Tetracycline: production, waste treatment and environmental impact assessment

A ocorrência frequente de fármacos no meio aquático exige a avaliação do seu impacto ambiental e seus efeitos negativos em seres humanos. Dentre os fármacos com maior potencial de impacto ambiental estão os antibióticos, que chegam ao meio ambiente através dos efluentes de indústrias químico-farmacêuticas e, principalmente, através de esgotos domésticos e criação de animais, visto que 25% a 75% dos fármacos são excretados em forma inalterada após passagem pelo Trato Gastrointestinal. Parcela significativa do consumo mundial dos antibióticos corresponde à classe das tetraciclinas, representando consumo humano de 23 kg/dia no Brasil, em 2007. Atualmente, há pesquisas de novas tetraciclinas que incorporam metais pesados (Hg, Cd, Re, Pt, Pd) às suas estruturas com o intuito de aumentar suas atividades bactericidas. As estações de tratamento de águas residuais convencionais não são capazes de degradar moléculas orgânicas complexas, diminuir a sua toxicidade e melhorar a sua biodegradabilidade. Por esta razão, as novas tecnologias, como, por exemplo, os processos oxidativos avançados, estão sendo desenvolvidos para lidar com esta demanda. Os objetivos deste trabalho são fazer uma revisão da literatura sobre os processos de obtenção de tetraciclinas, apresentar métodos de tratamento de seus resíduos e avaliar o seu impacto ambiental.The frequent occurrence of pharmaceuticals in the aquatic environment requires an assessment of their environmental impact and their negative effects in humans. Among the drugs with high harmful potential to the environment are the antibiotics that reach the environment not only, as may be expected, through the effluents from chemical and pharmaceutical industries, but mainly through the sewage and livestock; because around 25 to 75% of the ingested drugs are excreted in unchanged form after the passage through the Gastro-Intestinal Tract. Tetracycline has high world consumption, representing a human consumption of about 23 kg/day in Brazil in 2007. At the moment, researches are being made to develop new tetracycline that incorporate heavy metals (Hg, Cd, Re, Pt, Pd) to their structures in order to increase their bactericidal effect. The conventional wastewater treatment plants are not able to degrade complex organic molecules to reduce their toxicity and improve their biodegradability. For this reason new technologies, i.e., the advanced oxidation processes, are being developed to handle this demand. The objectives of this study are to review the literature on the processes of obtaining tetracycline, presenting its waste treatment methods and evaluation of their environmental impact

Influence of the drying process on the chemical profile of the extracts of Mikania laevigata and Mikania glomerata, chemical characterization,fractionation and evaluation of the extract biological activities through in vitro studies

Orientador: Alexandra Christine Helena Frankland SawayaTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: Este trabalho teve como objetivo avaliar como os métodos de secagem das folhas de Mikania glomerata Spreng. e Mikania laevigata Schultz influenciam no perfil de constituintes presentes nos extratos obtidos, avaliar a composição e atividade biológica de extratos hidroetanólicos e aquosos de ambas as espécies e obtenção de frações enriquecidas através de Cromatografia de Contra Corrente. Os processos de secagem avaliados foram secagem em estufa e secagem à sombra em temperatura ambiente. Todos os extratos obtidos foram analisados quanto à sua composição por Cromatografia Líquida de Ultra Alta Eficiência acoplada a Espectrometria de Massas (UHPLC-MS) para obter seu perfil químico. Os extratos foram avaliados quanto a sua citotoxicidade por: avaliação do efeito sobre células musculares e epiteliais de epiderme e de brônquios; avaliação do efeito sobre células tumorais; transativação por gene repórter; ensaio antidesnaturação em albumina sérica bovina. Sua atividade antioxidante foi avaliada por 2,2-difenil-1-picril-hidrazila (DPPH) e por ORACFL. E Atividade antimicrobiana foi avaliada por placa de ágar perfurado. Estes mesmos extratos foram submetidos ao processo de fracionamento por Cromatografia de Contra Corrente para a obtenção de frações enriquecidas e suas composições foram analisadas em corrida similar ao que os extratos originais foram submetidos em UHPLC-MS. Através dos resultados obtidos nesta tese, é nítida a impossibilidade que estas espécies sejam intercambiáveis para a produção de fármacos como é usualmente discutido, sendo as vias de ação em que cada espécie atua diretamente associada a sua composição. Os testes de citotoxicidade apontam que não há prejuízo para as células locais em concentração até 112,5 µg/mL e com o benefício de estimulação da proliferação celular, indicando potencial cicatrizante. A atividade antioxidante está relacionada ao tipo de extrato, principalmente à concentração de ácidos cafeoilquínicos e compostos fenólicos presentes. A maioria das cepas bacterianas estudadas foram inibidas em concentração dos extratos de até 5 mg/mL. Quando submetidos aos testes de secagem, o tecido vegetal apresentou-se mais próximo a composição original após a secagem em estufa, possivelmente devido a inativação de enzimas e micro-organismos responsáveis pela degradação. A obtenção de frações enriquecidas através da Cromatografia de Contra Corrente demostrou que, apesar da complexidade dos extratos, este método é reprodutível quanto a obtenção de compostos, mesmo em concentração extremamente baixa no extrato original, robusta quanto ao aumento de escala e de baixa perda quando realizado o balanço de massas ao final do processo, menor que 17%Abstract: This study aimed to evaluate how the drying methods of leaves of Mikania glomerata Spreng. and Mikania laevigata Schultz influence on the profile of constituents present in their extracts, to evaluate the composition and biological activity of hydroethanolic and aqueous extracts of both species and obtaining enriched fractions by Counter Current Chromatography. The drying processes evaluated were oven drying and shade drying at room temperature. All extracts were analyzed for their composition by Ultra High Efficiency Liquid Chromatography coupled to Mass Spectrometry (UHPLC-MS) to obtain their chemical profile. The extracts were evaluated for their cytotoxicity by: evaluation of the effect on muscle and epithelial cells of the epidermis and bronchi; evaluation of the effect on tumor cells; transactivation by gene reporter; immunoassay in bovine serum albumin. Their antioxidant activity was evaluated by 2,2-diphenyl-1-picryl-hydrazila (DPPH) and by ORACFL. Antimicrobial activity was evaluated by perforated agar plate. These same extracts were subjected to the fractionation process by Counter Current Chromatography to obtain enriched fractions and their composition was analyzed in a similar run as the original extracts were submitted to UHPLC-MS. Through the results obtained in this thesis, it is clearly impossible for these species to be interchangeable for the production of drugs as is usually discussed, as the pathways of action in which each species acts directly associated with its composition. Cytotoxicity tests indicate that there is no damage to cells in concentrations up to 112.5 ?g / mL and with the benefit of stimulation of cell proliferation, indicating potential healing. The antioxidant activity is related to the type of extract, mainly to the concentration of caffeoylquinic acids and phenolic compounds present. Most of the bacterial strains studied were inhibited at concentrations of extracts up to 5 mg / mL. When submitted to the drying tests, the plant tissue presented composition closer to the original composition than after drying in the greenhouse, possibly due to the inactivation of enzymes and microorganisms responsible for the degradation. The obtainment of enriched fractions through Counter Current Chromatography showed that, despite the complexity of the extracts, this method is reproducible in obtaining compounds, even when they are in extremely low concentration in the original extract, robust for scaling up and presented low loss when performed the mass balance at the end of the process, less than 17%DoutoradoFármacos, Medicamentos e Insumos para SaúdeDoutor em CiênciasCAPE

Cytotoxicity of Doxycycline Effluent Generated by the Fenton Process

This study aims at determining the Minimum Inhibitory Concentration with Escherichia coli ATCC 25922 and cytotoxicity to L929 cells (ATCC CCL-1) of the waste generated by doxycycline degradation by the Fenton process. This process has shown promise in this treatment thanks mainly to the fact that the waste did not show any relevant inhibitory effect on the test organism and no cytotoxicity to L-929 cells, thus demonstrating that the antibiotic properties were inactivated

FISCAL AND QUASI-FISCAL MEASURES IN FACE OF THE COVID-19 PANDEMIC: SELECTED INTERNATIONAL EXPERIENCES

In face of the Covid-19 pandemic, the active adoption of fiscal policy has been essential to mitigate both sanitary and socioeconomic effects. This paper aims to analyze and compare fiscal and quasi-fiscal measures implemented over 2020 by five countries: China, United States, Germany, United Kingdom and Brazil. In some cases, the amount of resources surpassed 20% of gross domestic product (GDP), showing the importance of such measures. In light of these selected experiences, the complementarity between sanitary and economic measures is highlighted, thus reinforcing the need for coordination between them for effectively coping with the pandemic impacts. Among countries, efforts in both sanitary policies and fiscal and quasi-fiscal resource allocation have been very diverse, directly affecting the evolution of their respective sanitary and socioeconomic conditions

Multidifferential study of identified charged hadron distributions in ZZ-tagged jets in proton-proton collisions at s=\sqrt{s}=13 TeV

Jet fragmentation functions are measured for the first time in proton-proton collisions for charged pions, kaons, and protons within jets recoiling against a $Z$ boson. The charged-hadron distributions are studied longitudinally and transversely to the jet direction for jets with transverse momentum 20 $< p_{\textrm{T}} < 100$ GeV and in the pseudorapidity range $2.5 < \eta < 4$. The data sample was collected with the LHCb experiment at a center-of-mass energy of 13 TeV, corresponding to an integrated luminosity of 1.64 fb$^{-1}$. Triple differential distributions as a function of the hadron longitudinal momentum fraction, hadron transverse momentum, and jet transverse momentum are also measured for the first time. This helps constrain transverse-momentum-dependent fragmentation functions. Differences in the shapes and magnitudes of the measured distributions for the different hadron species provide insights into the hadronization process for jets predominantly initiated by light quarks.Comment: All figures and tables, along with machine-readable versions and any supplementary material and additional information, are available at https://cern.ch/lhcbproject/Publications/p/LHCb-PAPER-2022-013.html (LHCb public pages

Study of the B−→Λc+Λˉc−K−B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-} decay

The decay $B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-}$ is studied in proton-proton collisions at a center-of-mass energy of $\sqrt{s}=13$ TeV using data corresponding to an integrated luminosity of 5 $\mathrm{fb}^{-1}$ collected by the LHCb experiment. In the $\Lambda_{c}^+ K^{-}$ system, the $\Xi_{c}(2930)^{0}$ state observed at the BaBar and Belle experiments is resolved into two narrower states, $\Xi_{c}(2923)^{0}$ and $\Xi_{c}(2939)^{0}$, whose masses and widths are measured to be $$m(\Xi_{c}(2923)^{0}) = 2924.5 \pm 0.4 \pm 1.1 \,\mathrm{MeV}, \\ m(\Xi_{c}(2939)^{0}) = 2938.5 \pm 0.9 \pm 2.3 \,\mathrm{MeV}, \\ \Gamma(\Xi_{c}(2923)^{0}) = \phantom{000}4.8 \pm 0.9 \pm 1.5 \,\mathrm{MeV},\\ \Gamma(\Xi_{c}(2939)^{0}) = \phantom{00}11.0 \pm 1.9 \pm 7.5 \,\mathrm{MeV},$$ where the first uncertainties are statistical and the second systematic. The results are consistent with a previous LHCb measurement using a prompt $\Lambda_{c}^{+} K^{-}$ sample. Evidence of a new $\Xi_{c}(2880)^{0}$ state is found with a local significance of $3.8\,\sigma$, whose mass and width are measured to be $2881.8 \pm 3.1 \pm 8.5\,\mathrm{MeV}$ and $12.4 \pm 5.3 \pm 5.8 \,\mathrm{MeV}$, respectively. In addition, evidence of a new decay mode $\Xi_{c}(2790)^{0} \to \Lambda_{c}^{+} K^{-}$ is found with a significance of $3.7\,\sigma$. The relative branching fraction of $B^{-} \to \Lambda_{c}^{+} \bar{\Lambda}_{c}^{-} K^{-}$ with respect to the $B^{-} \to D^{+} D^{-} K^{-}$ decay is measured to be $2.36 \pm 0.11 \pm 0.22 \pm 0.25$, where the first uncertainty is statistical, the second systematic and the third originates from the branching fractions of charm hadron decays.Comment: All figures and tables, along with any supplementary material and additional information, are available at https://cern.ch/lhcbproject/Publications/p/LHCb-PAPER-2022-028.html (LHCb public pages

Measurement of the ratios of branching fractions R(D∗)\mathcal{R}(D^{*}) and R(D0)\mathcal{R}(D^{0})

The ratios of branching fractions $\mathcal{R}(D^{*})\equiv\mathcal{B}(\bar{B}\to D^{*}\tau^{-}\bar{\nu}_{\tau})/\mathcal{B}(\bar{B}\to D^{*}\mu^{-}\bar{\nu}_{\mu})$ and $\mathcal{R}(D^{0})\equiv\mathcal{B}(B^{-}\to D^{0}\tau^{-}\bar{\nu}_{\tau})/\mathcal{B}(B^{-}\to D^{0}\mu^{-}\bar{\nu}_{\mu})$ are measured, assuming isospin symmetry, using a sample of proton-proton collision data corresponding to 3.0 fb${ }^{-1}$ of integrated luminosity recorded by the LHCb experiment during 2011 and 2012. The tau lepton is identified in the decay mode $\tau^{-}\to\mu^{-}\nu_{\tau}\bar{\nu}_{\mu}$. The measured values are $\mathcal{R}(D^{*})=0.281\pm0.018\pm0.024$ and $\mathcal{R}(D^{0})=0.441\pm0.060\pm0.066$, where the first uncertainty is statistical and the second is systematic. The correlation between these measurements is $\rho=-0.43$. Results are consistent with the current average of these quantities and are at a combined 1.9 standard deviations from the predictions based on lepton flavor universality in the Standard Model.Comment: All figures and tables, along with any supplementary material and additional information, are available at https://cern.ch/lhcbproject/Publications/p/LHCb-PAPER-2022-039.html (LHCb public pages

Degradation of doxycycline hydrochloride by the Fenton process

O aumento no consumo de antibióticos por seres humanos e animais tem elevado a sua concentração na sua forma inalterada ou de seus metabólitos, que chegam às estações de tratamento de efluentes, onde os tratamentos convencionais são incapazes de degradar estas moléculas, sendo liberadas diretamente nos corpos d\'água receptores. A liberação destas moléculas no meio ambiente tem proporcionado a seleção de organismos patogênicos resistentes, capazes de transmitir geneticamente esta característica a seus descendentes. Este trabalho tem como objetivo o estudo da degradação da molécula de cloridrato de doxiciclina, por ser um antibiótico de largo espectro da família das tetraciclinas, através do processo Fenton. Foram determinados neste trabalho as influências da temperatura, concentração inicial de peróxido de hidrogênio, concentração inicial de íon ferroso e do pH sobre a concentração residual de cloridrato de doxiciclina, concentração residual de peróxido de hidrogênio, concentração de íon ferroso e de carbono orgânico total (COT) em função do tempo de reação. Os métodos analíticos empregados foram baseados em espectrofotometria, análise instrumental para a determinação do carbono orgânico total (COT), titulometria e cromatografia líquida (CLAE) com detectores de UV e de massa. Testes preliminares mostraram que as melhores condições operacionais de temperatura, concentrações de íon ferroso e de peróxido de hidrogênio estariam ao redor de CFe2+ = 62,5 mg/L, CH2O2 = 500 mg/L e T = 20ºC. Estas condições foram utilizadas como ponto central do planejamento fatorial do tipo Delineamento Composto Central Rotacional (DCCR). Os resultados do planejamento tratados com o software Statistica® mostraram que as condições operacionais para a menor concentração residual de fármaco (0 mg/L) e redução de 40,9% da carga orgânica em solução deveriam estar ao redor de concentração de íon ferroso, CFe2+ = 25 mg/L, concentração de peróxido de hidrogênio, CH2O2 = 611 mg/L e temperatura = 35ºC, das quais a variável CH2O2 apresentou a maior importância estatística. Ao redor destas condições foi feita uma análise paramétrica para se verificar as influências individuais da temperatura, CFe2+, CH2O2 e da relação CFe2+/ CH2O2. Também foi testada a citotoxicidade dos resíduos gerados. Este estudo mostrou que a variável de maior importância sobre o processo foi a concentração de peróxido de hidrogênio, que tem relação direta com a razão CFe2+/ CH2O2 apresentado melhores resultados quando esta é mantida igual a 0,16. Houve melhora acentuada na mineralização da matéria orgânica com a inserção de microbolhas de ar, obtendo assim redução de até 44% da matéria orgânica em relação à concentração inicial de carbono orgânico de 55 mg/L. Apesar da baixa mineralização obtida pelo estudo, o processo Fenton se mostrou promissor na degradação do cloridrato de doxiciclina, devido principalmente ao fato de os resíduos de degradação não possuírem ação inibitória sobre o organismo teste Escherichia coli e nem citotoxicidade sobre as células L-929, evidenciando que as propriedades antibióticas da molécula foram inativadas, inibindo a capacidade de promover a resistência bacteriana a este antibiótico.The increasing consumption of antibiotics by humans and animals have increased their concentrations in the environment in their unchanged form or of their metabolites, that reach public wastewater treatment plants, which are unable to degrade these molecules, and are released into surface waters. The release of these molecules in the environment has caused the selection of resistant pathogenic organisms, genetically capable of transmitting this feature to their descendants. This work has as objective the study of the degradation of doxycycline hydrochloride, which is a common broad-spectrum antibiotic of the tetracycline family, by the Fenton process. In this work were determined the influences of temperature, initial hydrogen peroxide concentration, initial ferrous ion concentration and of pH on the final residual concentration of doxycycline, residual concentration of hydrogen peroxide, residual concentration of ferrous ion and total organic carbon (TOC) along the reaction time, using titrations and instrumental analytical techniques as spectrophotometry, TOC analyzer and high performance liquid chromatography (HPLC) with UV and mass detectors. Preliminary tests showed that the best operating conditions of temperature, concentrations of ferrous ion and hydrogen peroxide would be around CFe2+ = 62.5 mg/L, CH2O2 = 500 mg/L and T = 20°C. These conditions were use d as the central point of the Central Composite Rotational Design (DCCR). The results of the experimental planning were treated with the Statistica® software and it was showed that the operational conditions for the smallest residual drug concentration (0 mg/L) and 40.9% TOC reduction should be around ferrous ion concentration, CFe2+ = 25 mg/L, concentration of hydrogen peroxide, CH2O2 = 611 mg/L and temperature = 35°C. The hydrogen peroxide exhibited the highest statistical importance of the studied variables. It was accomplished a parametric study around the best operational conditions inferred from the statistical analysis to check the individual influences of temperature, CFe2+, CH2O2 and the ratio CFe2+/ CH2O2. It was also verified the cytotoxicity of the reaction mediums after the end of the experiments. This study showed that the process is highly dependent of the hydrogen peroxide concentration, directly related to the ratio CFe2+/ CH2O2 and presented the best results when this ratio was kept equal to 0.16. It was observed a marked improvement in the mineralization of the organic matter, up to 44% reduction of the initial TOC value of 55 mg/L, when air was pumped to the reaction medium. Despite the low mineralization obtained in this study, the Fenton process proved to be promising in the degradation of doxycycline hydrochloride, due mainly to the low cytotoxicity of the residues. It was observed neither inhibitory action on the test organism Escherichia coli nor cytotoxicity on L-929 cells, indicating that the antibiotic properties of the molecule had been inactivated and also its ability to stimulate bacterial resistance to this antibiotic

Phosphorus Shortage Induces an Increase in Root Exudation in Fifteen Eucalypts Species

A significant proportion of the carbon fixed by plants is transported to the roots and exuded to the rhizosphere. Exudates may have important roles in the rhizosphere, such as desorbing labile phosphorus (P) or mobilizing manganese (Mn) and other metal cations. This study evaluated the root exudation profiles of seedlings of 15 eucalypt species in response to a P shortage and if the ability to exude organic compounds was related to P and Mn accumulation in the shoots. The plants were grown on sand and were irrigated with nutrient solutions containing either sufficient P (500 µM) or low P (25 µM). Organic acids (OA), amino acids/polyamines, and phenolics were analyzed in the root exudates by UPLC-MS/MS. Plants with a low P level had low leaf P contents and growth reduction. A P shortage induced the exudation of the three groups of metabolites analyzed at higher levels than sufficient P availability. Despite that, the composition pattern of root exudates was similar among species under low or sufficient P concentrations. Citric and isocitric acids were the major OAs found in the exudates, followed by oxalic, malic, and succinic acids. Among the amino acids/polyamines identified, putrescine was the most abundant in all species, followed by glycine. Cinnamic acid was the predominant phenolic in the root exudates. Our results indicate that P limitation induces a conserved response genetic mechanism in eucalypts. Such results can be further investigated to adapt commercial clones to soils with low P availability