Impact of Delftia tsuruhatensis and Achromobacter xylosoxidans on Escherichia coli dual-species biofilms treated with antibiotic agents

Recently it was demonstrated that for urinary tract infections species with a lower or unproven pathogenic potential, such as Delftia tsuruhatensis and Achromobacter xylosoxidans, might interact with conventional pathogenic agents such as Escherichia coli. Here, single- and dual-species biofilms of these microorganisms were characterized in terms of microbial composition over time, the average fitness of E. coli, the spatial organization and the biofilm antimicrobial profile. The results revealed a positive impact of these species on the fitness of E. coli and a greater tolerance to the antibiotic agents. In dual-species biofilms exposed to antibiotics, E. coli was able to dominate the microbial consortia in spite of being the most sensitive strain. This is the first study demonstrating the protective effect of less common species over E. coli under adverse conditions imposed by the use of antibiotic agents.

Characterization of an in vitro fed-batch model to obtain cells released from S. epidermidis biofilms

Both dynamic and fed-batch systems have been used for the study of biofilms. Dynamic systems, whose hallmark is the presence of continuous flow, have been considered the most appropriate for the study of the last stage of the biofilm lifecycle: biofilm disassembly. However, fed-batch is still the most used system in the biofilm research field. Hence, we have used a fed-batch system to collect cells released from Staphylococcus epidermidis biofilms, one of the most important etiological agents of medical device-associated biofilm infections. Herein, we showed that using this model it was possible to collect cells released from biofilms formed by 12 different S. epidermidis clinical and commensal isolates. In addition, our data indicated that biofilm disassembly occurred by both passive and active mechanisms, although the last occurred to a lesser extent. Moreover, it was observed that S. epidermidis biofilm-released cells presented higher tolerance to vancomycin and tetracycline, as well as a particular gene expression phenotype when compared with either biofilm or planktonic cells. Using this model, biofilm-released cells phenotype and their interaction with the host immune system could be studied in more detail, which could help providing significant insights into the pathophysiology of biofilm-related infections.

Antibiofilm strategies in the food industry

Biofilms in food processing plants represent not only a problem to human health but also cause economic losses by technical failure in several systems. In fact, many foodborne outbreaks have been found to be associated with biofilms. Biofilms may be prevented by regular cleaning and disinfection, but this does not completely prevent biofilm formation. Besides, due to their diversity and to the development of specialized phenotypes, it is well known that biofilms are more resistant to cleaning and disinfection than planktonic microorganisms. In recent years, a considerable effort has been made in the prevention of microbial adhesion and biofilm formation on food processing surfaces and novel technologies have been introduced. In this context, this chapter discusses the main conventional and emergent strategies that have been employed to prevent bacterial adhesion to food processing surfaces and thus to efficiently maintain good hygiene throughout the food industries.

Role of biofilms in the oral health of animals

In humans the importance of biofilms in disease processes is now widely recognised together with the difficulties in treating such infections once established. One of the earliest and certainly most studied biofilm in humans is that of dental plaque which is responsible for two of the most prevalent human infections, namely dental caries and periodontal disease. However, comparable studies of dental plaque in animals are relatively limited, despite the fact that similar infections also occur, and in the case of farm animals there is an associated economic impact. In addition, biofilms in the mouths of animals can also be detrimental to human health when transferred by animal bites. As a result, an understanding of both the microbial composition of animal plaque biofilms together with their role in animal diseases is important. Through the use of modern molecular studies, an insight into the oral microflora of animals is now being obtained and, to date, reveals that despite differences in terms of microbial species and relative proportions occurring between humans and animals, similarities do indeed exist. This information can be exploited in our efforts to both manage and treat infections in animals arising from the presence of an oral biofilm. This Chapter describes our current understanding of the microbial composition of animal plaque, its role in disease and how oral hygiene measures can be implemented to reduce subsequent infection.

Antibacterial Ag/a-C nanocomposite coatings: The influence of nano-galvanic a-C and Ag couples on Ag ionization rates

Biofilm formation has been pointed as a major concern in different industrial applications, namely on biomedical implants and surgical instruments, which has prompted the development of new strategies for production of efficient antimicrobial surfaces. In this work, nano âgalvanic couples were created to enhance the antibacterial properties of silver, by embedding it into amorphous carbon (a-C) matrix. The developed Ag/a-C nanocomposite coatings, deposited by magnetron sputtering, revealed an outstanding antibacterial activity against S.epidermidis, promoting a total reduction in biofilm formation with no bacteria counts in all dilution. The open circuit potential (OCP) tests in 0.9% NaCl confirmed that a-C shows a positive \OCP\ value, in contrast to Ag coating, thus enhancing the ionization of biocidal Ag+ due to the nano-galvanic couple activation. This result was confirmed by the inductively coupled plasma-optical emission spectroscopy (ICP-OES), which revealed a higher Ag ionization rate in the nanocomposite coating in comparison with the Ag coating. The surface of Ag/a-C and Ag coatings immersed in 0.9% NaCl were monitored by scanning electron microscopy (SEM) over a period of 24 hours, being found that the Ag ionization determined by ICP-OES was accompanied by an Ag nanoparticles coalescence and agglomeration in Ag/a-C coating.

"Saber escrever, saber falar": a escrita criativa nas aulas de espanhol

Relatório de estágio de mestrado em Ensino de Português no 3º Ciclo do Ensino Básico e no Ensino Secundário e de Espanhol nos Ensinos Básico e Secundário

Candida tropicalis biofilms: biomass, metabolic activity and secreted aspartyl proteinase production

According to epidemiological data, Candida tropicalis has been related to urinary tract infections and haematological malignancy. Several virulence factors seem to be responsible for C. tropicalis infections, for example: their ability to adhere and to form biofilms onto different indwelling medical devices; their capacity to adhere, invade and damage host human tissues due to enzymes production such as proteinases. The main aim of this work was to study the behaviour of C. tropicalis biofilms of different ages (24120 h) formed in artificial urine (AU) and their ability to express aspartyl proteinase (SAPT) genes. The reference strain C. tropicalis ATCC 750 and two C. tropicalis isolates from urine were used. Biofilms were evaluated in terms of culturable cells by colony-forming units enumeration; total biofilm biomass was evaluated using the crystal violet staining method; metabolic activity was evaluated by XTT assay; and SAPT gene expression was determined by real-time PCR. All strains of C. tropicalis were able to form biofilms in AU, although with differences between strains. Candida tropicalis biofilms showed a decrease in terms of the number of culturable cells from 48 to 72 h. Generally, SAPT3 was highly expressed. C. tropicalis strains assayed were able to form biofilms in the presence of AU although in a strain- and time-dependent way, and SAPT genes are expressed during C. tropicalis biofilm formation.

As práticas de leitura e a sua relação com a compreensão leitora nas aulas de línguas

Relatório de estágio de mestrado em Ensino de Português no 3º Ciclo do Ensino Básico e no Ensino Secundário e de Espanhol nos Ensinos Básico e Secundário

Novel strategies to combat gram-negative bacterial pathogens using bacteriophage proteins

Dissertação de mestrado em Bioengenharia

Effect of pH on Candida tropicalis virulence factors

Dissertação de mestrado em Bioengenharia

Influence of oxygen conditions on bacterial interactions within biofilms related with cystic fibrosis

Dissertação de mestrado em Bioengineering

Antibacterial properties of multifunctional coatings Ag-ZrCN for biomedical devices: a novel approach

PhD in Sciences Specialty in Physics

The effects of sodium hypochlorite against selected drinking water-isolated bacteria in planktonic and sessile states

Chlorine is the most commonly used agent for general disinfection, particularly for microbial growth control in drinking water distribution systems. The goals of this study were to understand the effects of chlorine, as sodium hypochlorite (NaOCl), on bacterial membrane physicochemical properties (surface charge, surface tension and hydrophobicity) and on motility of two emerging pathogens isolated from drinking water, Acinetobacter calcoaceticus and Stenotrophomonas maltophilia. The effects of NaOCl on the control of single and dual-species monolayer adhered bacteria (2 h incubation) and biofilms (24 h incubation) was also assessed. NaOCl caused significant changes on the surface hydrophobicity and motility of A. calcoaceticus, but not of S. maltophilia. Planktonic and sessile S. maltophilia were significantly more resistant to NaOCl than A. calcoaceticus. Monolayer adhered co-cultures of A. calcoaceticus-S. maltophilia were more resilient than the single species. Oppositely, dual species biofilms were more susceptible to NaOCl than their single species counterparts. In general, biofilm removal and killing demonstrated to be distinct phenomena: total bacterial viability reduction was achieved even if NaOCl at the higher concentrations had a reduced removal efficacy, allowing biofilm reseed. In conclusion, understanding the antimicrobial susceptibility of microorganisms to NaOCl can contribute to the design of effective biofilm control strategies targeting key microorganisms, such as S. maltophilia, and guarantying safe and high-quality drinking water. Moreover, the results reinforce that biofilms should be regarded as chronic contaminants of drinking water distribution systems and accurate methods are needed to quantify their presence as well as strategies complementary/alternative to NaOCl are required to effectively control the microbiological quality of drinking water.

Insights into bacterial colony morphology evolution and diversification during infection development in cystic fibrosis

Tese de Doutoramento em Engenharia Biomédica.

Surface phenomena and hydrodynamic effects on the deposition of Pseudomonas fluorescens

Biofilm adhesion to metals (copper, aluminium and brass) was studied at two different velocities and pH values of 7 and 9. Both bacteria and metals showed negative surface charges at those values of pH, which tends to slow down adhesion. Film densities increased with the fluid velocity and were also affected by the pH and by the growth rate of the bacteria. Long duration tests based on heat transfer measurements were run at five different fluid velocities and at pH = 7, showing in general an asymptotic behaviour and a control of deposition by adhesion and growth phenomena.