Estudo para determinação da frequência de deformidade do mento em Chironomus sancticaroli (Diptera: chironomidae) em cultura de laboratório

The midge Chironomus sancticaroli (Diptera: Chironomidae) has been used in ecotoxicological tests because it is sensitive to a variety of inorganic pollutants. Among the parameters used to evaluate the toxicity of a substance is the frequency of mentum deformity, which is part of the oral system of this organism. However, there is still no consensus on the baseline level (percentage) of acceptable deformities in laboratory cultures not exposed to pollutants. The determination of this variable is important to ensure the validity of bioassays and to compare cultures from different research and teaching institutions. Once this value is established, it will also be used to monitor the quality of organisms cultured, since factors such as inbreeding could increase the frequency of mentum deformity. Thus, the objective of this study was to quantify the percentage of mentum deformity in the fourth instar of C. sancticaroli larvae from the culture of the Laboratory of Aquatic Ecosystems, at Embrapa Meio Ambiente. The average frequency of mentum deformity obtained was 6,63%. It is believed that factors such as the renewal of the culture with the inclusion of spawns from the laboratories of other institutions, as well as the control of the quality of the dilution water and the sediment of the breeding may have contributed to a low frequency of mentum deformity of the culture observed in this study.142sensível a uma variedade de poluentes inorgânicos. Um dos parâmetros utilizados para avaliar a toxicidade de uma substância é a frequência de deformidade do mento, que faz parte do sistema oral deste organismo. Entretanto, ainda não há consenso a respeito do nível basal (porcentagem) de deformidade aceitável em culturas de laboratório não expostas a poluentes. A determinação desta variável é importante para assegurar a validade de bioensaios e comparar culturas de diferentes instituições de pesquisa e ensino. Uma vez estabelecido, este número também será usado para o controle da qualidade dos organismos criados, já que fatores como o endocruzamento poderiam aumentar a frequência de deformidade do mento. Assim, o objetivo deste estudo foi quantificar a porcentagem de deformidade do mento em larvas de quarto instar de C. sancticaroli de cultura do Laboratório de Ecossistemas Aquáticos da Embrapa Meio Ambiente. A média de frequência de deformidade obtida para a cultura foi de 6,63%. Acredita-se que fatores como a renovação da cultura com a inclusão de desovas de laboratórios de outras instituições, assim como o controle da qualidade da água de diluição e do sedimento da criação, podem ter contribuído para uma baixa frequência de deformidade do mento da cultura

Predictive Factors and Morbidities Associated With Patent Ductus Arteriosus in Very Low Birth Weight Infants With Gestational Age 27 - 31 Weeks

A persistência de canal arterial hemodinamicamente significativo (PCAHS) é uma patologia frequente em recém-nascidos de muito baixo peso. O objectivo deste estudo foi identificar fatores de risco e morbilidades associadas à PCAHS no recém-nascido de muiot baixo peso com idade gestacional de 27 e 31 semanas. Estudaram-se os recém-nascidos(RN) com idade gestacional entre 27 e 31 semans e peso de nascimento inferior a 1500 gramas, admitidos numa unidade de cuidados intensivos neonatais entre 2010 e 2012. Realizou-se um estudo caso-coorte, tendo-se identificado os casos com diagnóstico ecográfico de PCAHS e uma amostra sistemática de RN sem diagnóstico de PCAHS (coorte de controlos). Foram explorados por regressão logística modelos preditivos da ocorrência de PACHS e da sua contribuição para a principal morbilidade neonatal. Nos três anos de estudo, a incidência de PACHS foi de 15%, intervalo de confiança (IC) 95% 11,3 - 19,5. A análise dos 44 RN com PACHS e dos 60 sem PACHS identificou como melhores preditores de PACHS a necessidade de ventilação venosa invasiva (odds ratio (OR) 3.65: IC95%1, 268 - 10,479: p=0,016) e a administração de surfatante (OR ajustado 4,52; IC95% 1,738-11,735; p=0,002); a PACHS mostrou ser significativa no modelo preditivo de leucomalácia periventricular (LPV) (OR ajustado 4,42: IC95% 1,621-12,045; p=0,004). Os resultados obtidos sugerem que estratégias preventivas e terapêuticas que permitam a redução da necessidade de administração de surfatante e de ventilação mecânica invasiva podem reduzir o risco de PCAHS. A PCAHS está positivamente associada à incidência de LPV

Development of multidrug resistance in staphylococci driven by efflux

[P282]Aim: To study the efflux driven response of two major staphylococcal pathogens, S. aureus and S. epi-dermidis to the challenge by non-antibiotic drugs. Methods: We adapted three reference strains to ethidium bromide (EtBr), a broad substrate of bacte-rial efflux pumps. The parental strains, S. aureus ATCC25923, S. epidermidis ATCC12228 and S. epider-midis RP62A were cultured in varying concentrations of EtBr, to obtain their EtBr-adapted derivatives; ATCC25923_EtBr; ATCC12228_EtBr and RP62A_EtBr. Susceptibility of parental and adapted strains to 10 antibiotics and 6 biocides was evaluated by microdilution MIC determination with or without efflux inhibitors. Efflux activity was established by fluorometric assays and the relative expression of the genes coding for the main efflux pumps (EPs) of each species quantified by RT-PCR.Results: For each strain tested, exposure to EtBr resulted in the development of a multidrug resistance (MDR) phenotype, which included resistance to fluoroquinolones and decreased susceptibility to bio-cides, including cetrimide, benzalkonium chloride and tetraphenylphosphonium bromide. Efflux inhib-itors such as verapamil reduced these resistance levels. The EtBr-adapted cultures showed increased efflux activity, which was accompanied by over-expression of distinct EP genes, in a temporal pattern. Conclusion: These results show that both S. aureus and S. epidermidis have the potential to develop efflux driven MDR phenotypes when exposed to a non-antibiotic substrate of multidrug EPs, which can be mediated by distinct efflux pumps, depending on the drug and the bacterial genetic background.publishe

Mycobacterium tuberculosis drug-resistance testing: challenges, recent developments and perspectives

Drug-resistance testing, or antimicrobial susceptibility testing (AST), is mandatory for Mycobacterium tuberculosis in cases of failure on standard therapy. We reviewed the different methods and techniques of phenotypic and genotypic approaches. Although multiresistant and extensively drug-resistant (MDR/XDR) tuberculosis is present worldwide, AST for M. tuberculosis (AST-MTB) is still mainly performed according to the resources available rather than the drug-resistance rates. Phenotypic methods, i.e. culture-based AST, are commonly used in high-income countries to confirm susceptibility of new cases of tuberculosis. They are also used to detect resistance in tuberculosis cases with risk factors, in combination with genotypic tests. In low-income countries, genotypic methods screening hot-spot mutations known to confer resistance were found to be easier to perform because they avoid the culture and biosafety constraint. Given that genotypic tests can rapidly detect the prominent mechanisms of resistance, such as the rpoB mutation for rifampicin resistance, we are facing new challenges with the observation of false-resistance (mutations not conferring resistance) and false-susceptibility (mutations different from the common mechanism) results. Phenotypic and genotypic approaches are therefore complementary for obtaining a high sensitivity and specificity for detecting drug resistances and susceptibilities to accurately predict MDR/XDR cure and to gather relevant data for resistance surveillance. Although AST-MTB was established in the 1960s, there is no consensus reference method for MIC determination against which the numerous AST-MTB techniques can be compared. This information is necessary for assessing in vitro activity and setting breakpoints for future anti-tuberculosis agents

Impact of efflux in the development of multidrug resistance phenotypes in Staphylococcus aureus

WOS: 000363382300003Background: Efflux has been recognized as a resistance mechanism to antimicrobials in Staphylococcus aureus; however its role on the development of clinically relevant resistance is still poorly characterized. This study aimed to examine the impact of efflux on development of resistance to fluoroquinolones and other antimicrobials in S. aureus strains representing relevant phenotypes in terms of antibiotic susceptibility and efflux activity. Methods: Two closely related methicillin- and ciprofloxacin-resistant Staphylococcus aureus clinical strains, with different efflux capacity and the pan-susceptible strain ATCC25923 were exposed to constant concentrations of the efflux pump (EP) substrates ciprofloxacin, ethidium bromide and cetrimide. Parental and exposed strains were tested regarding their susceptibility towards antibiotics, biocides and ethidium bromide, efflux capacity and levels of EP gene expression. Occurrence of resistance-associated mutations was screened by sequencing. Results: Multidrug resistance phenotypes emerged upon exposure, independently of the substrate or its concentration, which were correlated with increased efflux capacity of the exposed strains. The temporal pattern of EP gene expression disclosed an early-response with high expression of several genes, followed by a late-response, characterized by overexpression of specific genes. The overall cell response was more pronounced for strains with an initial basal efflux activity. Remarkably, detection of the IS256 element in the promoter regions of mgrA and norA, in some cases associated with increased gene expression, suggests that these genes may be hot spots for IS256 insertion events. The results obtained with exposure of ATCC25923 to ciprofloxacin were particularly striking, revealing a step-wise development of fluoroquinolone resistance, with a first efflux-mediated response, followed by the occurrence of a mutation in grlA that resulted in phenotypic resistance. Additionally, challenge by non-fluoroquinolone agents, particularly cetrimide, promoted cross resistance to fluoroquinolones, revealing the potential role of biocides as selective pressure for the emergence of resistance to these antibiotics. Conclusions: This study reveals efflux as a significant component of S. aureus resistance to fluoroquinolones and biocides and as a primary mechanism to withstand stress imposed by antimicrobials. This efflux-mediated response can result in the emergence of multidrug resistance in healthcare environments and should be taken into account in the management of this major pathogen.publishersversionpublishe

Cardiopulmonary complications in a patient with bezoar

We report a case of a 17-year-old institutionalised male with a medical history of fragile X syndrome, bilateral congenital glaucoma, cataracts and pica disorder. He was transferred to our paediatric intensive care unit owing to respiratory failure and hypotension. On transoesophageal echocardiogram, he presented left atrium compression. A CT of the thorax and mediastinum revealed an unknown heterogeneous material in the lumen of the stomach and oesophagus, with a lung parenchyma suggestive of alveolar foreign material. Endoscopic evaluation showed diaper fragments inside the oesophagus and stomach. Fragmentation and suction of diaper material was made. Medical treatment was performed with inotropic support, conventional mechanical ventilation and antibiotics

Revisiting susceptibility testing in MDR-TB by a standardized quantitative phenotypic assessment in a European multicentre study

Objectives Treatment outcome of MDR-TB is critically dependent on the proper use of second-line drugs as per the result of in vitro drug susceptibility testing (DST). We aimed to establish a standardized DST procedure based on quantitative determination of drug resistance and compared the results with those of genotypes associated with drug resistance. Methods The protocol, based on MGIT 960 and the TB eXiST software, was evaluated in nine European reference laboratories. Resistance detection at a screening drug concentration was followed by determination of resistance levels and estimation of the resistance proportion. Mutations in 14 gene regions were investigated using established techniques. Results A total of 139 Mycobacterium tuberculosis isolates from patients with MDR-TB and resistance beyond MDR-TB were tested for 13 antituberculous drugs: isoniazid, rifampicin, rifabutin, ethambutol, pyrazinamide, streptomycin, para-aminosalicylic acid, ethionamide, amikacin, capreomycin, ofloxacin, moxifloxacin and linezolid. Concordance between phenotypic and genotypic resistance was >80%, except for ethambutol. Time to results was short (median 10 days). High-level resistance, which precludes the therapeutic use of an antituberculous drug, was observed in 49% of the isolates. The finding of a low or intermediate resistance level in 16% and 35% of the isolates, respectively, may help in designing an efficient personalized regimen for the treatment of MDR-TB patients. Conclusions The automated DST procedure permits accurate and rapid quantitative resistance profiling of first- and second-line antituberculous drugs. Prospective validation is warranted to determine the impact on patient car

Project 8 Phase III Design Concept

We present a working concept for Phase III of the Project 8 experiment, aiming to achieve a neutrino mass sensitivity of $2~\mathrm{eV}$ ($90~\%$ C.L.) using a large volume of molecular tritium and a phased antenna array. The detection system is discussed in detail.Comment: 3 pages, 3 figures, Proceedings of Neutrino 2016, XXVII International Conference on Neutrino Physics and Astrophysics, 4-9 July 2016, London, U