In this thesis, I present an approach to a pipeline for time lapse photography using conventional digital images converted to HDR (High Dynamic Range) images (rather than conventional digital or film exposures). Using this method, it is possible to capture a greater level of detail and a different look than one would get from a conventional time lapse image sequence. With HDR images properly tone-mapped for display on standard devices, information in shadows and hot spots is not lost, and certain details are enhanced.

Many construction companies today utilize webcams on their jobsites to monitor and record construction operations. Jobsite monitoring is often limited to outdoor construction operations due to lack of mobility of wired webcams. A wireless webcam may help monitor indoor construction operations with enhanced mobility. The transfer time of sending a photograph from the wireless webcam, however, is slower than that of a wired webcam. It is expected that professionals may have to analyze indoor construction operations with longer interval time-lapse photographs if they want to use a wireless webcam. This research aimed to determine the maximum time interval for time-lapse photos that enables professionals to interpret construction operations and productivity. In order to accomplish the research goal, brickwork of five different construction sites was videotaped. Various interval time-lapse photographs were generated from each video. Worker?s activity in these photographs was examined and graded. The grades in one-second interval photographs were compared with the grades of the same in longer time interval photographs. Error rates in observing longer time-lapse photographs were then obtained and analyzed to find the maximum time interval of time-lapse photography for monitoring construction operations. Research has discovered that the observation error rate increased rapidly until the 60-second interval and its increasing ratio remained constant. This finding can be used to predict a reasonable amount of error rate when observing time-lapse photographs less than 60-second interval. The observation error rate with longer than 60-second interval did not show a constant trend. Thus, the 60-second interval could be considered as the maximum time interval for professionals to interpret construction operations and productivity.

Observation is a simple method of acquiring information and is a critical step in the scientific method for both developing and investigating testable hypotheses. Cameras have long played a role in observation, and as technology advances, new tools and methods for collecting, interrogating, and displaying large quantities of high-resolution images have evolved. We describe an automated digital time-lapse camera system and present an example field deployment to observe the temporal and spatial patterns of riparian use by humans and animals during a 3-mo period. We also describe software tools for image interrogation and visualization, as well as new information gathered through their use. The system was tested in 2014, in a 2.4-ha site within the ApacheSitgreaves National Forest in east central Arizona, United States where elk (Cervus elaphus nelsoni) and cattle grazed. Photographs were taken every 30 sec for 38 d, after which an electric fence was installed to restrict cattle access and the time step was increased to every 3 min. We observed that elk exhibited the unique behavior of standing in and traveling within the stream channel while grazing and tended to graze and lie in close proximity to the channel. Cattle drank from, but typically did not enter, the stream channel and tended to lie away from the channel. Recreational use by people had the distinct impact of dispersing elk from the riparian corridor. Zoomable time-lapse videos allowed us to observe that in contrast to the cattle, elk grazed while lying down. High-temporal resolution photography is a practical tool for observing phenomena that are important for local resource management. Published by Elsevier Inc. on behalf of The Society for Range Management.

Electrical methods of geophysical survey are known to produce results that are hard to predict at different times of the year, and under differing weather conditions. This is a problem which can lead to misinterpretation of archaeological features under investigation. The dynamic relationship between a ‘natural’ soil matrix and an archaeological feature is a complex one, which greatly affects the success of the feature’s detection when using active electrical methods of geophysical survey. This study has monitored the gradual variation of measured resistivity over a selection of study areas. By targeting difficult to find, and often ‘missing’ electrical anomalies of known archaeological features, this study has increased the understanding of both the detection and interpretation capabilities of such geophysical surveys. A 16 month time-lapse study over 4 archaeological features has taken place to investigate the aforementioned detection problem across different soils and environments. In addition to the commonly used Twin-Probe earth resistance survey, electrical resistivity imaging (ERI) and quadrature electro-magnetic induction (EMI) were also utilised to explore the problem. Statistical analyses have provided a novel interpretation, which has yielded new insights into how the detection of archaeological features is influenced by the relationship between the target feature and the surrounding ‘natural’ soils. The study has highlighted both the complexity and previous misconceptions around the predictability of the electrical methods. The analysis has confirmed that each site provides an individual and nuanced situation, the variation clearly relating to the composition of the soils (particularly pore size) and the local weather history. The wide range of reasons behind survey success at each specific study site has been revealed. The outcomes have shown that a simplistic model of seasonality is not universally applicable to the electrical detection of archaeological features. This has led to the development of a method for quantifying survey success, enabling a deeper understanding of the unique way in which each site is affected by the interaction of local environmental and geological conditions.

A divisão celular nas bactérias requer a formação do divisomo, um complexo protéico que tem como o primeira etapa a polimerização da proteína FtsZ, seguida pela associação de 15 outras proteínas conhecidas. Os mecanismos envolvidos na regulação espacial do divisomo são bem caracterizados, mas o controle temporal da divisão celular em relação a outros eventos do ciclo, como a replicação do cromossomo, segue controversa. Neste trabalho, aplicamos a metodologia de microscopia de série temporal para estudar duas questões fundamentais do processo de divisão: a montagem do complexo que executa a divisão e a possibilidade da oscilação periódica na expressão de um ou mais genes envolvidos em divisão possa participar do controle temporal da montagem do divisomo. Para investigar se há oscilação da expressão gênica, construímos inicialmente variantes instáveis GFP através da adição de sequências peptídicas C-terminais que encaminham para a degradação em B. subtilis e utilizamos estes repórteres para criar fusões transcricionais sob o controle de promotores de genes centrais do processo de divisão. Depois de otimizar as condições de microscopia de série temporal com fusões transcricionais usando a variante instável GFPAISV, observamos que a autofluorescência de B. subtilis interferia nas nossas quantificações. Como forma de contornar a autofluorescência, construímos então fusões transcricionais com duas variantes de YFP (proteína fluorescente amarela) e optamos por trabalhar com Ypet-AISV. A análise de filmes de células individuais, tanto com fusões a GFPAISV como a Ypet-AISV, indicou que apenas o promotor do operon ftsL-pbpB apresentava um padrão de oscilação significativamente diferente de um promotor artificial usado como controle negativo. Esta hipótese, no entanto, não foi confirmada por medidas estáticas de populações de células nas quais correlacionamos intensidade de fluorescência com posição no ciclo celular. Portanto, nossos dados não foram capazes de evidenciar flutuações na expressão dos genes ftsL-pbpB, minCD, ftsZ, ftsA e zapA ao longo do ciclo celular. Para estudar a cinética de montagem divisomo foram realizados experimentos de microscopia de série temporal de FtsZ-mCherry e Pbp2B-GFP, onde observamos que a associação de Pbp2B ao divisomo ocorre 3 minutos após a formação do anel de FtsZ em meio rico e 4 minutos em meio mínimo. Também realizamos experimentos de microscopia de série temporal com uma cepa contendo FtsZ-YFP e DivIVA-CFP, determinando que DivIVA é incorporado ao divisomo 16 minutos após a formação do anel de FtsZ em meio rico e 20 minutos em meio mínimo. Estes dados confirmam que a montagem do divisomo ocorre em três etapas, e não duas, como anteriormente proposto. / Cell division in bacteria requires the formation of the divisome, a protein complex that has as the first step polymerization of FtsZ, followed by the assembly of 15 other known proteins. The mechanisms that underlie spatial regulation of divisome assembly have been largely elucidated, but the temporal control that ties the timing of cell division to other cell cycle events, such as chromosomal replication, remains surrounded by controversy. In this work, we use time-lapse microscopy to address two issues in B. subtilis cell division: the timing of divisome assembly, and the possibility that a periodic oscillation in expression of one or more genes essential for divisome assembly may play a role in defining the timing of cell division. To study the possibility of oscilation in gene expression, we have first built unstable variants of GFP by adding to its C-terminus peptide sequences that target the protein for degradation and used those variants to build transcriptional fusions to access the promoter activity of core cell division genes. After optimizing time-lapse conditions with transcriptional fusions to cell divison genes with the unstable GFPAISV, we observed that B. subtilis autofluorescence was an issue to our quantifications. To improve our signal-to-noise ratio, we built transcriptional fusions with two variants of YFP (Yellow Fluorescent Protein), and decided to work with Ypet. In our single-cell analysis for GFPAISV and for Ypet-AISV, only the ftsL operon promoter presented an oscilating pattern different from our negative control. This was not confirmed, however, when we attempted to correlate fluorescence signal with cell cycle position in static single-cell measurements. Thus, we conclude that that there are no fluctuations in ftsL, pbpB, minCD, ftsZ, ftsA or zapA gene expression throughout the cell cycle. To study divisome assembly we performed time-lapse microscopy of FtsZ-mCherry and Pbp2B-GFP, and determined that the association of Pbp2B occurs 3 minutes after FtsZ polymerization in rich medium and 4 minutes in minimal medium. We also performed time-lapse microscopy with FtsZ-YFP and DivIVA-CFP, determining that DivIVA is incorporated to the divisome in 16 minutes after FtsZ polymerization in rich medium and 20 minutes in minimal medium. This data confirms the assembly of the divisome in three steps rather than two, as previously proposed.

The principle of time-lapse system is based on the continuous sensing of the development of the pre-implantation embryo and the assessment of its morphology. This is advantageous in assessing the quality and implantation potential of embryos, which are subsequently evaluated according to certain established criteria. This gives the opportunity to select the highest quality embryo to be implanted into the mother's womb during embryo transfer. This system also allows us to detect any abnormalities in embryo development. This is very important, because the occurrence of abnormalities in early embryonic cell division is quite common. Morphological evaluation of embryos indicates a high incidence of tripolar mitosis during this early embryo development. The result of this division is three blastomers instead of two, which is associated with an irregular chromosome separation, each of them may contain a different number of chromosomes. In the case where conventional embryo observation is used to observe embryo morphology at longer intervals, it may be that the changes associated with tripolar mitosis are not detected at all, and such embryos appear to be falsely prosperous and are often selected for transfer. This can have serious consequences in the case of implantation, because these embryos are not...

Abstract Kidney diseases affect an increasing number of people worldwide, and there is a growing demand to develop new treatments and increase the number of transplantable organs. New treatments can be designed when new knowledge is gained by studying the details of kidney development. The ex vivo culture techniques have been used for over a century to study the development of kidneys, but they are not optimal for long-term imaging and following the nephrogenesis process over time. Kidney organoids, which are cellular aggregates resembling the in vivo kidney, together with intact embryonic kidneys, present a platform for these studies. However, there are limitations when working with primary embryonic kidney cells. Primary embryonic metanephric mesenchymal cells are usually low in number and lose the ability to undergo nephrogenesis rapidly. New ways to culture, biobank, and transfect cells can offer ways for functional testing of the effects of different genes on the nephrogenesis. This study presents new tools for studying nephrogenesis. Time-lapse imaging of organ development may be enhanced by using a Fixed Z-direction (FiZD) culture system where the kidney explant is grown in a restricted 70μm space. The technique enables the segmentation of the individual cells in a two-dimensional image and a dynamic analysis of the time-lapse data. This study also presents a technique of dissociation and reaggregation of the uninduced kidney metanephric mesenchyme (MM). With this novel method of culturing the dissociated MM cells in a growth factor medium for 24 hours, the cells can keep their competence for nephrogenesis. This technique allows the genetic manipulation of the MM cells before the induction to form nephrons, allowing functional testing of genes in the metanephric mesenchyme. This study further presents different techniques for gene editing of MM cells and introduces biobanking of primary kidney cells. It is shown here that the MM and ureteric bud (UB) cells have the capability to remember their fates and build nephron-like structures or continue branching after the cryopreservation in the liquid nitrogen. The methods introduced here provide new ways to create kidney organoids, manipulate their genome, and biobank the primary embryonic kidney cells. The developed FiZD culture system enhances the imaging of kidney development compared to the previously used culture methods. Using this method, the morphogenesis of the developing kidney can be followed more precisely, even in a single cell level. This culture method may also be used to culturing other organs, such as ovary, and may help provide insights into the development of other tissues as well. / Tiivistelmä Munuaissairauksiin sairastuvien määrä on lisääntynyt maailmanlaajuisesti, ja se on aikaansaanut tarpeen uusien hoitokeinojen sekä siirtoelimien kehitykseen. Näiden kehittämiseksi tarvitsemme uutta tietoa munuaisen kehityksestä ja toiminnasta. Munuaisen kehitystä on tutkittu ex vivo -viljelyn avulla jo yli vuosisadan ajan, mutta nykyiset elinviljelytekniikat eivät ole kuitenkaan optimaalisia pitkäkestoiseen time-lapse-kuvaukseen. Tässä työssä käytetään munuaisen kehityksen tutkimiseen hiiren alkion munuaisia sekä munuaisorganoideja, jotka ovat munuaissoluista koostuvia ja aitoa munuaista mallintavia soluaggregaatteja. Primaaristen munuaissolujen käyttöön sisältyy rajoitteita, ja tämä luo tarpeen uusien organoiditekniikoiden kehitykseen ja optimointiin. Primaarisia munuaissoluja on yleensä käytettävissä pieniä määriä, ja ne eivät myöskään sovellu pitkäkestoiseen kasvatukseen, koska ne menettävät nopeasti kykynsä muodostaa nefroneita. Uusien tekniikoiden avulla voidaan parantaa näiden solujen kasvatusta, säilytystä ja transfektointia ja edistää eri geenien vaikutuksia tutkivat funktionaaliset testaukset. Tässä tutkimuksessa esitetään uusia työkaluja nefrogeneesin tutkimiseen. Elinten kehitystä seuraavan time-lapse-kuvauksen laatua voidaan parantaa käyttämällä tässä työssä esitettyä FiZD-kasvatusmenetelmää, jossa munuaiseksplantti kasvaa rajoitetussa 70μm:n tilassa. Kuvat ovat korkealaatuisia, ja se mahdollistaa 2D-kuvan yksittäisten solujen segmentoinnin ja solujen liikkeiden dynaamisen analyysin. Lisäksi tässä tutkimuksessa esitetään ei-indusoidun munuaismesenkyymin käsittelyyn kehitetty dissosiaatio- ja reaggregaatiomenetelmä. Munuaisen kehityksen alkuvaiheessa on mahdollistaerottaa nefroneja muodostava metanefrinen mesenkyymi (MM) sekä munuaisen kokoajaputkiston muodostava ureterin silmu. Metanefrinen mesenkyymi voidaan hajottaa yksisolususpensioksi, säilyttää 24 tuntia kasvutekijämediumissa ja tämän jälkeen reaggregoida ja indusoida muodostamaan nefroneita. Tämä tekniikka mahdollistaa MM-solujen geneettisen muokkauksen, ennen kuin munuaisen kehitys alkaa. Tämä tekniikka mahdollistaa myös dissosioitujen MM solujen geneettiset muokkaukset. Geenien yliekspression tai hiljentämisen avulla voidaan tehdä funktionaalisia kokeita näiden muutosten vaikutuksesta nefrogeneesiin. Lisäksi tässä työssä esitetään munuaisprogenitorisolujen säilömistä syväjäädytyksellä. Munuaisprogenitorisolut voidaan säilöä nestetyppeen, minkä jälkeen ne ovat edelleen kykeneviä muodostamaan nefronirakenteita tai haarautumaan. Tässä väitöskirjatyössä esitettyjen menetelmien avulla on tulevaisuudessa mahdollista saada lisätietoa munuaisten kehitysprosessista. Kehitetty FiZD-kasvatusmenetelmä parantaa munuaisen kehityksen kuvantamista ja mahdollistaa yksittäisten solujen seuraamisen. Tämä kasvatusmenetelmä sopii myös muiden elinten, kuten munarauhasten, ja kudosten kasvatukseen, ja sen avulla voidaan saada tietoa myös niiden kehityksestä.

A divisão celular nas bactérias requer a formação do divisomo, um complexo protéico que tem como o primeira etapa a polimerização da proteína FtsZ, seguida pela associação de 15 outras proteínas conhecidas. Os mecanismos envolvidos na regulação espacial do divisomo são bem caracterizados, mas o controle temporal da divisão celular em relação a outros eventos do ciclo, como a replicação do cromossomo, segue controversa. Neste trabalho, aplicamos a metodologia de microscopia de série temporal para estudar duas questões fundamentais do processo de divisão: a montagem do complexo que executa a divisão e a possibilidade da oscilação periódica na expressão de um ou mais genes envolvidos em divisão possa participar do controle temporal da montagem do divisomo. Para investigar se há oscilação da expressão gênica, construímos inicialmente variantes instáveis GFP através da adição de sequências peptídicas C-terminais que encaminham para a degradação em B. subtilis e utilizamos estes repórteres para criar fusões transcricionais sob o controle de promotores de genes centrais do processo de divisão. Depois de otimizar as condições de microscopia de série temporal com fusões transcricionais usando a variante instável GFPAISV, observamos que a autofluorescência de B. subtilis interferia nas nossas quantificações. Como forma de contornar a autofluorescência, construímos então fusões transcricionais com duas variantes de YFP (proteína fluorescente amarela) e optamos por trabalhar com Ypet-AISV. A análise de filmes de células individuais, tanto com fusões a GFPAISV como a Ypet-AISV, indicou que apenas o promotor do operon ftsL-pbpB apresentava um padrão de oscilação significativamente diferente de um promotor artificial usado como controle negativo. Esta hipótese, no entanto, não foi confirmada por medidas estáticas de populações de células nas quais correlacionamos intensidade de fluorescência com posição no ciclo celular. Portanto, nossos dados não foram capazes de evidenciar flutuações na expressão dos genes ftsL-pbpB, minCD, ftsZ, ftsA e zapA ao longo do ciclo celular. Para estudar a cinética de montagem divisomo foram realizados experimentos de microscopia de série temporal de FtsZ-mCherry e Pbp2B-GFP, onde observamos que a associação de Pbp2B ao divisomo ocorre 3 minutos após a formação do anel de FtsZ em meio rico e 4 minutos em meio mínimo. Também realizamos experimentos de microscopia de série temporal com uma cepa contendo FtsZ-YFP e DivIVA-CFP, determinando que DivIVA é incorporado ao divisomo 16 minutos após a formação do anel de FtsZ em meio rico e 20 minutos em meio mínimo. Estes dados confirmam que a montagem do divisomo ocorre em três etapas, e não duas, como anteriormente proposto. / Cell division in bacteria requires the formation of the divisome, a protein complex that has as the first step polymerization of FtsZ, followed by the assembly of 15 other known proteins. The mechanisms that underlie spatial regulation of divisome assembly have been largely elucidated, but the temporal control that ties the timing of cell division to other cell cycle events, such as chromosomal replication, remains surrounded by controversy. In this work, we use time-lapse microscopy to address two issues in B. subtilis cell division: the timing of divisome assembly, and the possibility that a periodic oscillation in expression of one or more genes essential for divisome assembly may play a role in defining the timing of cell division. To study the possibility of oscilation in gene expression, we have first built unstable variants of GFP by adding to its C-terminus peptide sequences that target the protein for degradation and used those variants to build transcriptional fusions to access the promoter activity of core cell division genes. After optimizing time-lapse conditions with transcriptional fusions to cell divison genes with the unstable GFPAISV, we observed that B. subtilis autofluorescence was an issue to our quantifications. To improve our signal-to-noise ratio, we built transcriptional fusions with two variants of YFP (Yellow Fluorescent Protein), and decided to work with Ypet. In our single-cell analysis for GFPAISV and for Ypet-AISV, only the ftsL operon promoter presented an oscilating pattern different from our negative control. This was not confirmed, however, when we attempted to correlate fluorescence signal with cell cycle position in static single-cell measurements. Thus, we conclude that that there are no fluctuations in ftsL, pbpB, minCD, ftsZ, ftsA or zapA gene expression throughout the cell cycle. To study divisome assembly we performed time-lapse microscopy of FtsZ-mCherry and Pbp2B-GFP, and determined that the association of Pbp2B occurs 3 minutes after FtsZ polymerization in rich medium and 4 minutes in minimal medium. We also performed time-lapse microscopy with FtsZ-YFP and DivIVA-CFP, determining that DivIVA is incorporated to the divisome in 16 minutes after FtsZ polymerization in rich medium and 20 minutes in minimal medium. This data confirms the assembly of the divisome in three steps rather than two, as previously proposed.

La capacité remarquable des neurones et des cellules gliales à migrer collectivement sur de longues distances assure l’architecture finale du cerveau. Ce processus est extrêmement dynamique et dépend non seulement de l’interaction entre les cellules mais aussi de la présence de facteurs de transcriptions spécifiques au sein de la cellule migrante. Les protéines d’adhésion comme les cadhérines et les chimioattractants/chimiorépulsifs sont connus pour réguler et guider la migration. Si le mode d’action de ces molécules a été extensivement étudié, les cascades de signalisation qui déclenchent le chimiotropisme sont loin d’être élucidées. Au cours de mon doctorat, j’ai analysé la régulation et le rôle d’un récepteur des chimioattractant au cours de la migration de la glie. Pour ceci j’ai utilisé le modèle du développement de la chaine gliale dans l’aile de la drosophile qui représente un outil de choix pour étudier les mécanismes moléculaires régulant la migration collective. / The remarkable ability of neurons and glia to undergo long distance and collective migration ensures the final architecture and function of the brain. This is an extremely dynamic process that not only depends on cell interactions, but also on the presence of specific transcription factors in the migrating cells. Adhesion molecules such as classic cadherins and chemoattractants/repellants are known to regulate directional migration, however, how are these pathways regulated is largely unknown. While the role of these molecules controlling cell interactions has been extensively investigated, the signaling cascades that trigger chemotropism are not understood. During the course of my PhD I have analyzed the role of an adhesion molecule and the impact of a chemoattractant receptor regulated by an early transcription factor in the process. The glial chain in a developing Drosophila wing provides an excellent tool to study the molecular pathway underlying collective migration.

Les bactéries forment des communautés spatiales adhérentes à des surfaces, appelées biofilms. Ces organisations bactériennes sont omniprésentes dans les milieux naturel, industriel et médical et peuvent porter atteinte à notre santé lorsqu’elles hébergent des agents pathogènes, parmi lesquels le médiatique Staphylococcus aureus sur lequel a porté l’ensemble de ce travail de thèse. Cette bactérie est l’une des principales causes d’infections chroniques, mais également d’infections nosocomiales, impliquant le plus souvent des biofilms. Il est aujourd’hui reconnu qu’une telle biostructure est un véritable bouclier à l’action des antimicrobiens et à celle du système immunitaire. Outre les résistances génétiques des bactéries pathogènes aux antibiotiques, l’hétérogénéité chimique et biologique de la structure tridimensionnelle des biofilms pourrait être à l’origine de ces phénomènes de tolérance et de chronicité d’infections. C’est à cette problématique que se rattache ce travail de thèse concernant l’action de la vancomycine sur des biofilms de S. aureus. Alors que les connaissances sur la réactivité de cet antibiotique clef avec S. aureus proviennent essentiellement d’études réalisées sur des cellules planctoniques, l’originalité de notre approche a été d’étudier la diffusion-réaction de la vancomycine in situ dans l’épaisseur des biofilms en utilisant en particulier des outils avancés de microscopie de fluorescence (Time-Lapse, FLIM, FRAP, et FCS). Nous avons ainsi évalué sa biodisponibilité dans la matrice d’exopolymères, ainsi que l’impact de la physiologie spécifique des bactéries incluses en biofilms sur l’activité de cet antibiotique, utilisé seul ou en association avec la rifampicine. Cette approche multidisciplinaire a permis une meilleure compréhension des mécanismes impliqués dans la singulière tolérance de ces biostructures à l’action des antibiotiques, et de souligner l’urgence de développer des approches préventives telles que le diagnostic précoce des infections impliquant des biofilms. / Bacteria form architecturally complex communities adherent to surfaces, known as biofilms. These structured living cells are ubiquitous and found in natural, industrial and medical environments. They can affect our health when they host pathogens as the well known Staphylococcus aureus species which constitute the main purpose of this thesis. This bacteria is one of the major causes of chronic and nosocomial infections, most often involving biofilms. It is now recognized that such biostructure is a true shield against the action of antimicrobial agents and the host immune system. In addition to the genetic resistance of pathogenic bacteria to antibiotics, the chemical and biological heterogeneity of biofilms could be the cause of these phenomena of tolerance and apparition of chronic infections. This work aimed at studying of the action of vancomycin on S. aureus biofilms. While the knowledge on the reactivity of this key antibiotic with S. aureus bacteria comes mainly from studies of planktonic cells, the originality of our approach was to study the diffusion-reaction processes of vancomycin in situ in the thickness of biofilms using particularly advanced fluorescence imaging tools (Time-Lapse, FLIM, FRAP and FCS). We thus assess its bioavailability in the exopolymeric matrix, and the impact of the cell physiology of bacteria included in biofilms on the activity of this antibiotic when used alone or in combination with rifampicin. This multidisciplinary approach has allowed a better understanding of the mechanisms involved in the particular tolerance of these biostructures to the action of antibiotics, and underlines the emergency to develop preventive approaches such as early diagnosis of infections involving biofilms.