All extension reactions were performed at least twice with independent RNA preparations and the reproducible peaks were selected. Animal cell cultures and invasion assay HeLa cell lines were obtained from ATCC (Manassas, VA). Cells were grown to a monolayer at 37°C, 5% CO2 in DMEM with 10% heat-inactivated fetal bovine serum. Cells were then infected at an MOI of 100 in 24-well plates. Bacteria were spun onto the HeLa cells and incubated at 4°C

for 30 min, then at 37°C for 1 hour. Extracellular bacteria were killed https://www.selleckchem.com/products/sn-38.html with 50 μg/ml gentamicin for 30 min. HeLa cells were then lysed with 0.1% Triton X-100 and plated for CFU determination. Mouse studies Food and water were withdrawn 4 h before inoculation of female BALB/c mice (weighing 16 to 18 g). Mice (10 for each strain) were inoculated with 106 bacteria by oral gavage using a 22-gauge feeding needle. Dilutions of the stationary-phase cultures were plated to determine the number of bacteria present in the inoculum. For virulence assays, time of death was recorded as days post-infection. Competition infection experiments were conducted as described above, except that the mutant strain was co-infected with a chloramphenicol marked wild

type strain (JSG224, phoN2 ZXX::6251dTn10-Cam). After plating bacteria on appropriate media from organs four days post-infection, the competitive index was calculated as the CFU mutantplate count from organ/wild find more typeplate count from organ divided by Cl-amidine in vivo mutantinoculum/wild typeinoculum. All experiments were reviewed and approved by the Ohio State

University Institutional Animal Care PtdIns(3,4)P2 and Use Committee. Motility assays 0.3% agar DMEM plates were made containing, where indicated, 10 or 20 μM autoinducer-2 (AI-2 was a gift from Dr. Dehua Pei, Department of Chemistry, The Ohio State University), 10 or 50 μM epinephrine, or equivalent amounts of acidified water as a control for epinephrine plates (epinephrine was solubilized in acidified water). Overnight cultures were grown in LB, 37°C with shaking, adjusted to an OD of 0.1 at 600 nm and incubated for 2 hours at 37°C with shaking. Plates were stab-inoculated and incubated at 37°C for 14 hours. The diameter of the motility circles were measured at various times and compared. Results Transcriptome of the PreA/PreB two-component system In previous experiments, we realized that the preAB TCS was not fully activated during growth in LB, as indicated by the absence of regulatory effects on the two known target genes (yibD, pmrCAB) when comparing a nonpolar mutation in the preA response regulator to the wild type strain [3]. This was confirmed in this study by microarray analysis co-hybridizing preA and wild type cDNA to a multistrain slide microarray of Salmonella enterica (data not shown).

3. Cribb PJ, Williams AD, Stathis CG, Carey MF, Hayes A: Effects of whey isolate, creatine, and resistance training on muscle hypertrophy. Medicine and science in sports and exercise 2007,39(2):298–307.check details PubMedCrossRef 4. Tipton KD, Rasmussen BB, Miller SL, Wolf SE, Owens-Stovall SK, Petrini BE, Wolfe RR: Timing of amino acid-carbohydrate ingestion alters anabolic response of muscle to resistance exercise. American journal of physiology 2001,281(2):E197–206.PubMed 5. White JP, Wilson JM, Austin KG, Greer BK, St John N, Panton LB:

Effect of carbohydrate-protein supplement timing on acute exercise-induced muscle damage. J Int Soc Sports Nutr 2008, 5:5.PubMedCrossRef 6. Willoughby DS, Stout JR, Wilborn CD: Effects of resistance training and protein plus amino acid supplementation 4SC-202 concentration on muscle anabolism, mass, and strength. Amino acids 2007,32(4):467–477.PubMedCrossRef 7. Faria IE: Applied physiology of cycling. Sports medicine (Auckland, NZ) 1984,1(3):187–204.CrossRef 8. Edge J, Bishop D, Goodman

C: Effects of chronic NaHCO3 ingestion during interval training on changes to muscle buffer capacity, metabolism, and short-term endurance performance. Journal of applied physiology 2006,101(3):918–925.PubMedCrossRef 9. Graef JL, Smith AE, Kendall KL, Fukuda DH, Moon JR, Beck TW, Cramer JT, Stout JR: The effects of four weeks of creatine supplementation and high-intensity interval training on cardiorespiratory fitness: a randomized controlled trial. Journal of the International Society of Sports Nutrition 2009,6(1):18.PubMedCrossRef 10. Kendall KL, Smith AE, Graef JL, Fukuda DH, Moon JR, Beck TW, HM781-36B Cramer JT, Stout JR: Effects of four weeks of high-intensity interval training and creatine supplementation on critical power and anaerobic working

capacity in college-aged men. Journal of strength and conditioning research/National Strength 4-Aminobutyrate aminotransferase & Conditioning Association 2009,23(6):1663–1669. 11. Smith AE, Walter AA, Graef JL, Kendall KL, Moon JR, Lockwood CM, Fukuda DH, Beck TW, Cramer JT, Stout JR: Effects of beta-alanine supplementation and high-intensity interval training on endurance performance and body composition in men; a double-blind trial. Journal of the International Society of Sports Nutrition 2009, 6:5.PubMedCrossRef 12. Smith AE, Moon JR, Kendall KL, Graef JL, Lockwood CM, Walter AA, Beck TW, Cramer JT, Stout JR: The effects of beta-alanine supplementation and high-intensity interval training on neuromuscular fatigue and muscle function. European journal of applied physiology 2009,105(3):357–363.PubMedCrossRef 13. Syrotuik DG, Game AB, Gillies EM, Bell GJ: Effects of creatine monohydrate supplementation during combined strength and high intensity rowing training on performance. Canadian journal of applied physiology = Revue canadienne de physiologie appliquee 2001,26(6):527–542.PubMed 14.

Initially, when the coating has 10 bilayers it is possible to appreciate well-separated AgNPs with a very low roughness of 5.8 nm. However, when the number of bilayers is increased, the roughness is changing from 10.2 nm (20 bilayers) to 23.9 nm (30 bilayers) and 28.7 nm (40 bilayers). It is important to remark that after a thermal treatment, the total

evaporation of the polymeric chains induces an agglomeration of the AgNPs without preserving their distribution along the films. This aspect is corroborated due to a color change from violet to orange in the resultant films. Figure 8 AFM images (25×25 μm) of PAH/PAA-AgNPs (violet coloration) after a thermal treatment as a function of number of bilayers (a) 10 bilayers; (b) 20 bilayers; (c) 30 bilayers and (d) 40 bilayers. In other words, the fact that a higher number of bilayers during the LbL fabrication process, and consequently, a higher Selleck Erastin thickness of the

resultant films, promote a better definition of the color, mostly in the green coloration (see Figure  9) because of a better entrapment of both initial clusters (hexagons with higher size) and nanometric spherical AgNPs in the multilayer assembly. Additionally, new PAH/PAA-AgNPs coatings of 80 bilayers at pH 7.5 have been fabricated in order to show clearly the final coloration onto the glass slides as a function of the initial synthesized multicolor silver nanoparticles (PAA-AgNPs). Figure 9 Final aspect almost of the PAH/PAA-AgNPs multilayer GANT61 cost assembly (violet, green, orange coloration) for a total number of 80 bilayers. Figure  10 shows the UV–vis www.selleckchem.com/products/Cisplatin.html spectra of the samples prepared with this thickness (80 bilayers) and the spectra

reveal that the position of the absorption bands is the same than previous spectra (Figures  3, 4 and 5) but with a considerable increase in intensity of the absorption peaks due to a higher number of the metallic silver nanoparticles that have been incorporated into the multilayer film. Therefore, when the thickness is increased, it is possible to corroborate the presence of the same aggregates species or AgNPs than the original colloidal solutions. In other words, when the thickness is increased, the final coloration of the resultant films (violet, green or orange) is similar than the color of the original colloidal PAA-AgNPs solutions. These results of coloration as a function of number bilayers indicate that a higher thickness leads to a better incorporation of higher size aggregates (clusters) in the resultant films. This is the first time that a study about colored AgNPs synthesis and their incorporation in multicolor films (violet, green or orange) is investigated using the LbL assembly. These multicolor LbL films can be used for optical fiber sensor applications [41].

, 2003). **mean of quantification by oprL qPCR tested in duplicate. NA: not applicable. P. aeruginosa Histone Methyltransferase antagonist isolation Ten μl of liquefied sputum pure and diluted into 1/1000, were inoculated and incubated onto several non selective and selective media for P. aeruginosa isolation, including Columbia blood agar supplemented with 5% defribinated horse blood (Oxoid, Dardilly, France), Columbia chocolate agar (Oxoid), and cetrimide agar (Oxoid).

All media were incubated aerobically at 37°C for five days and monitored daily. All different morphotypes of bacterial colonies were identified phenotypically with conventional screening methods (Gram coloration, oxidase test) followed by mass spectrometry identification (MicroFlex LT, Bruker Daltonics, Germany) [33, 34]. Quantification was conducted based on the colony forming unit (CFU) counts and the dilution ratio of the plate. P. aeruginosa detection and quantification by quantitative PCR (qPCR) DNA extraction For each Cobimetinib price isolate of the bacterial click here collection, 1 ml of a 0.5 McFarland suspension was extracted. For each sputum sample, one of the two 1 ml-aliquots was treated by 5 min of sonication using a bath sonicator (Elamsonic

S10, Singen, Germany). After a 10 min-centrifugation (5000 g), the pellet was suspended in 200 μl of DNA free water. Ten μl of the IC2, an internal control provided in the DICO Extra r-gene™ kit (Argène, Verniolle, France), were added in each sample and, for each batch of extraction, in 200 μl of DNA free water as a negative control. DNA was extracted using the QIAamp DNA Minikit® (Qiagen, Courtaboeuf, France) according to the instructions of the manufacturer (“Tissue protocol”)

with elution volumes of 100 μl. oprL qPCR oprL qPCR was performed using primers OPRL-F and OPRL-R and hydrolysis probe Dimethyl sulfoxide oprL-MGB, previously described by Joly et al. [30] (Table 2). The reaction mix comprised 12.5 μl of Qiagen Quantitect Probe Master Mix, 0.3 μM of each primer, 0.2 μM of hydrolysis probe and 4.5 μl of DNA extract, and was made up to a final reaction volume of 25 μl with water. A negative amplification control was used for each batch. For sputum samples, a standard curve provided a full concentration range of P. aeruginosa extending from 102 to 106 CFU/mL. Each qPCR assay was repeated twice, and the mean value of the quantification was calculated for each duplicate (Table 1). Cycling was performed on an ABI Prism 7300 Real Time PCR System (Applied Biosystem, Foster city, Californy), with an initial hold at 95°C for 15 min, followed by 50 cycles at 95°C for 15 s, and 60°C for 1 min. The oprL-MGB probe was labelled with carboxyfluorescein (FAM).

The etching rate of the silicon wall may be not the same as that of the silicon substrate under this Abemaciclib porous layer because of the different circumstance. To achieve the etching rate of the silicon substrate, i.e., the formation rate of the SiNWs, the samples were etched for a longer duration while keeping the other conditions the same as in the previously mentioned case wherein the etching was carried out for 10 min. Supposing a linear relationship between the SiNW height and the etching duration [14], the etching rate can be calculated by comparing the heights

of the SiNWs with those etched for 10 min; the results are shown in Figure 6. Clearly, a high etching rate (>250 nm/min) was obtained in the present conditions, and the etching rate increases with TSA HDAC order increasing thickness of the Au film. The etching was also performed at a solution temperature of 28°C. The same trend was observed with a higher etching rate of over 400 nm/min. Figure 5 SEM images of the SiNW arrays catalyzed using the Au mesh with different thickness. Cross-sectional (a, b, c) and the corresponding plan-view (d, e, f) SEM images of the vertically aligned

SiNW arrays catalyzed using the Au mesh with thicknesses of 15, 30, and 45 nm, respectively, for 10 min at 22°C. For the SEM observation, the samples were tilted by 15°. Figure 6 Relationship of the thickness of the Au film and the etching rate of the Si substrate. Mechanism for difference in the etching rate The result above GNS-1480 mw is the first to cite the difference in the silicon etching rate induced using a Au film with different thicknesses. The exact mechanism is not clear at the moment. The etching rate might

be controlled by the mass transfer process of the reagent and the by-product [13, 14]. A short diffusion path facilitating the rapid mass transfer of the reagent and the by-product is expected to result in a high etching rate. Figure 7a schematically illustrates the possible diffusion paths of the reagent and the by-product in the Si GBA3 etching process. In path I, the reagent and the by-product diffuse along the interface between the Au film and the Si, which signifies that the etching rate decreases with the increase in the lateral size of the Au catalyst because of the long lateral diffusion distance. In path II, the Si atoms underneath the Au are dissolved in the Au and then diffuse through the Au film to the Au/solution interface where the silicon atoms are oxidized and etched away [14, 20]. On one hand, if the etching rate is dominated by the mass transfer through path I during the chemical etching, a thick Au mesh should lead to a low etching rate because of the increasing lateral size of the Au catalyst caused by the shrinking of the holes induced by the closure effect (see Figure 2).

In order to get a deeper insight into the interference phenomenon, we have performed non-equilibrium Green’s function calculations using the ground-state electron density (of the molecules in gas phase) obtained from the density functional theory. In Figure 4, the calculated transmissions through the π-systems of both molecules are shown. At energies between the HOMO and LUMO levels, the transmission of the meta-OPV3 molecule is more than an order of magnitude smaller than that of a para-OPV3, with an anti-resonance occurring at 4.56 eV, where the transmission drops

substantially. This drop is Entospletinib clinical trial caused by the destructive interference between transmission coefficients of different selleck screening library orbitals. In the Landauer formalism, the charge propagation through molecules can be described as a transmission through different molecular orbitals [7]. Using the non-equilibrium Green’s function formalism, it is possible to separate the total transmission into contributions from the individual molecular orbitals. Since these contributions are complex (i.e., they have an amplitude and a phase), interference effects can arise when transmission through different orbitals are combined. Figure 4 Calculated transmissions through the π-systems Momelotinib clinical trial of both molecules. (a) Calculated transmission of para- and meta-OPV3 derivatives in gas phase. (b) Amplitude and phase of the

transmission through the HOMO and LUMO of a para-OPV3 molecule. (c) Amplitude and phase of the transmission through the HOMO and LUMO of a meta-OPV3 molecule. The amplitudes of the transmissions are approximately the same for both molecules, however, the phase of the

transmission through the LUMO differs by π from para- to meta-OPV3, while the phase selleck chemical of the HOMO is the same (see Figure 4b,c). This results in constructive interference for a para-OPV3 molecule when the transmission through the HOMO and LUMO are combined. For meta-OPV3 molecule, the phase shift results in destructive interference between the HOMO and LUMO transmission, as evident from the drop in the full transmission plot (Figure 4a). It should be noted that also the HOMO-1 and LUMO+1 orbitals contribute to the transmission within the HOMO-LUMO gap. The phase behavior of these orbitals is the same as for the HOMO and LUMO, i.e., constructive and destructive interference for para- and meta-OPV3 molecules, respectively. Note that the transmission of the meta-OPV3 does not go to zero at the anti-resonance due to the contributions from the HOMO-2 and HOMO-3 orbitals. This analysis therefore confirms the occurrence of constructive and destructive interferences in the molecules studied experimentally. Conclusion In conclusion, we have shown that the low-bias conductance through a single meta-OPV3 molecule is one order of magnitude smaller that through a para-OPV3 one.

BMC Genomics 2011, 12:261.PubMedCrossRef 27. Petersen L, Bollback J, Dimmic

M, Hubisz M, Nielsen R: Genes under positive selection in Escherichia coli. VX-689 cost Genome Res 2007,17(9):1336–1343.PubMedCrossRef 28. Farfán M, Miñana-Galbis D, Fusté M, Lorén JG: Divergent evolution and purifying selection of the flaA gene sequences in Aeromonas. Biol Direct 2009, 4:23.PubMedCrossRef 29. Jiggins F, Hurst G, Yang Z: Host-symbiont conflicts: positive selection on an outer membrane protein of parasitic but not mutualistic Rickettsiaceae. Mol Biol Evol 2002,19(8):1341–1349.PubMedCrossRef 30. Snijder H, Ubarretxena-Belandia I, Blaauw M, Kalk K, Verheij H, Egmond M, Dekker N, Dijkstra B: Structural evidence for dimerization-regulated activation of an integral membrane phospholipase.

Nature 1999,401(6754):717–721.PubMedCrossRef find more 31. Gancz H, Censini S, Merrell D: Iron and pH homeostasis intersect at the level of Fur regulation in the gastric pathogen Helicobacter pylori. Infect Immun. Infect Immun 2006,74(1):602–614. 32. Reid A, Pandey R, Palyada K, Whitworth L: E D, Stintzi A: Identification of Campylobacter jejuni genes contributing to acid adaptation by transcriptional profiling and genome-wide mutagenesis. Appl Environ Microbiol 2008,74(5):1598–1612.PubMedCrossRef 33. Tannaes T, Dekker N, Bukholm G, Bijlsma J, Appelmelk B: Phase variation in the Helicobacter pylori phospholipase A gene and its role in acid adaptation. Infect Immun 2001,69(12):7334–7340.PubMedCrossRef 34. Padhi A, Verghese Napabucasin supplier B, Otta S: Detecting the form of selection in the outer membrane protein C of Enterobacter aerogenes strains and Salmonella species. Microbiol Res 2009,164(3):282–289.PubMedCrossRef 35. Oleastro M, Cordeiro R, Ménard A, Yamaoka Y, Queiroz D, Mégraud F,

Monteiro L: Allelic diversity and phylogeny of homB, a novel co-virulence marker Suplatast tosilate of Helicobacter pylori. BMC Microbiol 2009, 9:248.PubMedCrossRef 36. Pride D, Blaser M: Concerted evolution between duplicated genetic elements in Helicobacter pylori. J Mol Biol 2002,316(3):629–642.PubMedCrossRef 37. Cao P, Lee K, Blaser M, Cover T: Analysis of hopQ alleles in East Asian and Western strains of Helicobacter pylori. FEMS Microbiol Lett 2005,251(1):37–43.PubMedCrossRef 38. Yamaoka Y, Orito E, Mizokami M, Gutierrez O, Saitou N, Kodama T, Osato M, Kim J, Ramirez F, Mahachai V, et al.: Helicobacter pylori in North and South America before Columbus. FEBS Lett 2002,517(1–3):180–184.PubMedCrossRef 39. Avasthi T, Devi S, Taylor T, Kumar N, Baddam R, Kondo S, Suzuki Y, Lamouliatte H, Mégraud F, Ahmed N: Genomes of two chronological isolates (Helicobacter pylori 2017 and 2018) of the West African Helicobacter pylori strain 908 obtained from a single patient. J Bacteriol 2011,193(13):3385–3386.PubMedCrossRef 40.

Figure 1 Total ion chromatogram of crude serum organic extract. (A) Total ion current of bulk serum following liquid/liquid extraction and HPLC-coupled mass spectrometry as explained in the methods. (B) Extracted mass spectra of all masses from (A). (C) Extracted ion chromatograms of GTAs 446, 448 and 450 from the total ion current shown in A. (D) Cell proliferation, as assayed by MTT, for SW620 cells treated with up to 80 ug/ml of the crude serum extract. Organic serum extract was next subjected to flash

column chromatography as described in the methods, resulting in 12 fractions which were subsequently analyzed by find more SP600125 molecular weight HPLC-MS to determine GTA content. Although other components were present in all the fractions, only fraction 9 out of the 12 was enriched for the C28 GTAs (referred to as the GTA+ve fraction). A GTA negative control fraction (fraction 8, lacking any detectable GTAs) was also selected PND-1186 cost for the studies described below. Representative total ion chromatograms, extracted mass spectra and selected ion chromatograms of the three C28 GTAs for the GTA-ve and GTA+ve fractions are shown in Figures 2A and 2B, respectively. By comparing the sums of the selected ion chromatograms of the three GTAs to the total ion currents, we estimated that the GTA+ve fraction contained approximately 21% C28 GTAs while the GTA-ve fraction had no detectable

levels (bottom panel of Figures 2A and 2B). The non-GTA background components for both fractions were similar, and the most abundant non-GTA components in the GTA+ve fraction were also the most abundant components in the GTA-ve fraction. Therefore, the two fractions were compositionally similar

other than the 21% GTA content of the GTA+ve fraction, which represented an approximately 143-fold enrichment Carnitine palmitoyltransferase II of the three C28 GTA metabolites over the crude organic serum extract (as shown in Figure 1A). These fractionations were repeated several times with consistent results. We therefore concluded that the fractions were sufficiently matched for investigating biological activity as described below. For comparison, the relative levels of the three C28 GTAs from 40 pooled CRC patients’ serum and serum from 40 matched control subjects is shown in Figure 2C. Figure 2 Mass spectrometry characterization of semi-purified GTA-ve and GTA+ve extracts. (A) Crude serum extract (as shown in Figure 1) was subject to flash column chromatography as described in the methods resulting in two adjacent eluates, one positive and one negative for the presence of GTAs. The total ion chromatogram (top), extracted mass spectra (middle), and extracted ion chromatograms for three GTAs (GTA446, 448 and 450; bottom) of the GTA-ve fraction. (B) Same as (A) for the GTA+ve fraction. (C) For comparison, the extracted ion chromatograms of GTA446, 448 and 450 from the extracts of serum pooled from 20 CRC patients and 20 controls is shown.

In Danish postmenopausal women, the

Ile568Asn loss-of-function polymorphism was associated see more with 10-year vertebral fracture incidence and increased rate of bone loss [16]. In contrast, we, like two other association studies [17, 20], did not find any association between the Ile568Asn loss-of-function polymorphism and BMD. However, only two women homozygous for the variant allele could be identified in this study. Since both the Arg307Gln and Ile568Asn were previously selleck products showed to be associated with either decreased BMD and/or fracture risk, the observed low prevalence of these SNPs in our fracture cohort is contrary to our expectations. The variant allele of the Gly150Arg polymorphism in our study was associated with decreased lumbar spine BMD, supporting the results found by Husted and colleagues [17], who observed reduced total hip BMD values in subjects carrying the 150Arg allele. This effect on BMD might be explained by its complete loss-of-function effect on the P2X7R [25, 32]. In line with several in vitro studies which showed that the variant allele of the Glu496Ala polymorphism was associated with a loss of receptor function [16, 23, 28, 33, 34], human cohort studies showed this polymorphism to be associated with decreased BMD values in both men and women [17] and increased fracture incidence over 10 years after menopause [16]. In concordance with these findings, we also found significantly decreased

BMD values at the total hip in women with at least one variant allele of the

Glu496Ala Napabucasin molecular weight polymorphism. Furthermore, analysis of haplotypes containing the Glu496Ala polymorphism (i.e. haplotype P2X7-3) also showed a significant association with decreased BMD values at the lumbar spine. This is in line with the results found by Stokes et al. [24], indicating that this haplotype is associated with decreased receptor function. The Suplatast tosilate studied P2X7R SNPs mostly affect the lumbar spine. Since bone turnover is primarily taking place on the bone surfaces and the changes in BMD due to the P2X7R SNPs are relatively small, one possible explanation for affecting this particular skeletal site could be that trabecular bone is lost more rapidly than cortical bone. As the amount of trabecular bone is higher in the vertebrae than in the hip, the bone loss will be most pronounced in the vertebral spine. The present study has several limitations. First, our study population is not population-based, as the recruitment strategy was based on the presence of a fracture. The prevalence of low BMD is, therefore, expected to be higher in our study population than in the general population. Furthermore, if the studied P2 receptor SNPs could affect fracture risk either directly or indirectly (independent of BMD) then the prevalence of this particular SNP would also be expected to be higher in our study sample than in the general population. This could potentially lead to bias in the results in the sense that extrapolation to the general population is compromised.

Moreover the low value of the standard error (0.2 pfu/g) of the phage titer after two days of treatment demonstrated that there

were small variations in the dose of phage that each bird received. Figure 4 Numbers of SGC-CBP30 datasheet Campylobacter jejuni 2140CD1 (a) and phages (b) in faeces from broilers orally administered a phage cocktail by gavage. Thirty day-old chicks were inoculated with Campylobacter jejuni 2140CD1. One week later the birds were randomly assigned to a treated group or an untreated group and were inoculated by oral gavage with antacid containing 1 × 106pfu of a phage cocktail, or antacid only respectively. Faecal samples were collected from all birds at intervals and Campylobacter and phages enumerated. Error bars represent the standard error of the mean. At 2 dpa, 4 dpa and 7 dpa there is a significant difference between control and infected group at P GSK2126458 purchase < 0.05. Figure 5 Numbers of Campylobacter coli A11 (a) and phages (b) in faeces from broilers orally administered phage by food or by oral gavage. Forty-five, day-old chicks were inoculated with Campylobacter coli A11. One week later the birds were randomly assigned to one of three groups, a non-treated group and two treated groups: a group receiving the phage cocktail by oral gavage; and a group receiving the phage cocktail in feed. Birds were inoculated with antacid only, antacid containing 1 × 106pfu

phage cocktail or antacid followed by feeding with the phage cocktail laced with 1.5 × 107pfu, respectively. Faecal samples were collected from all birds at intervals and Campylobacter and phages enumerated. Error bars represent the standard error of the mean. At 1 dpa, Vistusertib datasheet 2 dpa, 4 dpa and 7 dpa there is a significant difference between control and infected groups at P < 0.05. Table 1 Difference between the geometric means of the Campylobacter Leukocyte receptor tyrosine kinase titre from broilers with and without the phage cocktail administration Experiment Administration route Campylobacter titre (log10cfu/g)     Day 2 Day 4 Day 7 Experiment

1 Oral Gavage 1.74 2.34 2.18 Experiment 2 Oral Gavage 1.25 1.58 1.69   Feed 2.00 1.45 1.96 The phage titers from faecal samples of the chicks infected with C. jejuni and C. coli were log10 5.3 pfu/g and log10 3.4 pfu/g for Experiment 1 and Experiment 2 respectively. These values remained approximately constant throughout the experimental period showing that phages delivered to chicks (either by oral gavage or in feed) were able to replicate and therefore able to reduce the Campylobacter populations. Previous studies [40, 41] have used the number of Campylobacter in the caecal contents of the birds as a measure of Campylobacter colonisation levels in the GI tract of chickens [41, 34]. Although this may be a representative of colonisation levels, the animals must be killed and dissected to obtain the sample. This can lead to the use of an excessive number of birds when multiple time points are required to evaluate phage levels over the lifetime of the bird.