Browsing by Author "Duarte, Aida"
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- Activated carbons: efficient materials for the removal of multi- resistant bacteriaPublication . Silva, Sandra; Jordao, Luisa; Duarte, Aida; Carvalho, Ana PaulaThe rapid emergence of multi-resistant bacteria is occurring worldwide being hospital effluent a major source of these microorganisms. To remove these bacteria possible strategies are those involving activated carbon that due to their tunable properties can be suitable adsorbents for different types of microorganisms, for example Gram + or Gram – bacteria. The objective of this study was to test an activated carbon (AC) commercialized for water treatment purpose to reduce the content of bacteria commonly detected in hospital wastewater. The assays were made using the carbon in two different granulometries against multi-resistant strains: Klebsiella pneumoniae, Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus. The adsorption tests were performed following a procedure adapted from Naka et al. [1]. In any case in parallel with the adsorption assays, the growth of the bacteria was also assessed.The results displayed at Figure 1 show that carbon AC allows a continue decrease of bacteria amount in the liquid media, despite the continuous increase of bacteria demonstrated by the blank experiment. The images displayed in Figure 2 (b) and (c) show the cells adherent to carbon, after 90 minutes of contact. These preliminary results allow us to conclude that activated carbon can be used to increase the removal efficiency of multi-resistant bacteria present in hospital wastewater. Acknowledgments The financial support of FCT to CQB (PEst-OE/QUI/UI0612/2013) is acknowledged. [1] K. Naka, S.T. Watarai, K. Inoue, Y. Kodama, K. Oguma, T.Yasuda, H.J. Kodama, 2001, Veter. Med. Science., 63, 281-285.
- Adenoid bacterial colonization in a paediatric populationPublication . Subtil, João; Rodrigues, João Carlos; Reis, Lúcia; Freitas, Luís; Filipe, Joana; Santos, Alberto; Macor, Carlos; Duarte, Aida; Jordao, LuisaAdenoids play a key role in both respiratory and ear infection in children. It has also been shown that adenoidectomy improves these symptoms in this population. The main goal of the present study was to evaluate adenoid bacterial colonization and document a possible relation with infectious respiratory disease. A prospective observational study was designed to evaluate the proposed hypothesis in a paediatric population submitted to adenoidectomy by either infectious or non-infectious indications and compare these two cohorts. A total of 62 patients with ages ranging from 1 to 12 years old were enrolled in the study. Adenoid surface, adenoid core and middle meatus microbiota were compared. A close association between adenoid colonization and nasal infection was found, supporting that adenoids may function as bacterial reservoir for upper airway infection. The obtained results also contribute to explain the success of adenoidectomy in patients with infectious indications.
- Adenoids, friends or foes?Publication . Subtil, João; Lavado, Paula; Rodrigues, João; Rodrigues, Lúcia; Nogueira, Isabel; Duarte, Aida; Jordao, LuisaThe main goal of this study is to evaluate the existence of a link between biofilm assembly on adenoids and the incidence of recurrent infections within a paediatric population. Thirty-three different bacterial genera were isolated from 186 samples (nasal/adenoid scrubs and adenoid biopsies) being Haemophilus, Neisseria, Streptococcus and Staphylococcus the most frequent. Biofilms were present in 27.4% of the adenoid samples as assessed by SEM. Since the aim of the study is to assess the role of biofilms on adenoid colonization/ invasion and onset of infection a group of samples harbouring clinically relevant bacteria (H.influenzae, S. aureus, S. pyogenes, S. pneumococcus and Moraxella) both on the adenoid surface and core were selected for further studies. Based on biofilm assembly in vitro, assessed by crystal violet assay, bacteria were classified as weak, moderate and strong biofilm assemblers. No direct relation between the ability to assemble biofilms in vitro and the presence of biofilms on the adenoid (biofilm in vivo) was found. A similar result was obtained for antibiotic susceptibility with the majority of bacteria being antibiotic susceptible independently of its origin (sample with or without biofilms). This result might be explained at least partially, by the nature of the sample since the adenoidectomy can only be performed in individuals without infection. H. influenzae, the most isolated bacterium, is an opportunistic pathogen, highly adapted to colonize the upper respiratory tract and easily progresses to infection, especially in children. For this reason, virulence factors such as the capsular type were investigated by PCR. However, all strains were characterized as non-capsulated, which might explain adenoid colonization and biofilm formation, as have been also described in the literature. Further studies must be performed to validate the thesis that adenoids function as a reservoir of etiologic agents of respiratory and ear infections.
- Antimicrobial activity of prophage endolysins against critical Enterobacteriaceae antibiotic-resistant bacteriaPublication . Gonçalves, Tiago; Marques, Andreia T.; Manageiro, Vera; Tanoeiro, Luis; Vital, Joana S.; Duarte, Aida; Vítor, Jorge M.B.; Caniça, Manuela; Gaspar, Maria Manuela; Vale, Filipa F.Enterobacteriaceae species are part of the 2017 World Health Organization antibiotic-resistant priority pathogens list for development of novel medicines. Multidrug-resistant Klebsiella pneumoniae is an increasing threat to public health and has become a relevant human pathogen involved in life-threatening infections. Phage therapy involves the use of phages or their lytic endolysins as bioagents for the treatment of bacterial infectious diseases. Gram-negative bacteria have an outer membrane, making difficult the access of endolysins to the peptidoglycan. Here, three endolysins from prophages infecting three distinct Enterobacterales species, Kp2948-Lys from K. pneumoniae, Ps3418-Lys from Providencia stuartii, and Kaer26608-Lys from Klebsiella aerogenes, were purified and exhibited antibacterial activity against their specific bacterium species verified by zymogram assays. These three endolysins were successfully associated to liposomes composed of dimyristoyl phosphatidyl choline (DMPC), dioleoyl phosphatidyl ethanolamine (DOPE) and cholesteryl hemisuccinate (CHEMS) at a molar ratio (4:4:2), with an encapsulation efficiency ranging from 24 to 27%. Endolysins encapsulated in liposomes resulted in higher antibacterial activity compared to the respective endolysin in the free form, suggesting that the liposome-mediated delivery system enhances fusion with outer membrane and delivery of endolysins to the target peptidoglycan. Obtained results suggest that Kp2948-Lys appears to be specific for K. pneumoniae, while Ps3418-Lys and Kaer26608-Lys appear to have a broader antibacterial spectrum. Endolysins incorporated in liposomes constitute a promising weapon, applicable in the several dimensions (human, animals and environment) of the One Health approach, against multidrug-resistant Enterobacteriaceae.
- Assessing the role played by biofilms on adenoidal surface in a pediatric populationPublication . Rodrigues, João; Reis, Lúcia; Nogueira, Isabel; Duarte, Aida; Subtil, João; Jordão, LuisaAdenoids are a mass of lymphatic tissue located within the nasopharynge. This work aims assessing the relationship between the formation of bacterial biofilms on the adenoid surface and the incidence of infections in the pediatric age.
- Avaliação do risco para a saúde pública resultante do contacto com águas recreativas e ornamentaisPublication . Fernandes, Vera; Paulino, Sérgio; Costa, Clélia; Rodrigues, João Carlos; Reis, Lúcia; Nogueira, Isabel; Carvalho, Patricia; Duarte, Aida; Jordão, LuísaObjetivos: Este trabalho teve como objetivo caraterizar a população de microrganismos presente em águas recreativas (piscinas) e ornamentais (lagos), bem como avaliar o risco para a saúde pública do contacto com as mesmas.
- Bacterial biofilms, antibiotic resistance and healthcare-associated infections: a dangerous connectionPublication . Bandeira, Maria; Carvalho, Patrícia; Duarte, Aida; Jordão, LuísaIn 2012, were estimated 6.7 million cases of healthcare-associated infections (HAI) either in long-term care facilities or acute-care hospitals from which result 37,000 deaths configuring a serious public health problem [1]. The etiological agents are diverse and often resistant to antimicrobial agents. One of the mechanisms responsible for the emergence of drug resistance is biofilm assembly. Biofilms are defined as thin layers of microorganisms adhering to the surface of a structure, which may be organic or inorganic, together with the polymers that they secrete [2]. They are dynamic structures which experience different stages of organization with the ageing and are linked to an increase in bacterial resistance to host defense mechanisms, antibiotics, sterilization procedures other than autoclaving, persistence in water distribution systems and other surfaces. The understanding of bacteria organization within the biofilm and the identification of differences between planktonic and sessile forms of bacteria will be a step forward to fight HAIs. Bacterial isolates were grown in adequate medium. Antibiotic susceptibility was evaluated by broth microdilution method and interpreted according to NCCLS guidelines. A similar assay was performed to evaluate biofilm susceptibility to antibiotics. Bacteria ability to assemble biofilms was assayed by the microtiter-plate test [3] being tested in both abiotic (materials present in healthcare units) and biotic (Hella cells) surfaces. The biofilm structure was assessed by scanning electron microscopy (SEM) in either backscattered electron diffraction or secondary electrons mode. The kinetic of biofilm assembly depends on bacteria growth rate, incubation temperature and medium. Furthermore, the SEM analysis of planktonic and sessile forms of the same bacteria allowed the identification of structural differences which may be involved in virulence (Fig. 1). Bacteria ability to assemble biofilms seems to be independent of the abiotic structure (Fig.2). The same is not observed in biotic surfaces. This fact suggests that biofilm assembly in vivo is dependent of bacteria tropism. The minimum inhibitory concentration (MIC) determine for bacteria organized in biofilms is higher than for their planktonic forms. The increase ranges from 2 to 200 folds and is proportional to the ability of bacteria to assemble biofilms. Further studies will be conducted in order to prevent biofilm assembly within healthcare units which will result in a decrease of HAI and emergence of antibiotic resistant bacteria.
- Bacterial biofilms, antibiotic resistance and healthcare-associated infections: a dangerous connectionPublication . Bandeira, Maria; Carvalho, Patricia Almeida; Duarte, Aida; Jordao, LuisaIn 2012, were estimated 6.7 million cases of healthcare-associated infections (HAI) either in long-term care facilities or acute-care hospitals from which result 37,000 deaths configuring a serious public health problem. The etiological agents are diverse and often resistant to antimicrobial drugs. One of the mechanisms responsible for the emergence of drug resistance is biofilm assembly. Biofilms are defined as thin layers of microorganisms adhering to the surface of a structure, which may be organic or inorganic, together with the polymers that they secrete. They are dynamic structures which experience different stages of organization with the ageing and are linked to an increase in bacterial resistance to host defense mechanisms, antibiotics, sterilization procedures other than autoclaving, persistence in water distribution systems and other surfaces. The understanding of bacteria organization within the biofilm and the identification of differences between planktonic and sessile forms of bacteria will be a step forward to fight HAIs.
- Bacterial biofilms: a story of persistence and invasionPublication . Sousa, Sara; Bandeira, Maria; Carvalho, Patricia ALmeida; Duarte, Aida; Jordão, LuísaBiofilms are described as colonies of microorganisms that are attached to each other and to a surface, in an irreversible mode. These structures are virtualy everywhere: natural and humanized environments, as well as within living beings (humans and animals). For a long time biofilms where regarded as a bacterial survival strategie. Nowadays, in the industrialized world, the impact of acute bacterial infections caused by rapidly proliferating planktonic cells have gradually decreased in comparison to chronic infections owing to environmental organisms growing as biofilms. The major concern in this field is the healthcare-associated infections (HAIs). In 2012, HAIs estimated cases reached 6.7 million either in long-term care facilities or acute-care hospitals from which result 37,000 deaths configuring a serious public health problem [1]. The etiological agents are diverse being often resistant to antimicrobials and able to assemble biofilms both in abiotic and biotic surfaces. Here we evaluated the ability of different bacteria to assemble biofilms on a model surface and materials mimicking surfaces present either on healthcare units or medical devices. All bacteria were able to assemble biofilmls although following different kinetics and exhibiting different structural features assessed by electron microscopy. Additionally a link was established between bacteria ability to assemble biofilms and increased antibiotic resistance [2]. 1. ECDC Europe; (2012), Annual epidemiological report 2011 2. Bandeira M; (2014), Pathogens (doi:10.3390/pathogens3030720)
- Beach sand and the potential for infectious disease transmission: observations and recommendationsPublication . Solo-Gabriele, Helena M.; Harwood, Valerie J.; Kay, David; Fujioka, Roger S.; Sadowsky, Michael J.; Whitman, Richard L.; Wither, Andrew; Caniça, Manuela; Carvalho da Fonseca, Rita; Duarte, Aida; Edge, Thomas A.; Gargaté, Maria J.; Gunde-Cimerman, Nina; Hagen, Ferry; McLellan, Sandra L.; Nogueira da Silva, Alexandra; Novak Babič, Monika; Prada, Susana; Rodrigues, Raquel; Romão, Daniela; Sabino, Raquel; Samson, Robert A.; Segal, Esther; Staley, Christopher; Taylor, Huw D.; Veríssimo, Cristina; Viegas, Carla; Barroso, Helena; Brandão, JoãoRecent studies suggest that sand can serve as a vehicle for exposure of humans to pathogens at beach sites, resulting in increased health risks. Sampling for microorganisms in sand should therefore be considered for inclusion in regulatory programmes aimed at protecting recreational beach users from infectious disease. Here, we review the literature on pathogen levels in beach sand, and their potential for affecting human health. In an effort to provide specific recommendations for sand sampling programmes, we outline published guidelines for beach monitoring programs, which are currently focused exclusively on measuring microbial levels in water. We also provide background on spatial distribution and temporal characteristics of microbes in sand, as these factors influence sampling programs. First steps toward establishing a sand sampling program include identifying appropriate beach sites and use of initial sanitary assessments to refine site selection. A tiered approach is recommended for monitoring. This approach would include the analysis of samples from many sites for faecal indicator organisms and other conventional analytes, while testing for specific pathogens and unconventional indicators is reserved for high-risk sites. Given the diversity of microbes found in sand, studies are urgently needed to identify the most significant aetiological agent of disease and to relate microbial measurements in sand to human health risk.