Avaliação das propriedades antimicrobianas do Shiitake (Lentinula edodes) em isolados bacterianos responsáveis por Iinfeções associadas aos cuidados de saúde (IACS)

Dupla diplomação com a Universidad de SalamancaA resistência bacteriana constitui atualmente uma das mais graves ameaças à Saúde Pública, havendo bactérias que são resistentes a mais do que uma classe de antibióticos. Uma das maiores problemáticas das resistências bacterianas são as IACS (Infeções associadas aos cuidados de saúde) provocadas por bactérias como Staphylococus aureus resistentes à meticilina (MRSA) e Klebsiella pneumoniae resistentes aos carbapenemos. Este fenómeno de resistência tem levado à procura de alternativas, quer seja pela síntese de novos compostos antibacterianos, quer seja pela procura de produtos naturais com propriedades antibacterianas. Atendendo à composição química dos cogumelos em compostos bioativos, e variedade de trabalhos que demonstram as suas propriedades antibacterianas, surge então a hipótese de que estes compostos poderiam ser promissores aplicados à antibioterapia. Nesse contexto, este trabalho teve como principal objetivo avaliar o potencial antimicrobiano de extratos de cogumelos da espécie Lentinula edodes (Shiitake), uma das espécies de cogumelos mais comercializada a nível mundial, em isolados de bactérias Gram-positivo e Gram-negativo, frequentemente relacionadas com as infeções associadas a cuidados de saúde. Dois tipos de extratos (aquoso e metanólico) obtidos de duas variedades de cogumelo Shiitake (Donko e Koshin) foram avaliados quanto ao seu potencial efeito antibacteriano em isolados responsáveis por IACS. O estudo incluiu um screening da atividade antimicrobiana, a avaliação do efeito sinérgico entre extratos e antibióticos e ainda a determinação da concentração mínima inibitória (CMI) e concentração mínima bactericida (CMB). Os resultados mostraram que na avaliação da atividade antimicrobiana, houve um aumento do tamanho dos halos de inibição nos isolados de Gram-positivo com o aumento da concentração dos extratos aquosos de ambas as variedades, sendo a variedade Koshin a que apresentou melhores resultados. Na avaliação do efeito sinérgico observou-se que para os Gram-positivos, o aumento da concentração do extrato levou a que a ação de alguns antibióticos fosse potenciada. Contudo, nos isolados de Gram-negativo verificou-se a ocorrência de antagonismo com a adição de extrato ao meropenemo. Na determinação das CMIs, determinada em Gram-positivos, ambos os extratos foram eficazes, verificando-se inibição do crescimento a menores concentrações com o extrato aquoso da variedade Koshin. Estas diferenças entre as duas variedades e entre os dois solventes (aquoso e metanólico) podem ser explicadas pelo conteúdo em fenóis totais versus atividade antioxidante, que foi maior no extrato aquoso da variedade Koshin. Assim sendo, valoriza-se o Shiitake como uma promissora fonte de compostos bioativos.La resistencia bacteriana constituye actualmente una de las más graves amenazas en la Salud Pública, esto debido a la existencia de bacterias que son resistentes a más de una clase de antibióticos. Una de las mayores problemáticas a las resistencias bacterianas son las IAAS (infecciones asociadas a la atencion en salud), provocadas por bacterias como Staphylococus aureus, resistentes a la meticilina (MRSA) y Klebsiella pneumoniae resistente a los carbapenems. Este fenómeno de resistencia ha llevado a la búsqueda de alternativas, ya sea con la síntesis de nuevos compuestos antibacterianos, o por la introducción de productos naturales con propiedades antibacterianas. En cuanto a la composición química de las setas, compuestos bioactivos, y variedad de trabajos que demuestran sus propiedades antibacterianas; surge entonces la hipótesis de que esos compuestos podrían ser grandes prometedores en la terapia con antibióticos. En este contexto, este trabajo tuvo como principal objetivo evaluar el potencial antimicrobiano de extractos de setas de la especie Lentinula edodes (Shiitake), una de las especies de setas más comercializadas a nivel mundial, en aislados de bacterias Gram-negativas y Gram-positivas, de ambiente hospitalario. Dos tipos de extractos (acuosos y metanólicos) obtenidos de dos variedades de setas Shiitake (Donko y Koshin) se estudiaron con el fin de evaluar su efecto antibacteriano en aislados responsables de las IAAS. El estudio incluyó un screening de la actividad antimicrobiana, la evaluación del efecto sinérgico entre extractos y antibióticos, así como la determinación de la concentración mínima inhibitoria (CMI) y concentración mínima bactericida (CMB). Los resultados mostraron que en la evaluación de la actividad antimicrobiana, hay un crecimiento en el tamaño de los halos de inhibición en Gram-positivos al aumentar la concentración de extractos acuosos de ambas variedades, arrojando mejores resultados la variedad Koshin. En la evaluación del efecto sinérgico, se observó que para Gram-positivos, el aumento de la concentración del extracto condujo a la acción de algunos antibióticos mejorados. Sin embargo, en Gram-negativos, se produjo antagonismo con la adición de extracto a meropenem. En la determinación de las CMI, analizadas en Gram-positivos, ambos extractos fueron efectivos, con inhibición del crecimiento a concentraciones más bajas con el extracto acuoso de la variedad Koshin. Estas diferencias entre las dos variedades de y entre los dos solventes (acuoso y metanólico) pueden explicarse por el contenido total de fenol versus la actividad antioxidante, que fue mayor en el extracto acuoso de la variedad Koshin. Por lo tanto, Shiitake se valora como una fuente prometedora de compuestos bioactivos.Bacterial resistance is currently one of the most serious threats to public health, with bacteria that are resistant to more than one class of antibiotics. One of the major problems of bacterial resistance is the HCAIs (Health care-associated infections), caused by bacteria such as methicillin resistant Staphylococus aureus (MRSA) and carbapenem resistant Klebsiella pneumoniae. This phenomenon of resistance has led to the search for alternatives, either by synthesizing new antibacterial compounds or by looking for natural products with antibacterial properties. Given the chemical composition of mushrooms, bioactive compounds, and the variety of works that demonstrate their antibacterial properties, it is hypothesized that these compounds could be promisingly applied to antibiotic therapy. In this context, the main objective of this work was to evaluate the antimicrobial potential of mushroom extracts of the species Lentinula edodes (Shiitake), one of the most traded mushroom species in the world, in Gram-negative and Gram-positive bacteria isolates, frequently related. with healthcare-associated infections. Two types of extracts (aqueous and methanolic) obtained from two varieties of Shiitake (Donko and Koshin) mushroom were evaluated for their potential antibacterial effect on isolates responsible for HCAIs. The study included screening for antimicrobial activity, evaluating the synergistic effect between extracts and antibiotics, and determining the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC). The results showed that in the evaluation of antimicrobial activity, there was an increase in the size of inhibition halos in Gram-positive isolates with increasing concentration of aqueous extracts of both varieties, being Koshin variety the best results. In the evaluation of the synergistic effect, it was observed that for Gram-positive, the increased concentration of the extract led to the action of some antibiotics being enhanced. However, in Gram-negative isolates, antagonism occurred with the addition of extract to meropenemo. In the determination of CMIs, determined in Gram-positive, both extracts were effective, with growth inhibition at lower concentrations with aqueous extract of the Koshin variety. These differences between the two varieties and between the two solvents (aqueous and methanolic) can be explained by the total phenol content versus antioxidant activity, which was higher in the aqueous extract of the Koshin variety. Thus, Shiitake is valued as a promising source of bioactive compounds.A realização deste trabalho foi possível mediante o apoio financeiro das seguintes entidades: CIMO - Centro de Investigação de Montanha, Instituto Politécnico de Bragança CITAB - Centro de Investigação e Tecnologias Agroambientais e Biológicas, Universidade de Trás-os-Montes e Alto Douro Projeto FUNGITECH – Tecnologia e inovação na cadeia de produção do shiitake e de outros cogumelos com valor nutracêutico e medicinal, N.º 33788, financiado pelo Fundo Europeu de Desenvolvimento Regional (FEDER) através do NORTE 2020 (Programa Operacional Regional do Norte

Development of a bacteriophage based product to control colibacillosis in poultry

Tese de doutoramento em Engenharia Química e BiológicaEscherichia coli (E. coli) is part of the commensal microflora of the chicken intestinal tract, being commonly an opportunistic bacteria that causes disease in immunologic deprived chickens. However, there are extra-intestinal E. coli strains, the avian pathogenic Escherichia coli (APEC) that are able to cause colibacillosis by itself, due to its invasive ability. The colibacillosis and colisepticemia are responsible for significant economic losses in poultry industries worldwide, mainly due to the low feed conversion rate with consequent weight loss, high cost of treatments during production, poor carcass quality with consequent rejection at slaughter and high mortalities rates. The increasing high patterns of antibiotic resistance acquired by these bacteria, as well as the restriction to the antibiotic usage implemented by the European Union, have encouraged the search of new solutions to control severe infections ensuring good meat quality and minimizing environmental impact. Bacteriophages (phages), virus infecting exclusively bacteria, have been proposed as valuable alternatives to antibiotics based on their capacity to infect and destroy the bacteria, releasing in few minutes progeny that will infect the surrounding hosts. The presented work aimed at developing an efficient, safe and competitive phage based product to control colibacillosis in poultry. The work encompassed five different stages: firstly, different bacteriophages active for a wide range of APEC strains were isolated and characterized; secondly, an in vivo evaluation of the toxicity of the phage suspensions were performed, thirdly the effect of the route of administration and phage titre on phage dissemination in the chickens’ organisms was assessed; fourthly the efficacy of the phages presenting the wider lytic spectrum was evaluated through in vivo efficiency trials; finally, large animal trials were performed to validate the efficacy of the phage product. Phages were isolated from poultry sewage and tested against 148 O-serotyped APEC strains. The results showed that 70.5% of the tested strains were sensitive to a combination of three of the five isolated phages. Taxonomically, two of these three phages, phi F61E and phi F78E look like 16-19, T4-like phages (Myoviridae) and the other, phi F258E is a T1-like phage and belongs to the Syphoviridae family. All belong to the Caudovirales order. Restriction fragment length polymorphism (RFLP) patterns demonstrate that all phages were genetically different. The in vivo evaluation of the toxicity of the phage lysate revealed that the phage suspensions did not promote any decrease in feed and water intake, or body weight lost during the in vivo trial and the post mortem necropsies did not show any macroscopic lesions in the internal organs. These observations supported that the lysate was not toxic for chickens. The in vivo assessment of the effect of the route of administration and the dosage in the dissemination of the phages in the chicken’s organs, revealed that when administered orally and by spray, all the phages reached the respiratory tract, as well as the bloodstream. Intramuscular administration enabled the phages to reach all chickens’ internal organs. Results suggested that, besides the intramuscular administration (not feasible to use in flocks), the oral and spray administration can be considered promising administration routes to treat respiratory E. coli infections in poultry. The in vivo evaluation of the efficacy of phi F78E to control severe E. coli infections revealed that phage performance is dosage dependant and only a high concentration of 109 PFU/ml allowed a decrease in 25% and 43% in chickens’ mortality and morbidity, respectively. Interestingly, the phage cocktail (of phi F61E, phi F78E and phi F258E) administered in the water drinking and by spray in a single application, and composed by 5×107 PFU/ml of each bacteriophage, was able to control the mortality rate in naturally infected chicken flocks, refractive to antibiotherapy. The mortality felt from 2.2% in average, to under 0.5% in no more than three weeks, with no recidivism. In conclusion, with this work it was possible to obtain an antimicrobial product, comprised by a combination of three different lytic phages, which demonstrated to be safe for chickens and efficient against colibacillosis in the poultry industry.A Escherichia coli (E. coli) é uma bactéria que integra a flora intestinal das aves, e que frequentemente se comporta como oportunista, causando doença em aves imunodeprimidas. Contudo, existem estirpes extra-intestinais patogénicas (APEC) que são capazes, pelas suas capacidades invasivas, de causar infecções (colibacilose) que podem degenerar em septicemias. Estas infecções são responsáveis por perdas económicas importantes na indústria avícola, devido a baixas taxas de conversão alimentar com consequentes perdas de peso das aves, elevados custos de tratamento na produção, baixa qualidade da carcaça que leva a rejeições no matadouro e elevadas taxas de mortalidade. O aumento do padrão de resistências a antibióticos adquiridos por estas bactérias, bem como as restrições que a Comunidade Europeia tem vindo a impor no uso destes antimicrobianos, vem relançar a importância do desenvolvimento de alternativas terapêuticas. Os bacteriófagos (fagos), são vírus que parasitam exclusivamente bactérias e que têm vindo a surgir como alternativa valiosa à terapia tradicional. A sua capacidade de infectar e destruir bactérias hospedeiras, libertando nova geração com potencial para infectar os hospedeiros circundantes são características que corroboram a sua valia. O presente trabalho visou o desenvolvimento de um produto à base de fagos para o controlo de colibaciloses na indústria avícola, que se mostrasse eficiente e inócuo e que fosse competitivo. O trabalho compreendeu cinco fases diferentes: em primeiro lugar, bacteriófagos distintos que infectavam uma vasta gama de estirpes APEC foram isolados e caracterizados; seguidamente, foi efectuada a avaliação in vivo da toxicidade da suspensão de fagos; em terceiro lugar, foi testada a influência da via de administração e da concentração de fagos na sua disseminação no organismo das aves; posteriormente a eficiência dos fagos com o espectro de lise mais amplo foi avaliada, através de experimentação in vivo; finalmente foram realizadas experiências em aviários experimentais para validar a eficácia do produto à base de fagos. Os fagos foram isolados de camas de aviários e testados para 148 APEC serotipadas para o antigénio “O”. Destas, 70.5% revelaram-se sensíveis a pelo menos um de três dos fagos. Taxonomicamente, todos os fagos se inserem na ordem dos Caudovirales, sendo que dois deles, o phi F61E e o phi F78E são fagos 16-19, do tipo T4 (Myoviridae) e o outro, phi F258E é um fago do tipo T1 (Syphoviridae). Padrões de RFLP demonstraram que todos os fagos são geneticamente diferentes. A avaliação in vivo da toxicidade do lisado de fagos revelou que a suspensão de fagos não provocou qualquer diminuição na ingestão de alimento e água nem perda de peso, e a avaliação post mortem das carcaças não revelou lesões macroscópicas nos órgãos. Estas observações corroboram a inocuidade do produto, para as aves. As experiências in vivo para analisar o efeito da via de administração e da concentração da suspensão, na disseminação dos fagos no organismo das aves, indicou que quando administrados por via oral e por spray, qualquer dos tipos de fago em teste atingiram o tracto respiratório e a corrente sanguínea. A administração intra-muscular, permitiu que os fagos testados fossem recuperados em todos os órgãos analisados. Os resultados sugerem que, para além da administração intramuscular (modo que não é prático para o maneio em bandos), a administração oral e nasal deverão ser veículos eficientes de transporte de fagos para o tratamento de infecções respiratórias por E. coli em aves. A avaliação in vivo da eficiência do phi F78E para o controlo de infecções severas de E. coli, revelou que o desempenho do fago estava dependente da dose, e que apenas uma concentração elevada, de 109 PFU/ml, permitiu um decréscimo na mortalidade e morbilidade das aves, respectivamente de 25% e 43% em média. Curiosamente, a combinação de fagos (phi F61E, phi F78E e phi F258E), numa concentração de 5×107 PFU/ml cada um, administrado na água de bebida e por spray numa única aplicação, foi eficiente no controlo da mortalidade em bandos naturalmente infectados por APEC, e em que os antibióticos não tinham tido sucesso terapêutico. A mortalidade desceu, em média, de 2.2% para valores inferiores a 0.5% em não mais de 3 semanas, sem recidivas. Em conclusão, este trabalho possibilitou o desenvolvimento de um produto constituído por três fagos, que demonstrou ser inócuo e eficiente no controlo de colibacilose na indústria avícola.Fundação para a Ciência e a Tecnologia (FCT), SFRH/BDE/15508/200

In vivo efficiency evaluation of a phage cocktail in controlling severe colibacillosis in confined conditions and experimental poultry houses

Infections caused by avian pathogenic Escherichia coli (APEC) cause important economic losses to poultry industry. The studies presented herein, aimed at investigating the in vivo performance of a cocktail of three phages in treating severe respiratory E. coli infections in experimentally contaminated birds and naturally infected flocks. Three lytic coliphages, phi F78E (Myoviridae), phi F258E (Siphoviridae) and phi F61E (Myoviridae) were combined in a 5.0 × 107 PFU/ml cocktail to be used in naturally APEC infected flocks (refractive to antibiotherapy). Experimentally infected birds were treated with phi F78E at two different titres (107 PFU/ml and 109 PFU/ml). Phage administration was performed orally and by spray, in a single application. The morbidity, mortality and pathology scores were compared with control birds not receiving phage therapy. The results revealed that the success of phage therapy in experimental rooms was dosage dependent, being 107 PFU/ml not enough to treat the infected chickens whereas a concentration of 109 PFU/ml of phi F78E allowed a decrease of 25% and 43% in chickens’ mortality and morbidity, respectively. In the large scale experiments, the results obtained showed a remarkable efficacy of the low titre phage cocktail (107 PFU/ml) in decreasing the flocks’ mortality to levels below 0.5% in no more than 3 weeks, with no recidivism. Based on the results we can conclude that phage treatment is a valuable alternative to control APEC infections in poultry

Anti-biofilm effect: combining bacteriophages with Portuguese honey

Nowadays, the rising of multi resistant bacteria due to the misuse of antibiotics, together with the lack in the development of new antimicrobial molecules, represents a serious threat to Medicine. Many of these multi resistances bacteria are pathogens responsible for nosocomial infections causing problem in Human health and care.This work addresses the problematic of treating chronic wounds, lacerations and infected burns. In these types of infections, bacteria colonize the tissues and form biofilms, conditioning the effectiveness of antibiotics.Based on this reality, the development of news approaches to treat multi resistance organisms is essential. A combination of two antimicrobial solutions, bacteriophages (phages) and honey, is suggested herein. Bacteriophages (phages) are natural bacteria predators that specifically recognize hosts, destroying it. Honey is also able to destroy bacteria, but also to degrade biofilms, to improve the healing process, promoting tissue regeneration, and to decrease the inflammation. Both are efficient against antibiotic resistant microorganisms, including those present in chronic wounds. To date there is no study about this combinatorial therapy.In this study, the anti-biofilm effect of the combination of a Portuguese honey together with wellstudied lytic phages was enhanced, in Escherichia coli and Pseudomonas aeruginosa biofilms relatively to the same antimicrobials used individually. This result was confirmed by a decrease in the total biomass, and by a reduction, in average, of more than 3 log of cultivable cells

A new phage endolysin as a powerful tool to detect and kill Paenibacillus larvae

American foulbrood (AFB) is an infection caused by Paenibacillus larvae (P. larvae), a Gram-positive spore forming bacteria. This disease occurs in honeybee larvae, when spores germinate and proliferate in their midgut and subsequently penetrate into the hemolymph, causing sepsis and larval death. This work was motivated by the need of finding alternatives to antibiotics, that will leave residues in honey if used to treat this infection. Bacteriophages (phages) and/ or their endolysins might represent valuable tools to use in AFB control as have already proved to be powerful biological antimicrobials. Particularly in this work, endolysins were mainly focused. We have previously isolated and reported the first known P. larvae phage genome and by its in silico analysis we further identified, expressed and characterized the first P. larvae endolysin, PlyPl23. This enzyme has two functional domains: a catalytic domain (Amidase_2) and a totally new cell wall binding domain (CBD). The latter confers specificity to the enzyme, targeting specific bonds of the cell wall surface. The antimicrobial activity of PlyPl23 was tested in vitro against a panel of P. larvae strains and in vivo in bee larvae. PlyPl23 was effective in decreasing P. larvae infection yields in bee larvae experimentally infected with spores and no toxicity effects were encountered. In a complementary study, the cell wall binding domain (CBD) of PlyPl23, fused to a green fluorescence probe was heterologously expressed. The specificity of the PlyPl23 CBD was assessed through flow cytometry and epifluorescence microscopy. Overall the results demonstrate the potential of a phage endolysin for detection and control of P. larvae

Mechanical versus manual harvest of Pinus pinea cones

Umbrella pine cone production is an important forest non-wood product in Portugal,especially in the region of Alcácer do Sal, where it plays an important role to the local development. Traditionally umbrella pine cones are manually harvested, increasing production costs and, above all, with very high accident risk to the workers. The development of equipment for mechanical harvesting started in Italy in the 1980's. Studies report different values for harvesting efficiency and tree damage, the latter in terms of immature cones and branches detached. In this study a trunk shaker was used to evaluate mechanical harvesting both in terms of efficiency and tree damage induced by trunk vibration. In comparison to the manual process, time required for mechanised harvesting was about 1/15th of the time. The results revealed a mechanical harvesting efficiency higher than 86% with negligible tree damage. Inter-annual harvest efficiency variability was also observed

An innovative maturity model to assess supply chain quality management

Purpose: This paper aims to present and discuss an innovative maturity model (MM) to assess supply chain quality management (SCQM). The SCQM MM can be used to guide organizations in the development and improvement of quality in their supply chains (SCs). Additionally, this paper intends to better understand that integration and its impacts on organizational performance. Design/methodology/approach: The proposed MM was developed based on an exhaustive literature review of the most relevant MMs developed in the areas of quality management, supply chain management and other relevant domains. Findings: The proposed MM consists of a matrix with 100 criteria organized in five organizational dimensions and five maturity levels, embracing the most relevant SC issues and describing a progressive path towards a state of full maturity. Originality/value: It is an innovative tool useful for both academic experts and practitioners to integrate quality management across the SC, thus promoting and improving organizational performance from an integrated and sustainable perspective.This work has been supported by FCT – Fundação para a Ciência e Tecnologia within the R&D Units Project Scope: UIDB/00319/2020

Characterization of PlyPl23, an endolysin from a Paenibacillus larvae bacteriophage

EMBO Conference on Viruses of Microbes III: Structure and Function - from Molecules to Communities (Programme and Abstract Book)Bees pollination presents a great impact in a wide range of terrestrial ecosystems. The estimated world value for the contribution of pollinators, especially bees, to the production of crops used directly for human food was estimated in 153 billion. Yet, beekeeping is facing many challenges, as changes in agricultural practices, extensive use of pesticides and the presence of pathogens. One of the most widespread and the most destructive of the honeybee brood diseases is the American foulbrood (AFB), caused by Paenibacillus larvae, a Gram-positive bacterium wherein the spore is the infectious form. In most situations P. larvae is highly virulent and when contracted will kill a complete colony. This represents an economic threat to commercial beekeepers and apiculture worldwide. The European regulation limits the presence of antibiotics in honey, excluding its use for therapy. The antibacterial features of (bacterio)phages and endolysins were considered for the development of a therapeutic product for this disease. A P. larvae phage, phiIBB_Pl23, belonging to the Siphoviridae family was isolated and its genome has been sequenced and analyzed. Annotation of the phiIBB_Pl23 genome identified gp21 as an endolysin protein, designated as PlyPl23, which was further characterized and is presented herein. In silico analysis revealed that PlyPl23, is a single domain globular protein, which is a rare feature of Gram-positive endolysins. The catalytic domain was identified as an N-acetylmuramoyl-L-alanine amidase (pfam Amidase_2 family) with a molecular weight of 25.8 kDa and isoelectric point of 5.74. Activity tests were performed at 37oC, which is similar to the hive temperature (about 34-35ºC) and revealed that this is a broad-spectrum endolysin for P. larvae with an optimal pH for reactivity between 3 and 5. This matches the pH levels of honey, nectar, pollen, and royal jelly (pH 3 to 4). The enzyme also displays a high antimicrobial activity between 5 and 7 when supplemented with NaCl, matching also the intestinal pH of bee larvae (pH 6.8) and adult bees (pH 5.6 to 6.3). Furthermore, tests performed with honeybee larvae juice revealed a higher bacterial decrease reduction, which may be a good indicator for the in vivo effectiveness of the endolysin. The present work describes, to our knowledge, the first characterization of an endolysin from a P. larvae phage. Moreover, characterization of endolysin PlyPl23 showed that it presents high potential in the development of a commercial product to control the problematic AFB.info:eu-repo/semantics/publishedVersio

Synergistic antimicrobial interaction of honey and bacteriophage in Escherichia coli biofilms

I Simpósio de Bioquímica AplicadaWound colonization by biofilms-forming bacteria is one of the main obstacles to the treatment of chronic wounds, causing a number of biological and financial problems. Biofilms are structured communities of bacterial cells enclosed in a self-produced polymeric matrix, adhered to inert or living surfaces, blocking antibiotics and patients immune cells from reaching bacteria. Bacterio(phages), viruses infecting exclusively bacteria, and honey are being considered as valuable alternatives to treat a variety of infections (Table 1). Phages are harmless to mammalian cells, specific for target bacteria therefore not affecting commensal microflora, have the ability to selfreplicate as long as the host is present, and are effective against antibiotic resistant bacteria. Honey is a complex substance with broad spectrum antimicrobial activity, essentially attributed to the high sugar content, low pH, the presence of hydrogen peroxide and methylglyoxal that reacts with important biological molecules (RNA, DNA and proteins). Honey had yet the potential to promote tissue regeneration, cicatrization, decrease inflammation, improving wound healing. In this work the combination of those two antimicrobial agents was considered in the control E. coli biofilms.Hospital Escala Braga (Braga,Portugal) for providing the clinical isolate. This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope off the strategic funding of the UID/BIO/04469/2013 unit and COMPETE 2020(POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte 2020-Programa Operacional Regional do Norte and the Project RECI/BBB-EBI/0179/2012(FCOMP-01-0124-FEDER-027462) and the project PTDC/CVT-EPI/4008/2014(POCI-01-0145-FEDER-016598). Portuguese Foundation for Science and Technology (FCT) through the SFRH/BPD/69356/2010 and SFRH/BD/93078/2013. FCT (IF/01413/2013)info:eu-repo/semantics/publishedVersio

Exantema após vacinação do sarampo: análise laboratorial de casos notificados em São Paulo

OBJECTIVE: The clinical differential diagnosis of rash due to viral infections is often difficult, and misdiagnosis is not rare, especially after the introduction of measles and rubella vaccination. A study to determine the etiological diagnosis of exanthema was carried out in a group of children after measles vaccination. METHODS: Sera collected from children with rash who received measles vaccine were reported in 1999. They were analyzed for IgM antibodies against measles virus, rubella virus, human parvovirus B19 (HPV B19) using ELISA commercial techniques, and human herpes virus 6 (HHV 6) using immunofluorescence commercial technique. Viremia for each of those viruses was tested using a polimerase chain reaction (PCR). RESULTS: A total of 17 cases of children with exanthema after measles immunization were reported in 1999. The children, aged 9 to 12 months (median 10 months), had a blood sample taken for laboratory analysis. The time between vaccination and the first rash signs varied from 1 to 60 days. The serological results of those 17 children suspected of measles or rubella infection showed the following etiological diagnosis: 17.6% (3 in 17) HPV B19 infection; 76.5% (13 in 17) HHV 6 infection; 5.9% (1 in 17) rash due to measles vaccine. CONCLUSIONS: The study data indicate that infection due to HPV B19 or HHV 6 can be misdiagnosed as exanthema due to measles vaccination. Therefore, it is important to better characterize the etiology of rash in order to avoid attributing it incorrectly to measles vaccine.OBJETIVO: O diagnóstico diferencial de doenças exantemáticas causadas por vírus é geralmente difícil, e equívocos não são raros, especialmente depois da introdução da vacina contra o sarampo e a rubéola. Um estudo laboratorial foi conduzido com o objetivo de estabelecer o diagnóstico etiológico de casos de exantema em crianças que receberam a vacina contra o sarampo. MÉTODOS: Soros de casos de exantema em crianças que receberam vacina contra o sarampo, em 1999, foram analisados para anticorpos IgM contra os vírus do sarampo, da rubéola e do parvovírus humano B19 (HPV B19), por técnicas comerciais de Elisa, e o herpes vírus humano tipo 6 (HHV 6), por técnica comercial de imunofluorecência. A viremia para cada um desses vírus foi testada pela reação em cadeia da polimerase (PCR). RESULTADOS: Foram notificados, em 1999, 17 casos de crianças com exantema pós-vacinal. A idade das crianças era de nove a 12 meses (mediana, dez meses). Uma amostra de sangue colhida para investigação laboratorial foi obtida para cada criança. O tempo decorrido entre a aplicação da vacina e o aparecimento do exantema variou de um a 60 dias. Os resultados da sorologia das 17 crianças sugeriram o seguinte diagnóstico etiológico para o exantema: 17,6% (três em 17) infecção pelo HPV B19; 76,5% (13 em 17) infecção pelo HHV 6; 5,9% (um em 17) exantema originado pela vacina do sarampo. CONCLUSÃO: Os resultados indicaram que a infecção pelo HPV B19 ou pelo HHV 6 pode ser diagnosticada como sarampo de origem vacinal. Portanto, é fundamental incluir esses vírus no diagnóstico laboratorial para corretamente apontar a etiologia das doenças exantemáticas, evitando, assim, atribuir à vacina do sarampo efeito colateral