Serum starvation is as efficient as roscovitine on the cycle synchronization in G0/G1 of red-rumped agouti fibroblasts.

Fibroblast cycle synchronization in G0/G1 is an essential step for nuclear reprogramming by cloning or induced cells to pluripotency. Considering the diversity among rodents and the ecological and scientific importance of these animals, we compared the contact inhibition, serum starvation, and 10 µM of roscovitine as methods of synchronization of red-rumped agouti fibroblasts. The effects of each protocol were evaluated on the percentage of cycle phase, morphology, viability, and apoptosis levels. The results showed that culturing the cells to serum starvation for 24 h (75.9%), 48 h (81.6%), 72 h (86.2%), 96 h (84.0%), and 120 h (83.7%) yielded a significantly higher percentage of cells arrested in the G0/G1 (P < 0.05) phase than cells not subjected to any cell cycle synchronization method (31.4%). Also, this effect was not different between the times of 48 and 120 h (P > 0.05). A similar response was observed for cells cultured with roscovitine for 12 h (86.9%), 24 h (74.8%), and 48 h (81.7%), with a higher percentage of synchronized cells in G0/G1 compared to cells not submitted to any synchronization treatment (52.2%). Nevertheless, this effect was best evidenced at 12 h (P < 0.05). Also, the contact inhibition for 24-120 h could not synchronize cells in G0/G1, with values ranging from 70.9 to 77.9% (P > 0.05). Moreover, no difference was observed for morphology, viability, and apoptosis levels in any synchronization method (P > 0.05). Therefore, serum starvation is as efficient as roscovitine on cycle synchronization in G0/G1 of red-rumped agouti fibroblasts.

Oropharyngeal cavity floor morphology in Eretmochelys imbricata (Testudines: Cheloniidae) hatchlings and evolutionary implications.

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

Pronephros and mesonephros characterization during the embryonic development of the giant South American river turtle, Podocnemis expansa (Podocnemididae: Testudines).

This study aimed to describe pronephros and mesonephros morphology during the embryonic development of Podocnemis expansa. Eggs were collected on an artificial beach at Balbina, Amazonas, Brazil, during the entire incubation period (mean of 59 days). The kidney-gonad complex was processed using light microscopy and the mesonephros using transmission electron microscopy. The pronephros was present for the first time on stage 4, composed of external glomeruli devoid of a capsule, protruding into the coelomic cavity, and internally composed of a capillary network. The pronephros degenerated after development stage 15. The first sign of the appearance of the mesonephros occurred around stage 8, indicated by the early formation of renal corpuscles. The mesonephros comprised an renal corpuscles, neck segment, proximal tubule, intermediate segment, distal tubule, collector tubule, and collector duct. Ultrastructural analysis of the mesonephros brush border was done in the proximal tubule, and the presence of cells with structural characters indicative of secretory activity was detected in the juxtatubular region. Renal corpuscles and proximal tubules were the main components that underwent morphological alterations during mesonephros degeneration. The pronephros is a transient kidney, and the mesonephros became the functional embryonic kidney in P. expansa. Mesonephros degeneration occurs in the cranial-caudal direction, and histologically, the degeneration is identified by changes in the morphology of the renal corpuscle and proximal tubule. However, the mesonephros is still present after hatching.

Morphological characterization of the digestive tube of hawksbill sea turtle (Eretmochelys imbricata) hatchlings.

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

Structural and Ultrastructural Characteristics of the Spix's Yellow-Toothed Cavy (Galea spixii, Wagler, 1831) Tongue.

The tongue is a fundamental organ in feeding, vocalization, and grooming. It is characterized by evolutionary adaptations reflected by diet, habitat, and function. Rodents are a very diverse mammalian order and the tongue's morphology varies in size, form, and presence of papillae. This work aimed to describe the morphological and ultrastructural aspects of the tongue of Spix's yellow-toothed cavy (Galea spixii, Wagler, 1831). Tongues of Spix's yellow-toothed cavies were analyzed with light microscopy, scanning, and transmission electron microscopy. The results showed that the tongue was divided into apex, body, and root. There were different types of papillae, such as vallate, foliate, laterally placed fungiform, fungiform, filiform, and robust filiform. The epithelium was organized into layers, including keratinized, granulous, spinous, and basal, below were lamina propria, and musculature, which evolved mucous and serous gland clusters. The tongue of Spix's yellow-toothed cavy was structurally and ultrastructurally similar to other rodents and had papillae with similar morphologies to other Caviidae species. However, the presence of robust filiform papillary lines and laterally placed fungiform papillae showed the main differences from other species. This was the first description of the tongue of Spix's yellow-toothed cavy.

Effects of somatic tissue cryopreservation on puma (Puma concolor L, 1771) tissue integrity and cell preservation after in vitro culture.

Somatic resource banks play a crucial role in the conservation of genetic diversity, allowing for the preservation of biological samples from different populations. Puma somatic cells can be recovered from these banks and used in assisted techniques toward enhancing their multiplication and conservation. In response to the population reduction of this ecologically importance species, we aimed to evaluate the capacity of cryopreservation of somatic tissues on the maintenance of the integrity and quality of the cells recovered after culture, with the aim of establishing a somatic tissue bank that will allow for the safeguarding of a wide genetic sampling of pumas. Cryopreservation increased the thickness of the corneum layer in the tissues, and the number of perinuclear halos and empty gaps. Nevertheless, cryopreservation was able to maintain normal fibroblast patterns, even showing an increase in the percentage of collagen fibers. Cryopreservation maintained the proliferative potential of the tissues and the parameters evaluated during in vitro culture, mainly regarding the viability, proliferative activity, and apoptosis levels. Nevertheless, cells from cryopreserved tissues showed decreased metabolism and mitochondrial membrane potential when compared to cells from non-cryopreserved tissues. In summary, we demonstrated for the first time that puma somatic tissues subjected to cryopreservation are viable and maintain tissue integrity, featuring minimal changes after warming. Although viable somatic cells are obtained from these tissues, they undergo alterations in their metabolism and mitochondrial membrane potential. Improvements in the conservation conditions of somatic samples are needed to increase the quality of somatic tissue banks in this species.

Colocolic Intussusception Associated with Octangium sp. (Digenea: Microscaphidiidae) in a Green Sea Turtle Chelonia mydas.

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

Morphometry of the heart of greater rhea (Rhea americana americana, Linnaeus, 1758).

Heart diseases in birds are frequent and generate significant production disorders. Morphometry is a valuable tool to provide fundamental information about heart conditions. Few studies have addressed morphological aspects of the heart of ratite birds, such as the Greater rhea. The present study aimed to analyse rhea heart morphometry, comparing young and adult subjects, in order to provide relevant information for the diagnosis of heart disease in this species. Hearts of young (n = 10) and adult (n = 10) female rheas were used in this research. Heart length and width and sternum length were measured using a caliper. Heart length and width and sternum length in adults were approximately three times greater than in young individuals. The left ventricular wall (LVW) was thicker than the right ventricular wall (RVW) at all ages, and the RVW was thicker in adults when compared to young subjects. The basal and middle RVW regions thicken with advancing age, and the thickness of the interventricular septum (ISW) occupies an intermediate position between the LVW and RVW. In general, an increase in rhea heart thickness and size relative to age is observed. The morphometric variations between young and adult rhea hearts observed in the present study may serve as a comparative subsidy for the diagnosis of cardiac abnormalities observed in this species.

Establishment, characterization, and cryopreservation of cell lines derived from red-rumped agouti (Dasyprocta leporina Linnaeus, 1758) - A study in a wild rodent.

Somatic cells can be used for rescuing wild mammals of ecological and economic importance, such as red-rumped agouti, through their application in advanced technologies. Thus, appropriate cell isolation, culture, and storage through cryopreservation can ensure the future safe use of these cells. We aimed to establish and evaluate the effects of culture time (second, fifth, and eighth passages) and cryopreservation on the morphology, viability, metabolism, proliferative activity, reactive oxygen species (ROS) levels, mitochondrial membrane potential (ΔΨm), and apoptosis on somatic cells derived from red-rumped agouti skin. Initially, we identified six dermal fibroblast lines by morphology, immunophenotyping, and karyotyping assays. In vitro culture after the second, fifth, and eighth passages, as well as the cryopreservation conditions used did not affect the metabolism or level of apoptosis. Nevertheless, cells in the fifth passage featured a reduction in proliferative activity and an increase in ROS levels when compared to second and eighth passage cells. Moreover, cryopreservation resulted in reduced ΔΨm when compared to non-cryopreserved cells. Additionally, cryopreserved cells showed a reduction in viability immediately after thawing; nevertheless, the viability of these cells was re-established after 11 days of in vitro culture and was similar to that of non-cryopreserved cells. In conclusion, we have shown that viable fibroblasts can be obtained from red-rumped agouti skin, featuring minimal changes after eight passages in in vitro culture systems. Additionally, adjustments to the cryopreservation protocol are necessary to reduce cellular oxidative stress caused by low temperatures.

Isolation, characterization, and cryopreservation of collared peccary skin-derived fibroblast cell lines.

BACKGROUND: Biobanking of cell lines is a promising tool of support for wildlife conservation. In particular, the ability to preserve fibroblast cell lines derived from collared peccaries is of significance as these wild mammals are unique to the Americas and play a large role in maintaining the ecosystem. We identified collared peccary fibroblasts by immunofluorescence and evaluated their morphology, growth and adherence capacity. Further, we monitored the viability and metabolic activity of the fibroblasts to determine the effects of passage number and cryopreservation on establishment of cell lines. METHODS: Skin biopsies were collected from the peripheral ear region from five adult animals in captivity. Initially, cells were isolated from fragments and cultured in the Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum and 2% antibiotic-antimycotic solution under a controlled atmosphere (38.5 °C, 5% CO2). We evaluated the maintenance of primary cells for morphology, adherence capacity of explants, explants in subconfluence, cell growth and absence of contamination. Moreover, we identified the fibroblast cells by immunofluorescence. Additionally, to evaluate the influence of the number of passages (first, third and tenth passage) and cryopreservation on establishment of cell lines, fibroblasts were analysed for the viability, metabolic activity, population doubling time (PDT), levels of reactive oxygen species (ROS), and mitochondrial membrane potential (ΔΨm). RESULTS: All explants (20/20) adhered to the dish in 2.4 days ± 0.5 with growth around the explants in 4.6 days ± 0.7, and subconfluence was observed within 7.8 days ± 1.0. Moreover, by morphology and immunocytochemistry analyses, cells were identified as fibroblasts which presented oval nuclei, a fusiform shape and positive vimentin staining. No contamination was observed after culture without antibiotics and antifungals for 30 days. While there was no difference observed for cell viability after the passages (first vs. third: P = 0.98; first vs. tenth: P = 0.76; third vs. tenth: P = 0.85), metabolic activity was found to be reduced in the tenth passage (23.2 ± 12.1%) when compared to that in the first and third passage (100.0 ± 24.4%, P = 0.006). Moreover, the cryopreservation did not influence the viability (P = 0.11), metabolic activity (P = 0.77), or PDT (P = 0.11). Nevertheless, a greater ΔΨm (P = 0.0001) was observed for the cryopreserved cells (2.12 ± 0.14) when compared to that in the non-cryopreserved cells (1.00 ± 0.05). Additionally, the cryopreserved cells showed greater levels of intracellular ROS after thawing (1.69 ± 0.38 vs. 1.00 ± 0.22, P = 0.04). CONCLUSIONS: This study is the first report on isolation, characterization and cryopreservation of fibroblasts from collared peccaries. We showed that adherent cultures were efficient for obtaining fibroblasts, which can be used as donor cells for nuclei for species cloning and other applications.

How does Spix's yellow-toothed cavy (Galea spixii Wagler, 1831) face the thermal challenges of the Brazilian tropical dry forest?

The aim of this work was to investigate the thermal biology of the Spix's yellow-toothed cavy (Galea spixii) from the hot and dry environment of the Brazilian Caatinga by infrared thermography and biophysical equations. We monitored the rectal temperature, as well as the non-evaporative (radiative and convective pathways) and evaporative heat exchanges of males and females. The mean rectal temperature of females and males was 37.58 ± 0.02 and 37.47 ± 0.02 °C, respectively. We identified thermal windows by infrared thermography. The surface temperatures and the long-wave radiation heat exchanges were higher in the periocular, preocular, pinnae and vibrissae regions, in that order. The surface temperature of the periocular and preocular regions correlated positively with rectal temperature. Convective heat exchange was insignificant for thermoregulation by G. spixii. Evaporative heat loss increased when the thermal environment inhibited the radiative pathway. Females showed higher evaporative thermolysis than males at times of greater thermal challenge, suggesting a lower tolerance to heat stress. Therefore, infrared thermography identified the thermal windows, which represented the first line of defense against overheating in G. spixii. The periocular and preocular surface temperatures could be used as predictors of the thermal state of G. spixii.

Production of collared peccary (Pecari tajacu Linnaeus, 1758) parthenogenic embryos following different oocyte chemical activation and in vitro maturation conditions.

To optimize the protocols for assisted reproductive techniques (ARTs) in collared peccary (Pecari tajacu Linnaeus, 1758), we evaluated various conditions for oocyte in vitro maturation (IVM) and chemical activation. Initially, we assessed the IVM rates, cumulus-oocyte complex (COC) quality, and oocyte morphometry in the absence or presence of epidermal growth factor (EGF). There was no difference between the COCs matured in absence or presence of EGF for the expansion of cumulus cells (97.6% ±â€¯1.2 vs. 100% ±â€¯0.0), presence of first polar body (65.9% ±â€¯1.2 vs. 70.5% ±â€¯1.8), nuclear status in second metaphase (62.5% ±â€¯11.6 vs. 68.4% ±â€¯4.9), cytoplasmic maturation (100.0% ±â€¯0.7 vs. 75.0% ±â€¯0.7), reactive oxygen species levels (0.5 ±â€¯0.2 vs. 0.3 ±â€¯0.1), and mitochondrial membrane potential (1.1 ±â€¯0.2 vs. 1.1 ± 0.1). However, the zona pellucida thickness of matured COCs was reduced in the presence of EGF. Thus, the EGF group was used for further experiments. The oocytes were artificially activated with ionomycin and four secondary activator combinations [6-dimethylaminopurine (6D), 6D and cytochalasin B (6D + CB), cycloheximide (CHX), and CHX and CB (CHX + CB)]. The effect of immature COCs based on cumulus cell layers and cytoplasm homogeneity (GI and GII or GIII COCs) on embryonic development and quality was evaluated. There was no difference in the cleavage rates among the groups of secondary activators. The cleavage rates of embryos derived from GI/GII and GIII COCs were greater than 72.2% and 25.0%, respectively. Moreover, treatment with CHX showed a reduction in the cleavage rate of embryos derived from GIII COCs when compared to the cleavage rate of embryos derived from GI/GII COCs (P < 0.05). Nevertheless, higher rates of blastocyst/total GI and GII COCs were observed in the 6D group (27.6% ± 0.3) compared to CHX group (6.9% ± 0.3). Additionally, only 6D treatment resulted in the production of embryos derived from GIII COCs (25.0% ± 0.2). The percentage of the ICM/total cell ratio was also greater in blastocysts derived from 6D (42.5% ± 19.0), 6D + CB (37.9% ± 21.9), and CHX + CB (43.8% ± 19.6) groups when compared to CHX (3.6% ± 0.1) group. Thus, the combination of ionomycin and 6D could produce collared peccary embryos by activation of both GI/GII COCs and GIII COCs. These optimized IVM conditions using EGF and chemical activation using ionomycin and 6D in collared peccaries form the first steps for establishing ARTs to conserve this species.

Establishment of a protocol for the isolation of ovarian preantral follicles derived from collared peccaries (Pecari tajacu).

We compare the efficiency of mechanical or enzymatic methods, and their combination, for the isolation of ovarian preantral follicles (PFs) from collared peccaries. The ovaries from six females were subjected to the different methods investigated here. For the enzymatic method, ovary fragments were exposed to collagenase type IV in TCM-HEPES medium; the mechanical procedure was based on ovarian cortex dissociation by using a scalpel blade. The residual solution obtained after the mechanical isolation was subjected to the enzymatic procedure. The number of isolated PFs was quantified and classified as primordial, primary, or secondary; their viability was assessed using trypan blue dye assay. To confirm the results, PFs derived from the most efficient method were evaluated for integrity using scanning electron microscopy (SEM) and subjected to a 24 h in vitro culture for subsequent evaluation of viability by using fluorescent probes. A higher number of PFs (P < 0.05) was obtained from the enzymatic method (961.7 ± 132.9) in comparison with the mechanical method (434.3 ± 88.9), but no difference was observed between the two methods and their combination (743.2 ± 92.8). The trypan blue assay showed that the enzymatic method (98.7 ± 0.6%) provided the highest percentage of viable follicles (P < 0.05). Furthermore, SEM confirmed the ultrastructural integrity of the surface architecture of peccary PFs isolated by the enzymatic procedure; epifluorescence microscopy was used to confirm their viability (86.0%). In conclusion, we suggest that the enzymatic method investigated here is useful for the isolation of viable ovarian PFs from collared peccaries.

Effects of cryopreservation techniques on the preservation of ear skin - An alternative approach to conservation of jaguar, Panthera onca (Linnaeus, 1758).

Currently, it has been observed that a considerable segment of the jaguar population is declining mainly because of hunting, and destruction and fragmentation of habitat. Given this scenario, efforts of the scientific community have been concentrated on the development of conservation strategies, such as the formation and use of somatic sample banks. We aimed to assess the effects of cryopreservation techniques of the ear skin of jaguar [slow freezing (SF) or direct vitrification in cryovials (DVC) or solid-surface vitrification (SSV)] on the morphological analysis and cell ability during the culture. All cryopreserved fragments regardless of the technique used, showed a reduction in the dermis and total thickness of the skin. Although a collagen matrix similar to the control group (fresh) has been observed only for the fragments from SF and SSV groups, all cryopreserved techniques were able to maintain normal patterns of the fibroblasts. Moreover, DVC and SSV methods maintained the proliferative activity of the tissues even after warming. After the culture, SF and SSV techniques were efficient for the recovery of the somatic cells according to most of the evaluated parameters, especially with regard to the duration of culture and cell metabolic activity. In conclusion, SSV was found to be a more efficient technique for cryopreserving jaguar skin when compared to DVC and SF. These results are relevant for the formation of somatic resource banks of this species, directed at cryopreserving adequate samplings of different individuals and generations for future applications in regenerative medicine, and assisted reproductive technologies.

Ultrastructural description of fresh and frozen/thawed sperm derived from collared peccaries (Pecari tajacu Linnaeus, 1,758).

The aim was to describe, through scanning electron microscopy (SEM) and transmission electron microscopy (TEM), the ultrastructure of peccaries' fresh and frozen-thawed sperm. For that, semen derived from three mature males was obtained by electroejaculation and evaluated for motility, membrane integrity, membrane functionality, chromatin integrity, and morphology through light microscopy. Samples were frozen using a Tris extender plus egg yolk (20%) and glycerol (6%). Then, fresh and frozen-thawed semen samples were mixed in different sperm pools that were processed for SEM and TEM. Sperm motility, membrane integrity, and functionality were impaired (p < .05) by freezing-thawing procedures, but sperm morphology, and chromatin integrity evaluated by light microscopy were not significantly affected. The SEM revealed that peccaries' sperm presents a flattened head in a paddle format, measuring 6.07 µm in length and 3.84 µm in width, with a vastus acrosome (4.46 µm). Normal tails measure 38.11 µm, being formed by an extensive midpiece with 15.52 µm in length. In frozen-thawed samples, both SEM and TEM provide us information about damage undetected through light microscopy as the presence of vesicles in the acrosome, loose plasma membrane, vacuolized mitochondria, dense fibers disorganized, and decondensed chromatin. In conclusion, we provide the first description of the sperm ultrasctruture in collared peccaries. Moreover, SEM and TEM help us to identify some nanometric damage provoked by freezing-thawing procedures, thus providing valuable information for the improvement of such important protocols used for biobanking formation.

Morphology and anatomic-surgical segmentation of the spleen of Pecari tajacu Linnaeus, 1758 / Morfologia e segmentação anátomo-cirúrgica do baço de Pecari tajacu Linnaeus, 1758

The spleen is a secondary lymphoid organ with importance in clinical surgery. Due this and the lack of data in the literature, the current paper analyzes the organ´s morphology and segmentation in collared peccaries. Twenty animals were used at the Center for the Multiplication of Wild Animals of the Universidade Federal Rural do SemiÁrido, Mossoró RN Brazil. The spleen was removed with the splenic pedicle preserved, identifying the lineal artery and vein. Fragments from four spleens were harvested to be examined under light microscopy. They were fixed in a paraformaldehyde solution 4% and buffered with sodium phosphate 0.1M, pH 7.4. Routine histological techniques were performed: the spleens were dehydrated in increasing ethanol concentrations; diaphanized in xylol; soaked in paraffin; 7µm cuts were obtained and stained by hematoxylin-eosin or Gomori trichrome technique. The intraparenchemal vascularization of sixteen spleens were analyzed by latex or vinilite acetate perfusal of the lineal artery and vein and the organ fixed, respectively, in a water solution of formaldehyde 10% or immersed in a solution of sulfuric acid 30%. The collared peccary´s spleen had a tongue-like shape. Under the microscope, the spleen featured an intermediary type, with a great amount of white pulp, a predominance of red pulp and few trabeculae. The spleen´s segments had three different regions, namely dorsal, middle and ventral, in irrigation terms with a possible surgical removal of the dorsal region. Knowledge on the angio-architecture and segmentation of the spleen will be a contribution for surgical procedures in wild species, having a great relevance when partial splenectomy is required.

O baço é um órgão linfoide secundário e com importância na clínica cirúrgica. Diante da inexistência de dados na literatura, este artigo estudou a morfologia e segmentação deste órgão em catetos. Foram utilizados 20 animais que vieram a óbito por causas naturais no Centro de Multiplicação de Animais Silvestres da Universidade Federal Rural do Semi-Árido, Mossoró-RN, Brasil. O baço foi coletado preservando-se o pedículo esplênico, pelo qual identificavam-se a artéria e a veia lienais. Fragmentos de quatro baços foram coletados para análise em microscopia de luz. Estes foram fixados em solução de paraformaldeído a 4% tamponado com fosfato de sódio 0,1M, pH 7,4 e submetidos a técnicas histológicas de rotina. Os baços foram desidratados em concentrações crescentes de etanol; diafanizados em xilol; embebidos em parafina e obtidos cortes de 7µm e corados pela técnica de hematoxilina-eosina ou tricrômio de Gomori. A vascularização intraparenquimal de vinte baços foram analisadas através da perfusão de latex ou acetato de vinilite na artéria e veia lienais e logo o órgão foi fixado, respectivamente, em solução aquosa de formaldeído a 10% ou imersos em solução de ácido sulfúrico a 30%. O baço do cateto apresentou-se como uma estrutura longa com forma similar a uma língua. Microscopicamente, o baço caracterizou-se como do tipo intermediário, possuindo quantidade considerável de polpa branca, com predominância da polpa vermelha e poucas trabéculas. A segmentação do baço em termos de irrigação apresentou três regiões distintas: dorsal, média e ventral, com a região dorsal passível de remoção cirúrgica. O conhecimento sobre a angioarquitetura e segmentação do baço de cateto contribuirá para a realização de procedimentos cirúrgicos nesta espécie silvestre e de grande importância quando se fizer necessário a esplenectomia parcial.

Development of Adrenal Glands from the Embryonic Phase to Puberty in Spix's Yellow-toothed Cavies (Galea spixii, Wagler, 1831)

ABSTRACT Several authors have underscored the importance of establishing parameters in morphological development by gender comparison to establish clinical and pre-clinical assays through the use of experimental models. Current research compares the morphometry of right and left adrenal glands of males and females and describes differentiation of the cortex and medulla tissue during the embryonic, pre-puberty and puberty phases in Spix´s yellow-toothed cavies. Embryos aged 22 (22D), 25 (25D), 30 (30D), 40 (44D) and >50 (50D) days of gestation and neonates aged 15 (15DPN) (DPN= Days postnatal), 30 (30DPN) and 90 (90DPN) days after birth were analyzed. Comparisons included morphometric and histological analysis in all periods described. When compared the right and left adrenal glands, results show that the length and width have statistical differences (p<0.05). Statistical difference between right and left glans for weight occurred only after 30D in males and after 50D in females. When compared male and females, no statistical difference in the right and left glands was extant. In the case of tissue differences, the glomerular zone is the first to emerge after 22D, followed by the fasciculate zone after 25D and by the reticular zone during the post-natal period. Medullar tissue was spread between the cortical tissue at the onset of development, establishing itself at the center of the organ since the end of pregnancy (>50D) up to puberty. Considering tissue differentiation, there was no difference between the adrenal glands of male and female cavies or between the right and left adrenal glands.

Collateral branches of the aortic arch in the greater Rhea (Rhea americana americana Linnaeus, 1758) / Ramos colaterais do arco aórtico de emas (Rhea americana americana Linnaeus, 1758)

The greater rhea (Rhea americana americana) is a bird of the Rheidae family, and is known as a ratite for being a flightless bird. This animal has great reproductive and productive potential, according to the products and by-products that it can provide such as meat, leather, feathers and fat which are very popular in the world market. Given its economic importance and lack of information in the literature on its morphology, especially in regard to its cardiovascular apparatus, this study aimed to describe the collateral arteries of the aortic arch, in order to establish the origin and distribution of arteries and thus contribute information to the biology of the species. The bodies of 20 young and adult rheas of both sexes which had died from natural causes and were being stored in a freezer at CEMAS / UFERSA were used. The study was approved by CEUA /UFERSA (Opinion No. 09/2015, process No. 23091.004968 / 2015-23). The animals were thawed and had the cannulated thoracic aorta artery and the vascular system perfused with Neoprene 450 latex colored with yellow pigment. Subsequently, the animals were fixed in 3.7% aqueous solution of formaldehyde, and after 72 hours dissections were carried out, images were obtained and schematic drawings were prepared. The right and left brachiocephalic trunks emerged from the aortic arch in 100% of the specimens from the right brachiocephalic trunk origined a common trunk the thyroid arteries, syringotracheal trunk, vertebral artery, superficial lateral cervical artery, basecervical artery, and ascending esophageal artery. The left brachiocephalic trunk collaterally stemmed in the left common carotid artery, which in turn led to the left internal carotid and a common trunk which stemmed the thyroid arteries, the syringotracheal trunk, vertebral artery, superficial lateral cervical artery, basecervical artery and descending esophageal artery. At the end of its trajectory, the right and left brachiocephalic trunks give rise to the right and left subclavian arteries, which in turn, stem the sternoclavicular, axillary, and intercostal arteries, pectoral trunk, cranial pectoral arteries, pectoral caudal artery and collateral branches of the pectoral trunk. Based on the results, it was concluded that the aortic arch in rheas issued right and left brachycephalic trunks.

A ema (Rhea americana americana) é uma ave da família Rheidae e por isto denominada de ratita, por não apresentar aptidões para o voo. Este animal tem grande potencial reprodutivo e produtivo, em função dos produtos e subprodutos que podem fornecer como carne, couro, penas e gordura muito procurados no mercado mundial. Dada a sua importância econômica e pela falta de informação na literatura sobre sua morfologia, principalmente no que diz respeito ao seu aparelho cardiovascular, objetivou-se descrever os ramos colaterais do arco aórtico, de modo a estabelecer a origem e distribuição destas artérias e, assim, contribuir com informações para a biologia da espécie. Foram utilizadas 20 emas jovens e adultas de ambos os sexos, oriundas do CEMAS/UFERSA, as quais vieram a óbito por causas naturais e que se encontravam armazenadas em freezer. A experimentação foi aprovada pela CEUA/UFERSA (Parecer n° 09/2015, processo n° 23091.004968/2015-23). Os animais foram descongelados e tiveram a artéria aorta torácica canulada e o sistema vascular perfundido com látex Neoprene 450 corado com pigmento amarelo. Posteriormente, os animais foram fixados em solução aquosa de formaldeído a 3,7% e após 72 horas realizaram-se as dissecações e obtenção de imagens fotográficas e elaboração de desenhos esquemáticos. Em 100% dos espécimes, emergiram a partir do arco aórtico os troncos braquiocefálicos direito e esquerdo. O tronco braquiocefálico direito emitiu colateralmente a partir de um tronco comum as artérias tireoide, tronco siringotraqueal, vertebral, cervical superficial lateral, basecervical e esofageana ascendente. Já o tronco braquiocefálico esquerdo emitiu colateralmente a artéria carótida comum esquerda, que por sua vez, originou a carótida interna esquerda e um tronco comum que emitiu as artérias tireoide, tronco siringotraqueal, vertebral, cervical superficial lateral, basecervical e esofageana descendente. No final de seu percurso, os troncos braquiocefálicos direito e esquerdo, originaram as artérias subclávias direita e esquerda, que por sua vez, emitiram as artérias esternoclaviculares, axilar, intercostal, tronco peitoral, peitorais craniais e peitoral caudal e ramos colaterais do tronco peitoral. Com base nos resultados, concluiu-se que, em emas, o arco aórtico emitiu os troncos braquicefálicos direito e esquerdo.