Avaliação do antagonismo do efeito do sódio sobre o tratamento de água residuária de Charqueada com elevada salinidade em reator anaeróbio de manta de lodo (UASB)

As águas residuárias provenientes da indústria do charque são conhecidas por apresentarem elevado teor de cloreto de sódio, aliado a grandes concentrações de matéria orgânica proveniente do sangue liberado ao longo do processo industrial. Esse tipo de água residuária apresenta potencial para degradação biológica, contudo, o cloreto de sódio, em concentração elevada, pode inibir a atividade dos microrganismos e, em alguns casos, levar sistemas biológicos à falência. No presente trabalho, foi avaliada a viabilidade de degradação anaeróbia de efluente sintético de Charqueada contendo elevado teor de cloreto de sódio, em reator anaeróbio tipo UASB (Upflow Anaerobic Sludge Blanket), em escala de laboratório. Foram utilizados 4 reatores, alimentados com água residuária sintética com características similares à água residuária de Charqueada. O reator 1 foi utilizado como controle, o reator 2 recebeu NaCl e os demais (3 e 4) foram operados na presença de NaCl acrescidos de: betaína e potássio com cálcio, respectivamente. Os compostos citados são conhecidos como antagonizantes, por possuirem capacidade de minimizar o efeito inibitório do sódio sobre o processo de digestão anaeróbia. Os reatores foram inoculados com lodo de reator UASB e submetidos à concentração de 5000 mg/L de matéria orgânica, como DQO. A carga orgânica aplicada foi de 5 Kg/m3.d e os reatores não suportaram tal carga. Reiniciou-se a operação com aumento progressivo da DQO de 500 a 2000 mg/L resultando em cargas orgânicas de 0,5 a 2,0 Kg/m3.d, respectivamente. Após estabilização dos reatores, iniciou-se a fase de introdução de cloreto de sódio (1.500 a 13.500 mg/L) e antagonizantes com aumento progressivo a cada fase. Na presença ou ausência de antagonizantes, os reatores 2, 3 e 4 não tiveram o desempenho alterado até a concentração de NaCl de 6000 mg/L. Na presença de 9000 mg/L de NaCl, a betaína se mostrou pouco efetiva como soluto compatível no reator 3 e os antagonizantes do reator 4, potássio e cálcio, apresentaram efeitos estimulatórios. As morfologias encontradas ao longo do experimento foram cocos, víbrios, bacilos, sarcinas, além de morfologias semelhantes a Methanosarcina sp. e Methanosaeta sp. O aumento da concentração de cloreto de sódio provocou a redição da população de Arqueas.

Estado sanitário do jacaré-de-papo-amarelo (Caiman latirostris) em paisagens antropizadas no Estado de São Paulo

O jacaré-de-papo (Caiman latirostris) é considerado um crocodiliano de médio porte que apresenta uma ampla distribuição latitudinal na América do Sul. Possivelmente a espécie possui a situação mais complexa entre os crocodilianos brasileiros quanto ao aspecto da conservação, basicamente porque suas populações encontram-se fragmentadas em grande parte de sua área de distribuição original e por utilizarem áreas com fortes atividades antrópicas. Embora a espécie possua aparentemente um processo adaptativo frente a estas pressões, um aspecto fisiológico pode ainda sofrer rápida alteração quando submetido a elas: o estado sanitário. Desta maneira, este estudo objetivou determinar o estado sanitário do maior predador aquático, utilizando duas áreas distintas no Estado de São Paulo, através da determinação dos perfis hematológicos e bioquímicos do sangue, além da caracterização da microbiota oral, servindo de modelo à conservação da espécie em ambientes alterados. No primeiro capítulo foram determinados o perfil hematológico e bioquímico do sangue utilizando-se 29 indivíduos (19 machos e 10 fêmeas) capturados em Angatuba e 11 indivíduos (2 machos, 4 fêmeas e 5 filhotes) capturados em Cubatão. Diferenças estatísticas significativas foram encontradas nos valores de creatinina na comparação entre fêmeas de ambas as áreas de estudo (p = 0,033) e nos valores de alanina aminotransferase (p = 0,003), hemácias (p = 0,034), hemoglobina (p = 0,049) e volume corpuscular médio (p = 0,027) quando da comparação entre sexos. No segundo capítulo determinou-se a microbiota oral destes animais através do isolamento, identificação e caracterização bacteriana, em conjunto com o teste de perfil de susceptibilidade aos antimicrobianos. Foram determinados 14 diferentes tipos de bactérias na área de Angatuba, sendo uma classificada como baciliforme aeróbio Gram-positivo, uma como bacilo Gram-negativo e 12 classificadas como Enterobactérias Gram-negativas. Na área de Cubatão foram isolados cinco tipos de bactérias, sendo quatro classificadas como Enterobactérias Gram-negativas e uma como baciliforme aeróbio Gram-positivo. Considerando que o uso de antimicrobianos é um processo primário no tratamento de animais e pessoas, principalmente nos casos que envolvam infecções provocadas pela interação homem x animais, para ambas as áreas de estudo, as quinolonas Enrofloxacina e Norfloxacina, além do aminoglicosídeo Gentamicina, foram os antimicrobianos que apresentaram os menores índices de resistência frente aos isolados testados.

Novel particulate BCG-loaded delivery system for mucosal immunization against tuberculosis

Tese de doutoramento, Farmácia (Tecnologia Farmacêutica), Universidade de Lisboa, Faculdade de Farmácia, 2016

Die Unterscheidung der Tuberkelbazillus von den übrigen säurefesten Stäbchen mittels des Antiformins.

Inaug.-diss.--Hannover, 1912.

The postantibiotic effect of the carbapenem antibiotics on gram-negative bacteria

Postantibiotic effect (PAE) describes the suppression of microbial growth occurring after a short exposure to an antimicrobial agent. PAE appears to be a property of the majority of antimicrobial agents and is demonstrated by a wide variety of microorganisms. At present, carbapenems and penems are the only members of the -lactam group of antimicrobial agents that exhibit a significant PAE on Gram-negative bacilli. A standardised method was developed to evaluate the in vitro PAE of three carbapenems; imipenem, meropenem and biapenem on Gram-negative bacteria under reproducible laboratory conditions that partially mimicked those occurring in vivo. The effects on carbapenem PAE of the method of antimicrobial removal, concentration, exposure duration, inoculum size, inoculum growth phase, multiple exposures and pooled human serum were determined. Additionally, the reproducibility, susceptibility prior to and after PAE determination and inter-strain variation of carbapenem PAE were evaluated. The method developed determined PAE by utilising viable counts and demonstrated carbapenem PAE to be reproducible, constant over successive exposures, dependent on genera, concentration, duration of exposure, inoculum size and growth phase. In addition, carbapenem PAE was not significantly effected either by agitation, the antimicrobial removal method or the viable count diluent. At present, the mechanism underlying PAE is undetermined. It is thought to be due to either the prolonged persistence of the antimicrobial at the cellular site of action or the true recovery period from non-lethal damage. Increasing the L-lysine concentration and salinity at recovery decreased and increased the carbapenem and imipenem PAE of Pseudomonas aeruginosa, respectively. In addition, no apparent change was observed in the production of virulence factors by P.aeruginosa in PAE phase. However, alterations in cell morphology were observed throughout PAE phase, and the reappearance of normal cell morphology corresponded to the duration of PAE determined by viable count. Thus, the recovery of the penicillin binding protein target enzymes appears to be the mechanism behind carbapenem PAE in P. aeruginosa.

High susceptibility of MDR and XDR Gram-negative pathogens to biphenyl-diacetylene-based difluoromethyl-allo-threonyl-hydroxamate LpxC inhibitors.

OBJECTIVES: Inhibitors of uridine diphosphate-3-O-(R-3-hydroxymyristoyl)-N-acetylglucosamine deacetylase (LpxC, which catalyses the first, irreversible step in lipid A biosynthesis) are a promising new class of antibiotics against Gram-negative bacteria. The objectives of the present study were to: (i) compare the antibiotic activities of three LpxC inhibitors (LPC-058, LPC-011 and LPC-087) and the reference inhibitor CHIR-090 against Gram-negative bacilli (including MDR and XDR isolates); and (ii) investigate the effect of combining these inhibitors with conventional antibiotics. METHODS: MICs were determined for 369 clinical isolates (234 Enterobacteriaceae and 135 non-fermentative Gram-negative bacilli). Time-kill assays with LPC-058 were performed on four MDR/XDR strains, including Escherichia coli producing CTX-M-15 ESBL and Klebsiella pneumoniae, Pseudomonas aeruginosa and Acinetobacter baumannii producing KPC-2, VIM-1 and OXA-23 carbapenemases, respectively. RESULTS: LPC-058 was the most potent antibiotic and displayed the broadest spectrum of antimicrobial activity, with MIC90 values for Enterobacteriaceae, P. aeruginosa, Burkholderia cepacia and A. baumannii of 0.12, 0.5, 1 and 1 mg/L, respectively. LPC-058 was bactericidal at 1× or 2× MIC against CTX-M-15, KPC-2 and VIM-1 carbapenemase-producing strains and bacteriostatic at ≤4× MIC against OXA-23 carbapenemase-producing A. baumannii. Combinations of LPC-058 with β-lactams, amikacin and ciprofloxacin were synergistic against these strains, albeit in a species-dependent manner. LPC-058's high efficacy was attributed to the presence of the difluoromethyl-allo-threonyl head group and a linear biphenyl-diacetylene tail group. CONCLUSIONS: These in vitro data highlight the therapeutic potential of the new LpxC inhibitor LPC-058 against MDR/XDR strains and set the stage for subsequent in vivo studies.

Effect of experimental parameters on alginate/chitosan microparticles for BCG encapsulation

The aim of the present study was to develop novel Mycobacterium bovis bacille Calmette-Guérin (BCG)-loaded polymeric microparticles with optimized particle surface characteristics and biocompatibility, so that whole live attenuated bacteria could be further used for pre-exposure vaccination against Mycobacterium tuberculosis by the intranasal route. BCG was encapsulated in chitosan and alginate microparticles through three different polyionic complexation methods by high speed stirring. For comparison purposes, similar formulations were prepared with high shear homogenization and sonication. Additional optimization studies were conducted with polymers of different quality specifications in a wide range of pH values, and with three different cryoprotectors. Particle morphology, size distribution, encapsulation efficiency, surface charge, physicochemical properties and biocompatibility were assessed. Particles exhibited a micrometer size and a spherical morphology. Chitosan addition to BCG shifted the bacilli surface charge from negative zeta potential values to strongly positive ones. Chitosan of low molecular weight produced particle suspensions of lower size distribution and higher stability, allowing efficient BCG encapsulation and biocompatibility. Particle formulation consistency was improved when the availability of functional groups from alginate and chitosan was close to stoichiometric proportion. Thus, the herein described microparticulate system constitutes a promising strategy to deliver BCG vaccine by the intranasal route.

The cellular and mechanistic study of nisin inhibition of bacillus anthracis spore outgrowth, nisin alteration of infection, and native producer immunity

A critical step during Bacillus anthracis infection is the outgrowth of germinated spores into vegetative bacilli that proliferate and disseminate rapidly within the host. An important challenge exists for developing chemotherapeutic agents that act upon and kill B. anthracis immediately after germination initiation when antibiotic resistance is lost, but prior to the outgrowth into vegetative bacilli, which is accompanied by toxin production. Chemical agents must also function in a manner refractive to the development of antimicrobial resistance. In this thesis we have identified the lantibiotics as a class of chemotherapeutics that are predicted to satisfy these two criteria. The objective of this thesis was to evaluate the efficacy of nisin, a prototypical lantibiotic, in prevention of outgrowth of germinated B. anthracis spores. Like all lantibiotics, nisin is a ribosomally translated peptide that undergoes post-translational modification to form (methyl)lanthionine rings that are critical for antimicrobial activity. Our studies indicate that nisin rapidly inhibits the in vitro outgrowth of germinated B. anthracis Sterne 7702 spores. Although germination initiation was shown to be essential for nisin-dependent antimicrobial activity, nisin did not inhibit or promote germination initiation. Nisin irreversibly killed germinated spores by blocking the establishment of a membrane potential and oxidative metabolism, while not affecting the dissolution of the outer spore structures. The membrane permeability of the spore was increased by nisin, but germinated spores did not undergo full lysis. Nisin was demonstrated to localize to lipid II, which is the penultimate precursor for cell wall biogenesis. This localization suggests two possible independent mechanisms of action, membrane pore formation and inhibition of peptidoglycan synthesis. Structure-activity studies with a truncated form of nisin lacking the two C-terminal (methyl)lanthionine rings and with non-pore forming mutants indicated that membrane disruption is essential for nisin-dependent inhibition of spore outgrowth to prevent membrane potential establishment. Finally, utilizing an in vitro infection model, it was shown that nisin reduced the viability of B. anthracis spores within an infection resulting in increased survival of immune cells while reducing infection-mediated cytokine expression. Fluorescence microscopy indicated that nisin localizes with spores within phagosomes of peritioneal macrophages in germinating conditions. These data demonstrate the effectiveness of nisin, as a model lantibiotic, for preventing spore outgrowth. It is speculated that nisin targeting of lipid II, resulting in membrane perturbations, may be effective at inhibiting the outgrowth of spores prepared from bacteria across a number of species.

Hansen Neuropathy: Still a Possible Diagnosis in the Investigation of a Peripheral Neuropathy

INTRODUCTION: Leprosy is still one of the most frequent causes of peripheral neuropathy. Although regarded as eradicated in Portugal, is still documented in neuropathological study of patients with clinical peripheral neuropathy without proper diagnosis.

Um caso pediátrico de tuberculose miliar: achados raros e evolução atípica

Introdução: A tuberculose miliar resulta da disseminação linfohematogénica do Mycobacterium tuberculosis, sendo uma manifestação grave da infeção. Caso clínico: Criança de 9 anos, género feminino, com his¬tória de febre prolongada. O diagnóstico de tuberculose miliar foi colocado após telerradiografia torácica com infiltrado reticu¬lonodular difuso bilateral, e corroborado pelo achado de tubér¬culos coroideus no olho direito e visualização de bacilos álcool¬ ¬ácido resistentes em amostra de suco gástrico. Detetaram¬-se tuberculomas cerebrais na ressonância magnética. Isolou¬se Mycobacterium tuberculosis multissensível em amostras de suco gástrico. Após mais de 40 dias de tratamento, persistia a febre e baciloscopia positiva. Foi excluída infeção pelo vírus da imunodeficiência humana. Não foram detetadas complicações. Posteriormente, a evolução clínica foi favorável. Discussão/Conclusão: A tuberculose mantém-¬se um diag¬nóstico relevante na criança com febre prolongada. A associa¬ção da imagem torácica, baciloscopias positivas e tubérculos coroideus foram fundamentais para a celeridade do diagnóstico e implementação do tratamento. Reforça¬-se a importância de manter elevado índice de suspeição para uma patologia que tem tratamento.

Improving bioactive compounds extractability of Amorphophallus paeoniifolius (Dennst.) Nicolson

Elephant foot yam (Amorphophallus paeoniifolius (Dennst.) Nicolson) is an underground, unbranched deciduous plant that produces a large tubercle (rhizome) with recognized health effects. In this study, the influence of solvent nature (water, water/etanol (1:1) and absolute ethanol) and processing type (fresh, lyophilized and boiled) on the antioxidant activity and bioactive compounds extractability of elephant foot yam was evaluated. Extracts were compared for their contents in total phenolics, flavonoids and tannins. Moreover, their antioxidant capacity was assessed by the ferric reducing antioxidant power (FRAP) and the 2,2-diphenyl-1-picrylhydrazyl radical (DPPH▪) scavenging capacity assays. Phenolics (154 mg GAE/L) and tannins (109 mg GAE/L) were maximized in lyophilized samples extracted with the hydroalcoholic solvent, which attained also the highest FRAP value (711 mg FSE/L). In turn, flavonoids reached the highest yields in lyophilized samples (95 mg ECE/L) extracted with pure ethanol, as well as the highest DPPH▪ scavenging activity. These findings might have practical applications to define the best processing methodology regarding the enhancement of elephant foot yam, either for prompt consumption, as well as to develop food supplements or pharmaceutical related products.

Prevalence of colonizing bacteria and their association with primary bacteremias in hemodialysis of a university hospital

to be from 2.5 to 5.5 cases per 1,000 catheter-day. the clinical impact is relevant and increases the cost of the HD Unit. Methods: The present study is the irst of 2 phases. It was conducted from January to December of 2012, and included all patients and nurses who were in the HD Unit. The prevalence of Gramnegative bacilli (GNB) and methicillin-resistant Staphylococcus aureus (MrSA) colonizing the nasal passages and the skin is described. Also, phenotypic association was sought by genus, species and sensitivities between colonizing bacterial strains and blood cultures with GNB and MRSA. Results: the study included 70 patients and 10 nurses. the prevalence of nasal colonization in patients by GNB was 9% and 6% in the pericatheter, and no nursing GNB colonization was discovered. The prevalence of MRSA nasal colonization was 19% and 6% in the pericatheter for patients and in the nurses the nasal colonization was 50% and 10% in the hands. We identiied 29 cases of primary bacteremia. The primary bacteremia rate is 1.5 per 1,000 catheter-day or 0.4 episodes per patient per year. Conclusion: We demonstrated a high prevalence of MrSA colonization in patients and nurses in the HD Unit. No relationship was found between primary bacteremia by GNB and patients and nurses’ bacteria colonization by the phenotypic comparison.

Application of dental morphological characteristics for medical-legal identification: sexual diagnosis in a portuguese population

Licenced under a Creative Commons Attribution 3.0.

Evaluación del desempeño de pruebas diagnósticas de tuberculosis en pacientes VIH positivo en dos hospitales públicos de Bogotá

La tuberculosis TB es una de las principales causas de muerte en el mundo en individuos con infección por VIH. En Colombia esta coinfección soporta una carga importante en la población general convirtiéndose en un problema de salud pública. En estos pacientes las pruebas diagnósticas tienen sensibilidad inferior y la enfermedad evoluciona con mayor frecuencia hacia formas diseminadas y rápidamente progresivas y su diagnóstico oportuno representa un reto en Salud. El objetivo de este proyecto es evaluar el desempeño de las pruebas diagnósticas convencionales y moleculares, para la detección de TB latente y activa pacientes con VIH, en dos hospitales públicos de Bogotá. Para TB latente se evaluó la concordancia entre las pruebas QuantiFERON-TB (QTF) y Tuberculina (PPD), sugiriendo superioridad del QTF sobre la PPD. Se evaluaron tres pruebas diagnósticas por su sensibilidad y especificidad, baciloscopia (BK), GenoType®MTBDR plus (Genotype) y PCR IS6110 teniendo como estándar de oro el cultivo. Los resultados de sensibilidad (S) y especificidad (E) de cada prueba con una prevalencia del 19,4 % de TB pulmonar y extrapulmonar en los pacientes que participaron del estudio fue: BK S: 64% E: 99,1%; Genotype S: 77,8% E: 94,5%; PCRIS6110 S: 73% E: 95,5%, de la misma forma se determinaron los valores predictivos positivos y negativos (VPP y VPN) BK: 88,9% y 94,8%, Genotype S: 77,8% E: 94,5%; PCRIS6110 S: 90% y 95,7%. Se concluyó bajo análisis de curva ROC que las pruebas muestran un rendimiento diagnóstico similar por separado en el diagnóstico de TB en pacientes con VIH, aumentando su rendimiento diagnostico cuando se combinan

Descripción del análisis microbológico realizado mediante maldi-tof en muestras de la fundación santa fe de Bogotá.

Introducción: La rápida detección e identificación bacteriana es fundamental para el manejo de los pacientes críticos que presentan una patología infecciosa, esto requiere de métodos rápidos para el inicio de un correcto tratamiento. En Colombia se usan pruebas microbiología convencional. No hay estudios de espectrofotometría de masas en análisis de muestras de pacientes críticos en Colombia. Objetivo general: Describir la experiencia del análisis microbiológico mediante la tecnología MALDI-TOF MS en muestras tomadas en la Fundación Santa Fe de Bogotá. Materiales y Métodos: Entre junio y julio de 2013, se analizaron 147 aislamientos bacterianos de muestras clínicas, las cuales fueron procesadas previamente por medio del sistema VITEK II. Los aislamientos correspondieron a 88 hemocultivos (60%), 28 urocultivos (19%), y otros cultivos 31 (21%). Resultados: Se obtuvieron 147 aislamientos con identificación adecuada a nivel de género y/o especie así: en el 88.4% (130 muestras) a nivel de género y especie, con una concordancia del 100% comparado con el sistema VITEK II. El porcentaje de identificación fue de 66% en el grupo de bacilos gram negativos no fermentadores, 96% en enterobacterias, 100% en gérmenes fastidiosos, 92% en cocos gram positivos, 100% bacilos gram negativos móviles y 100% en levaduras. No se encontró ninguna concordancia en bacilos gram positivos y gérmenes del genero Aggregatibacter. Conclusiones: El MALDI-TOF es una prueba rápida para la identificación microbiológica de género y especie que concuerda con los resultados obtenidos de manera convencional. Faltan estudios para hacer del MALDI-TOF MS la prueba oro en identificación de gérmenes.