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Avaliando os efeitos das proteínas anticongelantes na viabilidade de embriões bovinos produzidos in vitro vitrificados

Mon, 29 Aug 2022 00:00:00 GMT

Título: Avaliando os efeitos das proteínas anticongelantes na viabilidade de embriões bovinos produzidos in vitro vitrificados Autor: SILVA JÚNIOR, Rafael Artur da Primeiro orientador: BATISTA, André Mariano Abstract: Embryo cryopreservation is an important tool that enables the logistics of storage and transfer of blastocysts of high genetic merit. There are currently two main distinct cryopreservation methodologies, slow freezing and vitrification, developed in order to avoid cryoinjuries. Vitrification is a rapid freezing technique that transforms the liquid into a glassy state, reducing the negative effects of ice crystal formation. However, the vitrification technique can still cause damage related to the formation of ice crystals. Some proteins have been identified as inhibitors of intracellular ice formation, called antifreeze proteins (AFP). These proteins have action in modifying the structure of the ice crystal, decreasing the freezing point and inhibiting the recrystallization activity. Based on the above, this study aimed to evaluate the cryoprotective effects of AFP extracted from the grass Lolium perenne (LpAFP) and AFP extracted from the larva of the Tenebrio molitor beetle (TmAFP) on the vitrification of bovine embryos. For this, in vitro produced bovine blastocysts were vitrified using the cryotop method in both experiments. In the first experiment, during the vitrification process, the blastocysts were randomly separated into two experimental groups, the control group (GC) containing 0 ng/mL LpAFP and the treatment group (GT) supplemented with 500 ng/mL LpAFP. Vitrification was carried out by transferring the blastocysts to the equilibrium solution: 7.5% ethylene glycol (EG) and 7.5% dimethyl sulfoxide (DMSO) for 2 min, then to the vitrification solution: 15% EG, 15 % dimethyl sulfoxide (DMSO) and then deposited on the cryotop rod and submerged in liquid nitrogen. Heating was carried out in three steps with decreasing concentrations of sucrose (1.0, 0.5 and 0.5 M sucrose). After heating, the blastocysts were cultured for 24 hours and analyzed for survival rate (expansion and/or hatching), stained with Hoechst to evaluate the total number of cells and analyzed under transmission electron microscopy for ultrastructural evaluation. In the second experiment, in vitro produced blastocysts were randomly separated into three experimental groups and vitrified with stabilization and vitrification medium supplemented with different concentrations of TmAFP: 0 ng/mL; 500 ng/mL and 1000 ng/mL. The vitrification and heating process took place in the same way as in the first experiment. After 24 hours of post-warming cultivation, the survival rate (expansion and/or hatching) was analyzed and ultrastructural analysis of the expanded embryos was performed. In experiment 1, the results showed that there was no significant difference in the reexpansion rate 24 hours after heating, however there was variation (P < 0.05) in the hatching rate in the GT. The total number of cells 24 hours after heating was significantly higher in the GT when compared to the GC (GT 114.87 ± 7.24 vs. GC 91.81 ± 4.94). The ultrastructural analysis showed a decrease in cytoplasmic lesions, mainly in mitochondria and rough endoplasmic reticulum in GT. In experiment two, the group supplemented with 500 ng/mL of TmAFP (500TmAFP) had a higher survival rate when compared to the other two groups, control (0 ng/mL of TmAFP) and 1000TmAFP (1000 ng/mL of TmAFP), also a higher rate of blastocoel expansion was observed in the 500TmAFP group. Ultrastructural lesions were observed in all vitrified embryos, however embryos from the 500TmAFP and 1000TmAFP groups showed less cytoplasmic lesions when compared to the control group. In conclusion, experiment 1 demonstrated that the addition of 500 ng/mL of LpAFP during the vitrification of in vitro produced bovine embryos proved to be favorable in improving the survival and development of blastocysts after heating. In addition, experiment 2 showed that TmAFP supplementation can mitigate cellular changes, which involve organelles and cellular components essential for proper functioning and viability after heating of vitrified in vitro produced bovine embryos. Instituição: Universidade Federal Rural de Pernambuco Tipo do documento: Tese

Caracterização do recebimento de jabutis-piranga (Chelonoidis carbonaria) pelo centro de triagem de animais silvestres de Pernambuco de 2016 a 2020 e desenvolvimento de prótese 3d para lesões traumáticas de carapaça

Mon, 25 Jan 2021 00:00:00 GMT

Título: Caracterização do recebimento de jabutis-piranga (Chelonoidis carbonaria) pelo centro de triagem de animais silvestres de Pernambuco de 2016 a 2020 e desenvolvimento de prótese 3d para lesões traumáticas de carapaça Autor: MELO, Thábata Morales Vieira de Primeiro orientador: SILVA, Jean Carlos Ramos da Abstract: The objective was to characterize the red-footed tortoises (Chelonoidis carbonaria) population received by the Pernambuco Wild Animal Screening Center in the period from 2016 to 2020. The survey resulted in 1,374 cataloged animals, mostly from the Metropolitan Region of Recife, Pernambuco State, Brazil, through voluntary surrender of individuals. Of these, 531 (n=38.6%) were in captivity, with inadequate handling, resulting in poor clinical conditions. Of the 132 (9.6%) hospitalized animals, 41.6% (n=55) had traumatic injuries, where the attack by other animals corresponds to 38.2% (n=21) of the cause of origin. These attacks are also identified as the most frequent reason for the death of these individuals within CETAS during this period. In view of these data, it is possible to plan inspection and educational actions aimed at raising awareness among the population, especially in the Metropolitan Region of Recife, about the importance of preserving animals and the consequences of inadequate captive management, highlighting the role of CETAS in this scenario, encouraging the delivery of animals, so that they can be rehabilitated and returned to nature. Among the animals received, a red-footed tortoise (Chelonoidis carbonaria), female, adult, with a history of burns after a fire in a cane field was rescued and sent to the Wild Animal Screening Center of Pernambuco. The animal had carapace lesions and loss of corneal and bone plaques. Aiming to protect the animal until its recovery, the goal of the study was to build a 3D full-shell prosthesis, discussing the methodology and its adaptations, thus allowing it to be applied by other veterinarians. For 3D modeling, images of the animal were obtained by photogrammetry, these were treated by the software Regard3D 0.95 and Blender 2.78, and the final file was exported to MatterControl for printing. Polylactic acid was used as material, and the final prosthesis was finished in acrylic resin mimicking the camouflage of these animals. It can be said that the prosthesis was efficient and fulfilled the requirements by providing protection to the corneal plate bed until healing. The methodology used requires basic knowledge of the tools for the 3D modeling stage, and familiarity with the materials used in the finishing of the hull, advising the formation of a multidisciplinary team in these cases. Instituição: Universidade Federal Rural de Pernambuco Tipo do documento: Tese

Dermatopatias em animais de produção no nordeste do Brasil

Fri, 30 Dec 2022 00:00:00 GMT

Título: Dermatopatias em animais de produção no nordeste do Brasil Autor: LIMA, Telma de Sousa Primeiro orientador: MENDONÇA, Fábio de Souza Abstract: In Brazil, animal production stands out considerably among the most varied zootechnical species; however, maintaining the health of the herds is still one of the main bottlenecks that the country needs to overcome in order to maintain its prominent position in the competitive environment of the market. The skin is a constant target of pathogens or environmental disturbances and is often the first to show signs of illness on the property, so that greater attention should be given to this organ as a tool to prevent the advancement and perpetuation of diseases and, consequently, reduce the impacts of disease on the farm. The objective of this work was to offer an update on the main dermatopathies in production animals in Brazil. Therefore, the study was divided into two parts: the first contains a general literature review, in which some of the skin diseases described in livestock in the Northeast are addressed, with emphasis on pathological findings. The second part is divided into three chapters and contains: a) a literature review called “skin diseases of donkeys and mules-an update”. This article was published in the international journal Animals and addresses some of the skin diseases that are described in donkeys and mules from Brazil and the world. In addition, he reports an outbreak of primary photosensitization by Froelichia humboldtiana in feral donkeys from Rio Grande do Norte, characterized by exuberant, ulcerated, exudative and pruritic lesions, especially in the face region, complicated by self-mutilation. b) A scientific article “Outbreaks of dermatopathy due to intoxication by Tephrosia noctiflora in Brazilian cattle herds”. This work will be submitted to the journal Toxicon and provides an update on cases of intoxication by Tephrosia noctiflora, a plant present in the forest zone of Brazil with the potential to cause skin, liver and kidney lesions. The outbreaks described here occurred in Pernambuco and Alagoas and affected both adult cattle and calves, characterized by weight loss and varying degrees of lichenification, hyperkeratosis, hyperpigmentation and secondary infection. It is possible that contact and consumption produce lesions in cattle that consume the plant even with adequate pasture supply. The last chapter (c) is a short communication entitled “Dorsal thermal necrosis in a Brazilian sheep herd”. This study will be submitted to the veterinary research journal and describes an outbreak of sun-induced skin necrosis in sheep raised in Alagoas. The animals developed lesions characterized by linear to serpiginous alopecia that progressed to skin detachment, showing the underlying dorsal musculature, from the cervical to the sacral region. The skin that fell off was dry, firm and black, while the musculature was dark red, covered with fibrin and fetid. These studies indicate that toxic and environmental disturbances are important for ruminants and horses in Northeastern Brazil and that management measures should be employed to reduce the impacts of these diseases on the herd. In all production spectrums, the negative repercussion of dermatopathies for the producer is evident. It is hoped, with these studies, that skin disorders receive due attention from producers and professionals who deal with species of zootechnical interest in Brazil in the field. Instituição: Universidade Federal Rural de Pernambuco Tipo do documento: Tese

Estudo morfológico da superfície ocular Bradypus variegatus (Schinz, 1825)

Mon, 28 Feb 2022 00:00:00 GMT

Título: Estudo morfológico da superfície ocular Bradypus variegatus (Schinz, 1825) Autor: SPINELLI, Taciana Pontes Primeiro orientador: EVÊNCIO NETO, Joaquim Abstract: Sloths (Bradypus variegatus) are directly or indirectly affected by human action, for example: car accidents with collisions, deforestation, hunting for consumption of exotic meat and pesticide poisoning occur frequently. Several studies on ecology, reproduction and nutritional aspects of wild animals have already been published, however, most anatomical characteristics are described in a generalist way or remain unknown. Veterinary ophthalmology is an expanding field and research in comparative ophthalmology produces a range of knowledge that allows the understanding of different evolutionary processes among species. The ocular surface is of vital importance for vision, being in direct contact with the environment, it is vulnerable to dryness, injuries and pathogens and therefore has a complex mechanism for maintaining its homeostasis. Sloths (Bradypus variegatus) may be affected by several ocular diseases, like humans, due to different etiologies, thus the importance of characterizing the normal tissues of the ocular surface in these animals since studies on the sloth ocular anatomy and physiology are still scarce. The present study aimed to characterize the normal anatomy of the ocular surface of this species. For the development of the histomorphometric study of the ocular surface, eyes were collected from animals that died for reasons unrelated to this research. The eyes were processed for inclusion in paraffin blocks and stained by the Hematoxylin-Eosin (HE), Schiff's Periodic Acid (PAS), alcian blue, tricromic masson, and then submitted to evaluation by light microscopy. Anatomical, histological, and histochemical characteristics of the lacrimal gland, the superficial and profound glands of the third eyelid, palpebral glands, cornea and limbus were described. Histologic examination revealed tarsus, tarsal glands and cilia were not present.Instead, well-developed sebaceous glands associated with accessory hairs along the palpebrae were present. The inner face of the eyelids is lined with the palpebral conjunctiva, which consists of a thin layer of loose connective tissue with a stratified epithelium. Mixed-secreting accessory lacrimal glands were noted in the deep stroma of both palpebrae. The lacrimal gland was situated in the dorsolateral angle of the orbit between the dorsal rectus and the lateral rectus muscles. Histologically, the gland is a tubuloalveolar type. The third eyelid protrudes from the medial canthus over the anterior surface of the globe, its cartilage resembles an anchor in shape. The stroma of the nictitans consists of loose to dense connective tissue that supports glandular and lymphoid tissue. Histological and histochemical examination revealed a superficial and a deep (harderian) glandular tissue. The deep gland is large and situated in the anteroventral region of the orbit. It is a compound branched, tubulo-alveolar, with a single layer or columnar cells. The cornea is elliptical in shape, with a horizontal diameter greater than the vertical. The anterior epithelium consists of a single cell layer of basal cells, which are columnar in shape and lie on a thin basement membrane; two three layers of polyhedral cells; and three layers of nonkeratinized squamous cells. The Bowman membrane is absent, and vessels are present in the stroma reaching the central cornea. Instituição: Universidade Federal Rural de Pernambuco Tipo do documento: Tese