Search

Conservation of the genetic diversity through skin and cartilage biobanks represents an essential strategy for maintaining biodiversity. Biobanks for the wild species of the order Rodentia have been little studied. Considering that the cryopreservation technique has specific relationships with the tissue and species of interest, we propose investigating different techniques for preserving tissue integrity and cell viability after cartilage and skin culture from Spix's yellow-toothed cavies. Subsequently, two techniques [solid-surface vitrification (SSV) vs. slow freezing (SF)] were used for cartilage and skin cryopreservation. Tissues not subjected to cryopreservation were used as controls. All tissues were evaluated for morphology and proliferation by histological techniques. Moreover, fragments were cultured, and cells were evaluated for viability, proliferation, metabolism, and apoptosis. Regardless of the cryopreservation technique, no differences were observed for the thickness of the epidermis, dermis, skin, spinous and basal layers, fibroblasts, and proliferative activity regarding the number of nucleolar organizer regions (NOR). SSV ensured better maintenance of epidermal cells, normal chondrocytes, filled gaps, collagen fibers, proliferative activity by NOR area/cell, and reduced perinuclear halos and empty gaps compared to SF. SF ensured the conservation of corneum thickness compared to the control. Although both techniques promoted cell recovery after culture, cells from SF resulted in better subconfluence time and day with cell growth around fragments compared to SSV. In conclusion, both cryopreservation techniques resulted in viable cells after culture. However, SSV promoted better maintenance of tissue morphological integrity, and SF ensured the preservation of all cell quality parameters in Spix's yellow-toothed cavies. [ABSTRACT FROM AUTHOR]

Phylogenetically, the yolk sac is an ancient structure in vertebrate evolution and crucial in the placentation of viviparous animals, playing an essential role in embryogenesis. Considering that the yolk sac persists until birth in rodents, forming an active placenta, this study aimed to describe the yolk sac development process in red-rumped agoutis (Dasyprocta leporina) during the early gestation phase and associating the progression to morphological changes in other structures. To this end, six female red-rumped agoutis obtained from CEMAS-UFERSA were monitored for 24 h. The first gestation day was defined as 24 h post-copulation. Samplings were carried out on the 13th, 14th and 15th pregnancy days. Pregnant uteri were fixed in 8% paraformaldehyde for 72 h and histologically processed, with 5 μm sections stained with Hematoxylin-Eosin (HE) and analyzed under a light microscope. On the 13th day, the blastocyst is implanted on the antimesometrial side of the uterus, surrounded by uterine stromal cells organizing themselves around this structure. On the 14th day, the intramural wall between the uterus and the embryo begins to break down and the parietal yolk sac exhibits spindle cells, while the visceral yolk sac presents eosinophilic cuboidal cells. On the 15th day, the intramural layer is almost completely ruptured, revealing a vascular bed between the embryo and the decidua. The visceral yolk sac did not yet contain its characteristic villi at this stage. Blastocyst formation and yolk sac differentiation take place between the 13th and 15th pregnancy days. The yolk sac inversion process is also described, taking place between the 14th and 15th gestation days, which had not yet been documented for red-rumped agoutis. [ABSTRACT FROM AUTHOR]

Establishing new somatic cell cultures has raised significant attention as an effective and convenient way to preserve genetic samples for different applications. Although many lines have been established in model animals, none derived from six-banded armadillo species is currently available. We report the successful isolation and characterization of fibroblasts from six‐banded armadillos, evaluating the cell quality after extended culture and cryopreservation. Initially, we collected ear skin from five captive adult individuals and identified fibroblast lines by morphology, karyotyping, and immunophenotyping assays. The isolated fibroblasts were evaluated after several passages (fourth, seventh, and tenth passages) and cryopreservation by slow freezing. Cell morphology, viability, metabolism, proliferative activity, mitochondrial membrane potential, and apoptosis levels were analyzed. The skin explants had great adhesion, and cell outgrowth could be seen after 3–6 d. The cells were verified as fibroblasts at the fourth passage by vimentin expression and normal karyotype (2n = 58). The viability remained high (> 87%) and constant from the fourth to the tenth passage (p > 0.05). The passages did not change the cell morphology and metabolic and growth rates. Moreover, cryopreservation did not affect most evaluated parameters; post-thawed cells maintained their viability, growth, metabolism, and apoptosis levels. Nevertheless, cryopreservation increased mitochondrial membrane permeability and cell population doubling time compared to non-cryopreserved cells (p < 0.05). In summary, viable fibroblasts can be obtained from six-banded armadillo skin while conserving their quality as the number of passages increases and featuring few changes after cryopreservation. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

The purpose of this study was to develop an edible coating containing babassu starch, babassu oil, and plasticizers to maintain egg quality. Firstly, it evaluated the effects of 5% babassu starch with 10% (SC-10%), 15% (SC-15%), and 20% (SC-20%) oil coating on egg quality. The effects of polyols such as mannitol and sorbitol alone or in combination on coating effectiveness were also assessed. For the first part, all formulations containing starch and/or babassu oil effectively preserved the internal quality characteristics of weight loss and Haugh units of stored eggs for 28 days. For the yolk and albumen percentages, the eggs for the treatments containing oil (SC-10%, SC-15%, and SC-20%) had the values closest (p < 0.05) to those of fresh eggs. Thus, according to these parameters, the presence of the oil in the coatings was more efficient in delaying egg quality. Despite the increase in albumen pH with the storage, the eggs stored with coatings kept the pH around 8.5. For the yolk color, the b* values of SC-10%, SC-15%, and SC-20% varied from 22.64 to 24.25 and did not differ (p > 0.05) from fresh eggs. Regarding the shell color, ΔE values of coated eggs were higher in the starch babassu treatment (6.97), followed by the treatments SC-10% (4.79), SC-15% (5.18), and SC-20% (5.17), and finally without coating (15.95). The formulation SC-10%, which contained 5% starch and 10% babassu oil, was selected to compare plasticizers. For the plasticizers, the babassu starch and oil in combination with 30% mannitol kept better the egg quality. Weight loss was lower (p < 0.05) in eggs only with 30% of mannitol (2.69%) and the Haugh Units were higher (60.04). For the b* value of yolk, 30% of mannitol had higher values (48.83) (p < 0.05) when compared to the control treatment (40.39). Thus, the presence of babassu starch (5%) and oil (10%) in combination with 30% mannitol maintains the egg quality during 28 days of storage. This study provides an approach that can potentially be applied to egg shelf life using products of the babassu industry. [ABSTRACT FROM AUTHOR]

The greater rhea, Rhea americana, is a wild ratite of high scientific importance and significant and zootechnical value, especially considering the current development state of Brazilian poultry production, where research aimed at increasing the productivity of these animals has become extremely relevant. Studies concerning fetal attachments and embryonic development are paramount, as they can provide essential information concerning reproductive and nutritional animal management. However, a lack of information on greater rhea fetal morphology is noted. Therefore, the aim of the present study was to establish a standard model for fetal attachments in this species. Greater rhea eggs were incubated from 0 to 36 days, and macroscopic and microscopic embryonic attachment characterizations were performed. Histologically, all embryonic annexes exhibit germ layers, namely the ectoderm (outer layer), mesoderm (middle layer) and endoderm (inner layer). The findings indicate that greater rhea development patterns are similar to other birds. [ABSTRACT FROM AUTHOR]

Animal cloning is a promising technology for biodiversity conservation, and its success depends on the recovery of nucleus donor cells. Specifically for collared peccaries, found sometimes in regions that are difficult to access, the storage at 4-6°C of skin tissues would be an alternative for the conservation of genetic material. Therefore, we aimed to evaluate different storage periods and the presence of a nutrient medium at 4-6°C on the recovery of somatic cells from the skin of collared peccaries. To analyze cell recovery rates, ear explants were distributed in non-refrigerated samples and samples refrigerated for 10, 30, and 50 d in the absence or presence of nutrient medium. All explants were analyzed by histologically and cultured. Only the fragments stored for 50 d without medium showed an increase in the total thickness of skin. Moreover, increased storage period, regardless of the presence of medium, increased the halo number and reduced the metabolic activity. After culture, only the fragments stored without medium for 50 d did not yield any somatic cells. Cells recovered from explants stored for 10 d showed similar characteristics to these recovered from non-refrigerated explants, regardless of the presence of medium, including the day at which explants achieved attachment and the total time to reach subconfluence. In conclusion, viable cells can be recovered from somatic tissues of collared peccaries stored for up to 50 d in the presence of medium, and tissues refrigerated for up to 10 d in the presence of medium yielded more viable cells. [ABSTRACT FROM AUTHOR]