Nevertheless, ZnO has one major drawback, which is the lack of stable and reproducible p-type ZnO with low resistivity, high carrier concentration, and high carrier mobility. Doping with the first group elements like Li, Na, K, and Cs in ZnO would substitute Zn2+ by the monovalent cations, thus making it possible to realize n-type conduction. The realization of n-type conduction is very important for ZnO applications in optoelectronic devices, and there are reports on the electrical property

of the first group element-doped ZnO thin-films [32–36]. buy PD173074 Various techniques such as pulsed laser deposition [37, 38], magnetron sputtering [39, 40], and molecular beam epitaxy [41] have been used to deposit thin-films of ZnO. The sol-gel method [42] has been receiving increased attention because

of its many advantages such as low cost, Alvocidib supplier simple deposition procedure, easier composition control, low processing temperature, and easier fabrication of large area films. Therefore, here, we demonstrate the improved performance of P3HT:PCBM and P3HT:ICBA-based inverted bulk-heterojunction solar cells see more through the appropriate interface modification by Cs2CO3-doped ZnO on the electron collecting ITO interface. Recently, Yang et al. has reported that a solution-processed Cs2CO3 is able to make interface dipoles layer on ITO. One may say that these two entities (ZnO and Cs2CO3)

are completely different but the most important thing is that these entities do improve the performance of the device. Moreover, we have seen a number of works on tuning the work function of ITO by adding an electron transport layer such as ZnO [43], TiO2 [44–46], Cs2CO3 [44–46], and poly(ethylene oxide) (PEO) [47]. The created dipole moment helps to reduce the work function of ITO, allowing ITO to serve as the cathode. The improved device performance is due to the reduction of series resistance, improved shunt performance, and enhanced open-circuit voltage of the cell which can be attributed to the improvement of the following aspects: (1) reduction of the contact resistance between the ZnO:Cs2CO3 and active organic buy Cobimetinib layer, (2) enhancement of the electronic coupling between inorganic ZnO:Cs2CO3 and active organic layer to mediate better forward charge transfer and reduce back charge recombination at the interface, and (3) affect the upper organic layer growth mode and morphology. Methods ZnO solution preparation ZnO solution was prepared using similar procedures to the one reported by Jang et al. [27]. Cs2CO3 solution was prepared by dissolving in ethanol in the ratio of 1.25 wt%. Organic solar cell fabrication Schematic diagram of organic solar cells is shown in Figure 1b, where the device is fabricated using pre-patterned ITO-coated glass substrate.

PubMedCrossRef 39.

Tseng T-T, Tyler BM, Setubal JC: Protein secretion systems in bacterial-host associations, and their description in the Gene Ontology. BMC Microbiology 2009,9(Suppl 1):S2.PubMedCrossRef 40. Lilley CJ, Atkinson HJ, Urwin PE: Molecular aspects of cyst nematodes. Mol Plant Pathol 2005, 6:577–588.PubMedCrossRef 41. Hahn M, Mendgen K: Signal and nutrient exchange at biotrophic plant fungus interfaces. Current Opinion in Plant Biology 2001, 4:322–327.PubMedCrossRef 42. Hardham AR: Cell biology of plant-oomycete interactions. Cellular Microbiology 2007,9(1):31–39.PubMedCrossRef 43. Lindeberg M, Biehl BS, Glasner JD, Perna NT, Collmer A, Collmer CW: Gene Ontology annotation highlights shared and divergent pathogenic strategies of type III effector proteins deployed by the plant pathogen

Pseudomonas syringae pv tomato DC3000 and animal pathogenic 8-Bromo-cAMP chemical structure Escherichia coli strains. BMC Microbiology 2009,9(Suppl 1):S4.PubMedCrossRef 44. Torto-Alalibo T, Collmer CW, Gwinn-Giglio M: The Plant-Associated Microbe Gene Ontology (PAMGO) Consortium: Community development of new Gene Ontology terms describing biological processes selleck kinase inhibitor involved in microbe-host interactions. BMC Microbiology 2009,9(Suppl 1):S1.PubMedCrossRef 45. Dangl JL, Jones JD: Plant pathogens and integrated defence responses to infection. Nature 2001,411(6839):826–833.PubMedCrossRef 46. Jia Y, McAdams SA, Bryan GT, Hershey HP, Valent BAY 63-2521 mw B: Direct interaction of resistance gene and avirulence gene products confers rice blast resistance. Embo J 2000,19(15):4004–4014.PubMedCrossRef 47. de Vries JS, Andriotis VM, Wu AJ, Rathjen JP: Tomato Pto encodes a functional N-myristoylation motif that is required for signal transduction Dichloromethane dehalogenase in Nicotiana benthamiana. Plant J 2006,45(1):31–45.PubMedCrossRef 48. Fu ZQ, Guo M, Jeong BR, Tian F, Elthon TE, Cerny RL, Staiger D, Alfano JR: A type III effector ADP-ribosylates RNA-binding

proteins and quells plant immunity. Nature 2007,447(7142):284–288.PubMedCrossRef 49. Armstrong MR, Whisson SC, Pritchard L, Bos JI, Venter E, Avrova AO, Rehmany AP, Bohme U, Brooks K, Cherevach I, et al.: An ancestral oomycete locus contains late blight avirulence gene Avr3a, encoding a protein that is recognized in the host cytoplasm. Proc Natl Acad Sci USA 2005,102(21):7766–7771.PubMedCrossRef 50. Lahaye T, Bonas U: Molecular secrets of bacterial type III effector proteins. Trends Plant Sci 2001,6(10):479–485.PubMedCrossRef 51. Kemen E, Kemen AC, Rafiqi M, Hempel U, Mendgen K, Hahn M, Voegele RT: Identification of a protein from rust fungi transferred from haustoria into infected plant cells. Mol Plant Microbe Interact 2005,18(11):1130–1139.PubMedCrossRef 52. Kanneganti TD, Bai X, Tsai CW, Win J, Meulia T, Goodin M, Kamoun S, Hogenhout SA: A functional genetic assay for nuclear trafficking in plants. Plant J 2007,50(1):149–158.PubMedCrossRef 53. Elling AA, Davis EL, Hussey RS, Baum TJ: Active uptake of cyst nematode parasitism proteins into the plant cell nucleus.

However, (i) it is considerably faster (especially if analysing more sequences at once), (ii) it shows only results relevant to potential enzybiotic activity and (iii) provides greater versatility for input formats. Figure 1 Sample output from phiBiScan CH5183284 cost program utility. Two domains corresponding to peptidoglycan hydrolytic activity (Pfam IDs CHAP and Glyco_hydro_25) were identified in the sequence of analysed protein. Ro 61-8048 cost To evaluate the overall accuracy of phiBiScan, we analysed protein sequences from known phage genomes in order to identify proteins with peptidoglycan hydrolytic activities. Phage genomes deposited in NCBI Genome database were used ( http://​www.​ncbi.​nlm.​nih.​gov/​sites/​genome).

Firstly, four groups of bacteriophages were excluded from the analysis: (i) phages lacking any peptidoglycan hydrolases, i.e. phages belonging to the families employing strategies for progeny release, which does not result in host cell lysis (Microviridae, Inoviridae, Leviviridae, Lipothrixviridae, Rudiviridae); (ii) unclassified phages and phages belonging to the novel phage families (e.g. Ampullaviridae); (iii) phages of Archaea; (iv) genomes, where no conventional peptidoglycan hydrolases were experimentally identified or predicted. Consequently the phiBiScan click here search was run

against 37 930 protein sequences from 444 phage genomes. The number Rolziracetam of positive and negative hits was recorded. Going through gene annotations manually, along with additional standard Pfam search in ambiguous cases, we distinguished true and false matches. 673 proteins tested positive in phiBiScan and indeed having domain(s) corresponding to the lytic activity were considered as true positives

(TP); 18 proteins tested positive, but obviously without any lytic activity were false positives (FP); 37 189 proteins tested negative and lacking lytic activity were true negatives (TN); 5 negative hits for proteins with confirmed lytic activity were considered as false negatives (FN). Solid prediction strength of phiBiScan was confirmed by high performance of binary classification test: sensitivity (99%), specificity (100%) and also positive predictive value (PPV, 97%) and negative predictive value (NPV, 100%). phiBiScan has identified 700 positive hits (567 proteins matched in one Pfam domain, 133 proteins in two Pfam domains) in 396 phages. In 48 phages no match with any applied profile was noted. Only 2 out of 18 false positive matches were assessed as significant positive hits, the rest were insignificant (Table  3). Table 3 Summary of statistical assessment of phiBiScan tool True positive (TP) 673 False positive (FP) 18 True negative (TN) 37 189 False negative (FN) 5 Sensitivity 99% Specificity 100% PPV 97% NPV 100% Correlation coefficient 0.

However, when these indices have been used to assess the ability of dietary supplements to enhance recovery after heavy eccentric

exercise, they have been unable to detect a treatment DNA Damage inhibitor effect [10, 13]. Muscle soreness, assessed by the subjective pain rating in the present study, is one of the most commonly used measures of exercise-induced muscle injury [2]. However, Warren et al. [2] suggested that soreness correlates poorly with muscle function. In the current study, the patterns of recovery for hanging joint angle, relaxed arm circumference, and subjective pain ratings were similar in the ANA and PLA conditions (Figure 3b). Therefore, the lack of effect of supplementation on the hanging joint angle and relaxed Dehydrogenase inhibitor arm circumference in these studies [10, 13], the poor correlation between muscle function and soreness [2], and the results of the present study have collectively suggested that these indicators of muscle damage may not be sensitive to dietary supplement interventions to improve recovery from eccentric-induced muscle learn more damage. Future studies may wish to consider these

findings when selecting outcome variables for assessing the efficacy of dietary supplementation for reducing muscle damage. A secondary objective of this study was to examine the effects of 10 days of ANA dietary supplementation on resting heart rate and blood pressure. As a minor alkaloid with a similar chemical structure to nicotine, we hypothesized that tuclazepam ANA would cause moderate decreases in systolic and diastolic blood pressure and small increases in resting heart rate. Previous studies [14, 26, 27] have shown that acute nicotine exposure causes an increase heart rate and blood pressure through stimulation of the sympathetic nervous system. Minami et al. [28] showed that smoking cessation caused a reduction in heart rate, which implied that chronic nicotine use may elevate heart rate. However, cross sectional studies [26,

29] have shown that systolic and diastolic blood pressures are lower in smokers than in non-smokers. The results of the present study indicated that, over a period of 10 days, ANA had no effect on heart rate or blood pressure compared to placebo. Thus, ANA supplementation may be safe regarding short-term use (10 days) on resting heart rate and blood pressure. However, future studies may wish to examine the acute and chronic effects of ANA consumption on blood pressure and heart rate to further discern its safety. Conclusions In conclusion, ANA supplementation had no effect on the recovery of muscle strength, hanging joint angle, arm swelling, or subjective pain ratings after a bout of maximal eccentric exercise in the forearm flexors. Therefore, ANA may not be beneficial for those seeking to improve recovery from heavy exercise training or competition.

After sufficient muscle drying, the samples were then placed in an ultra-low freezer at -80°C. Dried muscle

was powdered by grinding on a porcelain plate with a pestle. Connective tissue was removed and discarded, whereas powdered muscle was placed into pre-weighed microfuge tubes. Powdered muscle was extracted in a 0.5 M perchloric acid/1 mM EDTA solution on ice for 15-minutes, while periodically vortexing. Samples were then spun at 15,000 rpm at 4°C for 5-minutes. The Ivacaftor datasheet supernatant was transferred into a microfuge tube and neutralized with 2.1 M KHCO3/0.3 M MOPS solution and then centrifuged again at 15,000 OICR-9429 rpm for 5-minutes. In order to determine muscle total creatine concentration, supernatant from the above reaction was combined with ddH2O and 0.4 N HCl and heated at 65°C for 10-minutes to hydrolyze phosphate groups. The solution was then neutralized with of 2.0 N NaOH and the samples were allowed to incubate at room temperature allowing for color formation, which was detected by a spectrophotometer at 520 nm. Then the samples were run in

duplicate against a standard curve of known creatine concentrations. The mean correlation coefficient of variation between duplicates was 1.53%. The standard curve correlation coefficient between plates for total muscle creatine was 0.998. Dietary intake records and supplementation compliance Throughout the course of the study, participants’ dietary intake was not supervised; BTSA1 however, it was required that all participants keep detailed dietary records and not Cytidine deaminase change their routine dietary habits throughout the course of the study. As such, participants were required to keep weekly physical activity records and four-day dietary records (three weekdays

and one weekend) prior to each of the four testing sessions. The four-day dietary recalls were evaluated with the Food Processor dietary assessment software program (ESHA Research, Salem, OR) to determine the average daily macronutrient consumption of fat, carbohydrate, and protein. The participants were instructed to turn in their dietary records during each testing session. Each participant returned all of their dietary evaluations at the required time points for a 100% compliance rate. In an effort to ensure compliance to the supplementation protocol, participants were supplied with the appropriate amount of supplement to be ingested during the time between last three testing sessions. Upon reporting to the lab for each testing session at days 6, 27, and 48, participants returned the empty containers they had acquired between testing sessions Reported side effects from supplements At the last three testing sessions, participants reported by questionnaire whether they tolerated the supplement, supplementation protocol, as well as report any medical problems/symptoms they may have encountered throughout the study.

Anal Biochem 1996, 236:302–308.CrossRefPubMed 26. Storey JD, Tibshirani R: Statistical significance for genome wide studies. Proc Natl Acad Sci USA 2003, 100:9440–9445.CrossRefPubMed 27. Xia Q, Wang T, Park Y, Lamont RJ, Hackett M: Differential CHIR98014 quantitative proteomics of Porphyromonas gingivalis by linear ion trap mass spectrometry: Non-label methods comparison, q -values and LOWESS curve fitting. Int J Mass Spectrom 2007, 259:105–116.CrossRefPubMed 28. Peng J, Elias JE, Thoreen CC, Licklider LJ, Gygi SP: Evaluation of multidimensional chromatography coupled with tandem mass spectrometry (LC/LC-MS/MS) for

large-scale protein analysis: the yeast proteome. J Proteome Res 2003, 2:43–50.CrossRefPubMed 29. Elias JE, Gibbons FD, King OD, Roth FP, Gygi SP: Intensity-based protein identification by machine learning from a library of tandem mass spectra. Nat Biotechnol 2004, 22:214–219.CrossRefPubMed 30. human.protein.faa[http://​www.​ncbi.​nlm.​nih.​gov/​Ftp/​] 31. Hendrickson EL, Kaul R, Zhou Y, Bovee D, Chapman P, Chung J, Conway de Macario E, Dodsworth JA, Gillett W, Graham DE, et al.: Complete genome sequence of the genetically tractable hydrogenotrophic methanogen Methanococcus maripaludis. J Bacteriol

2004, 186:6956–6969.CrossRefPubMed 32. Tumbula DL, Makula RA, Whitman WB: Transformation of Methanococcus maripaludis and identification of a Pst I-like restriction system. FEMS Microbiol Lett 1994, 121:309–314.CrossRef Authors’ contributions QX and TW performed protein biochemistry, 2-D capillary HPLC separations, mass spectrometry and data analysis. ELH performed data analysis and bioinformatics. Lenvatinib datasheet TJL

assayed expression of the Na+-alanine symporter gene. MH and JAL supervised the research. JAL wrote the manuscript.”
“Background Bacteriophages (phages) are viruses that specifically infect bacteria. They can be found in almost all ecosystems and it is estimated that approximately 1031 phages exist globally (108 phage species predicted), making them the most prominent biological system on earth [1–5]. Despite these enormous numbers it is estimated that less than 1% of all phage species have been detected by the plaque assay because of undersampling, which is often attributed to the use of classical bacteriophage propagation procedures [4, 5]. The selleck chemical ability of a phage to lyse its host bacterium, producing a plaque ZD1839 mouse within a bacterial lawn, led to the discovery of phages in 1915 by Frederick W. Twort and is the basis of the classic plaque assay, the double-layer agar (DLA) technique, which has been used ever since [6–8] to identify and enumerate phages and isolate mutants. In recent years, interest in phages has increased not only because of their potential use as alternatives to antibiotics (phage therapy) but also because of their applications in many other fields (phage display, immunology, microbial genetics, diagnostics, vaccine development, biosensors, etc.).

Journal of Clinical Microbiology 2006,44(5):1859–1862.PubMedCrossRef Authors’ contributions RCLM: Study design, primers design, fieldwork and data collection, laboratory tests, data analysis, manuscript writing; ASR: Study design, primers design, laboratory tests, data analysis, manuscript writing; FFM: Primers

design, laboratory test, data analysis, manuscript writing; MASC: Fieldwork, data collection and analysis, manuscript writing; KDE: Fieldwork and data collection; ADF: Fieldwork and data collection; LMSM: Diagnostic laboratorial tests; MSL: Data interpretation and analysis, manuscript writing; BW: Coordination, study design, fieldwork and data collection, data analysis, manuscript writing. All authors read an approved the final draft.”
“Background The animal gastrointestinal Poziotinib ic50 tract harbors a complex microbial network and its composition reflects the constant co-evolution of these microorganisms with their host environment [1]. Uncovering the taxonomic composition and functional capacity within the animal

gut microbial consortia is of great importance to understanding the roles they play in the host physiology and health. Since animal feces can harbor human pathogens, understanding the genetic composition NU7441 price of fecal microbial communities also has important implications for food and water safety. The structure and function of the gut microbial community has received significant attention for decades, although most of the work was restricted by the use of culture-based techniques. Recently, sequence Branched chain aminotransferase analysis of the 16S rRNA gene has shed new light on the diversity and composition of microbial communities within several animal gut systems [2]. While 16S rRNA gene-based techniques have revealed impressive microbial diversity within gut environments, this approach offers only limited information on the physiological role of microbial consortia within a given gut environment. Random sequencing of metagenomes has allowed scientists to reveal significant differences in metabolic potential within different environments [3], including microbial populations associated with host-microbial partnerships. Specifically,

the publicly available Idasanutlin research buy database IMG/M [4] contains 596 Mb of sequencing data, representing 1,424, 000 genes from 17 different gut microbiomes. Studying gut metagenomes has particularly helped in uncovering several important biological characteristics of these microbiomes. For example, when 13 human gut metagenomes were compared, Kurokawa et al [5] found that adult and infant type gut microbiomes have enriched gene families sharing little overlap, suggesting different core functions within the adult and infantile gut microbiota. This study also demonstrated the presence of hundreds of gene families exclusively found in the adult human gut, suggesting various strategies are employed by each type of microbiota to adapt to its intestinal environment [5].

Stenotrophomonas maltophilia K279a had a putative Major Facilitator Superfamily (MFS) efflux pump that usually function as specific exporters for certain classes of antimicrobial agents. This is related to the emrAB system from E. coli [60]. P. aeruginosa UCBPP-PA14 has a predicted Pritelivir chemical structure czc [Cd/Zn/Co] efflux system similar to those in D. acidovorans SPH-1 and C. testosteroni KF-1. P. aeruginosa PACS171b contains a homolog of UspA- the Universal

Stress Protein. The UspA protein is important for survival during cellular growth arrest in E. coli, but the exact physiological role of the protein is unknown [61]. Thioalkalivibrio sp. HL-EbGR7 has a set of genes with approximately 88% aa identity to the putative KdpFABC system in P. aeruginosa PA7. This variability is suggestive that this region may be a hotspot for insertion or recombination where insertion clearly does not disrupt or affect the expression of neighbouring genes. The variation in predicted gene function, GSK458 clinical trial size and lack of homology between elements is suggestive of this region contributing a number of different adaptive traits to hosts containing these ICEs. Following this variable region is encoded a putative transcriptional regulator protein TraR and a homologue of the type IV coupling protein TraG [similar to those in IncP plasmids]. TraG is responsible for DNA transfer during conjugation and is a putative DNA binding protein [62]. Interestingly

the gene order of this region and the order of genes preceding it are also suggestive of an insertion [of the variable

Methamphetamine region just discussed] into a primordial transfer module. The putative DNA binding gene traG is followed by a group of genes encoding proteins [TrbBCDEJLFGI] with similarity to the mating-pair formation [mpf] apparatus or type IV secretion system closely related to IncP and Ti plasmids. This system presumably mediates the DNA transfer of the ICE to recipient cells [63, 64]. These genes show similarity to those required for conjugative transfer of the Agrobacterium Ti plasmid, pNGR234a and RP4, except that two genes, trbK and trbH, found on these plasmids are missing [65]. In the Tn4371-like elements the gene order was trb BCDEJLFGI in all the characterised elements found in this study and similar to the molecular organisation in ICEMlSymR7A [[19], Fig. 1]. The TrbB, TrbC, TrbE, TrbG, and TrbL proteins are involved in the creation of the mpf apparatus, TrbC is involved in pilus formation and TrbE displays ATPase activity [65]. The novel ICEs detected in this study are integrated into various locations in the genomes of the host bacteria where they were discovered. In Acidovorax sp. JS42 other partial copies of Tn4371-like elements were also found in addition to the full element reported here. Two elements were discovered and characterised in D. acidovorans SPH-1. A Vactosertib further partial element was found in B. petrii this however lacked the int Tn4371 gene. This situation is similar to that found in R.

2-DEST-Plk1) was verified

according to the reference sequence. PLK-1 (GenBank accession no. NM_005030) siRNAs, targeting regions of the Plk-1 transcript at positions 362-384, were also used in this study. HeLa cells were transfected at 70% to 90% confluency using PLK-1 plasmid DNA (up to 4 μg) mixed with Lipofectamine 2000 (Invitrogen) at a DNA (μg)/lipid (μL) ratio of 1:2.5. Similarly, PLK-1 silencing was performed by transfecting HeLa cells with PLK-1 siRNA plasmids. At 4-6 h post-transfection, the plasmid- or siRNA-containing medium was replaced with normal culture medium containing 10% FCS, and the cells were incubated Blasticidin S molecular weight in a 5% CO2 incubator at 37°C. Transfected cells were then cultured in fresh medium for up to 12-36 h and harvested for gene expression and other assays. For cisplatin treatment, cisplatin (4 μg/ml) was added to HeLa cells, with DMSO as see more control. The time point chosen for the addition of cisplatin to the transfected cells was 24 h after transfection, and was based on preliminary experiments (data not shown). Quantitative RT-PCR analysis for mRNA levels Real-time RT-PCR was performed as detailed in our previous report [14]. Briefly, total RNA was extracted with TRIzol reagent (Invitrogen),

following the manufacturer’s instructions. Reverse transcription (RT) was performed, and the cDNA was synthesized from 2 μg of total RNA by using an oligo (dT)18 primer and M-MLV reverse transcriptase (TAKARA, Syuzou, Shiga, Japan) for quantitative PCR. Expression of mRNA was determined using the ABI PRISM 7300 Detection System (Applied Biosystems, Foster City, CA) https://www.selleckchem.com/products/ag-881.html and SYBR Premix Taq™ (TAKARA). The sequences of the primers were as follows: PLK1 (NM_005030) forward: 5′-GGA CTA TTC GGA Sclareol CAA GTA CG-3′; PLK1 reverse: 5′-CGG AAA TAT TTA AGG AGG GTG A-3′; β-actin (NM_001101) forward: 5′-AAG ATG ACC CAG ATC ATG TTT GAG ACC-3′; β-actin reverse:

5′-AGC CAG GTC CAG ACG CAG GAT-3′. The mean value of the replicates for each sample was calculated and expressed as cycle threshold (Ct). The amount of gene expression was then calculated as the difference (ΔCt) between the Ct value of the target gene and the Ct value of β-actin. Assessment of cell viability by MTT Assay Treated or untreated cells were seeded into 96-well plates at 1 × 103 cells per well overnight and incubated with different concentrations of cisplatin (0 or 4 μg/ml) per treatment. After culture for 24 h, 20 μl MTT dye solution (5 mg/ml) was added to each well and samples were incubated at 37°C for 4 h. The formazan product was dissolved by adding 200 μL of DMSO to each well. The plates were read at 570 nm. Immunoblotting analysis Immunoblotting was performed as previously described [14]. Briefly, treated and untreated HeLa cells were collected and the protein concentrations of lysates were determined by the Bradford method (Pierce, Rockford, IL).

Cultivation on selective media indicates a slight dominance of Pseudomonas GANT61 manufacturer spp. in air-stored samples at low temperatures while molecular based methods, both 16S rRNA cloning analysis and t-RFLP, indicate a high dominance of P. phosphoreum in both air and MA packaging. Analysis of volatiles produced during Cisplatin mw Storage at -2°C supported the dominance of P. phosphoreum showing intense TMA production. The species diversity was higher after short storage of less than one week, especially

in air packaging, but with time, P. phosphoreum reached a high dominance, depending on the storage conditions. Discrepancy was observed between the conventional cultivation and molecular methods and requests a further investigation to elucidate this Sepantronium nmr matter. Nevertheless, combined strategy of cultivation and cultivation independent methods might be the key for deeper understanding of bacterial population developments during the spoilage process of food. Methods Raw material The fish used for the shelf life experiments was captured by trawl in October 2006 in the North of Iceland, gutted onboard, washed with excessive seawater and stored iced in tubs until filleted, providing a temperature around 0°C. The sea temperature was 8.5-9°C on the day of capture. The raw material was 2-3 or

4-5 days old when it was filleted, deskinned, cut into loins and packaged for the shelf life experiment. Storage conditions Earlier to packaging, a part of the many fish was filleted and stored in 4% brine for two days at around 1°C while the other part was processed and cooled down in a 4% brine for 8 min prior to trimming and packaging. These treatments resulted in two groups with a final salt (NaCl) concentration of 2.5 ± 1.0% (HS) and 0.4 ± 0.2% (LS). The fish was stored in air (open bags in styrofoam boxes) and in modified atmosphere packaging (50% CO2, 5% O2, 45% N2) at 0°C

(only LS group), -2°C and -4°C resulting in 10 treatments (Table 4). Temperatures were monitored with loggers placed in packages at the bottom recording the temperature every 90 s. The gas composition was monitored using a CheckMate 9900 instrument (PBI Dansensor, Ringsted, Denmark). Sampling was performed in duplicate periodically during the storage time. Aerobic samples were stored for 12 (0°C) and 15 days (-2°C), MA-packed samples at 0°C for 21 days but 28 days for superchilled products. Table 4 Overview of fish treatments tested Treatments Temperature (°C) Atmosphere Salt content Sampling time (days) 1 0 Air LS 6, 13 2 -2 Air LS 6, 15 3 -4 Air LS 6, 15 4 0 MAP LS 7, 21 5 -2 MAP LS 7, 28 6 -4 MAP LS 7, 21 7 -2 Air HS 6, 15 8 -4 Air HS 6, 15 9 -2 MAP HS 13, 21 10 -4 MAP HS 7, 28 R Raw material 0 Cultivation Viable microbial developments were done essentially as described before [16].