It is a circular-mapping DNA molecule of 28 601 bp with a low GC content of 25%. It contains PARP inhibitor the usual set of mitochondrial protein and RNA genes characteristic of the majority of sequenced filamentous fungi mitochondrial genomes (Table S1). RNA-encoding genes include 27 tRNA genes and genes for large and small ribosomal RNA (rnS, rnL), as well as a predicted rnpB gene encoding the subunit of mitochondrial RNase P (mtP-RNA), known to be responsible for tRNA processing (Seif et al., 2003). Protein-encoding genes include those for ATP-synthase subunits 6, 8 and 9 (atp6, atp8 and atp9), subunits of cytochrome oxidase (cox1, cox2 and cox3), apocytochrome b (cob), one ribosomal protein

(rps5) and NADH dehydrogenase subunits (nad1, nad2, nad3, nad4, nad4L, nad5 and nad6). Group I or group II introns, frequently interrupting yeast and filamentous fungi mitochondrial genes (Lang et al., 2007), are not found. Two open reading frames (ORFs) located between cox2 and tRNA-R, and between tRNA-H and atp9 could encode for hypothetical proteins without apparent homology to any known proteins in the

GenBank database. All genes are located on one strand and apparently selleck kinase inhibitor transcribed in one direction (Fig. 1). To extend our analysis of mitochondrial genome organization to other members of the Penicillium/Aspergillus clade, we included mitochondrial genomes that have already been sequenced in whole genome sequencing programs, such as the mitochondrial genomes of P. chrysogenum, A. terreus and A. oryzae. These genomes are available from GenBank as partially annotated or unannotated

contigs. The general features of all compared genomes are summarized in Table 1. It is evident that all compared Penicillium and Aspergillus species possess conserved features of mitochondrial genome organization, including gene content. Genome size variation is low and is explained by the length of intergenic regions and the presence of one intron in the A. oryzae and P. digitatum mitochondrial genomes. The majority of P. solitum mitochondrial tRNA genes are organized into two dense gene clusters, a feature common to many sequenced mitochondrial genomes of filamentous fungi. This Nintedanib (BIBF 1120) set of 27 tRNA genes is sufficient to decode all codons present in the predicted ORFs, alleviating the need for tRNA import into the mitochondria from the cytoplasm (Kolesnikova et al., 2000), as is the case for some yeast, plant and protist mitochondrial genomes. The presence of tRNA-W (anticodon UCA) recognizing the TGA codon, as well as the TGG codon, and the absence of abnormal tRNA-T (anticodon CUN) indicate that P. solitum mitochondrial protein-encoding genes are translated according to genetic code 4 (Fox, 1987), as shown for other Pezizomycotina mitochondrial genomes. All protein-encoding sequences start with the ATG codon, except cox1, which starts with the codon TTG.

Murine innate defence against A. fumigatus is sufficient to prevent infection, even in heavily infected animals, and immunosuppression is required to establish

infection (Lewis & Wiederhold, 2005). In the McDonagh study, both macrophage and neutrophil cell populations were chemotherapeutically targeted, using hydrocortisone acetate and cyclophosphamide, respectively, the former drug administered in a single dose 1 day before infection and the latter periodically administered throughout the duration of the experiment (Lewis & Wiederhold, 2005). Phagocytosis by macrophages harvested from hydrocortisone-treated A. fumigatus-infected mice is known to occur, but fungal killing is compromised (Philippe et al., 2003). It is likely, therefore, that host Verteporfin purchase cells, predominantly macrophages, are encountered in the alveolar and bronchial spaces (Fig. 2b and c), and encountered macrophages are compromised in their ability to kill A. fumigatus spores. Conversely, the encapsulated facultative intracellular pathogen C. neoformans can establish infection in immunocompetent mice. Moreover, the interaction between macrophages and C. neoformans is critical for containing the dissemination of this pathogenic yeast, whose success is subverted by C. neoformans-derived

factors. Cryptococcus neoformans is capable of replication within the macrophage phagolysosome, a process that ultimately leads to host cell lysis or phagosome extrusion (Tucker & Casadevall, 2002; Alvarez & Casadevall, find more 2006; Ma et al., 2006). As in vitro studies indicate that the time taken to extrude a C. neoformans-containing phagolysosome can be as little as 2 h (Tucker & Casadevall,

2002; Alvarez & Casadevall, 2006; Ma et al., 2006), it is likely that multiple macrophage encounters occurred during the experimental time frame, and, contrary Rho to the A. fumigatus infection model, noninfected macrophages were completely proficient with respect to killing ability. Carbon metabolism was, to varying degrees, commonly implicated among all of the mammalian pathogen datasets with acetyl-CoA synthetase and isocitrate dehydrogenase featuring in all four upregulated genesets. Combined with extant data on fungal carbon-metabolizing gene products and virulence, considerable insight can be gained from our comparative analysis. Firstly, the differential roles of glyoxylate cycle enzymes in virulence, which has been studied in multiple mammalian fungal pathogens, could not have been predicted from our comparative transcriptomic analysis. Glyoxylate cycle gene products are required for full virulence in C. albicans (Lorenz & Fink, 2001; Wang et al., 2003; Barelle et al., 2006) and M. grisea (Wang et al., 2003), but not in A. fumigatus (Schobel et al., 2007; Olivas et al., 2008) or C. neoformans (Rude et al., 2002). Indeed, based on our analysis, one might have predicted the necessity of glyoxylate pathway functionality in C. neoformans and A. fumigatus and nonrequirement in M. grisea (Table 2).

2f). Mosquera and colleagues targeted invasive hyphae (Fig. 2f) as their sampled population in order to avoid filamentous necrotrophic hyphae characteristic of late-stage infection. Invading hyphae were harvested from leaf sheaths at 36 h postinfection, obtaining a relatively synchronous cell population in which most hyphae were inside first-invaded cells. Leaf sheaths were

manually dissected in order to remove uninfected plant material and infected material was snap frozen before RNA extraction. RNA amplification was integral to the labelling protocol, with 500 ng of total RNA generating 10–15 μg of labelled cRNA. All of the studies captured significant numbers of differentially expressed genes, where GSK1120212 up/downregulated gene sets consisted of 1281/897 [9075] (McDonagh et al., 2008), 58/50 [85% of arrayed spots] (Walker et al., 2009), 255/221 [787] (Thewes et al., 2007); 1120/781 [15152] (Thewes et al., 2007) and 713/423

[6750] (Kamper et al., 2006), where square parentheses indicate the numbers of assayable spots per experiment. The C. neoformans SAGE analysis returned data on 84 gene tags (normalized to every 20 000 of the tag population sequenced), showing a higher representation relative to previously documented in vitro SAGE libraries, including a low-iron Protein Tyrosine Kinase inhibitor medium (LIM) SAGE library (Hu et al., 2007) against which most comparisons were made. We used several strategies to derive multispecies information on the co-ordinate regulation of homologous genes (Table 2) including best hit blast (Altschul et al., 1990) analysis, applied in a unidirectional sense, using peptides derived from the translation of species-specific differentially regulated transcript sequences. We also matched text descriptors from gene annotations in instances where spot annotations could not be readily matched to publicly accessible annotation databases or where significant redundancy of function among

multiple gene identifiers might be expected (e.g. oxidoreductases and alcohol dehydrogenases). Despite the variance among the size of datasets and disparate infection models, some interesting observations can be drawn from the comparison. We found impressive concordance between upregulated A. fumigatus and C. neoformans genes (Table 2). Such a similarity of the transcription profile is even more remarkable, given Baricitinib the varying immunosuppressive regimens used and different morphogenetic programmes of the two species (yeast vs. filamentous fungus). This intriguing finding may therefore reflect the similarity of nutrient sources and physiological conditions (such as temperature, iron limitation and oxygen tension) in the mammalian pulmonary niche and the dominance of such factors over immune status and species-specific traits. Despite the similarities in infection modelling procedures, the progression of infection would have differed significantly between the McDonagh and Hu studies in respect of the differential pathogenic strategy adopted by A. fumigatus and C.

Phages infecting S. thermophilus showed closed, but distinguishable patterns and slightly related to Φ936, ΦP335 and ΦSPP1. Escherichia coli phages also clustered together,

except ΦSOM1. Finally, S. epidermidis phages were also grouped, vB_SepiS-phiIPLA7 being the exception. This clustering was not surprising because of the phylogenetic relations among phages. As it has been described previously, phages infecting distantly related bacterial hosts typically share little or no nucleotide Y-27632 molecular weight sequence similarity, while phages infecting a specific bacterial host are more similar (Hatfull, 2008). Moreover, module exchanging could be the reason why phages vB_SepiS-phiIPLA7, ΦC2 and ΦSOM1 were grouped into a different cluster than the other phages infecting the same bacterial host. Phage morphology did not correlate with the RAPD-PCR clustering as phages belonging

to different morphological families selleck inhibitor were grouped together. This is the case of ΦX174 (Microviridae), ΦP1 (Podoviridae), ΦSOM8 and ΦSOM2 (Myoviridae), which were clustered with the rest of the phages belonging to the Siphoviridae family. The classification in families is mostly based on virion morphology and nucleic acid type, and bacteriophages belonging to different families may have similar DNA sequences (Ackermann, 2003). Thereby, similar RAPD-PCR profiles can be found among families. A similar discrepancy has already been reported when using fRFLP for bacteriophage typing (Merabishvili et al., 2007). It remains

Selleckchem Depsipeptide to be confirmed whether RAPD typing using phage lysates is also a feasible technique when using phages infecting high G+C bacterial hosts as those were not included in this study. However, based on the use of DMSO in the reaction buffer and the availability of enhanced DNA polymerases and buffers active on high G+C DNA templates, it is reasonable to speculate that this approach may also be useful. RAPD-PCR on phage suspensions is a suitable approach to quickly assess the genetic diversity among newly isolated bacteriophages infecting the same species while circumventing the need for DNA extraction and purification. Using this assay, genomic fingerprints from different phages infecting Staphylococcus, Bacillus, E. coli, Lactococcus and Streptococcus were distinct and showed variations in the number of bands, fragment size and intensity. This work was supported by grants AGL2009-13144-C02-01 from the Ministry of Education of Spain, IB08-052 from FICYT (Regional Government of Asturias) and PIE200970I090 (CSIC, Spain). Thanks are due to M. Muniesa, M.A. Álvarez, J.E. Suárez and S. Ayora for kindly providing E. coli, S. thermophilus, L. lactis, L. casei and B. subtilis bacteriophages used in this study. P.G. and B.M. contributed equally to this work.

Phages infecting S. thermophilus showed closed, but distinguishable patterns and slightly related to Φ936, ΦP335 and ΦSPP1. Escherichia coli phages also clustered together,

except ΦSOM1. Finally, S. epidermidis phages were also grouped, vB_SepiS-phiIPLA7 being the exception. This clustering was not surprising because of the phylogenetic relations among phages. As it has been described previously, phages infecting distantly related bacterial hosts typically share little or no nucleotide Selleckchem Y-27632 sequence similarity, while phages infecting a specific bacterial host are more similar (Hatfull, 2008). Moreover, module exchanging could be the reason why phages vB_SepiS-phiIPLA7, ΦC2 and ΦSOM1 were grouped into a different cluster than the other phages infecting the same bacterial host. Phage morphology did not correlate with the RAPD-PCR clustering as phages belonging

to different morphological families NU7441 price were grouped together. This is the case of ΦX174 (Microviridae), ΦP1 (Podoviridae), ΦSOM8 and ΦSOM2 (Myoviridae), which were clustered with the rest of the phages belonging to the Siphoviridae family. The classification in families is mostly based on virion morphology and nucleic acid type, and bacteriophages belonging to different families may have similar DNA sequences (Ackermann, 2003). Thereby, similar RAPD-PCR profiles can be found among families. A similar discrepancy has already been reported when using fRFLP for bacteriophage typing (Merabishvili et al., 2007). It remains

http://www.selleck.co.jp/products/Staurosporine.html to be confirmed whether RAPD typing using phage lysates is also a feasible technique when using phages infecting high G+C bacterial hosts as those were not included in this study. However, based on the use of DMSO in the reaction buffer and the availability of enhanced DNA polymerases and buffers active on high G+C DNA templates, it is reasonable to speculate that this approach may also be useful. RAPD-PCR on phage suspensions is a suitable approach to quickly assess the genetic diversity among newly isolated bacteriophages infecting the same species while circumventing the need for DNA extraction and purification. Using this assay, genomic fingerprints from different phages infecting Staphylococcus, Bacillus, E. coli, Lactococcus and Streptococcus were distinct and showed variations in the number of bands, fragment size and intensity. This work was supported by grants AGL2009-13144-C02-01 from the Ministry of Education of Spain, IB08-052 from FICYT (Regional Government of Asturias) and PIE200970I090 (CSIC, Spain). Thanks are due to M. Muniesa, M.A. Álvarez, J.E. Suárez and S. Ayora for kindly providing E. coli, S. thermophilus, L. lactis, L. casei and B. subtilis bacteriophages used in this study. P.G. and B.M. contributed equally to this work.

In stage 2, the questionnaire was piloted to determine its validity and reliability. Finally, the questionnaire was sent to a random sample of community pharmacists to test the generalizability of the findings of the focus group interviews. The design (sequential) and the rationale for choosing mixed-methods approach were clearly described. The use of the mixed-methods approach provided a rich and generalizable

description of pharmacist prescribing in Canada by overcoming the limitations of qualitative (generalizability) and quantitative (in-depth understanding) methodology. Complementarity seeks elaboration, enhancement, illustration and clarification of the Cobimetinib mouse results from one method with the results from the other method.’[1] Bruhn et al. reported a pilot randomized controlled trial which was complemented with qualitative interviews to evaluate the effectiveness of pharmacist-led management of chronic pain in primary care (the PIPPC study).[6, 7] The patients were randomized to

one of three arms: (1) pharmacist Pifithrin-�� molecular weight medication review with pharmacist prescribing, (2) pharmacist medication review with feedback to GP and (3) treatment as usual. The qualitative component consisted of face-to-face interviews with the pharmacists, GPs and patients to explore their experiences. It is noteworthy that the qualitative interviews did not contribute towards answering the effectiveness question (the primary aim of the study); rather, they helped to understand and explain how the intervention might have worked. The two datasets were described separately in two different conference proceedings and were therefore not integrated. Integration of the

two datasets may have allowed researchers to draw more meaningful inferences from the findings and authors may do so in a full report. However, if the purpose of a mixed-methods study is to answer different research questions within the same study (embedded design), as in this example, the authors may choose to present findings separately.[8] Again, neither the rationale nor the design was reported. Initiation seeks the discovery of the paradox and contradiction, new perspectives of frameworks, the recasting of questions or selleck inhibitor results from one method with questions or results from the other method.’ It generates ideas by initiating new interpretations, highlighting areas for additional investigation and reshaping the entire research question. Initiation is predominantly used in the disciplines of social sciences and psychology. We were unable to find an example in the area of pharmacy practice to illustrate initiation. It should be noted that in these examples we have tied each example to only one reason or rationale for choosing a mixed-methods design, which in practice is not always true, as researchers might use a mixed-methods approach for more than one reason.

Using quantitative real-time PCR, the suitability of the HSP30 promoter to specifically drive stationary-phase expression of the native FLO5 and FLO11 ORFs in BM45 and VIN13 transgenic strains under synthetic MS300 wine fermentation conditions has been demonstrated in our recent research study (Govender et al.,

2010). In this study, transgenic yeast strains (BM45-F5H, BM45-F11H, VIN13-F5H and VIN13-F11H) in which an ORF of a dominant chromosomal flocculation gene (FLO5 or FLO11) was placed under the transcriptional control of the stationary-phase inducible HSP30 promoter displayed metabolic fermentation profiles in natural Merlot must that were almost indistinguishable from their parental host wine yeast strains. Considering that wines are regarded VE-822 nmr as dry if their residual sugar content is <5 g L−1, it is clearly evident that Merlot wines (≤1.95 g L−1 residual

sugars) produced by both parental host wine yeast strains and their HSP30p transgenic descendants were fermented almost equally well to dryness. Moreover, HSP30p transgenic wine yeast strains produced Merlot wines that displayed almost identical volatile and aroma compound profiles. Thus, it can be suggested that introduction of promoter replacement cassettes designed for induction of late fermentation flocculation does not compromise the desirable oenological properties of original nonflocculent host wine yeast strains under authentic red wine-making conditions. Alectinib The BM45-F5H and VIN13-F5H transformants displayed almost identical Flo1-type flocculation MK0683 mw intensity in both synthetic MS300 and Merlot wine fermentations (Govender et al., 2010). Only

the BM45-F5H strain was capable of generating compacted or ‘caked’ lees fractions, thereby providing a distinct separation of the fermented wine product and lees fractions. The benefit of this attractive property is that it facilitates simpler and faster recovery of wines and it also promotes a greater volume recovery of fermented wine product. This improvement has significant financial cost-saving implications and can be directly attributed to the superior flocculent ability of the BM45-F5H transgenic strain. The BM45-F11H and VIN13-F11H transgenic wine yeast strains yielded strong flocculent phenotypes that displayed a combination of both Ca2+-dependent and Ca2+-independent flocculation characteristics under authentic red wine-making conditions. In addition, no flocculent phenotype was displayed by the same transgenic yeast strains in aerobic shake-flask MS300 batch fermentations supplemented with an individual red wine fermentation component (pectin, potassium bitartrate, diatomaceous earth, gallic acid, caffeic acid, catechin or a tannin). As such, these individual components seem not to aid in the development of the novel FLO11-mediated flocculation phenotype observed under authentic red wine fermentations conditions.

As miRNAs generally bind to numerous target mRNAs, and many mRNAs are regulated by multiple miRNA species, the possibilities for fine orchestration of translation are enormous. Primary miRNA transcripts are processed in the nucleus by the RNAase III endonuclease Drosha to generate short-hairpin precursors of ∼70–100 nucleotides, which are exported from the nucleus and further processed by another RNAase family enzyme, Dicer, to produce a mature miRNA of ∼22 nucleotides in length. The activity of miRNAs may therefore be modulated at multiple steps in the biogenesis pathway as

well as through regulation of the miRNA-bound RISC (Ashraf et al., 2006; Kosik, 2006; Presutti et al., 2006; Kye et al., 2007; Winter et al., 2009). miRNAs play coordinating roles in a variety of cellular selleck kinase inhibitor processes, selleck including cell specification and apoptosis (Bartel, 2004; Chang et al., 2007). In neurons, recent studies have established roles for specific miRNAs in neurogenesis and dendritic spine morphogenesis (Vo et al., 2005; Krichevsky et al., 2006; Schratt et al., 2006; Cao et al., 2007; Fiore et al., 2009; Siegel et al., 2009). In the marine snail Aplysia, expression of miR-124 is linked to synapse-specific long-term sensitization (Rajasethupathy et al., 2009). In flies, degradation of the protein Armitage, a component of the miRNA-RISC,

promotes synaptic protein synthesis during long-term memory. Despite the advances in understanding neuronal miRNAs, little is known about miRNA regulation during activity-dependent synaptic plasticity in the adult mammalian brain. We therefore examined miRNA expression following induction of long-term potentiation (LTP) by high-frequency stimulation (HFS) of the perforant path input to the dentate gyrus of anesthetized rats. Using

miRNA expression profiling and quantitative oxyclozanide reverse transcription polymerase chain reaction (RT-PCR), we identified mature miRNAs with significantly increased (miR-132, miR-212) or decreased (miR-219) expression during LTP. Analysis of the primary and precursor transcripts demonstrated massive metabotropic glutamate receptor (mGluR)-dependent transcription of miR-132 and -212 in dentate granule cells that is functionally correlated with depotentiation rather than LTP. In contrast, activation of N-methyl-d-aspartate receptors (NMDAR) during LTP induction selectively downregulated mature miR-132, -212 and -219 levels, indicating stimulation of mature miRNA turnover. Animal experiments were carried out in accordance with the European Community Council Directive of 24 November 1986 (86/609/EEC) and approved by the Norwegian Committee for Animal Research. Experiments were performed on 45 adult male Sprague–Dawley rats. The electrophysiological procedures have been detailed elsewhere (Messaoudi et al., 2002; Panja et al., 2009).

gov/Blast.cgi), and the search for specific domains was performed using interproscan (http://www.ebi.ac.uk/Tools/InterProScan/). Alignments were generated using clustalx

1.81 or clustalw (http://www.ebi.ac.uk/Tools/clustalw2/index.html). The sequence of the SpHtp1 has been deposited in GenBank under accession number GU345745. RTG-2 cells were grown as a confluent monolayer in 75 cm2 in cell culture flasks (Nunc) and challenged with 5 × 104 zoospore/cysts at 24 °C. At several time points, media were discarded, except for time point 0, and 5 mL of Qiazol (Qiagen) or Trizol reagent (Invitrogen) was added to each flask. Cells were scraped loose with a cell scraper (Fisher) and the suspension was aliquoted as 1 mL portions into 2-mL screw-cap tubes containing 10–35 glass beads of 1 mm diameter (Biospec). Samples check details GKT137831 concentration were frozen immediately in liquid N2. Frozen cells were homogenized in a Fastprep machine (ThermoSavant) and shaken several times at speed 5.0 for 45 s until defrosted and homogenized. RNA was isolated with Trizol (Invitrogen) according to the manufacturer’s protocol, modified with an extra 1 : 1 (v/v) phenol : chloroform extraction after first chloroform addition. A similar approach was used for RNA isolation from zoospores/cysts and germinated cysts. RNA was isolated from mycelium and sporulating mycelium using the Qiagen RNeasy kit, according

to the manufacturer’s protocol for filamentous fungi. RNA was treated with Turbo DNA-free DNase (Ambion) according to the manufacturer’s protocol and checked for genomic DNA contamination by PCR with the primers used for quantitative RT-qPCR (Q-PCR). The concentration and purity of RNA were determined spectrophotometrically with Nanodrop at 260 and 260/280 nm ratios, respectively. Samples with a 260/280 nm ratio lower than 1.7 were discarded. Subsequent cDNA synthesis was performed using a First strand cDNA synthesis Cyclooxygenase (COX) kit (GE Healthcare) with 3–5 μg of RNA per 33-μL sample using the pd(N)6 random hexamers according to the manufacturer’s protocol. Transcript levels of SpHtp1 were analysed with a LightCycler® 480 (Roche),

using the LightCycler® 480 SYBR Green I Master mix (Roche), with 1 μL of cDNA in a total of 10 μL and according to the manufacturer’s protocol. The reaction was performed with an initial incubation at 95 °C for 5 min, followed by 45 cycles of 95 °C for 10 s, 58 °C for 10 s and 72 °C for 5 s, respectively. A dissociation curve as described in the LightCycler® 480 SYBR Green I Master mix (Roche) was performed to check the specificity of the primers. The amplicon length and optimized concentrations of the primers were 104 bp and 250 nM for SpHtp1, respectively, and 129 bp and 400 nM for SpTub-b, respectively. To correct for differences in the template concentration, several reference genes suggested by Yan & Liou (2006) were tested initially (Supporting Information, Fig.

, 2009). Putative

mutants were selected on NA with Km at 50 μg mL−1, and verified by Southern blot. Loss of swimming motility was confirmed in soft agar (0.3%) plates and under the microscope (not shown). MFCs were fabricated as described previously (De la Fuente et al., 2007b). Briefly, the chamber body was constructed with polydimethylsilioxane and consisted of two parallel channels measuring 80 μm wide, 3.7 cm long and 50 μm high, separated by a 50 μm wide polydimethylsilioxane ridge. Chamber bodies were then sandwiched between a cover glass and a supporting glass microscope slide. Teflon tubes were attached to inlet and outlet channels, and media were introduced into the channels using syringes controlled by pumps (Pico Plus, Harvard Apparatus). The chambers were mounted on a Nikon Enzalutamide supplier Ti/U E20L80 microscope (Nikon Co.) using 40 × phase-contrast and differential interference contrast optics. Time-lapse images were recorded using a DS-Qi1Mc digital camera and analyzed using nis elements software (Nikon Co.). The adhesion abilities of bacterial cells were evaluated using a modification of a described procedure (De La Fuente et al., 2007b): (1) cells were introduced from side channels, while the flow in the

main channels was stopped, allowing cells to attach; (2) introduction of cells from the side channels was www.selleckchem.com/products/Roscovitine.html stopped and medium flow in the main channels was resumed at a rate of 0.25 μL min−1 to remove unattached PAK6 cells; and (3) the flow rate in the main channels was gradually increased from 0.25 to 0.5, 1, 2, 4, 8, 16, 32 and 64 μL min−1, each rate being maintained

for one minute. Time-lapse movies were captured during the course of the assay and cells attached to the glass surface were quantified using nis elements software. Each repetition of steps 1–3 was considered a replicate. For each strain, at least three replicates in different locations along the channels were measured. For each flow rate, the amount of cells washed from the field of view was calculated as a function of the total number of cells present at the beginning of the assay. At the end of each flow rate, the number of attached cells was determined by averaging the amount of attached cells in the last three frames of that time period (corresponding to the last 15 s of the corresponding flow rate). Adhesion forces were determined according to De La Fuente et al. (2007b). Biofilm formation was monitored inside the MFCs by maintaining a flow rate of 0.25 μL min−1 in the main channel and capturing images at 30-s intervals for a period of 6–24 h. Swimming and twitching were assessed for all strains inside the MFCs. Twitching motility rates were calculated for six bacterial cells according to De La Fuente et al. (2007a). All experiments were repeated at least three times and data were subjected to the Tukey–HSD test using jmp in v3.2.1 (SAS Institute Inc.). For comparison of adhesion forces, one-way anova were performed using statistix 8.