Mestrado em Ciência Animal (EVZ)
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Navegando Mestrado em Ciência Animal (EVZ) por Por Orientador "Borges, Lígia Miranda Ferreira"
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Item Respostas eletrofisiológicas das Porossensilas das quelíceras de Rhipicephalus microplus frente à fagoestimulantes e soros de bovinos(Universidade Federal de Goiás, 2013-09-20) Ferreira, Lorena Lopes; Silva, Andrea Caetano da; Louly, Carla Cristina Braz; Borges, Lígia Miranda Ferreira; http://lattes.cnpq.br/2591106734434348; Borges, Lígia Miranda Ferreira; Lorenzo, Marcelo Gustavo; Fernandes, Everton Kort KampTicks are external parasites distributed in the world. Its parasitism causes direct and indirect damage to their hosts. Rhipicephalus microplus is the principal ectoparasite of the livestock causing economic and sanitary implications in the tropics regions. Control strategies are still a challenge and new alternatives are being sought to avoid the use of chemical methods that are reprehensible in relation to animal and human health, and also the environment. It is known that Bos indicus are more resistant to ticks than Bos taurus, while their crosses have a moderate resistance. The olfactory and gustatory senses allow these parasites to seek hosts, mating partners and distinguish between food and harmful substances. The receptors of these sensory organs are present throughout the body and appendages of arthropods, being pore sensilla in the chelicerae important when it comes to ticks taste receptors. Electrophysiology assists in the discovery of active principles that attract or repel these parasites and this will be proven after a behavioral test. This work was performed in order to characterize R. microplus electrophysiological responses to phagostimulants in different concentrations like salts (NaCl and KCl, 10-3 a 10-1M), glucose (10-4 a 10-1M), adenosine triphosphate (ATP, 10-6 a 10-2M), reduced glutathione (GSH, 10-6 a 10-2M), a mixture of substances (MIX, glucose1M+ATP10- 2M+GSH10-3M) and sera of susceptible and resistant cattle to ticks. A single cell electrophysiology technique was used in the taste sensila of R. microplus. Engorged nymphs of R. microplus were collected from naturally infested bovines. Females ticks were fed on rabbits for a period of three to five days. The females were removed and taken directly to the electrophysiology room in order to be prepared for the test. At least 10 females were tested per substance. The shape, frequency and amplitude of spikes were analyzed with the program Autospike. The statistical analysis was performed with the software R applying ANOVA followed by Tukey HSD post-hoc (5%). In relation to the phagostimulants it was observed that the taste receptors of the chelicerae were responsive for all substances tested, having a more intense responses at higher concentrations. The frequency of responses to salts ranged from 11.09 ± 0.98 to 28.50 ± 3.49 spikes/s; to glucose from 25.40 ± 2.33 to 37.15 ± 4.47 spikes/s, to ATP from 17.91 ± 1.28 to 37.30 ± 4.77 spikes/s; to GSH from 19.10 ± 2.84 to 56.6 ± 5.08 spikes/s and the MIX 50.60 ± 4.42 spikes/s. The responses were uni or multicellular depending on the concentration. Sera from resistant cattle xiii triggered a higher frequency of spikes/s (average of 53.48 ± 1.89 spikes/s, ranging from 48.45 ± 4.28 to 61.05 ± 5.14 spikes/s) when compared with the susceptible cattle (average of 40.33 ± 1.63 spikes/s, ranging from 33.30 ± 3.02 to 49.05 ± 5.20 spikes/s). The records analyzed showed that R. microplus was more stimulated by sera from resistant cattle than those from susceptible animals. It is known that in the serum has phagostimulants which probably triggered spikes in both groups. On the other hand, the greater number of spikes to resistant bovine serum might be due to the presence of deterrents compounds. Both sera showed a multicellular response. It was also observed that there were variations in the spikes frequency between animals of the same group according to the characteristic of spikes recorded. It was concluded that R. microplus respond in a dose dependent pattern to the phagostimulants tested. And also that, this tick was able differentiated the sera of susceptible to sera of resistant bovines.Item Reconstrução tridimensional do lobo olfativo do carrapato Amblyomma sculptum (Acari: Ixodidae)(Universidade Federal de Goiás, 2017-03-07) Menezes, Karolina Martins Ferreira; Linhares, Guido Fontgalland Coelho; http://lattes.cnpq.br/6261928164195145; Welber Daniel Zanetti Lopes, Welber Daniel Zanetti c; http://lattes.cnpq.br/4480577038117234; Borges, Lígia Miranda Ferreira; Borges, Lígia Miranda Ferreira; Famadas, Kátia Maria; Monteiro, Caio Márcio de OliveiraAmblyomma sculptum belongs to Amblyomma cajennense complex and parasites human beings as well domestic and wild animals. This tick species has a world recognized importance in public and animal health due blood expoliation and transmission of some pathogens such as Rickettsia rickettsia, causal agent of Rocky Mountain Spotted Fever in Brazil. Ticks oriented themselves using chemical stimuli perceived by their sensorial organs. The Haller organ, which is localized in the first leg, has olfactory sensilla which perceive volatile molecules, as the odors. Inside of the sensilla there are neurones that project themselves to olfactory lobes in the synganglion, the central nervous system. In ticks, there is a gap in how the perception of the olfactory information is integrated in the synganglion and this comprehension would allow obtain details that could be used to the development of new control strategies. This work aims to verify the integration between the Haller organ and the synganglion by determining its neuronal projection patterns until the olfactory lobes as well accomplish its three-dimensional reconstruction. Males and females of A. sculptum were immobilized and the Haller organ was excised. The olfactory nerves were filled with one drop of dextran tetramethylrhodamine 1%. The synganglions were dissected in saline, fixed in 4% paraformaldehyde, washed in saline and triton X-100, dehydrated in alcohol series and mounted in glicerol. The synganlions were examined in a fluorescence microscope (Axio Scope.A1) to confirm filling from the peripheral neurons to the synganglion. The olfactory lobes stained were digitalized in a confocal microscope TCS SP8 (Leica) and the series were examined in the Reconstruct software. The olfactory lobes of one male and three females were selected to tracing the edges of each glomerulus in successive optical sessions in order to obtain the 3D reconstruction. The volume was calculated using the same software and their correlation was analyzed by Pearson correlation. The neuronal projections from the Haller organ were confined in the olfactory lobes, which were organized in glomeruli. The number of glomeruli in the males was 26 and 30 in each olfactory lobe examined and in the females varied between 24 and 29. Male and females showed similar glomeruli in both olfactory lobes, meaning they are homologues, when compared intra and intersexually, with a positive correlation between 72% and 95%. Considering the volume, the glomeruli were also similar and no macroglomerulus was identified, as any glomerulus in the male and female were two times bigger than the average volume. The number of specimens evaluated does not allow inferring if the lower number of glomeruli in the females was statistically significant. This work shows, for the first time, the 3D reconstruction of the olfactory lobe in a tick species. The results obtained contribute to understand how ticks process their olfactory information.Item Avaliação do papel repelente de voláteis isolados no odor de cães da raça Beagle contra o carrapato Rhipicephalus sanguineus (Acari: Ixodidae)(Universidade Federal de Goiás, 2014-02-14) Oliveira Filho, Jaires Gomes de; Silva, Andrea Caetano da; Louly, Carla Cristina Braz; Borges, Lígia Miranda Ferreira; http://lattes.cnpq.br/2591106734434348; Borges, Lígia Miranda Ferreira; Pinto, Mara Cristina; Linhares, Guido Fontgalland CoelhoTicks are arthropods of great importance in public health, due to the transmission of many pathogens. Rhipicephalus sanguineus parasites mainly dogs. It is known that Beagle dogs are less susceptible to R. sanguineus than English Cocker Spaniel, and still have resistance patterns linked to immunity. The lower susceptibility of Beagle dogs is due to chemical compounds they produce, which act in reducing the parasitic load of R. sanguineus on these animals. Given that this research aimed to verify the chemical repellency against R. sanguineus of compounds that have been identified in the odor of resistant dogs, and synthetic repellent compounds to other tick species. For testing chemical repellency two tests were used, the Petri dish and the Y-olfactometer. Five compounds identified in Beagle odors were selected and tested: 2 - hexanone, nonane, decane, undecane and benzaldehyde, the compound of botanical origin β-citronellol and the standard repellent DEET (N,N-diethyl-3-methylbenzamide). These compounds were tested alone and also two mixtures were evaluated: benzaldehyde and 2-hexanone (1:1), and benzaldehyde, 2-hexanone and undecane (1:1:1). All these compounds were prepared in a stock solution of 7.2 % ( ≈ 0.200 mg/cm²) and two lower concentrations were also tested: 0.100 and 0.050 mg/cm². DEET was repellent in the two highest concentrations and 68.33 to 86.67% of the ticks were found in untreated areas in different time points. At the concentration of 0.050 mg/cm² ticks were distributed similarly (without significant difference) in both areas. β-citronellol was the only compound tested that was repellent at all times and concentrations tested. The percentage of ticks in untreated areas ranged from 73.33 to 93.33 %. When compared β-citronellol with DEET the first compound was more effective at repelling the adults of R. sanguineus. For benzaldehyde, 30 minutes at a concentration of 0.200 mg/cm² a percentage of 63.33 % of ticks was observed in the untreated area. Percentage of 76.67 was observed after 5 min. at a concentration of 0.050 mg/cm². 2-hexanone was repellent at 30 min. at a concentration of 0.200 mg/cm² at 10 min. at a concentration of 0.100 mg/cm² and 30 minutes at a concentration of 0.050 mg/cm². For undecane a rate of 65.00% of ticks was observed in the untreated area in one moment of testing, the highest concentration during the first 5 min. of the evaluation. Nonane and decane not were not repellent at any of the concentrations and time intervals. The blend benzaldehyde + 2-hexanone was repellent in the three highest concentrations in almost all evaluated times, including in the olfactometer testing. During the whole test this mixture showed statistical similarity when compared with DEET. The addition of undecane to this blend did not potentiate its repellent power. The main result was the finding of natural repellents in resistant dogs to the tick R. sanguineus.Item Atividade repelente e carrapaticida de compostos derivados do óleo de coco contra Amblyomma sculptum(Universidade Federal de Goiás, 2021-03-03) Santos, Mayara Macêdo Barrozo; Cotta, Viviane Zeringóta Rodrigues; http://lattes.cnpq.br/4269635925947346; Monteiro, Caio Márcio de Oliveira; http://lattes.cnpq.br/2496419306394657; Borges, Lígia Miranda Ferreira; http://lattes.cnpq.br/2591106734434348; Fernandes, Éverton Kort Kamp; Ferreira, Lorena Lopes; Borges, Lígia Miranda FerreiraAmblyomma sculptum can parasitize several mammals, including humans, and is the main vector of the Rickettsia rickettsii bacterium in Brazil, the etiological agent of Brazilian spotted fever. Several studies have been carried out with products of plant origin to control ticks. In addition, the use of repellents is an important tool to prevent infestations. The present study aimed to evaluate the repellency of unfed nymphs of A. sculptum against substances derived from coconut oil (C12 dodecanoic acid methyl ester; C10 decanoic acid methyl ester; and coconut free fatty acid in lavender oil formulation) and catnip oil (Nepeta cataria); through modified Petri dish bioassays and, to evaluate the tick activity of these substances by means of the larval packet test. For the Petri dish test, a semicircle of filter paper containing the substance to be tested at a concentration of 10% diluted in ethanol and another semicircle containing the solvent (ethanol), were placed inside the dish; six non-fed nymphs were placed in the center of each plate and their location was assessed at 1, 3, 5, 10, 15 and 30 minutes and at 24, 48, 72, 96 and 168 hours after the start of the test. For the larval packet test, the substances were used in five concentrations (2.5; 5; 10; 15 and 20 mg / mL) on non-fed larvae of A. sculptum to assess mortality after 24 hours of treatment. As a result, all substances showed repellency, with the fatty acid free of coconut oil conferring repellency for 7 days, while N. cataria oil and C10 methyl ester showed repellency for 4 and 3 days, respectively. The concentration of the C12 methyl ester, effective against more than 93% of the tested larvae, was 15 mg / mL and the concentration, of that same substance, of 20mg / mL conferred 100% mortality. Thus, our results indicate that compounds derived from coconut oil have a repellent and tick effect on immature stages of A. sculptum and can be formulated to protect humans and other animals from infestation, as well as against pathogens transmitted by this species of tick.Item Análise bioquímica do produto de secreção/excreção de larvas de Cochliomyia hominivorax (Coquerel, 1858) (Diptera, Calliphoridae)(Universidade Federal de Goiás, 2015-07-03) Teixeira, Denise Gonçalves; Mastrangelo, Thiago de Araújo; http://lattes.cnpq.br/2335644825420420; Monteiro, Valdirene Neves; http://lattes.cnpq.br/8264822485508916; Borges, Lígia Miranda Ferreira; http://lattes.cnpq.br/2591106734434348; Borges, Lígia Miranda Ferreira; Aguiar, Valéria Magalhães; Ulhoa, Cirano JoséThe species Cochliomyia hominivorax, popularly known as screwworm fly, is a Diptera belonging to the family Calliphoridae. Its larvae are obligated parasites of warm-blooded animals and its geographic range extends throughout Latin America. Some studies accomplished with flies that produce myiasis concluded that the production of digestive enzymes secreted and / or excreted by the larvae has crucial contribution to the establishment and survival of these parasites in the host. Due to the lack of information approaching this subject on C. hominivorax, this project aimed to characterize and classify the enzymes in the secretion and excretion (S/E) of the larva of this fly, analyzing its biochemical profile.A colony of this species was established in laboratory conditions in order to accomplish this study. The colony was established by obtaining third instar larvae (L3)on naturally infested animals, and from the cultivation of these was obtained first (L1), second (L2) and third (L3) larvae instars,. The profile of proteins of S/E was obtained by polyacrylamide gel electrophoresis and proteolytic activity was analyzed using gelatin, azocasein and Na-benzoyl-arginine-nitroanilide as substrate. In S/E of the three instars proteins were detected with an apparent molecular weight ranging between 116 and 20 kDa. In the azocasein assay, at different pH ranges, it was observed that the major proteolytic activity occurs at pH 7.5 for all larvaeinstars. For characterization of the enzyme, assays were performed using these same substrates in which the samples were treated with inhibitors Benzamidine, Pepstatin A, AEBSF, TLCK, TPCK, EDTA, leupeptin and E-64. Proteinases present in the E/S of L1 are mostly serine proteases trypsin and chymotrypsin, whereas for L2 and L3 the presence of serine proteases and aspartyl proteases were observed.Item Comparação dos métodos de controles estratégico e seletivo na carga parasitária de Rhipicephalus microplus em rebanhos bovinos leiteiros(Universidade Federal de Goiás, 2020-03-02) Vieira, Luciano da Silva; Cotta, Viviane Zeringóta Rodrigues; http://lattes.cnpq.br/4269635925947346; Monteiro, Caio Márcio de Oliveira; http://lattes.cnpq.br/2496419306394657; Borges, Lígia Miranda Ferreira; http://lattes.cnpq.br/2591106734434348; Borges, Lígia Miranda Ferreira; Cotta, Viviane Zeringóta Rodrigues; Monteiro, Caio Márcio de Oliveira; Louly, Carla Cristina Braz; Lopes, Wélber Daniel ZanettiRhipicephalus microplus is one of the main ectoparasites of economic importance for cattle in tropical and subtropical regions, causing drop in milk production and transmitting pathogenic agents such as Anaplasma and Babesia. Therefore, constant studies on treatment schemes for parasite control are necessary to minimize the damage and losses that follow infestations by this arthropod. In this way, the present study aimed to compare two methods for R. microplus control, the strategic method and the selective method, in dairy cattle herds, aiming to provide support to the technical guidelines on the most efficient control method of the parasitic load. In the strategic method, seven treatments with spray acaricides were carried out every 21 days, starting in November. In the selective method, treatments with acaricides were performed only on animals that had an infestation equal to or greater than 40 engorged females. Only 5% of animals submitted to the selective method received treatment in November. The average of tick counts was lower in the strategic method compared to the selective method, in December. The maximum peak of infestation occurred in January, in both control methods, with no significant difference, persisting until February. No animal of the strategic method had an infestation greater than 40 engorged females in March. However, the parasitic load in both control methods was similar in the second evaluation in March. The averages of counts in the strategic method were lower in the months of April (10.9) and May (16.9) compared to the averages observed in the selective method (56.3 and 48.4). Yet, it was necessary to carry out tactical treatments in the strategic method, after the treatment scheme, in up to 16.7% of the animals in April and up to 50.0% in May, while in the selective method, the treatments varied from 42.8 to 88.8% in April and in May it varied from 28.6 to 75.0%. The total count of R. microplus was 5,036 engorged females in the strategic method, while in the selective method there were 13,829 engorged females in the study period. Therefore, it is concluded that the treatment scheme for control of R. microplus significantly interfered in the parasitic load of dairy cows, favouring the maintenance of acceptable levels of parasitism when the strategic method of tick control was implemented compared to the adoption of the selective method.