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Dissertations |
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1
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ODAIR JOSE QUEIROZ DA SILVA
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EVALUATION OF TRYPANOSOMA CRUZI INFECTION IN TRIATOMINE BUGS IN MUNICIPALITIES CONSIDERED AT HIGH RISK OF TRANSMISSION IN RIO GRANDE DO NORTE, BRAZIL
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Advisor : ANTONIA CLAUDIA JACOME DA CAMARA
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COMMITTEE MEMBERS :
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THALLYTA MARIA VIEIRA
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ANTONIA CLAUDIA JACOME DA CAMARA
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PAULO MARCOS DA MATTA GUEDES
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Data: Feb 20, 2025
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Show Abstract
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The state of Rio Grande do Norte (RN) is endemic for Chagas disease, with 36 municipalities at high risk of Trypanosoma cruzi transmission. The aim of this study was to assess the occurrence of T. cruzi infected triatomines domiciliary units (UDs) in municipalities at high risk of the parasite transmission in RN, as well as the species diversity in the wild environment. Triatomines were collected in DUs and wild environments, identified, and their intestinal content analyzed by using direct examination, xenoculture, and kDNA PCR for parasite detection. A total of 22 DUs were investigated, 14 in São Tomé and six in Sítio Novo, with the presence of triatomines recorded in 21.4% (3/14) and 16.7% (1/6), respectively. The municipality São Tomé showed a high percentage of infected triatomines, corresponding to 14.2%. In the DUs, 40 triatomines were captured, and the only species identified was Triatoma brasiliensis in both São Tomé and Sítio Novo. The natural infection rate for T. cruzi was 7.5%, with the highest rate in T. brasiliensis (8.8%). In the wild environment, the highest species diversity was found in the ESEC-Seridó, where specimens of T. brasiliensis, Triatoma pseudomaculata, Panstrongylus lutzi, and Triatoma sp. were captured. Triatoma brasiliensis was the most frequently captured species in both ESEC-Seridó and Sítio Novo. The natural infection rate for T. cruzi was 3.5%, with the highest rate in P. lutzi (100% - 1/1), followed by T. pseudomaculata (50% - 1/2) and T. brasiliensis at 2.8% (4/141). These data highlight the importance of continuous surveillance and control in high-risk municipalities to prevent T. cruzi transmission to humans and animals.
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2
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LORENA SOUZA SILVA
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FROM ENVIRONMENT TO HOST: RESISTANCE IN YEASTS FROM A UNIQUE HEALTH PERSPECTIVE
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Advisor : RAFAEL WESLEY BASTOS
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COMMITTEE MEMBERS :
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DANIELLE LETÍCIA DA SILVA
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JULLIANA RIBEIRO ALVES DOS SANTOS
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RAFAEL WESLEY BASTOS
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Data: Mar 13, 2025
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Show Abstract
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The resistance to antifungals has been increasing in fungi, especially in the groups that cause invasive mycoses. Usually, resistance selection occurs due to prior treatment with antifungals. However, there are reports of infections caused by fungi that are already resistant, without prior exposure to antifungals. This finding raises the hypothesis that the process of acquiring resistance may occur in the environment, possibly selected by antimicrobials with antifungal effects, such as environmental fungicides and hospital disinfectants. Thus, the objective of this study was to investigate the drug resistance profile with antifungal activity in medically important yeasts found infecting/colonizing patients, in animals, and in the environment, testing the hypothesis of cross-resistance between clinical and environmental antifungals. Yeasts were isolated from different sources and identified by mass spectrometry (MALDI-TOF MS). A total of 135 isolates were identified, with a prevalence of the following genera: (i) Candida (101; 74.8%), (ii) Kodamaea (17; 12.6%), (iii) Cryptococcus (7; 5.2%), and (iv) Trichosporon (5; 3.6%). After identification, the sensitivity profile of the isolates was determined through the Minimum Inhibitory Concentration (MIC) test using the broth microdilution method with clinical antifungals, environmental antifungals, and hospital disinfectants. In total, the Candida genus exhibited 9.6% resistance to clinical antifungals. The highest resistance was attributed to Micafungin (6.9%) and occurred in Candida glabrata. Four multi-resistant isolates to Amphotericin B and Micafungin were also found, and one isolate exhibited multi-resistance to Fluconazole and Amphotericin B. The highest efficacy among environmental antifungals was attributed to Tebuconazole, inhibiting 77% of the isolates. An isolate exhibiting resistance to both Fluconazole and Tebuconazole was identified, in addition to demonstrating high MIC values for hospital disinfectants. In summary, this study provides data on antifungal resistance in yeasts collected from different environments in the State of Rio Grande do Norte (RN), contributing to the clarification of the multifactorial relationship of the emergence of resistant yeast isolates of clinical importance.
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3
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ADAILSON DE SOUZA ASSUNÇÃO
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Atenuating effect of the synthetic N-methyl analogue of the alkaloid caulerpine in the inflammatory process in murine models of inflammation.
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Advisor : JANEUSA TRINDADE DE SOUTO
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COMMITTEE MEMBERS :
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DEYSIANE OLIVEIRA BRANDAO
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GEORGE JOAO FERREIRA DO NASCIMENTO
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JANEUSA TRINDADE DE SOUTO
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Data: Mar 14, 2025
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Show Abstract
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Inflammation is a response of the organism to different types of injuries, aimed at repairing damaged tissues and eliminating dead cells. Plant-based substances are potent in the treatment of various inflammatory disorders, and alkaloids are among the chemical classes reported to have anti-inflammatory activity. Caulerpine is an alkaloid extracted from the marine algae Caulerpa racemosa with anti-inflammatory properties previously demonstrated in the literature. Through structural modifications in the indole core and the ester group of classic caulerpine (CLPc), an analogue was generated, the N-methyl (CLPNmt), which enhances its lipophilicity. Thus, the present study aimed to evaluate the anti-inflammatory action of the CLPNmt analogue in different murine models of acute inflammation, comparing it with CLPc and dexamethasone. Initially, a peritonitis model was performed to establish the best dose of CLPNmt for exerting anti-inflammatory activity. For this, male Swiss mice were treated orally with CLPc (2 mg/kg), CLPNmt (4, 2, and 1 mg/kg), and intraperitoneally with dexamethasone (1 mg/kg), and one hour later, they were administered zymosan (40 mg/kg) intraperitoneally. After 24 hours, the mice were euthanized, peritoneal lavage was performed, centrifuged, leukocyte counts were determined in a Neubauer chamber, and the supernatant was used for the measurement of Interleukin (IL)-1β, IL-6, and Tumor Necrosis Factor (TNF)-α by ELISA. For the ear edema model, Swiss mice were treated with CLPNmt at a dose of 2 mg/kg and other treatments as mentioned earlier, followed by topical application of xylene (40μL) to the right ears. One hour after treatment, the left and right ears were removed and weighed on a precision balance to calculate the percentage of edema inhibition and then stored in PBS/formalin 10% for histological analysis. In the acute lung injury (ALI) model, animals were treated orally with CLPc or CLPNmt (2 mg/kg) or intraperitoneally with dexamethasone (1 mg/kg). One hour later, the animals received 50 μl of LPS (100 μg/kg) intranasally. After 24 hours, the mice were euthanized, bronchoalveolar lavage (BAL) was collected, centrifuged, the cell pellet was used for total and differential leukocyte count, and the supernatant was used for measuring inflammatory cytokine levels and nitrite levels by the Griess reaction. Lung samples were processed for histological analysis and RNA extraction to evaluate the expression of inflammatory markers by real-time polymerase chain reaction (qPCR). The results showed that treatment with CLPNmt was effective in reducing cell migration and IL- 6 secretion in the peritoneal cavity, with the dose of 2 mg/kg being the most efficient in this inhibition. Due to this result, this dose was used as the standard for the other experiments. Thus, it was observed that CLPNmt significantly inhibited edema formation in the xylene-induced model, similar to the effects observed for CLPc and dexamethasone, in addition to inhibiting leukocyte recruitment to the pulmonary cavity, predominantly of polymorphonuclear cells, and decreasing the levels of IL-1β, IL-6, TNF-α, Interferon (IFN)-γ, and nitrite in BAL, and aiding in the reduction of lung damage in the ALI model. Therefore, methylation of CLP made it more soluble in the solvent used, maintaining its anti-inflammatory potential, similar to what was observed for CLPc and compared to the data obtained for dexamethasone, making this analogue a potential pharmacological target.
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4
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ISABELLE LISIANY DE LIMA TAVARES
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Prevalence of Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus colonizing the mouth of bedridden and homebound elderly people, and associated factors
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Advisor : MARIA CELESTE NUNES DE MELO
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COMMITTEE MEMBERS :
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DEYSIANE OLIVEIRA BRANDAO
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GILMARA CELLI MAIA DE ALMEIDA
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MARIA CELESTE NUNES DE MELO
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Data: Mar 21, 2025
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Show Abstract
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The elderly population is the fastest growing demographic in the world. As a consequence, there is an increase in elderly people living at home or even bedridden, with varying degrees of dependence. With aging, the immune system becomes less efficient, generating systemic impairments in the elderly, who become more vulnerable to possible bacterial infections. Therefore, the objective of this study was to evaluate the prevalence of Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus (MRSA) colonizing the mouth of bedridden and homebound elderly people, as well as evaluating factors associated with this colonization. For this, 263 elderly people were visited at their homes in the city of Natal/RN, samples of the oral environment were collected using swabs, which were placed in saline solution and sent to the research laboratory. The samples were pipetted into BHI (Brain Heart Infusion Broth) enrichment broth and after incubation, they were seeded on the salt mannitol agar selective culture medium. Colonies suggestive of Staphylococcus aureus were reisolated with subsequent identification through conventional laboratory tests and mass spectrometry (MALDI-TOF). The susceptibility profile and identification of the MRSA lineage were performed using the disk diffusion method. Data related to sociodemographic factors and self-perception of health were collected through the application of a questionnaire. The prevalence of elderly people colonized by S. aureus was 13.3% (n=35) and for MRSA it was 1.5% (n=4). MRSA was resistant only to penicillin, erythromycin and clindamycin. Statistical tests showed a significant association between the presence of S. aureus and the variables male sex (p= 0.033), bedridden elderly (p=0.020), poor self-perceived general health (p=0.001) and difficulty eating (p=0.004). The presence of S. aureus and especially the MRSA lineage colonizing elderly people may represent a risk to the health of this population, especially due to their greater immunological vulnerability.
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5
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SARAH RUTH ALVES DA SILVA
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Rosemary essential oil (Lippia grata Schauer): insecticidal and repellent potential against Chrysomya megacephala in laboratory conditions
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Advisor : RENATA ANTONACI GAMA
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COMMITTEE MEMBERS :
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RENATA ANTONACI GAMA
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TACIANO DE MOURA BARBOSA
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VERUSKA CAVALCANTI BARROS
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Data: Mar 21, 2025
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Show Abstract
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5. Resumo em inglês (abstract): The species Chrysomya megacephala (Fabricius, 1794) is notable for its wide distribution and medical-veterinary importance, serving as a mechanical vector for various pathogens. The use of synthetic insecticides is the primary method of controlling these insects, but it has led to the development of resistance and environmental problems. Consequently, there has been an increased interest in research into alternative control strategies, including the use of essential oils. Literature records the insecticidal potential of the species Lippia grata (alecrim-do-mato) against some insects of medical significance, but its application in combating muscoid diptera, such as C. megacephala, remains largely unexplored. This study aimed to evaluate the insecticidal and repellent potential of L. grata essential oil against C. megacephala. Bioassays were conducted under laboratory conditions, using L. grata essential oil obtained through hydrodistillation. The insecticidal potential was tested using concentrations of 3.125%, 6.25%, 12.50%, 15%, 17.50%, 25%, 50%, 75%, and 100% on adults, and 12.5%, 25%, 50%, 75%, and 100% on larvae, with topical applications (3 µL) in both tests. For the repellent bioassays, concentrations of 6.25%, 12.5%, 25%, 50%, and 100% were used, administered (1 mL) onto filter paper within a custom-designed system. In the adult insecticidal bioassay, mortality was observed at various time intervals: 15, 30, 45, 60, 120 minutes, 24, and 48 hours. For the third-instar larval insecticidal assays, observations were made 24 and 48 hours post-application. Repellency assays involved analyses every 10 minutes, totaling three observations (30 minutes). The lethal and repellent concentrations for 50% (LC50 and RC50) and 90% (LC90 and RC90) were calculated using Probit analysis. Carvacrol (51.4%) was identified as the major component of the oil. Adulticidal bioassays revealed dose-dependent mortality with significant differences across concentration ranges (p<0.001), where 100%, 75%, 50%, and 25% doses exhibited the highest lethality. The LC50 and LC90 for adults were 6.45% and 22.61%, respectively, after 48 hours. Larvicidal bioassays showed significant differences between concentration ranges (p<0.0051), with the 100% oil concentration having a significantly different effect (p<0.005) on immature mortality. Other concentration comparisons showed no significant differences. The LC50 and LC90 for L3 larvae were 25% and 927%, respectively, after 48 hours. In repellency bioassays, the RC50 was 31.76%, and the RC90 was 115.51%. The results demonstrated that the essential oil was effective as an insecticide, with dose-dependent mortality indicating high toxicity to C. megacephala adults even at relatively low concentrations. The oil also exhibited larvicidal activity, though less potent than against adults. The 100% concentration was significantly more effective than others, suggesting that higher doses are necessary for satisfactory larval control. Furthermore, the assays present promising results regarding the oil's potential as a natural repellent against C. megacephala. Thus, this study indicates that the oil may be a promising alternative to synthetic insecticides for controlling this insect of medical-veterinary importance.
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6
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XIMENYA GLAUCE DA CUNHA FREIRE LOPES
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MPOX: EPIDEMIOLOGICAL ASPECTS IN THE STATE OF RIO GRANDE DO NORTH
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Advisor : JOSE VERISSIMO FERNANDES
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COMMITTEE MEMBERS :
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JOSE VERISSIMO FERNANDES
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DEBORA DE ALMEIDA ALOISE
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THALES ALLYRIO ARAUJO DE MEDEIROS FERNANDES
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Data: Mar 25, 2025
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Show Abstract
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Monkeypox is a zoonosis of viral etiology, caused by the Monkeypox virus (MPXV), classified in the Poxviridae family, Orthopoxvirus genus whose natural reservoir has not yet been identified. In 2022, MPXV emerged as a public health problem, spreading across several countries around the world, affecting thousands of people. The World Health Organization declared a Public Health Emergency of International Concern and recommended the use of the name Mpox to replace monkeypox or monkeypox. The capacity for diagnosis, prevention and treatment of human cases of the disease has been expanded in order to contain the spread of the virus. The first confirmed case of the disease in Brazil occurred in São Paulo, in June 2022. In July of the same year, the presence of the disease was also confirmed in Rio Grande do Norte (RN). The objective of this study was to analyze the epidemiological aspects of mpox in RN. A total of 140 confirmed cases of MPOX were studied in this work. Here, we describe the analyzes and correlations involving characteristics of nationality, age group, sex, occupation, location, signs and symptoms, clinical evolution, treatment, sample collection for research, history of vaccination for human smallpox, form of transmission, risk factors, cases in the same household, among others. The results of this study can help in the implementation of more effective policies, aiming at the prevention and early diagnosis of the disease, reducing its impact on the population.
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7
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LARISSA ESTEVES MESQUITA
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Antibacterial potential and resistance modulation of cocrystals obtained from Momordica Charantia
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Advisor : DEYSIANE OLIVEIRA BRANDAO
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COMMITTEE MEMBERS :
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DEYSIANE OLIVEIRA BRANDAO
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MARIA CELESTE NUNES DE MELO
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LUANNE EUGÊNIA NUNES
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Data: Mar 26, 2025
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Show Abstract
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Natural substances, especially those from medicinal plants, contain compounds capable of enhancing drugs, and are essential for the development of new medicines due to their low cost and easy access. Momordica charantia has proven antibacterial activity and potential for therapeutic applications. In the industrial setting, extracts can be used in cocrystals to enhance therapeutic effects. This study evaluated the resistance-modulating activity and antimicrobial potency of cocrystals obtained from M. charantia extracts. During the methodological stage, the fruits were collected in São José da Mata (PB) and used to obtain five hydroalcoholic extracts in different proportions of water:ethanol (E1, E2, E3, E4, and E5). Antibacterial, phytochemical, and cytotoxicity analyses of the extract were performed. The cocrystals were formed by evaporation under reduced pressure and characterized by X-ray Diffraction (XRD), Differential Scanning Calorimetry (DSC) and Fourier Transform Infrared Spectroscopy (FTIR). The modulating activity was evaluated by microdilution, with calculation of the Fractional Inhibitory Concentration Index (FICI). As a result, Escherichia coli demonstrated greater sensitivity to extracts E1, E2 and E3. Phytochemical analysis revealed a higher concentration of tannins in relation to polyphenols and flavonoids, with E5 being the richest in these compounds, followed by E4 and E3. Extract E3 stood out for its Minimum Inhibitory Concentration (MIC), balanced phytochemical composition and ideal solvent ratio, being selected for the formation of the cocrystal. In the assays with L929 fibroblasts and P815 mast cells, all concentrations tested reduced cell viability, but the moderate cytotoxicity of E3 does not preclude its therapeutic use. Structural analyses of E3 associated with Carboxylic Salicylic Acid (EX-SA) suggest the formation of a cocrystal. The compound showed antimicrobial activity against E. coli, S. aureus, K. pneumoniae and P. aeruginosa, with a MIC of 1.04 mg/mL and synergism with Oxacillin, Amoxicillin+Clavulanate, Ceftriaxone and Meropenem. The results indicate that the EX-SA cocrystal not only improves the antimicrobial action of the extract, but also enhances the efficacy of conventional antibiotics, allowing the reduction of clinical doses. This finding reinforces the potential of M. charantia as a basis for new therapeutic formulations, opening perspectives for the development of more effective alternatives in combating bacterial resistance.
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8
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JESSICA KAROLINE DE LIMA SILVA
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Microevolution of antifungal resistance in Candida (Candidozyma) auris select by exposure to fungicide
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Advisor : RAFAEL WESLEY BASTOS
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COMMITTEE MEMBERS :
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RAFAEL WESLEY BASTOS
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DÉBORA CASTELO BRANCO DE SOUZA COLLARES MAIA
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LYSANGELA RONALTE ALVES
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Data: Dec 16, 2025
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Show Abstract
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Candida (Candidozyma) auris is an emerging pathogenic yeast frequently associated with hospital outbreaks. Certain clades within the species exhibit high rates of antifungal resistance and multidrug resistance, with fluconazole (FCZ) resistance being the most prevalent, reaching up to 77% of isolates. One of the main challenges regarding C. auris is understanding how its resistance mechanisms have evolved. One hypothesis is that the extensive use of agricultural azoles in the environment may select resistant isolates that subsequently infect humans. In this context, the present study aimed to investigate whether exposure to the agricultural azole tebuconazole (TBZ) could select for cross-resistance to FCZ, while also evaluating the role of temperature in this adaptive process. Initially, susceptibility tests to FCZ and TBZ were performed on thirty-nine isolates of C. auris, according to CLSI (2017) guidelines. The results revealed 41% resistance to FCZ, while 38.5% of isolates exhibited reduced susceptibility to TBZ. Notably, 94% (15/16) of FCZ-resistant isolates also displayed elevated MIC values for TBZ, indicating cross-resistance in 38.5% of cases. All isolates with reduced TBZ susceptibility were resistant to FCZ, reinforcing the hypothesis that environmental azole use may contribute to the emergence of clinical resistance. To further investigate this relationship, adaptation experiments were conducted with three isolates susceptible to FCZ and TBZ, originating from different geographic regions. The isolates were serially exposed, in duplicate, to fixed concentrations of TBZ (16× MIC) under two temperature conditions (30 °C and 37 °C). This serial exposure led to a progressive increase in MIC values for TBZ and, more prominently, for FCZ, particularly at 37ºC, suggesting that thermal stress enhances the selection of resistant phenotypes. The stability of this phenotype was evaluated by subsequent passages in drug-free medium, revealing transient cross-resistance in all isolates. Additionally, adaptation experiments with increasing TBZ concentrations (up to 256 μg/mL) showed that some colony groups acquired permanent cross-resistance even after drug withdrawal. Taken together, these findings demonstrate that exposure to TBZ can induce resistance to FCZ in C. auris, particularly under thermal stress, highlighting the role of the environment in the selection and maintenance of resistant isolates of clinical importance.
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