abstract: This dissertation studies the larger issue of antibiotic resistance with respect to how antibiotics are being introduced into the environment, focusing on two major anthropogenic pathways: animal husbandry for human consumption, and the recycling of wastewater and municipal sludge generated during conventional biological sewage treatment. For animal production on land (agriculture) antibiotics are often used for growth enhancement and increased feed efficiency. For animal production in water (aquaculture) antibiotics are often used as a prophylactic. I found that the same antibiotics are being used in both industries and that the same strains of human pathogens have also been isolated from both sources, expressing identical resistance mechanisms. In U.S. seafood, five out of 47 antibiotics screened for were detected at levels of 0.3 to 7.7 ng/g fresh weight. Although compliant with FDA regulations, the risk for resistance still exists, as even low antibiotic concentrations have been shown to exert selective pressure on bacteria. Similarly low concentrations of antibiotics were found in U.S. biosolids at levels of 0.6 to 19.1 ng/g dry weight. Of the five antibiotics detected, two have never been reported before in biosolids. Three have never been reported before in U.S. biosolids. Using the raw numbers obtained from antibiotic screenings in biosolids, I assessed the impact of employing four different LC-MS/MS methods, concluding that analysts should experimentally determine the most appropriate quantitation method based on the analyte targeted, matrix investigated, and research goals pursued. Preferred quantitation approaches included the isotope dilution method with use of an analogous standard and, although time and resource demanding, the method of standard addition. In conclusion, antibiotics introduced into the environment via agriculture, aquaculture, and wastewater recycling pose a combination of chemical and biological threats. Aside from exerting outright chemical toxicity to non-target organisms, antibiotic residues can promote the development of multi-drug resistance in human pathogens. Public health protection approaches to stem the risks posed by animal husbandry may include reserving drugs for exclusive, human use, decreasing their usage altogether, improving reporting efforts, reevaluating existing regulations on agricultural and aquacultural antibiotic usage, and improved risk assessment for biosolids application on land. / Dissertation/Thesis / Doctoral Dissertation Engineering 2015

Orientador: Ana Maria Lima de Azeredo Espin / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-23T10:46:46Z (GMT). No. of bitstreams: 1 Lopes_AlbertoMouraMendes_M.pdf: 2739543 bytes, checksum: 13578f8ab684d7f2c1af2020305008a8 (MD5) Previous issue date: 2013 / Resumo: O Brasil possui o maior rebanho bovino comercial do mundo, sendo este um dos setores de geração de renda mais importantes para o país. As infestações por ecto e endoparasitas estão entre os principais problemas que levam à diminuição da produtividade. Dentre eles, a Cochliomyia hominivorax (Coquerel), conhecida como mosca-da-bicheira, destaca-se como uma das principais causadoras de miíases, gerando prejuízos de aproximadamente US$ 1,7 bilhões/ano ao país. Produtos sistêmicos como as lactonas macrocíclicas/macrolactonas, ativadoras, dentre outros, do canal de cloreto (GLUCL?), vêm sendo utilizados intensivamente como medida preventiva e de tratamento nas infestações por C. hominivorax e diversos parasitas, e a intensa/prolongada utilização das lactonas macrocíclicas tem levado ao surgimento da resistência a tais produtos. A identificação dos mecanismos moleculares da resistência às lactonas macrocíclicas em outras espécies indica a ocorrência de possíveis alterações no canal de cloreto associados à resistência em C. hominivorax. Utilizando-se de sequências do canal de cloreto de outras espécies, previamente depositadas no banco de dados, além de sequências obtidas na caracterização do transcriptoma de C. hominivorax, o objetivo deste projeto é amplificar e sequenciar a região codificante do gene da subunidade desse canal de cloreto dependente de L-glutamato (GluCl?) em C. hominivorax. Além disso, serão comparadas as sequências de indivíduos resistentes e suscetíveis às macrolactonas visando à identificação da(s) mutação (ões) que confere(m) resistência a tais produtos e, consequentemente, o diagnóstico molecular da resistência / Abstract: Brazil has the largest commercial cattle herd in the world, being one of the most important sectors of income generation for the country. Infestations by ecto- and endo parasites are among the major problems that lead to low productivity. Among them, the Cochliomyia hominivorax (Coquerel), known as the New World Screwworm fly, stands out as a major cause of myiasis, resulting in losses of approximately US$ 1.7 billion each year to the country. Systemic products such as macrocyclic lactones/macrolactones (MLs), mainly the MLs member ivermectin, have been extensively used as a preventive measure and treatment in infestations by C. hominivorax and various parasites, and the intense/prolonged use of macrocyclic lactones have led to the emergence of resistance to such products. The macrolactones target-sites are the cys-loop ligand-gated ion channels receptors, specially the glutamate-gated chloride channel subunit (GLUCL?). The identification of the molecular mechanisms of macrocyclic lactones resistance in other species indicates that possible alterations in chloride channels and GLYR? and nAchR?7 are associated with macrolactones resistance in C. hominivorax. Using the glutamate-gated chloride channel subunit sequences from other species, previously deposited in the database, and sequences obtained in the characterization of the transcriptome of C. hominivorax, the aim of this project was to amplify and sequence the coding region of the glutamate-gated chloride channel subunit gene (GluCl?) in C. hominivorax. In addition, sequences will be compared among putative ivermectin resistant and susceptible individuals in order to identify mutation(s) conferring resistance to such product and thereby the molecular diagnosis of resistance / Mestrado / Genetica Animal e Evolução / Mestre em Genética e Biologia Molecular

The diploma thesis deals with the creation of a business concept to diversify the activities of a private farmer, operating in the region Pisecko, to the field of tourism, specifically to the agrotourism. At first, the surveyed area, accessibility, attractiveness and competitiveness were analyzed using secondary sources. In addition, primary sources were obtained with the help of a guided interview with the owner of a farm and a questionnaire survey conducted in travel agencies, tour operators and tourist information centers. Based on all the findings, the business concept of Jihočeské království was assembled. It includes a structural and technical solution, all services and the way of running the whole business, staffing and last but not least, the financial plan that demonstrates the viability of the project.