with the nucleolar organizer region (NOR), was seen on the long arms of chromosome pair 17. The C-banding technique revealed heterochromatin in the centromeric region of all the chromosomes and the NOR was C-banding positive. The giant otter presented the same diploid number as most mustelids, although its karyotype is quite species-specific.