Doutorado em Ciência Animal (EVZ)

URI Permanente para esta coleção


Submissões Recentes

Agora exibindo 1 - 20 de 210
  • Item
    Desenvolvimento de sorvete funcional simbiótico com polpa de cagaita (Eugenia dysenterica)
    (Universidade Federal de Goiás, 2021-12-17) Pfrimer, Renata Teixeira; Cordeiro, Clarice Gebara Muraro Serrate;; Coelho, Karyne Oliveira;; Nicolau, Edmar Soares;; Nicolau, Edmar Soares; Fonseca, Leorges Moraes da; Lage, Moacir Evandro; Antunes, Veridiana de Carvalho; Cavicchioli, Valéria Quintana
    Bio-inspired technological innovation in ice cream manufacturing has driven the market for dairy products. In this way, increasing the nutritional density and approving ice cream, with the use of whey, buttermilk, inulin, probiotic and fruit from the Cerrado are viable alternative for the sector. The objective was the production and physicochemical, microbiological and sensory characterization of different ice creams, varying those due to the milk/cream ratio, whey and buttermilk defined by the Simplex Centroide Design, and subsequently added a probiotic strain of Lactobacillus acidophilus, inulin and cagaita pulp (Eugenia dysenterica). Nine ice creams were produced with whey and buttermilk contents, ranging from 5% to 15%, and milk/cream ratio from 70% to 90%, as stipulated by the Simplex Centroide Design. All formulations result within the standards defined by current legislation. Ice creams showed higher content of palmitic, oleic and stearic fatty acids. They stood out as formulations with the highest concentration of co-products for presenting the best nutritional composition, considering the high levels of protein, protein fractions, lipids and lactose. The high concentration of whey and buttermilk concentration, as well as the control formulations, totaling Lactobacillus acidophilus six, were increased with 0.5% of, being three with the addition of 20% of cagaita pulp and three without the addition of pulp. Subsequently, all formulations were increased with 5% of inulin. Physical-composition, microbiological, fatty acid profile, quantitative descriptive analysis, rheology, texture profile analysis and confocal microscopy were performed. All formulations result within the standards defined by current legislation. Available probiotic and symbiotic ice creams were categorized into standard and premium ice creams by fat content. Formulations with cagaita pulp presented higher acidity, moisture and lower melting speed and higher instrumental hardness. The ice creams exhibited equivalence in relation to the semiquantification of image pixels, accompanied by confocal microscopy analysis, with the quantification of lipids and proteins. Ice cream with the addition of pulp, presented higher intensity of yellow color and citrus aroma. All ice creams are categorized as probiotic foods and source count 8 Log10 CFU / g, being great vehicles to provide probiotic culture and all its benefits when consumed.
  • Item
    Estudo macroscópico do telencéfalo, da vascularização da base e seios da dura-máter em Alouatta belzebul
    (Universidade Federal de Goiás, 2020-11-11) Pereira, Dayane Kelly Sabec; Pereira, Kleber Fernando;; Vulcani, Valcinir Aloisio Scalla;; Vulcani, Valcinir Aloisio Scalla; Franzo, Vanessa Sobué; Silva, Ana Paula Sousa Paixão Barroso da; Ramos, Luana dos Anjos; Birck, Arlei José
    Primates of the species Alouatta belzebul, also known as howler monkeys, guariba and barbado are endemic to Brazil, of large size, slow movements and quadrupedal locomotion. The knowledge of the morphology of this species is scarce and, in some aspects, as the brain is non-existent. The objective was to describe the macroscopic anatomy of Alouatta belzebul's telencephalon, as well as the vascularization of the base of the brain, nuclei of the base and the venous sinuses of the dura mater. Twenty specimens of Alouatta belzebul were studied, where the brain gyres and grooves were dissected, the base nuclei were identified and the arterial system and venous sinuses were perfused with the injection of colored latex. The telencephalon of A. belzebul presented lysencephalic characteristics, corroborating with several other species of non-human primates and differing from genera such as Pan and Homo. The nuclei of the base were quite evident and the caudate nucleus, putamen, pale medial globe and pale lateral globe, cloister and black substance have been described, which are functionally related to the motor behavior of the species. In the analysis of the encephalization index, it was observed that the Alouatta belzebul is phylogenetically closer to Sapajus and Macaca and more distant from genera such as Brachyteles and Callithrix, showing expressive cognition and intelligence. Regarding the inclination of the central groove, it was observed that in Alouatta belzebul the upper extremity is posterior to the lower extremity, data that corroborate with man, baboons and chimpanzees and reveal a large frontal lobe when compared to Sapajus libidinosus, showing the maximum development in these primates. The arterial circle of the Alouatta belzebul consists of two vascular systems: the vertebro-basilar and the carotid system, which anastomose to close the arterial circuit. In the caudal portion of the arterial circle are the vertebral arteries and their branches: the rostral spinal artery and the caudal inferior cerebellar artery. Anastomosis of the vertebral arteries gives rise to the basilar artery. This presented an anatomical variation with the formation of a double basilar artery, called the arterial island. Nine venous sinuses were observed in the dura mater: dorsal sagittal sinus, ventral sagittal sinus, transverse sinus, straight sinus, sigmoid sinus, temporal sinus, parietal sinus, basilar sinus and cavernous sinus, with morphological similarities in the origin, path and destination of the flow blood to the internal jugular vein, helping in the venous drainage function of the brain in this species. Of the venous sinuses observed, the cavernous sinus was of considerable clinical and surgical importance in Alouatta belzebul due to its topographic arrangement next to the internal carotid artery and pituitary gland, data similar to those described for Sapajus libidinosus, Macaca fascicularis, Macaca mulatta, Papio ursinus, Cercopithecus pygerithrus and Galago senegalensis. The morphological study of the telencephalon, as well as the mechanisms of lining, arterial irrigation and drainage of the venous sinuses, generated information about the brain organization of the primate Alouatta belzebul, which had not been previously described and many of these data generate subsidies for understanding other áreas ethological investigations.
  • Item
    Avaliação histológica, microestrutural e de biocompatibilidade in vivo de cartilagens auriculares de bovino tratadas em solução alcalina
    (Universidade Federal de Goiás, 2021-03-05) Ferreira, Kamilla Dias; Silva, Luiz Antônio Franco da;; Vulcani, Valcinir Aloisio Scalla;; Vulcani, Valcinir Aloisio Scalla; Silva, Danilo Conrado; Martins, Apóstolo Ferreira; Franzo, Vanessa Sobue; Oliveira, Kellen de Sousa
    With the advancement of techniques for the correction of tissue defects or losses, the use of biomaterials that promote repair with a minimum of inflammatory response is sought. Previous studies carried out used different means of preparing and obtaining elastic bovine cartilage, aiming at the use as biomaterial. However, one of the major challenges related to the use of these membranes, refers to the occurrence of unwanted immunogenic reactions at the implantation site. This study aimed to characterize the physical-chemical, microstructural and histological properties of elastic bovine cartilages treated in alkaline solution and their biocompatibility in vivo. Therefore, physical-chemical analyzes were carried out to characterize this material. The thermal analyzes performed, Thermogravimetry (TGA) and Differential Exploratory Calorimetry (DSC), verified the possible changes in the material in view of the temperature variations in which it was submitted. The TGA curves showed the variation in the mass of treated and untreated auricular cartilages as a function of temperature and in the DSC the temperature of the collagen denaturation of the cartilage samples was evaluated. The analyzes showed that the treated cartilage had physicochemical characteristics similar to untreated cartilage. In the microstructural evaluations, 2D and 3D computed microtomography and confocal laser scanner microscopy were performed. Two-dimensional Micro-Ct showed that the cartilage region showed higher density in relation to the perichondrium, and the alkaline treatment was effective in decellularization, due to the presence of gaps observed in the extracellular matrix interspersed with the collagen structure. The three-dimensional Micro-Ct showed that the cartilage has less porosity and pores with a larger diameter and in the laser scanner microscopy it was noted that the treated cartilage has considerable roughness, factors that can contribute to cell proliferation and adhesion. In this study, histological processing of cartilage was also performed, demonstrating that the alkaline treatment promoted tissue decellularization, with the maintenance of the architecture of the extracellular matrix and the structure of elastic and collagen fibers. It was concluded that the alkaline treatment was efficient to promote decellularization in the auricular cartilage. The last stage of the study consisted of evaluating the in vivo biocompatibility of elastic cartilage implants treated with alkaline solution (CA) compared to polypropylene (TP) mesh in rabbits. It was found that the group (CA) had a less intense inflammatory process than the group (TP), in which the formation of a fibrous capsule around the implants was observed. It was noted in the group (CA) the presence of calcification promoted by osteoinduction of osteoblasts due to alkaline processing, which can be considered a bias of interest for further studies, involving the regeneration of bone tissues, in which the cell-type applicability observed is feasible.
  • Item
    Aspectos clínico-epidemiológicos e tratamento da tripanosomose bovina por trypanosoma vivax no estado de Goiás
    (Universidade Federal de Goiás, 2019-03-01) Bastos, Thiago Souza Azeredo; Linhares, Guido Fontgalland Coelho;; Lopes, Welber Daniel Zanetti;; Lopes, Welber Daniel Zanetti; Felippelli, Gustavo; Krawczak, Felipe da Silva; Ferreira, Lorena Lopes; Silveira Neto, Osvaldo José da
    This study reports the first case of bovine trypanosomiasis caused by Trypanosoma vivax in Goiás, Brazil, as well as other cases that occurred within a period of 24 months, demonstrating epidemiological aspects such as needle sharing and commercialization of infected cows influencing the spread of this disease. Furthermore, it was observed that T. vivax was able to infect and develop the disease in calves when they were inoculated by the intradermal, subcutaneous, intramuscular or intravenous route. The presence of Anaplasma marginale latent in calves with trypanosomiasis interfer directly in the percentage of mortality of the animals. Finally, with regard to efficacy, only animals that received isometamidium were able to remain without new parasitaemia after 180 days of study. Treatment with diminazene allowed new parasitemia when reinfected 50 days after treatment and, finally, imidocarb was not effective in the treatment of bovine trypanosomosis. This is the first study that describes the presence of T. vivax, as well as its epidemiology in the State of Goiás, evaluated the infection capacity of this parasite by different pathways, clinical signs, and diagnostic methods and also evaluated the efficacy of the treatment with imidocarb, diminazene and isometamidium in cattle experimentally infected with T. vivax in Brazil.
  • Item
    Aspectos epidemiológicos e clínico-patológicos das poxviroses de bovinos no Estado de Goiás (2010-2018) e soroprevalência de vaccínia bovina em bovinos no Distrito Federal
    (Universidade Federal de Goiás, 2020-12-16) Silva, Lorena Ferreira; Cunha, Paulo Henrique Jorge da;; Sant’Ana, Fabiano José Ferreira de;; Sant’Ana, Fabiano José Ferreira de; Noronha Filho, Antônio Dionísio Feitosa; Sonne, Luciana; Flores, Eduardo Furtado; Borges, José Renato Junqueira
    Based on the importance and in the scarce literature about poxviruses in cattle in Brazilian Midwestern, the current study aimed to determine the seroprevalence of vaccinia virus (VACV) in cattle of Distrito Federal (DF) (2015-2016) and to describe the epidemiological, clinical and pathological aspects of officialy notified cases of poxviruses in cattle of Goiás State (GO), Brazil (2010-2018). The samples used in this study from DF and GO were kindly provided by Secretaria de Agricultura, Abastecimento e Desenvolvimento Rural (Seagri-DF) from DF and by Agência Goiana de Defesa Agropecuária (Agrodefesa), respectively. In the first study, samples of sera of 312 cows from 64 herds were tested by virus-neutralizing test (VN) for bovine vaccinia (BV) antibodies. Estimated prevalences of 33.3% (herds) and 10.6% (animals) were observed. No risk factor with biological relevance was associated with seropositivity to BV. In the second study, during the evaluation period, 33 suspected cases/outbreaks of vesicular diseases in cattle were notified. Twenty-five out of these cases were confirmed as associated with poxviruses: 13 BV, 6 pseudocowpox, 5 bovine papular stomatitis and 1 coinfection (VACV and Orf virus-like parapoxvirus). Most cases occurred in the dry season of the respective years, between the months of June and September. Main lesions included vesicles, ulcers, crusts, papules and scars. These lesions affected mainly the teats and udder of dairy cows. Main lesions in calves consisted of ulcers in the mouth and muzzle. There were concomitant cases with similar lesions in humans that worked closely with the infected cattle. The results of this study demonstrate that different poxviruses (mainly BV) infect cattle in part of the Midwestern Brazil, with zoonotic potencial, mainly in dairy farms, but also present on beef herds. As a result, these diseases require vigilance by the oficial veterinary and local public health service.
  • Item
    Ultrassonografia e morfometria do processo de involução dos componentes umbilicais de cordeiros sadios e com onfalopatia
    (Universidade Federal de Goiás, 2020-11-12) Pereira, Cristiane da Silva; Borges, Naida Cristina;; Sant’Ana, Fabiano José Ferreira de;; Sant’Ana, Fabiano José Ferreira de; Noronha Filho, Antônio Dionísio Feitosa; Queiroz, Paulo José Bastos; Sonne, Luciana; Ximenes, Fábio Henrique Bezerra
    Despite the vast literature on the systemic consequences of umbilical disorders, little has been published about the use of ultrasound to diagnose omphalopathies in small ruminants. Therefore, the objectives of this PhD thesis were: (i) to describe the physiological regression of umbilical structures in healthy sheep - from their first day of life until the completion of this research - using ultrasound imagery of umbilical structures in Santa Inês breeds from natural conception (SMN) (n = 15 ), Dorper from natural conception (DMN) (n = 15) and Dorper from embryo transfer (DTE) (n = 18); (ii) to compare the evolution in the diameters of the measured structures between the two breeds (Dorder and Santa Inês); (iii) to evaluate lambs with omphalopathies - from first clinical signs, or initial sonographic changes, until the end of trials - and compare the data with those from healthy animals; and (iv) sonographically and histotologically describe the clinical cases, as well as related systemic changes such as death or recovery, of five lambs affected by omphalopathies, which were followed throughout the research, and (v) validate ultrasound as an early diagnostic method for this species. In healthy animals, all intra-abdominal umbilical structures could be evaluated in their longitudinal direction. Among all groups of structures being analysed, the urachus (P2) was the first one to finish its involution. Natural conception groups (SMN and DMN) presented similar regression, after the urachus, the umbilical vein at (P5), in days very close to regression the umbilical arteries (P3 and P4) and finally, the hepatic portion of the umbilical vein, in the positions (P6 and P7). In the group (DTE), shortly after the regression of the urachus, the regression of the umbilical vein in the abdominal (P5) and hepatic (P6) portions have followed. Subsequently, after one to two days, the involution of the portion in (P7) has taken place and, only then, around the 14th day, occurred the regression of the umbilical arteries (P3 and P4). There were no great differences in the process of umbilical involution between the breeds Dorper and Santa Inês nor between the methods of conception. We have thus concluded that healthy lambs present a pattern of umbilical structures involution in which 80% of the animals finish the process between their 12th and 14th day of age. In this research, the cases of five lambs with omphalopathies (three from the Santa Inês breed and two from the Dorper breed) were described. Out of these unhealthy animals, three lambs - which were diagnosed with omphaloarteritis by ultrasonography and later in histopathology - have died naturally. Two of them were recovered after treatment, one had omphalophlebitis and the other one had omphaloarteritis. The lesions of the umbilical structures, monitored by ultrasonography, are described in the thesis. The observation and evaluation of these five reported cases have shown that the use of ultrasonography in the evaluation of lambs which present clinical signs suggesting alterations of umbilical structures, with or without systemic involvement, is promising. The results of this study demonstrate that ultrasonography can be used in ovines for describing the regression of the remaining umbilical structures in healthy animals and that ultrasound imagery can be used in routine exams of lambs that present anomalies in their physical tests as a way of diagnosing umbilical alterations and preventing the progression of diseases caused by umbilical infections.
  • Item
    Fotoimunoinativação do alfaherpesvírus bovino 1 em sêmen
    (Universidade Federal de Goiás, 2020-09-29) Oliveira, Taise Maria dos Anjos; Gonçalves, Pablo José;; Cunha, Paulo Henrique Jorge da;; Souza, Guilherme Rocha Lino de;; Borsanelli, Ana Carolina; Almeida, Luciane Madureira de; Souza, Márcio Adriano Rodrigues; Meirinhos, Maria Lúcia Gambarini
    Bovine alphaherpesvirus I (BoHV-1) poses a potential risk of infection to herds around the world, causing numerous economic losses related to both respiratory and reproductive infections. In the reproductive sphere, semen is an important source of spread of the virus to the herd. Thus, aiming at semen disinfection, photodynamic inactivation (PDI) was used as a viable alternative to conventional treatments. This technique involves the combination of a photosensitizing compound (PS), molecular oxygen and light, resulting in the generation of reactive oxygen species, capable of causing cell death by apoptosis, necrosis or autophagy. Several photosensitizing compounds are available on the market, however, new compounds are developed with the aim of improving efficiency against specific biological targets. Thus, this study evaluated the efficacy of palladium-bound porphyrins, halogenated sulfonated porphyrins, hematoporphyrins (HP) and tetracarboxy-phthalocyanine zinc (ZnPc), in vitro photodynamic inactivation BoHV-1. As well as, the development of immunoconjugates, composed by the direct conjugation between immunoglobulins (IgY anti-BoHV-1) and photosensitizers (ZnPc and HP) for PDI in semen. Among the photosensitizers tested, palladium porphyrins (dppf and dppe) at 10 μM showed better results in vitro, inactivating the virus completely with up to five minutes of irradiation. The photosensitizers HP and ZnPc were tested and proved to be safe for PDI in semen after evaluation of the parameters of quality and sperm viability. These PS were conjugated to IgY-anti-BoHV-1 and tested in semen infected experimentally with BoHV-1, being efficient for the complete inactivation of the virus within five minutes of irradiation.
  • Item
    Efeito da fotobiomodulação e de células-tronco derivadas do tecido adiposo na reparação de feridas por queimadura térmica em ratos
    (Universidade Federal de Goiás, 2020-07-31) Ribeiro, Maisa; Moraes, Julia de Miranda;; Vulcani, Valcinir Aloisio Scalla;; Vulcani, Valcinir Aloisio Scalla; Prado, Rodrigo Paschoal; Araújo, Gustavo Henrique Marques; Saturnino, Klaus Casaro; Rita, Ricardo de Mattos Santa
    Introduction: Injuries caused by full-thickness burns are considered a severe form of trauma with a high worldwide morbidity and mortality rate capable of promoting destruction of skin tissue and morphologically compromising the organism. The use of stem cells derived from adipose tissue (ADSC) and low-level laser (LLL) are promising therapeutic alternatives explored by regenerative medicine. The aim of this study was to establish an effective isolation model for more homogeneous ADSC cells; to evaluate the effects of ADSC and LBP on the skin healing process and to investigate the association of techniques (LBP and ADSC) in the treatment for full-thickness burn in an experimental rat model. Material and methods: The obtaining of the ADSC was carried out by means of imonumagnetic isolation by CD49d protein and conventional culture. A total of 150 Wistar rats were divided into a control group and groups treated with ADSC CD49d positive, ADSC CD49d negative, conventionally isolated stem cells (CULT), LLL, ADSC+LLL, ADSC-LLL. Scald burns were induced on the back of each animal, with subsequent surgical debridement. LLL with a wavelength of 660 nm, 30 mW of power at five points of the wound was used, under total fluence of 450 Joules / cm2 and total application of 5 Joules. Macroscopic and histopathological analyzes were performed according to biopsies collected on days 5, 14 and 21 post-injury, processed and stained using the techniques of Hematoxylin and Eosin (HE) and Gomori's Trichrome (GT). Statistical analyzes were performed using Tukey's test and analysis of variance (ANOVA), Kruskal-Wallis test (non-parametric) and T-Paired test. For all analyzes, a significance level of 5% (p <0.05) was used. Results: The results showed a statistical difference in morphometry between the groups with the best degree of reepithelization and reduction in the lesion area presented by the ADSC+ and CULT treatments at 21 days of biopsy. Histopathological analysis showed that ADSC+, LLL and ADSC+LLL treatments significantly influenced the healing process, showing improvement in the increase of mononuclear inflammatory infiltrates, greater angiogenesis, decreased edema, increased granulation tissue, in the number of fibroblasts at the beginning of tissue repair, in addition to promoting re-epithelialization in a more organized manner with greater deposition of collagen and formation of epithelial attachments at 21 days after the injury. Conclusion: low-power laser photobiomodulation of ADSC cells enhanced the repair process in full-thickness burn wounds, demonstrating a significant interaction for the remodeling of the extracellular matrix and skin tissue. Additional studies are needed to measure the effects of LBP and the activity of ADSCs on skin tissue repair.
  • Item
    Efeitos dos extratos de folhas de campomanesia adamantium e hymenaea martiana hayne sobre células de osteossarcoma canino e células endoteliais humanas
    (Universidade Federal de Goiás, 2020-02-28) Vieira, Vanessa de Souza; Cruz, Vanessa de Sousa;; Araújo, Eugênio Gonçalves de;; Araújo, Eugênio Gonçalves; Pfrimer, Gabriel de Abreu; Pereira, Kleber Fernando; Arnhold, Emmanuel; Oliveira, Gerlon de Almeida Ribeiro de
    Osteosarcoma (OSA) is a very aggressive tumor in dogs, with low survival rate and ineffective treatment. In the search for alternative sources of therapy, the Brazilian biome becomes a scientific hope, presenting a diversity of medicinal plants of popular knowledge and use. However, they can have controversial biological functions. The aim of this study was to verify the bioactivity of the leaves of Campomania adamantium and Hymenaea martiana Hayne on canine osteosarcoma (OC) cells from cell cultures and human umbilical vein endothelial cells (HUVEC). As cells were cultured and subjected to treatment with C. adamantium (1 μg / mL, 10 μg / mL, 100 μg / mL, 1000 μg / mL) and H. martiana Hayne (10μL / mL, 100μL / mL, 1000μL / mL , 2000μL / mL and 5000μL / mL), in the 24h, 48h and 72h exposure periods in normoxia and 24h in cells exposed to oxidative stress, induced by hydrogen peroxide. The results were analyzed by analyzing cell viability and cytotoxicity using the tetrazolium reduction method (MTT). This study brings unprecedented results in relation to extractive effects, as it was possible to demonstrate that they have little cytotoxic action. In addition, it was observed that the higher the dosage and the longer the exposure time, the greater the proliferative activity, with increased cell viability in the 72-hour group. In endothelial cells, similarly, there was no cytotoxic activity, in addition to the occurrence of increased cell viability, even after submission to oxidative stress. C. adamantium leaf extract increased viability and did not show cytotoxic action in canine osteosarcoma cells and endothelial cells under oxidative stress. The crude ethanolic extract of the leaves of Hymenaea martiana Hayne, increased the cell viability of canine osteosarcoma cells and human umbilical vein endothelial cells, submitted to oxidative stress.
  • Item
    Detecção dos genes spvC e prot6E e avaliação da infecciosidade de Salmonella sp. em poedeiras comerciais
    (Universidade Federal de Goiás, 2019-03-06) Figueira, Samantha Verdi; Rezende, Cíntia Silva Minafra e;; Moraes, Dunya Mara Cardoso;; Andrade, Maria Auxiliadora;; Andrade, Maria Auxiliadora; Faria, Adriana Marques; Rocha, Fernanda Rodrigues Taveira; Stringhini, José Henrique; Teixeira, Weslen Fabrício Pires
    The pathogenesis of Salmonella includes different factors, such as plasmidial genes, which because they are mobile genetic mechanisms can increase the bacterial genetic diversity contributing to the modification of virulence and adaptation to the hosts. The present work was developed to detect the presence of the virulence genes spvC and prot6E in different serovars of Salmonella enteric present in the metropolitan region of Goiânia and to verify the ability of these serovars to express their pathogenicity in embryonated eggs, neonates and laying hens and to cause contamination in eggs in experimental models. In Experiment 1, isolates of Salmonella Enteritidis, Gallinarum, Heidelberg, Infantis, Schwarzengrund and Typhimurium were obtained from organs of sick bird, eggs and environment of poultry chain and were investigated the presence of spvC and prot6E genes by real-time PCR. Of the 47 isolates, 15 (31.9%) were positive for the spvC gene and five (10.6%) for the prot6E gene. It is concluded that Salmonella Enteritidis, Typhimurium and Gallinarum, isolated from the bird and eggs contain the spvC genes. The prot6E gene was detected in the serovars Salmonella Enteritidis, Heidelberg and Typhimurium from egg samples. The serovars of Salmonella Heidelberg, Infantis, Schwarzengrund and Typhimurium from environmental samples do not have the plasmid genes spvC and prot6E. In experiment 2, two isolates of Salmonella Gallinarum, one positive and one negative for the spvC gene, were inoculated into embryos and neonates of laying birds to investigate whether the presence of the spvC gene in the Gallinarum serovar is able to determine embryonic mortality, affect the quality of the neonate and chick, production parameters and cause systemic disease in young birds. It was observed high mortality of the groups inoculated with Salmonella Gallinarum in alantoid route with one and 14 days of incubation, independent of the presence of the spvC gene. No embryonic mortality, changes in neonatal weight / egg weight and changes in neonatal quality were observed for birds inoculated through a 19-day in air chamber. Salmonella Gallinarum without and with the spvC gene was recovered from the heart, spleen, liver and ceca of birds inoculated in the air chamber, whereas for the oral inoculated group, the pathogen without the gene was recovered only in the ceca and the pathogen with the gene was recovered from spleen and cecum. Salmonella inoculation led to decreased weight gain and altered biometry of the heart and intestine. It is concluded that Salmonella Gallinarum is capable of causing embryonic mortality in embryos inoculated with one and 14 days in the allantoic cavity. The presence of the spvC gene in Salmonella Gallinarum does not affect embryo mortality, incubation parameters and systemic dissemination for birds inoculated in the air chamber. For birds inoculated by the oral route, the presence of the spvC gene determines enteric and systemic infection, whereas the isolate without the gene remains restricted to the cecum. In Experiment 3, the Salmonella Heidelberg isolate positive for the prot6E gene in experiment 1 was inoculated in laying hens in the oral, intravaginal and intravenous routes in order to investigate their ability to cause clinical disease, egg and gastrointestinal tract contamination. The pathogen was isolated in the excreta, only 12 hours after inoculation in 33% of the birds inoculated from oral route and 66% of the birds inoculated intravaginal route. In the eggs, the pathogen was isolated at 24 and 48 hours, seven and 15 days. It is concluded that the presence of the prot6E gene in Salmonella Heidelberg is not sufficient to cause clinical salmonellosis in laying hens, but it determines contamination of the eggs and the gastrointestinal tract of chickens, regardless of the route of inoculation, oral, intravaginal and intravenous.
  • Item
    Fatores de risco para mastite bovina e avaliação fenotípica de resistência antimicrobiana
    (Universidade Federal de Goiás, 2020-02-28) Almeida, Thamara Venâncio de; Cordeiro, Clarice Gebara Muraro Serrate;; Zacaroni, Ozana de Fátima;; Nicolau, Edmar Soares;; Nicolau, Edmar Soares; Alves, Fernanda Antunha de Freitas; Souza, Fernando Nogueira de; Rezende, Cíntia Silva Minafra e; Silva, Marco Antônio Pereira da
    Bovine mastitis is the most common disease that causes the most economic losses in dairy herds around the world. The objective of this study was to evaluate the risk factors for subclinical mastitis in dairy herds in the State of Goiás, to identify the gram-positive microorganisms that cause mastitis, as well as their antimicrobial resistance. Milk samples were collected from 1,034 lactating cows from 27 dairy farms located in the State of Goiás and analyzes of somatic cell count (SCC), bacterial examination and antimicrobial sensitivity test were performed. A checklist was applied to each participating property to obtain data on the animals, the properties and the hygienic and sanitary management of the herds. Risk factors for mastitis were assessed using a mixed generalized linear model. Of the 27 properties, 11 (40.74%) had SCC of the tank above 500,000 cells/mL, therefore above the maximum limit established by Brazilian legislation. The prevalence of subclinical mastitis in herds (SCC> 200,000 cells / mL) was 54.93%. The main risk factors associated with the occurrence of subclinical mastitis in dairy cows were old age, pendulous udder and advanced lactation stage. 216 microorganisms were identified, gram-positive cocci. Staphylococcus spp. (53.10%), particularly S. aureus (30.09%), stood out as one of the most common agents involved in the etiology of bovine subclinical mastitis, followed by Streptococcus spp. (22.12%) and Enterococcus sp. (16.81%). The highest rates of antimicrobial resistance were Staphylococcus aureus to penicillin (76%) and ampicillin (74%), Streptococcus spp. to neomycin (65.08%), gentamicin (46.03) and tetracycline (43.66%) and Enterococcus spp. tetracycline (52.38%). Isolates of Streptococcus spp. multiresistant (18.06%).
  • Item
    Caracterização físico-química e atividade da Curcuma longa L. em células endoteliais sob estresse oxidativo
    (Universidade Federal de Goiás, 2020-02-28) Teles, Juliana Carvalho de Almeida Borges; Saturnino, Klaus Casaro;; Cruz, Vanessa de Sousa;; Araújo , Eugênio Gonçalves de;; Araújo, Eugênio Gonçalves de; Iglesias, Ana Paula; Damasceno, Adilson Donizete; Beretta, Daniel Cortês; Arantes , Diego Antônio Costa
    Ischemic tissue injury is quite harmful, however, reperfusion causes an exacerbated level of reactive oxygen species (ROS), which directly interfere in the cell signaling cascade, with cell deleterious effects such as lipid peroxidation, protein oxidation and DNA damage. Researches had shown the beneficial effects of antioxidants present in vegetables and among them is curcumin, an antioxidant pigment found in Curcuma longa L. rhizomes, known as turmeric or saffron. This plant has demonstrated pharmacological activities, which have been reported since antiquity, as antioxidant, anti-inflammatory, antibacterial, antifungal, antitumor, antiparasitic, antiviral, healing, hypoglycemic, neuroprotective and immunomodulatory. Therefore, the aim of this research was to evaluate the possible antioxidant action of curcumin on human umbilical vein endothelial cells (HUVEC), in culture, submitted and not submitted to oxidative stress (EO). This study was divided into three phases. In the first time, the action of standard curcumin (Sigma Aldrich, Reference C1386) was evaluated at different concentrations (0 μmol, 20 μmol, 50 μmol, 100 μmol and 1000 μmol) and for three different times of continuous exposure (24 h, 48 h h and 72 h). The EO assay was induced through cellular exposure to 1% hydrogen peroxide and through the MTT (methyl-thiazolyl-tetrazolium) reduction assay, cell viability and cytotoxicity could be assessed. In the second phase, we determinated the presence and content of antioxidant compounds in the ethanol extract of Curcuma longa L. (EEC) from Mara Rosa-GO. EEC samples were subjected to Folin-Ciocalteau tests, capture of the DPPH radical (2,2-diphenyl-1-picryl-hydrazil) and the HPLC-Fluorescence method. In the third phase of this research, the effect of EEC in HUVEC cultures was evaluated under the same experimental conditions as the first phase, in addition with the possible signaling pathways of both survival and cell death. The results showed that the cells treated with 20 μmol of standard curcumin and at 24 h and 48 h showed better cell viability and, consequently, less cytotoxicity. The occurrence of oxidative stress was validated by the decrease in cell viability within the three times variables used, but an antioxidative protection was found at 50 μmol of curcumin. In EEC, we identified gallic acid, curcumin (16.7%) and the amount of 18.1 μg / mL of extract, to reduce the DPPH radical by 50%. Results from the third phase indicated that the EEC possibly alleviated cell damage by oxidative stress at the dosage of 100 μg / ml, by decreasing the immunoexpression of caspase3 and by increasing the signaling of the SIRT1 survival route. It was concluded that both pure curcumin and EEC can improve the cell viability of HUVEC under oxidative stress.
  • Item
    Salmonella sp. em excretas, carcaças e ovos de Gallus gallus comercializados em feiras de Goiânia, Goiás
    (Universidade Federal de Goiás, 2019-12-20) Cardozo, Stanislau Parreira; Andrade, Maria Auxiliadora;; Jayme, Valéria de Sá;; Jayme, Valéria de Sá; Nunes, Iolanda Aparecida; Santin, Ana Paula Iglesias; Laboissière, Michele; Braga, Ísis Assis
    The production and way of selling free-range chickens in fairs are important factors in the transmission of infections such as salmonellosis to humans. This work was carried out with the aim of investigating the occurrence of Salmonella enterica in poultry products sold at free fairs in Goiânia from which 100 samples of chicken carcasses, 100 samples of excretes and 50 dozen eggs were collected. In this study were used bacteriological research, serotyping and determination of antimicrobial resistance profile, and analysis of the genetic profile of the isolates. There was isolation in six excrete samples; five in the carcasses and three in the eggs. Regarding serotyping, there were identified in the carcass Salmonella Cerro, Salmonella Panama, Salmonella Rugosa, Salmonella Corvallis, besides the antigenic samples O:4,5; O:3,10 in the samples of excretes were identified Salmonella Typhimurium, Salmonella Agona, Salmonella Rugosa, Salmonella Anatun and a rough strain; and Salmonella Corvallis in the egg samples. In the analysis of the Resistance Profile 12 antibiotics were tested. For the isolates from the carcasses two did not present resistance to any antibiotics, two showed resistance to two antibiotics, one to just one antibiotic and another one presented multiresistance to drugs. The isolates from excretes three were sensitive to all antibiotics, one isolated to only one antibiotic and one with multiresistance (three antibiotics), while the three isolates from the eggs did not present resistance to any of the antibiotics tested. The result of polymerase chain reaction – PCR, besides confirming the presence of Salmonella in the positive samples from conventional bacteriology also identified the presence in three excrete samples, two carcasses and three of eggs. In addition, PCR also detected the presence of the blaTEM gene in four samples, the qnrS gene in five samples and aadA2 in any of the samples initially positive for Salmonella spp. Using Electrophoresis in Pulsed Field - PFGE, it was possible to identify that four Salmonella Corvallis isolates are part of the same clonal group as well as the two of S. Anatum, while the other identified serotypes belong to different clonal groups among themselves and those previously mentioned. From these results, it is infered that the system of free-range chicken productions, represents an important role in the epidemiology of salmonellosis, either as maintainer or as a disseminator of several Salmonella Enterica serovars, representing a concern for industrial poultry farming and public health.
  • Item
    Resposta humoral e clínica à vacinação contra leishmaniose em cães sororreagentes
    (Universidade Federal de Goiás, 2019-12-13) Bezerra, Lucilandia Maria; Junqueira-Kipnis, Ana Paula;; Moura, Veridiana Maria Brianezi Dignani de;; Moura, Veridiana Maria Brianezi Dignani de; Leandro, Mirian Dorta; Paiva, Monique; Matos, Moema Pacheco Chediack; Paz, Gustavo Fontes
    In endemic regions for visceral canine leishmaniasis (CVL), the biggest challenge currently faced is the detection of infected and asymptomatic animals, as it may be the biggest risk factor for maintenance of the infection. Therefore, the diagnosis based on more sensible and specific tests may be the possible solution to control this disease. Serological tests that can identify exposed, infected or diseased animals, such as Elisa IgG anti-Leishmania, stand out as excellent tools for detecting animals in this clinical situation in high-risk areas because of their high influence in these situations. Among the pathological clinical markers that may contribute to the detection of Leishmania infection, hematological and biochemical tests were performed. As well as the early and safe diagnosis, the treatment of infected or infectious dogs using vaccines has been of interest to researchers. In this sense, two experiments were performed. In the first, the immunotherapeutic response of a double dose anti-Leishmania vaccine (Leish-Tec®) in dogs from an endemic area for CVL was evaluated. For this purpose, 85 serum samples from asymptomatic, non-reactive and low titrated reagents (1/40 cutoff point) for RIFI tests negative to the parasitological bone marrow test were analyzed. The immune response of treated dogs was evaluated by the anti-rA2 IgG ELISA whose antigen (rA2) is part of the vaccine composition. The second evaluated the performance of a recombinant protein rK28, aiming to verify its sensitivity in detecting infected animals in endemic area. In this sense, a clinical intervention and control trial was designed to analyze the positive and negative outcomes of the animals under study. According to serological screening tests (RIFI) the animals were divided into five groups, including NRNV - unvaccinated nonreactive, n = 23; RNV - unvaccinated reagent, n = 7; NRVS - vaccinated nonreactive 1 ml, n = 22; NRVD - non-vaccinated reagent 2 ml, n = 16; and RVD - 2 ml vaccinated reagent, n = 15. After grouping, the animals were vaccinated with three applications at 21-day intervals and followed at six points of care every three months through clinical and laboratory examinations for a period of 18 months. To assess the presence or absence of parasites, bone marrow and PCR parasitological tests were performed at six and nine months after vaccination, respectively. The animal’s serological response to infection was assessed by anti-rA2 and anti-rK28 IgG levels. Hepatic and renal enzymes were tested as a condition of possible tissue damage in these organs, no changes compatible with infection or inflammation were observed, in the average of the groups in the last following point. Regarding the evaluation of responses to anti-rA2 IgG antibodies, there was a high response in the single-dose (NRVS) vaccinated groups and low response in the double dose (NRVD and RVD) vaccinated groups. When the groups were compared, there was a statistical difference between the vaccinated groups and the unvaccinated nonreactive control. The rK28 protein performed well in all groups, showing a high sensitivity in detecting positive and negative infected parasitological tests and PCR. It was concluded that the anti-leishmania vaccine (Leish-Tec®) is immunogenic by inducing high levels of anti-rA2 IgG antibodies. Double-dose vaccination reduces anti-rA2 antibody levels compared to single-dose vaccination. However, single or double dose vaccination of seropositive animals for IFAT does not provide protection against Leishmania infection. Also, detection of anti-rK28 IgG presents good performance for diagnosis of CVL in asymptomatic animals exposed in endemic area.
  • Item
    Caracterização fenotípica e genotípica de Salmonella enterica de origem avícola e atividade antimicrobiana de extratos de própolis
    (Universidade Federal de Goiás, 2019-11-21) Benicio, Cristyene Gonçalves; Machado, Bruna Aparecida Souza;; Andrade, Maria Auxiliadora;; Rezende, Cíntia Silva Minafra e;; Rezende, Cíntia Silva Minafra e; Borges, Liana Jayme; Teixeira, Weslen Fabricio Pires; Prado, Cristiano Sales; Lage, Moacir Evandro
    The objective of this study was to characterize phenotypically and genotypically Salmonella isolates from food and environmental samples, as well as to evaluate the in vitro antibiotic action of green and red propolis extracts. The 128 isolates of Salmonella enterica were obtained and analyzed for antimicrobial susceptibility by disk diffusion method. The antimicrobials and microgram concentrations tested were: amoxicillin-clavulanic acid (30μg), cotrimoxazole (trimetropim-sulfamethoxazole) (25μg), ciprofloxacin (5μg), enrofloxacin (5μg), ceftiofur (30μg), chloramphenicol (30μg), chloramphenicol (30μg) and doxycycline (30μg). Salmonella Enteritidis (ATCC 13076), Salmonella Typhimurium (ATCC 14028) and Escherichia coli (ATCC 25922) strains were used as a quality control of the sensitivity tests. Two genes of virulence and nine of resistance were searched. Among the 128 isolates, 40 were tested against propolis action. Propolis extracts diluted in ethanol were evaluated for their antimicrobial activity ranging from 0.2 to 100 mg/mL-1 by determining the minimum inhibitory and bactericidal concentration. We found 27 different serotypes, the most common was Salmonella Heidelberg, Saintpaul, Typhimurium and Minnesota. It was found that 85.2% (109/128) of the isolates were resistant to one or more antimicrobial agents used and only five serotypes showed no resistance. We identified 54 multiresistant isolates. Of the eight antimicrobials evaluated, for two of them (chloramphenicol and phosphomycin) all isolates were susceptible and the one with the largest number of resistant isolates was ciprofloxacin. Among the resistance genes evaluated (blaCTX-M and gyrA), about 70% (89/128) of the isolates harbored it. Regarding virulence genes, three (invA, hilA and sseD) were identified in all isolates tested. For the other six, spvR presented the highest number of isolates in which it was not identified. Of the 18 different serotypes tested against antibiotic action of propolis, the most susceptible were Salmonella Senftenberg and the most resistant: Agona, Braenderup, Heidelberg, Children, Minnesota, Schwarzengrund, Newport, Orion, Saintpaul and Anatum. In general, the lowest inhibitory and minimum bactericidal concentration values were 3.1 mg/mL and 6.25 mg/mL, respectively. The best antimicrobial action was for red propolis, regardless of the type of extraction used to obtain the extracts. Thus, propolis may be a viable alternative as antimicrobial under Salmonella sp because it it has expressive action potential. Seeking alternatives for the treatment and control of pathogenic microorganisms has been important through the emergence of multiresistant strains.
  • Item
    Desenvolvimento e avaliação de filmes incorporados de extratos naturais e determinação da oxidação em filés de tilápia
    (Universidade Federal de Goiás, 2019-12-13) Turella, Camila Cheker Brandão; Silveira, Miriam de Araújo Fontes;; Rezende, Cintia Silva Minafra e;; Lage, Moacir Evandro;; Souza, Adriana Régia Marques de; Tenório, Clarice Genabra Muraro Serrate Cordeiro; Oliveira, Raphael Rocha de; Bueno, Claudia Peixoto
    This study aimed to produce active biofilms with antioxidant action in the conservation of tilapia (Oreochromis niloticus) fillets. For this, by completely randomized design, natural extracts of pequi bark, araticum, cajazinho and saffron were extracted. All extracts were evaluated for phenolic compounds content and antioxidant capacity. The extract of pequi bark obtained the highest antioxidant capacity (1,874 μmol.g-1), and its phenolic profile showed that the main compounds responsible for its high antioxidant power were ellagic, gallic and protocatechic acids. Pequi bark, green tea / rosemary extracts and BHA were incorporated into cellulose acetate films. These films were analyzed for maximum supported stress, tensile strength, perforation strength, elongation, elasticity, CO2 permeability, migration of phenolic compounds in fat and water simulators and electron microscopy. Pequi bark extract films have good mechanical strength, with a rupture stress of 32.28MPa, a gradual and maximum migration of phenolic compounds to the simulating of two days, and also show to be porous and gas permeable. Films of four treatments (control, pequi bark, green tea / rosemary and BHA) were used to pack tilapia fillets, which were stored for 15 days at 4°C. The experiment was conducted in a randomized block design, consisting of four treatments and four replication. During the study period, the analysis of lipid (TBARS) and protein (carbonyl and sulfhydryl) oxidation were performed at times 0, 4, 7, 10, 12 and 15 days. At the end of this period, films with BHA and pequi bark extract were able to maintain the fillets with better oxidative rates, proving to be effective in preventing oxidation. Therefore, the extract produced from pequi bark showed potential antioxidant action and high effectiveness in combating oxidation, and can be used not only in fish but in various foods.
  • Item
    Tipagem molecular, detecção de genes de virulência e determinação do potencial patogênico de isolados de Escherichia coli de patos (Cairina moschata), perus (Meleagris gallopavo), galinhas (Gallus gallus)
    (Universidade Federal de Goiás, 2019-02-28) Almeida, Ana Maria de Souza; Rezende, Cintia Silva Minafra e;; Jayme, Valéria de Sá;; Andrade, Maria Auxiliadora;; Andrade, Maria Auxiliadora; Moraes, Dunya Mara Cardoso; Cervi, Renato Clini; Nunes, Iolanda Aparecida; Porto, Regiane Nascimento Gagno
    The general objective of this work is to evaluate the phylogenetics and to detect virulence genes from Escherichia coli isolates from different bird species, as well as to investigate their pathogenic potential in fertile broiler eggs. In the present study three experiments were carried out to analyze, in different aspects, E. coli isolates from different bird species. In experiment 1, E. coli isolated from ducks and turkeys were inoculated into broiler chicks one day to evaluate the pathogenicity of the strains. Fifty isolates of E. coli originating from ducks and 50 isolates from turkeys were tested for the genes iss, iucD, papC and tsh, all commonly found in APEC. Of the 100 isolates of E.coli analyzed, 42% (42/100) detected one or more genes and only 14.2% (6/42) had the four genes studied. After PCR, the positive strains were selected for the four genes common to APEC. Three isolates from ducks and three turkeys were purified separately and six inoculants were inoculated at 107 CFU / mL and inoculated via air sac in one - day - old chicks to evaluate the pathogenicity of these six positive isolates for the four genes. The chicks were evaluated twice a day for ten days in relation to mortality rate, clinical signs and macroscopic findings. The challenged chicks presented respiratory signs (sneezing, nasal discharge and rales) and macroscopic lesions such as aerossaculitis, persistence of the yolk sac, pericarditis and exudate in the nasal sinuses. In view of the results, it is concluded that E.coli originating from both ducks and turkeys are potentially pathogenic for day-old challenged cuttlefish chicks. In the experiment 2, embryos of broiler chickens were challenged with E. coli isolates positive for the iss, tsh, papC and iucD genes from hen, broiler, duck and turkey to analyze the pathological and hematological conditions , besides the performance of broiler chickens kept up to 42 days of age. Embryos with 18 days of incubation were challenged, via air chamber, with 0.1 mL of four different inoculants, according to the treatment, (T2 - E.coli of hen chicken, T3 - E.coli of broiler, T4 - E.coli of duck and T5 - E. coli of turkey) with concentration of 107 CFU / mL and with sterile saline solution (T1 - control group). Embryonic and post - hatch mortality rates, clinical signs, hematological alterations, interference of zootechnical performance and macroscopic findings during the incubation period and rearing up to 42 days of life were evaluated. At 14, 28 and 42 days, one bird from each of the four replicates of the five treatments. Regardless of the species of origin, E.coli isolates were not able to cause significant mortality in embryos challenged at 18 days of incubation. At birth and up to 42 days of age, broiler chickens developed respiratory disease, worsened zootechnical performance and there were no significant haematological changes. In the experiment 3, 33 E. coli isolates from ducks, turkeys, chickens and pigeons were evaluated by PFGE and antimicrobial susceptibility test to characterize their respective phylogenetic profiles and susceptibility to antimicrobials commonly used in poultry farming. E. coli originated from ducks, turkeys, chickens and pigeons grouped into a group with 41.1% similarity. Resistance to different antimicrobials tested (ampicillin, amoxicillin, ceftiofur, doxycycline, enrofloxacin, neomycin, sulfametazole, sulfonamide, tetracycline, cotrimoxazole and chloranfericol) was observed among the 33 isolates of E. coli, mainly tetracyclines, followed by fluoroquinolones and sulfonamides. It is concluded that E.coli originating from birds have different profiles and behaviors.
  • Item
    Microbioma do leite de bovinos das raças Curraleiro Pé-Duro e Pantaneiro
    (Universidade Federal de Goiás, 2019-08-22) Soares, Nayana Ribeiro; Tenório, Clarice Gebara Muraro Serrate Cordeiro;; Zacaroni, Ozana de Fátima;; Rezende, Cíntia Silva Minafra e;; Rezende, Cíntia Silva Minafra e; Carmo, Adriana Santana do; Andrade, Maria Auxiliadora; Fioravanti, Maria Clorinda Soares; Lage, Moacir Evandro
    The objective of the present study was to describe the microbiota of raw milk from naturalized bovine breeds, such as Curraleiro Pé- Duro and Pantaneiro and to increase the value of these herds, through their use as specific genetic banks of microorganisms. Attention is also given to the possible technological application of microbial populations present in the udder and milk of these breeds. Six samples of milk from the Curraleiro Pé-Duro herd were collected at the State University of Mato Grosso do Sul, University Unit of Aquidauana, while 15 samples from the Pantaneiro herd were collected from the Aquidauana Center for Pantaneiro Cattle Conservation also belonging to the State University of Mato Grosso do Sul, University Unit of Aquidauana .From the results found in this study, the Somatic Cell Count and the centesimal composition were determined to identify lactic acid bacteria present in the raw milk and to recognize those isolated lactic acid bacteria both by phenotype and genotype that produce bacteriocins with inhibitory activity against food pathogens and their susceptibility to antimicrobials. In addition, the genes present and protein sequences were described. The results suggest that, even though it is a herd of low milk production, it presents quality. The microbiota that makes up the Curraleiro Pé-Duro and Pantaneiro breeds is specifically interesting with regard to the functional use of lactic acid bacteria. Enterococcus faecium was the dominant species, with phenotypic variability for the production of bacteriocins and resistance to vancomycin and penicillin. Pantaneiro cattle isolates were known to produce bacteriocins anti- Listeria monocytogenes and with genome elements that show double suitability for agroindustrial use, as well as risk to animal and human health.
  • Item
    Capacidade de colonização e expressão imunomoduladora de Salmonella Heidelberg e Salmonella Typhimurium em poedeiras livres de patógenos específicos
    (Universidade Federal de Goiás, 2019-06-28) Rauecker, Ursula Nunes; Andrade, Maria Auxiliadora;; Rezende, Cíntia Silva Minafra e;; Rezende, Cíntia Silva Minafra e; Tenório, Clarice Gerbara Muraro Serrate Cordeiro; Sola, Marília Cristina; Teixeira, Weslen Fabrício Pires; Pascoal, Lívia Mendonça
    Laying hens and broilers are potential reservoirs for Salmonella enterica and the zoonotic risk composes predictive analyses, regarding the attributes for infection and multiplication in poultry flocks and, consequently, food contamination. The aim of this study was to evaluate the potential of internal organ colonization, fecal shedding patterns and egg contamination by Salmonella Heidelberg and Salmonella Typhimurium, and evaluate the influence of infection by Heidelberg and Typhimurium serovars on TLR-4, IL1β, IL6 and IL10 expression in spleen, magnus and uterus of experimentally inoculated layers. 28-week-old White Leghorn laying hens were selected, free of specific pathogens, divided into three groups kept in isolation units. In the first group, SH, 1.2 x 10 7 CFU of Salmonella Heidelberg was inoculated orally in each bird, in the second group, ST, 1.0 x 10 7 CFU of Salmonella Typhimurium. In the control group, birds received equal volume of sterile saline solution. At seven, 14 and 21 days after inoculation, five layers / group were euthanized and necropsied. Spleen, liver, ovaries, magnum, isthmus, uterus, small intestine and caecum were collected and destined for histopathological and bacteriological tests. Eggs were collected daily and excreted every three days, before and during the period of the experiment and analyzed. qPCR was performed for expression analysis of TLR-4, IL6, IL10 and IL1B genes in spleen, magnus and uterus. Results were normalized using constitutive genes RPLP-1 and β-actin and expression was calculated by the ratio of the mean differences between the control group and the target gene in challenge groups. None of the layers showed clinical signs after oral inoculation. Fecal excretion was intermittent and lasted 12 days for both serovars. Heidelberg serovar was identified in all tissues evaluated seven DPI, as well as Salmonella Typhimurium, identified in all tissues evaluated, except ovary, seven DPI. Salmonella Heidelberg persisted 14 DPI in the cecum and spleen, while Typhimurium serovar was not identified in any tissue over the same period. Experimental inoculation generated infection with 14 DPI resolution in ST layers and 21 DPI in SH layers. Salmonella Heidelberg persisted longer in the host, with microbiological isolation in the caecum and spleen associated with discrete induction of inflammatory response in the evaluated organs. Both serovars were able to contaminate the eggs produced in the post inoculation period, being isolated in shell from three DPI and yolk up to 12 DPI. The histopathological alterations found were of discrete score, without alterations in the spleen. There are interactions between the TLR4, IL1B, IL6 and IL10 genes, evidencing the presence of five functional partner genes STAT3, IL10RA, TRAF6, TICAM1 and LY96. The results show differences in antigen recognition and immune response determined by mRNA expression of TLR-4, IL1B, Il6 and IL10 genes in laying hens infected with Salmonella Heidelberg and Salmonella Typhimurium.
  • Item
    SNPs e microssatélites na caracterização racial e de resistência a enfermidades em bovinos da raça Curraleiro Pé-Duro
    (Universidade Federal de Goiás, 2018-12-05) Freitas, Thais Miranda Silva; Carmo, Adriana Santana do;; Moura, Maria Ivete de;; Fioravanti, Maria Clorinda Soares;; Fioravanti, Maria Clorinda Soares; Landi, Vincenzo; Cruz, Aparecido Divino da; Jayme, Valéria de Sá; Badr, Kareem Rady
    The Brazilian cattle breed Curraleiro Pé-Duro is an important genetic resource that has as characteristics of rusticity, adaptability to environmental conditions, and resistance to diseases and intoxications. This study aims to analyze the genetic diversity of Curraleiro Pé-Duro populations and verify the presence of SNPs and microsatellite polymorphisms for breed characterization and disease resistanceto Brucella abortus, Leptospira spp, Neospora caninum, leukosis virus, Infectious Bovine Rhinotracheitis, and Bovine Viral Diarrhea. A total of 1.182 serum and blood samples of Curraleiro Pé-Duro breed belonging to the Pro-Centro-Oeste Network database were used. Amplification and genotyping of 28 microsatellite markers of breed characterization and 13 markers related to the bovine leukocyte antigen (BoLA) and SLC11A genes were used. Twenty-four single-base polymorphisms (SNPs), detected by Kompetitive Allele-Specific PCR, were investigated in genes related to cytokines expression. Univariate analysis was performed for the association of genotypes with phenotypes, followed by logistic regression considering a significance level of 5%. Bayesian analysis grouped the animals into three breed clusters, indicating the presence of subpopulation, with similar prevalence for infections. Heterozygosity and Fst values indicated little differentiation between clusters and increased homozygosity. On BoLA gene, 42 allels associated to infections were identified. SNP haplotypes in BTA6 and BTA15 were significant in seronegative animals while BTA17 haplotype 2 was presented in 70% of Neospora caninum positive animals. The results showed that Curraleiro Pé-Duro cattle breed has alleles of resistance to infections that can be lost due to loss of genetic diversity.