The MIC was defined as the lowest concentration of metal that allowed no bacterial growth. For each metal and bacterial strain, at least three independent experiments were carried out. β-galactosidase assay Enzyme activities were measured from bacteria grown overnight in LB or in LB supplemented with different metal salts (concentrations are specified in Results). β-galactosidase activity was assayed according to a previously described protocol [68]. Western blotting Cell lysates were prepared from bacteria grown overnight in LB or in LB supplemented with either 0.6 mM ZnSO4

or 0.15 mM FeSO4. Equal amounts of total protein (3 μg) were separated by Tricine-SDS-PA gel electrophoresis, followed by protein transfer to a nitrocellulose membrane. For Western blotting, the membranes were probed with ColR-specific polyclonal antibodies, followed by treatment check details with alkaline phosphatase-conjugated goat anti-rabbit immunoglobulin G. The blots

were developed using bromochloroindolyl phosphate/nitro blue tetrazolium (BCIP/NBT). Acknowledgments We are grateful to Hedvig Tamman, Andres Ainelo and Hanna Hõrak for critically reading the manuscript. We thank Külliki Holtsmann for assistance in the cloning and Peeter Hõrak and Riho Teras for advice in the statistical analysis. This work was supported by the grant 7829 from the learn more Estonian Science Foundation, by Targeted Financing Project TLOMR0031 and by Institutional buy PX-478 Research grant IUT20-19.

Electronic supplementary material Additional file 1: Table S1: Bacterial strains and plasmids. (DOCX 56 KB) Additional file 2: Table S2: The oligonucleotides. (DOCX 22 KB) Additional file 3: Table S3: The oligonucleotide pairs used in two sequential PCRs for site-directed mutagenesis of colS. (DOCX 18 KB) References 1. Andreini C, Bertini I, Cavallaro G, Holliday GL, Thornton JM: Metal ions in biological catalysis: from enzyme databases to general principles. J Biol Inorg Chem 2008,13(8):1205–1218.CrossRef 2. Touati D: Iron and oxidative stress in bacteria. Arch Biochem Biophys 2000,373(1):1–6.PubMedCrossRef 3. Imlay JA: Iron-sulphur cAMP clusters and the problem with oxygen. Mol Microbiol 2006,59(4):1073–1082.PubMedCrossRef 4. McDevitt CA, Ogunniyi AD, Valkov E, Lawrence MC, Kobe B, McEwan AG, Paton JC: A molecular mechanism for bacterial susceptibility to zinc. PLoS Pathog 2011,7(11):e1002357.PubMedCentralPubMedCrossRef 5. Outten CE, O’Halloran TV: Femtomolar sensitivity of metalloregulatory proteins controlling zinc homeostasis. Science 2001,292(5526):2488–2492.PubMedCrossRef 6. Changela A, Chen K, Xue Y, Holschen J, Outten CE, O’Halloran TV, Mondragon A: Molecular basis of metal-ion selectivity and zeptomolar sensitivity by CueR. Science 2003,301(5638):1383–1387.PubMedCrossRef 7. Nies DH: Efflux-mediated heavy metal resistance in prokaryotes. FEMS Microbiol Rev 2003,27(2–3):313–339.PubMedCrossRef 8.

Interestingly, reads assigned to Gardnerella were only identified in 3/8 urine samples, even though this genus was the 3rd most abundant group in the pooled sequence

AMN-107 mw dataset for both the V1V2 and V6 regions (Figure 2A). Three other genera and a group of 5 genera were identified by reads belonging to 3 or 2 urine samples, respectively. 24 genera were only detected in 1 out of the 8 samples. Species richness and diversity estimates of the female urine microbiota Bacterial taxonomic richness and diversity varied greatly among urine samples investigated in this study. Community richness and diversity were determined using rarefaction plots, Chao1 and Shannon index estimations (Figure 3 and Table 2). Figure 3 Number of OTUs as function of the total number of sequences. A and B: Rarefaction curves of individual samples for the V1V2 (A) and the V6 datasets (B). Curves were generated at 3% genetic difference using MOTHUR v1.17.0 [39]. C and D: Rarefaction curves of the pooled dataset for both V1V2 reads (C) and V6 reads (D). OTUs with ≤3%, ≤6% and ≤10% pairwise sequence Emricasan difference generated using MOTHUR v1.17.0 [39] are assumed to belong to the same species, genus and family, respectively. Rarefaction curves were generated for 3% genetic difference level (e.g., at the species level).

The number of OTUs calculated for the eight individual samples ranged from 20-504 and 63-499 OTUs for the V1V2 and V6 regions, this website respectively Glycogen branching enzyme (Figure 3A, B and Table 2). OTU numbers of the total bacterial

community in the female urine at 3% difference for the V1V2 sequence pool was calculated to 1209 OTUs and to 1435 OTUs for the V6 sequence pool (Figure 3C, D and Table 2). Furthermore, total unique OTUs for the V1V2 pooled reads were 1354 and for the V6 pooled reads 2069 (Table 2). To compare the diversity between the eight different urine samples, the Shannon diversity index was determined both with the original, and with normalized numbers of sequences (Table 2). There was no substantial difference between the two Shannon indices calculated for the same sample. Discussion In this work we sequenced two different variable regions of 16S rDNA isolated from eight culture-negative urine samples. Urine samples are at risk of contamination by the bacterial flora of the female urogenital system [82, 83], therefore sampling of mid-stream urine was performed by the clean catch method, under guidance of an experienced urotherapy nurse. To avoid further bacterial growth, which could skew the results, the samples were kept on ice and analyzed within an hour. Amplicon lengths used here exceed the typical fragment size (150-200 bp) of circulating cell-free DNA in urine [84], thus reducing the frequency of such DNA in our analyses.

BioMetals 2010, 23:431–439.PubMedCrossRef 43. Schägger H: Tricine–SDS-PAGE. Nat Protoc 2006, 1:16–22.PubMedCrossRef 44. Iwatani S, Zendo T, Yoneyama F, Nakayama J, Sonomoto K: click here Characterization and structure analysis of a novel bacteriocin, lacticin Z, produced by Lactococcus lactis QU 14. Biosci Biotechnol Biochem 2007, 71:1984–1992.PubMedCrossRef Competing interests The authors declare that

they have no competing interests. Authors’ contributions XH carried out the reference collection and analysis, most experimental running of whole expermental work; RM participated partial experimental design, method improvement and partial paper writing; YZ was charge of expression and mainly fermentor JNJ-26481585 clinical trial running; DT was charge of codon optimization and all materials preparation; XW was charge of partial DNA cloning and PCR techinque, and partial result analysis; DX participated partial peptide purification; JH corrected partial techincal design on microbiological methods; JW participated Selleck A 1331852 and coordinated all sections of this work, design and running, results analysis and disscussion, paper writing and correction. All authors read and approved the final manuscript.”
“Background The Gram positive bacterium Streptococcus

pneumoniae frequently colonizes the nasopharynx but can also invade the host causing serious illnesses such as pneumonia, meningitis or bacteraemia [1]. A principal virulence factor of S. pneumoniae is the polysaccharide capsule protecting it from host immune defences by interfering with the deposition of complement and therefore opsonophagocytosis [2-4]. The capsule is the target of all currently available pneumococcal vaccines including the 13-valent pneumococcal

conjugate vaccine (PCV13) for children. The biochemical structure and linkage of repeating polysaccharide subunits determines the serotype of encapsulated strains. So far, more than 90 different serotypes have been identified [5-11] which differ in the type and number of genes encoding the proteins responsible for transcription, Bcl-w polymerization, elongation and export of the capsule. For almost all serotypes the capsule-encoding operon is located between non-capsule genes dexB and aliA [6,12,13]. The first four genes cpsA, cpsB, cpsC and cpsD are thought to play a role in regulation of capsular production and are largely conserved between serotypes [14,15]. Despite the importance of the capsule as a virulence factor, nonencapsulated pneumococci occur and in the nasopharynx may represent around 15% of pneumococcal isolates [16]. Nonencapsulated pneumococci are generally considered not to be virulent but are associated with outbreaks of conjunctivitis [17-19]. Although lacking the protection from opsonophagocytosis which a capsule affords, the absence of capsule may confer advantages.

Sections were examined with a Philips CM 100 TEM (Eindhoven, Holland) and images were recorded with an OSIS Veleta 2 k × 2 k CCD camera at the Core Facility for Integrated Microscopy of the University of Copenhagen, Denmark. Statistical analysis A Student’s t-test (run with

Excel software) was used to compare the experimental groups that were subjected to various stresses and the non-stressed controls. P-values of <0.05 were considered statistically significant. Acknowledgements This study was supported in part by the Pathos Project funded by the Strategic Research Council of Denmark (ENV 2104-07-0015) and Otto Mønsted Foundation, and in part by the Natural Sciences and Engineering Research Council of Canada (RGPIN 240762–2010 to Dr. Creuzenet). We thank Dr. Valvano for sharing the tissue culture facility and microscopes, and Dr. Koval for the use of her microscope. We GSK1210151A concentration also thank R. Ford for critical reading of this manuscript. References 1. Newton JM, Surawicz CM: Infectious gastroenteritis and colitis diarrhea. In Diarrhea, clinical gastroenterology.

selleck chemicals llc Edited by: Guandalini S, Vaziri H. New York: Humana Press; 2011:33–59. 2. Domingues AR, Pires SM, Halasa T, Hald T: Source attribution of human campylobacteriosis using a meta-analysis of case–control studies of sporadic infections. Epidemiol Infect 2012, 140:970–981.PubMedCrossRef 3. Beery JT, Hugdahl MB, Doyle MP: selleck kinase inhibitor Colonization of gastrointestinal tracts of chicks by Campylobacter jejuni. Appl Environ Microbiol 1988, 54:2365–2370.PubMed 4. Candon HL, Allan BJ, Fraley CD, Gaynor EC: Polyphosphate kinase 1 is a pathogenesis determinant in Campylobacter jejuni. J Bacteriol 2007, 189:8099–8108.PubMedCrossRef 5.

Friedman CR, Neimann J, Wegener HC, Tauxe RV: Campylobacter jejuni infections in the United States and other industrialized nations. In Campylobacter. vol. 2, 2 edition. Edited by: Nachamkin I. Washington, DC: ASM Press; 2000:121–138. MJB 6. Klančnik A, Guzej B, Jamnik P, Vučković D, Abram M, Možina Rebamipide SS: Stress response and pathogenic potential of Campylobacter jejuni cells exposed to starvation. Res Microbiol 2009, 160:345–352.PubMedCrossRef 7. Jackson D, Davis B, Tirado S, Duggal M, van Frankenhuyzen J, Deaville D, Wijesinghe M, Tessaro M, Trevors J: Survival mechanisms and culturability of Campylobacter jejuni under stress conditions. Antonie van Leeuwenhoek 2009, 96:377–394.PubMedCrossRef 8. Alter T, Scherer K: Stress response of Campylobacter spp. and its role in food processing. J Vet Med B Infect Dis Vet Public Health 2006, 53:351–357.PubMedCrossRef 9. Fields JA, Thompson SA: Campylobacter jejuni CsrA mediates oxidative stress responses, biofilm formation, and host cell invasion. J Bacteriol 2008, 190:3411–3416.PubMedCrossRef 10. Ma Y, Hanning I, Slavik M: Stress-induced adaptive tolerance response and virulence gene expression in Campylobacter jejuni. J Food Safety 2009, 29:126–143.CrossRef 11.

Similarly, in case of cucumber plant’s endogenous GAs, endophytic fungal application selleck inhibitor have rescued GAs biosynthesis as the level of bioactive GAs were much

pronounced compared to sole NaCl treated plants. Phytohormones, like GAs have been widely known for their role in plant growth and various developmental processes during plant’s life cycle [1, 3, 57]. Normal response of a plant to stress is to reduce growth by inter alia increasing ABA content and reducing GAs [56, 57]. GA-deficient plants are more susceptible to stress than those with higher levels of this hormone [56]. The higher amount of GA12 in endophyte-treated plant under salinity stress elucidates the activation of GAs biosynthesis pathway, while higher production of GA3 and GA4 confirm plant growth maintenance during stress condition. Thus, by maintaining GAs and, therefore, growth under stressful conditions, the endophyte is having a detrimental effect on the plant long-term survival. There Selleck PD0332991 are many previous reports

showing the ameliorative effects of exogenous application of GAs (GA3/GA4) and IAA on cucumber growth under LDC000067 molecular weight abiotic stress [58–60], while very little or no information’s are available on the regulation of plant endogenous hormones in association with phytohormones producing endophytic fungi under abiotic stress conditions. Some physiological evidence suggests that plants infected with endophytic fungi often have a distinct advantage against biotic and abiotic stress over their endophyte-free counterparts [61]. Beneficial features have been offered in infected plants; including drought acclimisation [62, 63] enhanced tolerance to stressful factors such high salinity [12]. Foliar application of GAs has been known for its role in plant stem elongation and mitigation of abiotic stress [54–60] while the same was observed in current study that endophytes producing GAs triggered the adverse effect of salinity stress. Conclusion P. formosus LHL10 produced many physiologically active and inactive GAs and IAA, which helped the Waito-C and Dongjin-byeo rice plants to

grow well and significantly mitigated the negative impacts of salinity stress on cucumber plants. The P. formosus LHL10 also minimized the lethal effects of salt stress on cucumber leaf tissues as evidenced from EL, RWC, photosynthesis rate, Dipeptidyl peptidase nitrogen assimilation, antioxidant activity and lipid peroxidation. The cucumber plants inoculated with P. formosus LHL10 have ameliorated their growth by possessing lower levels of stress responsive endogenous ABA and elevated GAs contents. Current study reveals that such endophytic fungal interactions can improve the quality and productivity of economically important crop species. However, the favourable role of this fungus still needs to be investigated under field conditions. Acknowledgements The present research work was funded by the Eco-Innovation Project, Korean Government’s R & D program on Environmental Technology and Development.

Patient with GCS ≤ 8 4. Gunshot wound to the head, neck, or torso 5. Need for blood transfusion en route to hospital or in the ED In order to assess the efficiencies and human resource implications of trauma activations not focusing on traditional thoracoabdominal injuries, a retrospective review of trauma patient resuscitations with head injuries requiring intubation or with a GCS < 13 in whom a CT scan was obtained. Patients were identified from the FMC Trauma Registry as having been admitted between April 01 2008 and March 31, 2009. To qualify for the trauma registry a patient must have an

Injury Severity Score (ISS) > 12 and be admitted to the trauma centre or die in the emergency department of the trauma centre. From the eligible cohort (186 TBI patients who met the inclusion criteria), a convenience sample of 101 charts was selected by medical records click here for review. Demographic data reviewed included age, gender, emergency department (ED) admission date, ED admission time, injury description, Maximum Abbreviated Injury Scale (MAIS) Head, Injury Severity Score (ISS), scene GCS, trauma centre GCS, patient intubation status at the time of the GCS was calculated, whether FTA was activated, time of trauma team activation, trauma surgeon, intensive care unit (ICU) admission, ICU length of stay (LOS), and discharge status. The following

data was collected directly from the charts: whether patient had a CT done at previous hospital, arrival time of trauma IDO inhibitor surgeon at FTA, CT head date and time, picture archiving and communication (PACS) time of CT head, electronic medical record time of CT Head, whether there was a reason for CT delay, and if there was a reason for delay then which interventions were done, interventions date, interventions time, and any comments about the patient. We initially sought to study the times until completion

of the CT head. However review of the time imprints embedded with the CT images in PACS was found to be non-sensical clinically, and a subsequent review of the electronic clocks in the CT scanners found them to be significantly inaccurate. Thus, the charted time the patient left the trauma bay for the CT scanner Non-specific serine/threonine protein kinase was used instead. The “Time from ED admission to CT head (TTCTH-unqualified)” was defined as the unqualified number of minutes from ED admission until the patient left for the CT scan. The “Time in ED after this website airways were secure (TTCT-after airways secure)” was defined as either the time in the ED until leaving for CT head if intubated pre-hospital or never intubated, or as the time in the ED after ED intubation until leaving for CT head. For those re-intubated in ED, the time from re-intubation until leaving for CT was used for this designation.

cruzi strains. Amastin amino acid sequences from CL Brener, Esmeraldo and Sylvio X-10 strains were used to generate a tree rooted with an α-amastin sequence from Crithidia sp. Bootstrap values followed by branch length are shown in the major basal nodes. In spite of the sequence divergence, an alignment of polypeptide sequences belonging to all amastin sub-families shows increased amino acid conservation within the putative hydrophobic Repotrectinib in vivo transmembrane domains. Within the predicted extracellular domains, two highly conserved cysteine and one tryptophan residues, that are part of the 10 amino acid “amastin signature” [8], may be critical for amastin function (Additional file 1: Figure S1B). On the other hand, the more variable

sequences present in the two predicted extracellular, hydrophilic domains suggest that this portion of the protein, which, in amastigotes, are in contact with the host cell cytoplasm, may interact with distinct

host cell proteins. Because the assembly of CL Brener genome does not include its complete sequence, we conducted a read-based analysis to estimate the total number of amastin genes in this strain of the parasite. It is well known that the assembly of the CL Brener genome is only accurate for non-repetitive regions, and AR-13324 for tandemly repeated genes, misassembles frequently occurred since most repetitive copies usually collapse into one or two copies. Therefore, we used the entire dataset of reads generated by the Tri-Tryp consortium to select reads 3-oxoacyl-(acyl-carrier-protein) reductase containing sequences homologous to amastin and, based on a 13 × genome coverage [13], we estimated a total number of 14 copies of amastin genes, 2 βATM Kinase Inhibitor research buy -amastins and 12 δ-amastins in the CL Brener genome. Similar analyses performed with sequencing reads generated by Franzen et al. (2011) [14] from the genome of Sylvio X-10 indicated a comparable number of copies in the genome of this

T. cruzi I strain. In the current assembly of the CL Brener genome, amastin genes are shown to be organized in three loci on chromosomes 26, 32 and 34. Forty one pairs of homologous chromosomes (corresponding to the Esmeraldo-like and non-Esmeraldo haplotypes) have been assembled using the majority of the contigs and scaffolds generated by the Tri-Tryp consortium and inferences from synteny maps with the fully assembled T. brucei genome [15]. Based on the chromosome assemblies described by Weatherley et al. [15], three copies of δ-amastins are presented on chromosome 34 as a tandem array with alternating copies of tuzin genes. Interestingly, the divergent copy of δ-amastin (which has the Esmeraldo-like δ-Ama40 allele and the non-Esmeraldo allele δ-Ama50) is found as a single sequence linked to one tuzin pseudogene on chromosome 26. In a third chromosome, two copies of β-amastins are linked together without the association with tuzin genes. This gene organization is consistent with the analyses described by Jackson (2010) [9], who found tuzin genes associated only with δ-amastins.

Figure 1 Expression of S. aureus protein A (SPA) on the cell surface of L. monocytogenes strain Δ trpS,aroA,inlA/B,int ::P hly – spa × pFlo- trpS (Lm-spa + ). (a) Western blot analysis with polyclonal goat-anti-Protein

A antibody of protein extracts from ΔtrpS,aroA,inlA/B × Fedratinib pFlo-trpS (Lm-spa-, lanes 1 and 2) and Lm-spa+ (lanes 3 and 4); lanes 1 and 3: cell surface protein extracts; lanes 2 and 4: internal protein extracts. The arrow indicates the position Quisinostat solubility dmso of SPA in the SDS-PAGE. (b) Immunofluorescence micrographs showing specific binding of antibody Fc-part to SPA on the surface of Lm-spa+. Lm-spa+ were incubated with polyclonal anti-OVA antibody and stained with OVA-FITC protein (vii-ix). Lm-spa- Smoothened Agonist cell line stained with antibody and OVA-FITC (i-iii) and Lm-spa+ stained without antibody but with OVA-FITC protein (iv-vi) were used as negative controls. Phase contrast pictures are shown in the left column; FITC-stained images in middle column; picture overlays in the right column.

(c) Flow cytometry quantifying the specific Fc-mediated antibody binding to SPA on the surface of L. monocytogenes strains. Mid-logarithmic grown bacteria were stained with polyclonal FITC-conjugated rabbit-anti-goat immunoglobulin G (H+L). Grey area indicates strain Lm-spa-, while the white area indicates strain Lm-spa+. (d) Western blot analysis was used for indirect quantitation of protein A on the surface of Lm-spa+. 5 × 108 bacteria were incubated simultaneously with antibody directed against native albumin and an excess of albumin. After incubation bacteria were washed and the amount of albumin bound to the bacteria via antibody was quantified by Western blot analysis with a primary antibody directed against denatured albumin. In the right lane 10 ng of pure serum albumin was applied as control. Fc-mediated binding of antibodies to SPA on the surface of L. monocytogenes The functionality of Fc-mediated binding of antibodies to SPA on the surface of Lm-spa+ was first tested by immunofluorescence microscopy of Lm-spa- and Lm-spa+ after incubation of the bacteria with polyclonal rabbit antibodies directed against ovalbumin (OVA). After addition of FITC-conjugated OVA

no fluorescence was detected with Lm-spa-, while the else Lm-spa+ strain showed a strong fluorescence (Figure 1B). A more quantitative analysis of SPA expression was performed by flow cytometry after staining Lm-spa+ and Lm-spa- with FITC-conjugated rabbit-anti-goat-antibodies. Lm-spa- bacteria showed no staining while the Lm-spa+ bacteria were stained almost completely (Figure 1C). In addition, the number of SPA molecules per bacterial cell was determined indirectly. For this goal Lm-spa+ was incubated simultaneously with a primary antibody against native albumin as model protein in the presence of an excess of albumin. The bacteria bound the albumin-loaded antibody to their surface via SPA and later on the amount of bound protein was quantified.

showing a statistically significant increase in PFS, resulting in a reduced risk of progression of about 30%. In the meta-analysis conducted by Valachis et al, improved PFS was statistically significant only in the subgroup of patients receiving taxanes (or anthracyclines in a part of the study RIBBON-1) in combination with Bevacizumab [33], this advantage not seem to get in combination with capecitabine, although the latter are grouped in heterogeneous populations with regard to the treatment line. In the meta-analysis conducted by Lee et al, with populations more correctly grouped by line of treatment rather than medication, the benefit of

the addition of Bevacizumab in PFS is restricted to first-line treatment [32]. Moreover, this analysis shows MM-102 cost a marginal but statistically significant benefit in overall survival in first line. At the last ESMO meeting, a meta-analysis of 530 elderly patients (older than 65 years) enrolled in the randomized trials ECOG 2100, AVADO and RIBBON-1, was presented [34]. Although that represent a subgroup analysis, even in these featured advanced breast cancer patients’ sample, bevacizumab in combination with chemotherapy was associated with significantly improved PFS versus chemotherapy alone (HR 0.67, p = 0.0030). Hypertension was more frequent with the addition of bevacizumab,

as expected; besides, no differences according to age were found. Another relevant issue that emerges from our analysis is that the prior exposure to treatments containing taxanes does not affect the efficacy of bevacizumab (Table

4). Indeed, the meta-regression analysis for either PFS FG-4592 ic50 or OS learn more clearly indicates that no significant correlation exists between the efficacy of bevacizumab and taxanes pre-treatment (p = 0.96 and p = 0.45, respectively). This finding is consistent with the ECOG-2100 and AVADO previous release [14, 15], and with the recently presented meta-analysis of patients from studies ECOG-2100, AVADO and RIBBON-1, previously treated with taxanes (paclitaxel, docetaxel or paclitaxel protein-bound) [35]. This analysis included only 311 patients from the group of patients Endonuclease treated with taxanes of the RIBBON-1 and AVADO who received bevacizumab 15 mg/kg. The addition of bevacizumab led to an improvement in PFS from 6.2 to 10.6 months (HR 0.50, 95% CI 0.36-0.69). In line with the data of the single trials and our analysis, the authors conclude that patients pretreated with taxanes are good candidates for retreatment with bevacizumab and taxane [35]. With regard to serious adverse events, the main significant toxicity against the addition of bevacizumab was hypertension (Table 3); this represents a common finding in all disease setting when this monoclonal antibody is adopted. Our analysis shows that a weighted average of 4.5% difference between the control arm and patients undergoing bevacizumab was found, corresponding to 22 patients to be treated for one harmed (Table 3).

fellah control worker (A) and Rifampin treated worker midguts (B). The bacteriocytes of treated worker are hardly visible. Figure 2 Endosymbiont number estimation in worker midguts, after 3 months of antibiotic treatment. find more workers from treated groups present a mean number of bacteria significantly lower than the control group (Mann-Whitney’s U-test = 179.00, Z = -3.48, p < 0.001). The bars represent the mean number of 16S rDNA molecules ± semi-quartile range. Evaluation of colony development Each colony was composed of at least one larva, pupa or worker and queen. Colonies composed only with the queen or colonies with a dying queen during the experiment

were excluded. After seven months, seven control colonies and nine treated colonies were kept for further analysis. Workers, larvae and pupae numbers were not significantly different during the first three months after the JNJ-26481585 mw beginning of the experiments. After this time, untreated colonies displayed more accentuated larvae production and had a higher number of adult workers (Fig 3a and 3c, see table 1, for all statistical results). Pupae number varied significantly throughout the time of the experiment but no difference between treated and control colonies was observed

(Fig 3b). The variation in workers numbers was significatively different IDO inhibitor between treated and control colonies with untreated colonies having more workers (Fig 3c). Table 1   ANOVA main effects Mean number Antibiotic × control Time Interaction larvae F1,112 = 10.12** F7,112 Y-27632 2HCl = 6.08*** F7,112 = 0.26 pupae F1,112 = 2.79 F7,112 = 2.52* F7,112 = 1.20 workers F1,112 = 5.53* F7,112 = 1.69 F7,112 = 0.75 Mean number of larvae, pupae and workers analysed by ANOVA. Significance levels are *P < 0.05, **P < 0.01 and ***P ≤ 0.001. Figure 3 Mean number of larvae (a), pupae (b) and workers (c), square-root transformed (± SE), for control and antibiotic-treated colonies. N = 7 and 9, respectively. Amount of Blochmannia endosymbiont versus encapsulation response When expressing encapsulation rate versus 16S rDNA molecules amount (as measure of Blochmannia amount in individual midgut), control

and treated colonies displayed different patterns of immune response. We found a significant positive correlation between encapsulation rate and bacteria amount in the ants from control colonies: the bacteria did facilitate the encapsulation response (Pearson’s r, p = 0.003, n = 27, Fig. 4). On the contrary, ants from treated colonies did not display a correlation between the amount of bacteria in the midgut and the encapsulation response (Pearson’s r, p = 0.92, n = 29, Fig. 4). Thus, it seems that antibiotic treatment eliminated the bacterial effects on the immune encapsulation response. An ANCOVA analysis with the encapsulation rate as independent variable showed that treated workers present a significant increase in encapsulation rate (F1,53 = 8.61, p = 0.005).