Characteristics of Filiform, Fungiform and Vallate Papillae and Surface of Interface Epithelium-Connective Tissue of the Maned Sloth Tongue Mucosa (Bradypus torquatus, Iliger, 1811): Light and Scanning Electron Microscopy Study

The study of lingual surfaces and the surface of interface epithelium-connective tissue of the tongue of Bradypus torquatus was performed by employing the light and scanning electron microscopy (SEM) techniques. The results revealed that the rostral part of the tongue presents a round apex and covered by filiform and fungiform lingual papillae and a ventral smooth surface. It was observed that the epithelial layer of the dorsal surface possesses the basal, spinosum, granular and cornified epithelial cells. The lamina propria is characterized by a dense connective tissue forming the long, short and round papillae. Numerous typical filiform papillae are located especially in the rostral part intermingled for few fungiform papillae, which were revealed in three-dimensional SEM images. Usually, the fungiform papillae are located in the border of rostral apex of the tongue exhibiting the rounded form. They are covered by keratinized epithelial cells. In the fungiform papillae, several taste pores were observed on the surface. The vallate papillae presented numerous taste buds in the wall of epithelial cells, being that the major number of taste buds is located on the superior half of vallate papilla. The taste pores are surrounded by several laminae of keratinized epithelial cells. The samples treated with NaOH solution and examined by SEM revealed, after removal of the epithelial layer, the dense connective core in original disposition, presenting different sizes and shapes. The specimens stained with Picrosirius and examined by polarized light microscopy revealed the connective tissue, indicating the collagen fibres type I and type III.

Collection and evaluation of semen from the three-toed sloth (Bradypus tridactylus)

Sloths (Bradypus sp.) are extremely sensitive animals that suffer with the destruction and fragmentation of forests. They present a low population growth rate and need to be further studied for the preservation of the specie. Thus, the aim of this study was to establish an efficient semen collection protocol as well as characterize sperm concentration, motility and morphology in order to contribute with information about the reproductive traits of this specie, which has never been described in the literature before. For that, nine Bradypus tridactylus males were captured during the wet season and six during the dry season, in Manaus (AM), located in the north region of Brazil, semen was collected by electroejaculation with shocks given in sequences of progressive intensities (minimum 20 mA and maximum 60 mA). All animals ejaculated small volumes of semen and in some of them, the volume ejaculated was not enough for a complete spermiogram. Physical characteristics observed on the collections of the wet season were different from those seen in the specimen collected in the dry season. Motility an vigor was very low and did not show forward progression, only oscillatory movement. After Spermac stain, spermatozoa presented a wide variety of defects; however, the differences in morphology were not significant between seasons. The morphology assessed by scanning electron microscopy shows that the head in both groups could be elongated, short or could have a base narrower than the apex and the midpiece narrowed abruptly, forming a nip in its transition to the tail. Although further studies are necessary to verify our preliminary findings concerning seasonal variation in sperm quality, these results demonstrate that semen can be safely collected from sloths by electroejaculation and provide the first reports of semen characteristics in this species. (C) 2008 Elsevier Ltd. All rights reserved.

Investigações anatomicas sobre os Bradypódidas: o nervo vago no Bradypus tridactylus

I - No Bradypus tridactylus os X, direito e esquerdo, reunem-se em alça pré-esophageana. II - O nivel de formação desta alça variou nos dois casos estudados pelo autor, assim como a disposição dos ramos que della partiam. III - Ambos devem, pois, concorrer á constituição das rêdes anastomoticas terminaes vago-orthosympathicas. IV - Foi impossivel ao autor, mesmo com o auxilio de lentes, individualizar fibras vagaes na continuidade da intricada malha nervosa aortico-visceral, dependente do plexo solar.

Investigações anatomicas sobre os Bradypódidas: o Orthosympatico no Bradypus tridactylus

I - No Bradypus tridactylus foram encontrados 2 ganglios cervicaes e 13 thoracicos. II - O sympathico abdominal é constitutido inicialmente por um ganglio triangular que representa a fuzão do primeiro ganglio catenario lombar e do ganglio semi-lunar classico. Nota-se tambem na cadeia lombar uma massa ganglionar unica. III - A cadeia pelviana é representada por 7 ganglios, gradativamente reduzidos de tamanho. IV - No Bradypus tridactylus os dois ultimos ganglios pelvianos direito e esquerdo se reunem por uma anastomose transversal. V - No Bradypus tridactylus foi encontrada em dois exemplares origem thoracica rudimentar de um unico esplanchnico em cada lado; nos demais exemplares notou-se ausencia de individualisação anatomica dos nervos esplanchnicos. VI - No Bradypus tridactylus além de um ganglio triangular situado latero-ventralmente á aorta e caudalmente á emergencia da arteria celiaca, encontrou-se um ganglio aortico renal e um ganglio hypogastrico. VII - No Bradypus tridactylus verificou-se a existencia dos nervos esplanchnicos pelvianos de Delmas e do plexo inter-iliaco ou pré-sacro.

Sôbre um Nematódeo spirurídeo parasito de Bradypus Tridactylus L

The genus Paraleiuris has a single species P. lochii Vaz & Pereira, 1929. The new species here described is different from it mainly by the aspect of the bucal capsule which is rectangular and its longer spicule differs in size and form.

Caracterització de la població de Bradypus variegatus i Choloepus hoffmanni a INBioparque, Costa Rica

Estudi centrat en la caracterització de la població de peresosos (Choloepus Hoffmanni i Bradypus variegatus) d’INBioparque (Costa Rica) per tal de donar continuïtat al projecte de Casalé i Delclòs, 2010. S’ha determinat en 15 el nombre d’individus amb els que compta el parc i que les seves estructures poblacionals s’han vist alterades degut a la disminució d’individus respecte al 2010. S’han definit diferències significatives en la localització dels individus, essent els més freqüentats els hàbitats de bosc V.Central i bosc Humit, i detectat una gran variabilitat d’espècies arbòries tant ingerides com freqüentades, destacant el gènere Cecròpia. Pel que fa als registres d’activitat, tot hi la gran variabilitat i diferències entre espècies i sexes, és majoritàriament nul·la. També s’ha estudiat la població d’Iguana iguana com a limitadora de la població de peresosos d’INBioparque, sense obtenir uns resultats que permetin assegurar cap tipus de competència interespecífica.

Citologia vaginal de preguiça-de-coleira (Bradypus torquatus)

As preguiças-de-coleira (Bradypus torquatus) são mamíferos arborícolas da família Bradypodidae. Podem ser encontradas nos trechos de Mata Atlântica do Brasil e a maior diversidade genética de suas populações ocorre em matas do sul da Bahia. A observação desses animais na natureza é muito difícil, pois passam a maior parte da vida escondidos no denso emaranhado das copas, por isso, dados sobre aspectos reprodutivos são escassos e não existem informações sobre ciclo estral dessa espécie. Este trabalho teve por objetivo identificar as células do epitélio vaginal da preguiça-de-coleira (Bradypus torquatus) como forma de viabilizar o uso dessa técnica para estudar as fases do ciclo estral desses animais. As amostras para citologia vaginal foram obtidas de quatro preguiças de coleira que habitavam áreas de Mata Atlântica do sul da Bahia. Após captura manual do animal, procedeu-se a coleta de material biológico, introduzindo uma escova ginecológica estéril, na comissura dorsal da vulva. Para cada amostra foram feitos dois esfregaços rotacionando a extremidade da escova sobre cada lâmina de vidro, fazendo-se em geral três impressões lineares. O esfregaço foi imediatamente corado pelo método Panótico rápido (Laborclin®). Nas preguiças BT033, BT065 e BT042 foi possível identificar respectivamente 30%, 33% e 7% de células parabasais (PB); 56%, 22% e 10% de células intermediárias pequenas (IP); 6%, 18% e 6% de células intermediárias grandes (IG); 2%, 13% e 24% de células superficiais nucleadas (SN); 6%, 14% e 53% de células superficiais anucleadas (SA). Na preguiça BT464 foi possível fazer duas coletas com intervalo de 13 meses. Os dados da primeira e segunda coleta foram, respectivamente: 6% e 17,5 de células PB, 5% e 25% de células IP, 11% e 15,5% de células IG, 8% e 19,5% de células SN e 70% e 22,5% de células SA. Enfatiza-se que as porcentagens de células do epitélio vaginal variaram entre indivíduos e também na mesma preguiça. Isto sugere que a citologia vaginal possa ser uma ferramenta de avaliação do ciclo estral em preguiça-de-coleira.

Análise morfológica e morfométrica das gônadas de preguiça (Bradypus variegatus Schinz, 1825)

A preguiça pode ser designada, zoologicamente, como um mamífero da classe Eutheria, ordem Edentata, subordem Xenarthra e família Bradypodidae. O gênero Bradypus é constituído de três espécies distintas: a preguiça-de-bentinho (B. tridactylus), restrita à região amazônica; a preguiça-comum (B. variegatus), de ampla distribuição, ocorrendo nas Américas Central e do Sul e a preguiça-de-coleira (B. torquatus), única seriamente ameaçada de extinção. Percebe-se a necessidade de uma investigação mais aprofundada sobre a espécie B. variegatus, a fim de contribuir de forma efetiva com a elaboração de tratados de anatomia da fauna silvestre, além de buscar mais informações sobre a anatomia do sistema reprodutor do bicho preguiça (Bradypus variegatus Schinz, 1825) e, desta forma, aplicar os novos conhecimentos na sua preservação. Utilizamos quatro indivíduos machos e três fêmeas, pertencentes ao banco de espécies da Área de Anatomia do Departamento de Morfologia e Fisiologia Animal da Universidade Federal Rural de Pernambuco (DMFA/UFRPE), as quais foram dissecados e evidenciamos a vascularização das gônadas, bem como suas localizações e aspectos morfológicos e morfométricos dos testículos e ovários. Como resultados, observou-se que o macho possui testículos localizados no interior do espaço intraabdominal, ligados às glândulas adrenais por um ligamento de peritônio. A vascularização não é feita por um plexo pampiniforme, mas por uma artéria e uma veia testicular. Os testículos possuem, em média, volume igual a 1,42cm³ e espessura, largura e comprimento, respectivamente iguais a 0,98, 1,23 e 1,45cm. Os ovários também estão no interior do espaço intra-abdominal, no fundo do útero, protegidos por uma bolsa ovariana, composta por mesovário e mesossalpinge. A vascularização é realizada por um plexo arteriovenoso que irriga e drena o útero, e suas ramificações na parede uterina atingem os ovários. Os ovários possuem, em média, espessura, largura e comprimento, respectivamente iguais a 0,37, 0,73 e 0,62 cm.

Morfologia e morfometria da traqueia da preguiça (Bradypus variegatus): conhecimentos para procedimentos de emergência

Por ser uma espécie pouco estudada, principalmente do ponto de vista morfológico, objetivou-se conhecer a anatomia da traqueia da preguiça (Bradypus variegatus) a fim de fornecer informações para facilitar a eleição de tubo endotraqueal adequado, máscara laríngea ou cânula de traqueostomia para anestesia e procedimentos de emergência, uma vez que a mesma revelou-se possuidora de uma morfologia especial. Foram investigados 11animais jovens de idades diferentes, sendo quatro machos e sete fêmeas, provenientes do Museu Emilio Goeldi e doados a UFRA. Os exemplares foram perfundidos via intramuscular com solução aquosa de formol a 10% para fins de conservação e posteriormente foram dissecados em nível cervico-torácico, por meio de mesoscopia, expondo-se desde a laringe até os brônquios principais direito e esquerdo ao nível do hilo pulmonar. As traqueias foram divididas em cinco regiões (região cervical, região da primeira flexura, região da segunda flexura, região da terceira flexura e região da carina) as quais foram mensuradas altura e largura, assim como o comprimento traqueal total e parte do material foi submetido à técnicas de rotina histológica. Macroscopicamente, destacou-se a presença de sinuosidades acentuadas em porção médio caudal, contemplando a carina. O comprimento médio traqueal foi de 14,6cm. Microscopicamente, a traqueia era constituída por placas separadas de cartilagem hialina constituindo cada anel, sendo revestido por epitélio estratificado ciliado. Apesar da traqueia da preguiça comum apresentar revestimento padrão encontrado na traqueia de outros animais, na literatura não há registros de outras espécies que tenham morfologia macroscópica nas condições descritas, o que nos leva a sugerir, quando necessário acesso para ventilação de emergência, a prática da IOT e não a de traqueostomia.

Circadian rhythms in blood pressure in free-ranging three-toed sloths (Bradypus variegatus)

Blood pressure (BP) profiles were monitored in nine free-ranging sloths (Bradypus variegatus) by coupling one common carotid artery to a BP telemetry transmitter. Animals moved freely in an isolated and temperature-controlled room (24ºC) with 12/12-h artificial light-dark cycles and behaviors were observed during resting, eating and moving. Systolic (SBP) and diastolic (DBP) blood pressures were sampled for 1 min every 15 min for 24 h. BP rhythm over 24 h was analyzed by the cosinor method and the mesor, amplitude, acrophase and percent rhythm were calculated. A total of 764 measurements were made in the light cycle and 721 in the dark cycle. Twenty-four-hour values (mean ± SD) were obtained for SBP (121 ± 22 mmHg), DBP (86 ± 17 mmHg), mean BP (MBP, 98 ± 18 mmHg) and heart rate (73 ± 16 bpm). The SBP, DBP and MBP were significantly higher (unpaired Student t-test) during the light period (125 ± 21, 88 ± 15 and 100 ± 17 mmHg, respectively) than during the dark period (120 ± 21, 85 ± 17 and 97 ± 17 mmHg, respectively) and the acrophase occurred between 16:00 and 17:45 h. This circadian variation is similar to that observed in cats, dogs and marmosets. The BP decreased during "behavioral sleep" (MBP down from 110 ± 19 to 90 ± 19 mmHg at 21:00 to 8:00 h). Both feeding and moving induced an increase in MBP (96 ± 17 to 119 ± 17 mmHg at 17:00 h and 97 ± 19 to 105 ± 12 mmHg at 15:00 h, respectively). The results show that conscious sloths present biphasic circadian fluctuations in BP levels, which are higher during the light period and are mainly synchronized with feeding.

Cardiovascular responses to locomotor activity and feeding in unrestrained three-toed sloths, Bradypus variegatus

Heart rate (HR) and systolic (SBP), diastolic (DBP) and mean (MBP) blood pressure were recorded by biotelemetry in nine conscious unrestrained sloths for 1 min every 15 min over a 24-h period. The animals were allowed to freely move in an acoustically isolated and temperature-controlled (24 ± 1ºC) experimental room with light-dark cycle (12/12 h). Behavior was closely monitored through a unidirectional visor and classified as resting (sitting or suspended), feeding (chewing and swallowing embauba leaves, Cecropia adenops), or locomotor activity around the tree trunk or on the room floor. Locomotor activity caused statistically significant increases in SBP (+8%, from 121 ± 22 to 131 ± 18 mmHg), DBP (+7%, from 86 ± 17 to 92 ± 10 mmHg), MBP (+8%, from 97 ± 19 to 105 ± 12 mmHg), and HR (+14%, from 84 ± 15 to 96 ± 15 bpm) compared to resting values, indicating a possible major influence of the autonomic nervous system on the modulation of cardiac function during this behavior. During feeding, the increase in blood pressure was even higher (SBP +27%, from 119 ± 21 to 151 ± 21 mmHg; DBP +21%, from 85 ± 16 to 103 ± 15 mmHg; MBP +24%, from 96 ± 17 to 119 ± 17 mmHg), while HR remained at 14% (from 84 ± 15 to 96 ± 10 bpm) above resting values. The proportionally greater increase in blood pressure than in HR during feeding suggests an increase in peripheral vascular resistance as part of the overall response to this behavior.

Electrocardiographic studies of the three-toed sloth, Bradypus variegatus

Electrocardiograms (ECG) obtained with standard limb leads and augmented unipolar limb leads were recorded from 17 unanesthetized adult sloths. The animals were held in their habitual position in an experimental chair. We determined heart rate and rhythm from the R-R intervals, the amplitude and duration of each wave, and the duration of the segments and intervals of the ECG. The mean electrical axes of P and T waves and QRS complex were calculated on the basis of the amplitude of these waves in leads I, II, III, aV R, aV L, and aV F. The P wave appeared positive in most tracings with low amplitude in lead II, the QRS complex was generally negative in leads aV R, III and aV F, and no arrhythmias were observed. With a mean ± SD heart rate for all recordings of 81 ± 18 bpm, the duration of P and T waves, QRS complex, and PR, QT and RR intervals averaged 0.05 ± 0.02, 0.15 ± 0.05, 0.07 ± 0.02, 0.13 ± 0.02, 0.38 ± 0.04, and 0.74 ± 0.17 s, respectively. The ECG shape had a definite configuration on each lead. The angles of the mean ± SD electrical axes for atrial and ventricular depolarization and ventricular repolarization in the horizontal plane were +34 ± 68º, -35 ± 63º, and -23 ± 68º, respectively. All electrical axes showed great variations and their mean values suggest that, when the sloth is in a seated position, the heart could be displaced by the diaphragm to a semi-horizontal position.

Fecal estradiol and progesterone metabolite levels in the three-toed sloth (Bradypus variegatus)

The present study was carried out to assess the possibility of measuring fecal steroid hormone metabolites as a noninvasive technique for monitoring reproductive function in the three-toed sloth, Bradypus variegatus. Levels of the estradiol (E2) and progesterone (P4) metabolites were measured by radioimmunoassay in fecal samples collected over 12 weeks from 4 captive female B. variegatus sloths. The validation of the radioimmunoassay for evaluation of fecal steroid metabolites was carried out by collecting 10 blood samples on the same day as defecation. There was a significant direct correlation between the plasma and fecal E2 and P4 levels (P < 0.05, Pearson's test), thereby validating this noninvasive technique for the study of the estrous cycle in these animals. Ovulation was detected in two sloths (SL03 and SL04) whose E2 levels reached 2237.43 and 6713.26 pg/g wet feces weight, respectively, for over four weeks, followed by an increase in P4 metabolites reaching 33.54 and 3242.68 ng/g wet feces weight, respectively. Interestingly, SL04, which presented higher levels of E2 and P4 metabolites, later gave birth to a healthy baby sloth. The results obtained indicate that this is a reliable technique for recording gonadal steroid secretion and thereby reproduction in sloths.

Morphology, molecular phylogeny, and taxonomic inconsistencies in the study of Bradypus sloths (Pilosa: Bradypodidae)

This study focuses on morphological and molecular data analyses, misidentifications, and phylogenetic inconsistencies regarding Bradypus variegatus (the brown-throated sloth) and B. tridactylus (the pale-throated sloth). Misidentifications were recorded on 75 of 313 museum specimens of Bradypus. Almost 90% of the misidentified specimens were B. variegatus from north-central Brazil, erroneously attributed to B. tridactylus. These misidentified specimens are reported in taxonomic reviews as the southernmost records of B. tridactylus. A history of confusing nomenclature regarding sloth species exists, and these particular misidentifications could be attributable to the similarity in face and throat color between B. variegatus from north-central Brazil and B. tridactylus. The molecular phylogeny of morphologically confirmed sloth specimens exhibits 2 monophyletic lineages representing B. variegatus and B. tridactylus. The split time between these 2 lineages was estimated at 6 million years ago (mya), contradicting previous studies that estimated this divergence to be 0.4 mya. Taxonomic inconsistencies were detected when comparing the molecular phylogeny to previously published DNA sequences ascribed to B. tridactylus. Misidentification or introgression could underlie such phylogenetic incongruities. Regardless of their causes, these discrepancies lead to misstatements regarding geographic distribution, phylogeny, and taxonomy of B. variegatus and B. tridactylus.

A PCR-RFLP assay for gender assignment in the three-toed sloths (Bradypus, Pilosa, Bradypodidae)

The three-toed sloths (Bradypus) are slow-moving arboreal neotropical mammals. Understanding demographic variables (such as sex ratio) of populations is a key for conservation purposes. Nevertheless, gender assignment of Bradypus is particularly challenging because of the lack of sexual dimorphism in infants and in adults, particularly B. torquatus, the most endangered of the three-toed sloths, in which sex is attributed by visual observation of the reproductively active males. Here, we standardized a method for sexing Bradypus individuals using PCR-RFLP of sex-linked genes ZFX/ZFY. This assay was validated with known-gender animals and proved accurate to assign gender on three Bradypus species.