, 2001). Mcf toxins cause damage to the insect midgut after injection into larvae with loss of body turgor and a ‘floppy’ phenotype of the caterpillars (Dowling et al., 2004). Injection of PVCs destroys insect hemocytes, which undergo dramatic actin cytoskeleton condensation (Yang et al., 2006). Pir toxins act as binary proteins. Both PirA and PirB proteins are necessary for insecticidal activity. Injection of either PirA or PirB alone into caterpillars of Galleria is not associated with any mortality, and mixture of individual PirA and PirB preparations exhibits full activity against this insect (Waterfield et al., 2005). Histological examination of Plutella xylostella larvae fed

with recombinant Escherichia coli expressing PirA and PirB proteins reveals gross abnormalities of the midgut epithelium, with profound swelling and shedding of the apical membranes (Blackburn et al., 2006). PirAB toxins SB203580 clinical trial also show larvicidal activity

against mosquito larvae (Aedes aegypti and Aedes albopictus; Ahantarig et al., 2009). Binary toxins have also been reported in several other bacteria, including Clostridium botulinum C2 toxin, Clostridium difficile toxin (CDT), Clostridium perfringens iota (ι) toxin, Clostridium spiroforme toxin (CST), Bacillus anthracis edema and lethal toxins, as well as the Bacillus cereus vegetative insecticidal proteins (VIP; Barth et al., 2004). Normally, binary toxins consist of binding component and enzymatic component. The binding component recognizes a cell surface receptor and allows the internalization of the enzymatic component into the cytosol, and the enzymatic component catalyzes the reaction and induces Cabozantinib cell line the toxicity (Carman et al., 2011). Recently, a new binary toxin gene xaxAB from Xenorhabdus nematophila, a bacterial species closely related to P. luminescens, was cloned and sequenced. XaxAB toxin exhibited both

necrotic and apoptotic activities in both insect and mammalian cells in vitro. Incubations of sheep red blood cells with XaxAB showed that maximum hemolytic activity was obtained with equimolar concentrations of XaxA and XaxB. This binary toxin cannot be classified in any known family of cytotoxins on the basis of amino acid Reverse transcriptase sequences, locus organization, and activity features. The putative hemolysin loci, containing two closely linked genes similar to xaxAB, were also found to be present in the chromosome of Photorhabdus, Pseudomonas, and Yersinia (Vigneux et al., 2007). Analysis of the genomic sequence of P. luminescens TT01 (Duchaud et al., 2003) revealed that amino acid sequences encoded by plu1961 and plu1962 showed 76.8% and 74.9% similarity to XaxA and XaxB, respectively. To evaluate the biological activity of this potential binary toxin, plu1961 and plu1962 were cloned and expressed in E. coli. Both oral and injectable toxicities of Plu1961/Plu1962 were assayed against insect larvae. Cytotoxic effect of binary toxin was tested against insect midgut CF-203 cells and mammalian cell lines.

These two classes of HMGR share only 14–20% sequence identities. Class I HMGR differs from class II HMGR by having a ‘cis-loop,’ which is strictly conserved in class I HMGR and is involved in substrate binding. Recently, a number of reports have been published on the isolation of Actinobacteria from marine organisms. Screening of these marine-derived Actinobacteria has led to the discovery of many new bioactive metabolites. One typical example is the novel compound salinosporamide A (Feling et al., 2003), which is produced selleck chemicals by members of the genus Salinispora, and

has been identified as a proteasome inhibitor possessing anticancer activity. More than 70% of the Earth’s surface is covered by oceans inhabited by a high and as yet unexplored diversity of marine organisms. Marine sponges are of special interest as they are filter feeders and

assimilate bacteria during the filtration process. These marine sponges and seawater itself may support a number of undiscovered Actinobacteria, as is evident from culture-independent approaches such as denaturing gradient gel electrophoresis and 16S rRNA gene clone libraries (Zhang et al., 2006). Therefore, these uncultured marine Actinobacteria present a major resource for the discovery of new bioactive metabolites. Our group has recently engaged in the isolation of microorganisms, including fungi and Actinobacteria, from marine OSI906 sources. Some of the isolated microorganisms have been found to produce novel compounds, namely, JBIR-27, -28 (Motohashi et al., 2009a), JBIR-15 (Motohashi et al., 2009b), JBIR-37, -38 (Izumikawa et al., 2009), and JBIR-31 (Izumikawa et al., 2010). Among Actinobacteria, many novel members of the genus Streptomyces have been isolated, and these strains have been found to produce a number of novel compounds (S.T. Khan, T. Tamura,

M. Takagi & K. Shin-ya, unpublished data; S.T. Khan, H. Komaki, K. Motohashi, I. Kozone, A. Mukai, M. Takagi & K. Shin-ya, unpublished data). Thus, in the present study, we attempted to isolate Actinobacteria from marine organisms and sediments, screened the oxyclozanide strains for the presence of the hmgr gene as the marker of the mevalonate pathway, and isolated isoprenoid compounds from the cultures of these Actinobacteria. We collected 18 marine sponges, two marine sediments, and a tunicate sample from the sea near Tateyama, Chiba Prefecture, and from areas near Ishigaki Island, Okinawa Prefecture, Japan (Table 1). Samples were retrieved by scuba diving using sterile spades and were collected in plastic bags. The samples collected were processed on the same day as described below. Sponges and tunicate were rinsed three times with sterile natural seawater to remove the bacteria attached to the surface. These samples (wet weight: 20 g) were then either homogenized in a blender or cut into very small pieces using sterile scissors. Homogenized samples were resuspended in 30 mL of sterile seawater.

A laboratory-adapted

UPEC, strain 536 (Knapp et al., 1986), for which the genome sequence is available, was used as the primary experimental strain. Escherichia coli MG 1655 (Guyer et al., 1981) was used as a nonpathogenic control. Fresh clinical UTI isolates (prefix OF 5409, 6636, 5862, 6020, 6786, 6860, 6762, 6703, 5179, 5625, 5325, and 6869) were obtained from Auckland Hospital. A single colony was used to inoculate an overnight culture [RPMI 1640+10 μM ferric chloride (FeCl3)] and a portion of the culture was stored in 25% v/v glycerol at −80 °C. All subsequent testing was performed using UPEC strains recovered from −80 °C storage. Cells were grown in RPMI 1640 (Invitrogen, abbreviated as R) and R supplemented to a final concentration of 10 μM FeCl3 find more (abbreviated as RF), where stated. Other metal supplements were diluted as indicated from 10-mM stock solutions of MnSO4, ZnSO4, CuCl2, or NiCl2. Biorelevant iron Natural Product Library supplements were added as haemin (Acros Organics) at 10 μM, haemoglobin (Sigma) at 10 μM, ferritin (Sigma) at 0.5 μM, apo-transferrin

(Sigma), holo-transferrin (Sigma), and holo-lactoferrin (Sigma) at 0.6 μM. The following enzymes were added to cultures: amylase (Sigma) at 1600 U mL−1, cellulase (Sigma) at 13.8 U mL−1, and DNAse 1 (Sigma) at 90 U mL−1. All incubations were performed at 37 °C with shaking at 200 r.p.m. Inhibition of RNA synthesis was performed by the incubation of cultures with rifampicin at 100 μg mL−1 for 30 min before the addition of iron. Inhibition of new protein synthesis was achieved by the incubation of cultures with chloramphenicol at 35 μg mL−1 for 30 min before the addition of iron. Overnight cultures in RF were diluted 1 : 100 in the medium indicated and divided into 10-mL aliquots in V-bottomed polypropylene

50-mL tubes (Sarstedt). The tubes were incubated at 37 °C with shaking at 200 r.p.m. At given time intervals, one tube was used to measure aggregation. Bacterial aggregates were pelleted MRIP at 610 g for 2 min and the OD of the planktonic phase was measured at 600 nm (OD600 nm [planktonic]). The planktonic cells were returned to the original tube and all cells were pelleted at 2450 g for 5 min. The supernatant was discarded and cells were resuspended by vortexing in 10 mL of 0.3 M NaCl (Malik & Kakii, 2003). A homogenous suspension of bacterial cells was not produced unless this wash was performed. The total OD (OD600 nm [total]) was then measured. The AI was calculated as AI=(OD600 nm [total]–OD600 nm [planktonic])/OD600 nm [total]. The overall dispersion, induced by a given treatment, from an aggregated culture over a time period is measured as a relative AI reduction calculated as: (AImax–AIfinal) × 100/AImax.

All vaccines were administered by nurses in the immunization clinic and all medications were dispensed from the campus pharmacy. Institutional review board (IRB) approval was obtained prior to initiating the study. Basic characteristics of the travelers and the frequencies (or the average numbers) of the pretravel recommendations between the PTC and the PCP groups were compared by using chi-square test (or Fisher’s exact test) for categorical variables, and two-sample t-test or Wilcoxon–Mann–Whitney test (non-parametric version of independent-samples t-test) for continuous variables, if the normality assumptions

underlying the t-test were violated. The primary outcomes for vaccines and medications were (1) indicated and ordered, (2) indicated

and not ordered (excluding refused/declined), (3) not indicated and ordered, (4) and ordered and received (excluding refused/declined). The univariate and multivariate logistic selleck kinase inhibitor regressions (results not shown in tables) were performed to help to rate the findings according to their importance as risk/protective factors. All variables that showed an association with pretravel recommendations in the univariate models having p values below 0.10 were entered into the more comprehensive multiple logistic regression models, which included visit type (PTC or PCP), trip duration, purposes of travel (study abroad and volunteer work), and destination (Southeast Asia). All statistical significance was assessed using an alpha level of 0.05. Statistical analysis was performed find more using SAS 9.2. In 2007, 513 travelers were identified, 172 were seen by a PCP and 341 were seen in the PTC. Travelers who were seen in the PTC were more often prescribed antibiotics for self-treatment of travelers’ diarrhea when indicated (96% vs 50%, p < 0.0001), while

travelers seen by Selleck Venetoclax a PCP were more likely to be prescribed antibiotics not consistent with guidelines (not ordered when indicated 49% vs 6%, p < 0.0001 and ordered when not indicated 21% vs 3%, p < 0.0001) (Table 1). Furthermore, patients who were seen in the PTC were more likely to pick up their antibiotic from the pharmacy than those who were prescribed antibiotics by a PCP (75% vs 63%, p = 0.04). Travelers seen in the PTC were also more often prescribed antimalarials when indicated (98% vs 81%, p < 0.0001), while those seen by a PCP were more frequently prescribed antimalarials not consistent with guidelines (not ordered when indicated 15% vs 1%, p < 0.0001 and ordered when not indicated 19% vs 2%, p < 0.0001). There was no statistically significant difference in antimalarial pickup rates from the pharmacy between the two groups (Table 1). Results regarding the ordering and receipt of vaccines were similar to those of antibiotics and antimalarials. To account for multiple vaccines ordered at the same time, the primary outcomes for vaccines were calculated per patient and were used for comparison purposes.

M. Tsankova et al. (2006)Nat. Neurosci., 9, 519–525], but did not affect histone deacetylase 9. Exercise elevated the phosphorylated forms of calcium/calmodulin-dependent protein kinase II and cAMP response element binding protein, implicated in the pathways by which neural activity influences the epigenetic regulation of gene transcription, i.e. Bdnf. These results showing the influence of exercise on the remodeling of chromatin containing the Bdnf gene emphasize the importance of exercise on the control of gene transcription in the context of brain function and plasticity. Reported information about the impact of a behavior, inherently involved in the daily human routine, on the epigenome opens exciting new directions and therapeutic

opportunities in the war against neurological and psychiatric disorders. “
“Major

depressive disorder is a chronic disabling disease, often triggered and exacerbated by stressors of a social nature. Hippocampal Selleckchem Epigenetic inhibitor volume reductions have been reported in depressed patients. In support of the neurogenesis theory of depression, in several stress-based animal models of depression, adult hippocampal neurogenesis was reduced and subsequently rescued by parallel antidepressant treatment. Here, we investigated whether repeated social defeat and subsequent individual housing for 3 months induces long-lasting changes in adult hippocampal neurogenesis in rats, and whether these can be normalized by late antidepressant treatment, as would match human depression. Neurogenesis was analysed by stereological quantification of the number of immature doublecortin (DCX)-immunopositive cells, in particular young (class I) and more mature buy MG-132 (class II) DCX+ cells, to distinguish differential effects of stress or drug treatment on these subpopulations. Using this social defeat paradigm, the total DCX+ cell number was significantly reduced. This was most profound for older (class II) DCX+ cells with Carnitine dehydrogenase long apical dendrites, whereas younger, class I cells remained unaffected. Treatment with the broad-acting tricyclic antidepressant imipramine, only during the last 3 weeks of the 3-month period after social defeat,

completely restored the reduction in neurogenesis by increasing both class I and II DCX+ cell populations. We conclude that despite the lack of elevated corticosterone plasma levels, neurogenesis is affected in a lasting manner by a decline in a distinct neuronal population of more mature newborn cells. Thus, the neurogenic deficit induced by this social defeat paradigm is long-lasting, but can still be normalized by late imipramine treatment. “
“In the rodent nucleus accumbens (NAc), cocaine elevates levels of brain-derived neurotrophic factor (BDNF). Conversely, BDNF can augment cocaine-related behavioral responses. The latter could reflect enhancement of α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR) transmission, because AMPARs in the NAc mediate some cocaine-induced behaviors.

[9] The two cases of HCV infection

occurred in travelers to Vietnam and Thailand on short holiday trips. Screening for HCV in blood products is not universal in many developing countries and reuse of injection equipment without sterilization Protein Tyrosine Kinase inhibitor is common in Southeast Asia.[10] Neither Vietnam nor Thailand has mandatory reporting of HCV infection. Prevalence estimates for Thailand vary from 0.41% to 7.5%. In Vietnam prevalence estimates vary between 2 and 2.9% and up to 21% in studies of blood donors.[10] The one case of HBV infection occurred during a short trip to China, which is known to have an HBV prevalence of greater than 8%.[11] HCV transmission generally results from parenteral exposure to contaminated blood[11]: travelers who are exposed to contaminated blood or undertake medical procedures while abroad are at risk.[5] Transmission of HBV occurs through percutaneous or mucosal exposure to infected buy AZD5363 blood or bodily fluids. HBV acquisition in travelers has been associated with: duration of travel, immune status, VFR, casual sex, medical therapy, and the destination HBV prevalence.[2, 3] Both HBV and HCV may

have prolonged incubation periods (up to 6 months). A limitation of our study is the inability to exactly determine the date of HBV or HCV exposure. However, the travel duration together with the time to collection of post-travel serum makes it very likely that these infections were acquired abroad in countries with high endemic rates for both HBV and HCV infection. Despite limitations of this retrospective study, including inability to elucidate risk behaviors as relevant questions were not included in the traveler questionnaire, quantifying the risk of these infections among travelers is crucial in facilitating informed decision making regarding

the importance of vaccination and other preventative strategies. HCV infection prevention requires education and avoidance of high-risk activities. For HBV, the World Health Organization, Centers for Disease Control and Prevention, and Australian Guidelines recommend that HBV vaccination should be considered buy CHIR-99021 in nonimmune travelers to countries with a moderate to high prevalence of HBV (HBsAg ≥ 2%). Allowing sufficient time for pre-travel vaccination is crucial. For hepatitis B, an accelerated HBV vaccine schedule (doses on days 0, 7, 21, and 12 months) is safe and efficacious.[12] In this cohort, 59% (100/159) of travelers with an anti-HBs <10 mIU/mL attended a pre-travel clinic at least 21 days prior to departure to Asia providing sufficient time for HBV vaccination. The traveler diagnosed with HBV seroconversion attended clinic 32 days prior to travel and represents a potentially missed opportunity for vaccination.

piricola and Rhizoctonia solani– indicating that it does not lyse fungal cells (Ngai &

Ng, 2006). Similarly, schizolysin does not have a lytic action on fungal cells. Hemolysin production is related to virulence of microorganisms like bacteria, resulting in septicemia and diarrhea (Raimondi et al., 2000). Hemolysin causes lysis in various kinds of cells including erythrocytes, mast cells, neutrophils and polymorphonuclear cells, and increases virulence by inflicting tissue damage or by dissolving materials that would inhibit pathogens from spreading throughout the tissue. The expression of ostreolysin is undetectable during mycelial growth; it proceeds during formation of primordia and fruiting bodies, but declines during maturation LBH589 ic50 (Vidic et al., 2005). Schizolysin probably regulates fruiting initiation in the split gill mushroom, as suggested for the

oyster mushroom by Vidic et al. (2005). Aspergillus hemolysin (Sakaguchi et al., 1975) is expressed during sporulation. Whether hemolysin plays similar roles in bacteria, ascomycete fungi such as Aspergillus species, and basidiomycete fungi, including mushrooms, awaits clarification. This work was financially supported by National Grants of China (nyhyzx07-008, 2007BAD89B00 and 2010CB732202). Fig. S1. Ion-exchange chromatography of fraction C3 derived from fraction D3 adsorbed on DEAE-cellulose column, which was subsequently fractionated on CM-cellulose to yield C3 on a Q-Sepharose column (1×10 cm) in 10 mM phosphate buffer (pH 7.0). Fig. S2. FPLC-gel filtration Angiogenesis inhibitor of fraction Q2 adsorbed on Q-Sepharose on Superdex 75 in 10 mM phosphate buffer (pH 7.5) containing 0.15 M NaCl in the buffer. Table S1. Purification of hemolysin from 100 g fresh fruiting bodies of Schizophyllum commune. Table S2. Effects of compounds on hemolytic activity of Schizophyllum commune hemolysin (schizolysin).

Please note: Wiley-Blackwell is not responsible for the content or functionality of any supporting materials supplied by the authors. Any queries (other than missing material) should be directed to the corresponding author for the article. “
“Resistance to carbapenems in enterobacteria is mediated Protein kinase N1 by the production of several types of carbapenemases or by the decreased permeability of the outer membrane, combined with the expression of extended-spectrum β-lactamases (ESBLs) or AmpC-like cephalosporinases. The objective of this study was to characterize carbapenem-nonsusceptible (C-NS) isolates of Klebsiella pneumoniae in the University Hospital in Plzeň (Czech Republic) and compare them with carbapenem-susceptible (C-S) K. pneumoniae isolates from the same patients. Six C-NS K pneumoniae isolates from different patients were collected between January 2007 and June 2008, and from three of these patients, C-S isolates were available for the study as well. The isolates were typed by pulsed-field gel electrophoresis and multilocus sequence typing.

, 1991; Nakajima et al., 1994). Further, data from our previous work and other studies have established a close linkage between enolase and SS2 virulence (Esgleas et al., 2008; Feng et al., 2009; Zhang et al., 2009a). Similarly, pyruvate kinase (05SSU0544) is a key

enzyme involved in pneumococcal fermentative metabolism and thereby contributes to the virulence of S. pneumoniae (Yesilkaya et al., 2009). Recent work by Burall et al. (2009) also suggests that the reduced virulence of the ovine pathogen Chlamydia abortus live vaccine strain results from disrupted metabolic activity owing to altered pyruvate kinase expression. Additionally, 5′-nucleotidase is involved in various functions, such as cell–cell communication, nucleic acid repair, the purine salvage pathway for nucleotides synthesis, Pexidartinib clinical trial signal transduction and membrane transport (Hunsucker et al., 2005). In S. suis serotype 9 (SS9), 5′-nucleotidase is recognized as a putative virulence-associated factor based on comparative proteomics analysis (Wu et al., 2008). It should also be noted that many subunits of the F0F1-type ATP synthase locus were less efficiently expressed in the absence of VirR/VirS. However, the role of this enzyme complex in the pathogenesis of SS2 requires further investigation.

Finally, it is notable that the expression of many proteins involved in the stress response is repressed, such as membrane GTPase (05SSU0468), heat shock protein (HSP) 70 (DnaK, 05SSU0300), DnaJ (05SSU0302) and ATP-dependent caseinolytic

proteases (Clp, 05SSU0389 and 05SSU0390). These Selleckchem Ixazomib proteins play fundamental roles in stress tolerance and virulence in many pathogenic bacteria (Bukau & Horwich, 1998; Takaya et al., 2004; Ibrahim et al., 2005; Tu le et al., 2007; Kajfasz et al., 2009; Zhang et al., 2009b). To validate the proteomic data, the relative ability of the ΔvirRS mutant to survive H2O2-induced oxidative stress was examined. We found that the mutant was significantly more susceptible to the H2O2 treatment than WT, suggesting that VirR/VirS mafosfamide plays a crucial role in the oxidative stress response in S. suis 05ZYH33. In conclusion, the present study provides initial insight into the role of the VirR/VirS system in the physiology and virulence of SS2. Our results demonstrate that although the VirR/S systems of S. suis and C. perfringens are orthologous, the target proteins regulated by these systems are not identical in these two phylogenetically distinct bacteria. This may reflect the adaptation of these pathogens to the specific environments that they encounter during the course of infection. This work was supported by the National Natural Science Foundation of China (No. 30971574 and 30901282) and the Pre-Research Foundation of Third Military Medical University (No. 2009XYY02). H.W. and X.S. contributed equally to this work.

1% Coomassie brilliant blue R250 (CBB), 40% (v/v) methanol, and 1% glacial acetic acid solution, followed by destaining in 50% (v/v) methanol. Total proteins of the cells were solubilized for 2 h at 4 °C in a buffer containing 50 mM Tris–HCl (pH 7.4), 0.15 M NaCl, 2.5% (w/v) sodium deoxycholate, 2% (v/v) NP-40, 1 mM N, N, N′, N′-ethylenediaminetetraacetic acid

(EDTA), protease inhibitors (1 mM PMSF, 1 μg mL−1 aprotinin, 1 μg mL−1 leupeptin, and 1 μg mL−1 pepstatin), and phosphatase inhibitors (1 mM Na3VO4 and 1 mM NaF). The phosphoproteins were trapped on Phos-tag agarose phosphate-affinity beads and isolated according to the methods reported by MLN0128 mouse Kinoshita-Kikuta et al. (2009). For LC-MS/MS analysis, the blots used in Phos-tag detection assays were incubated in 1 N aqueous NH3 for 15 min three times to remove the biotinylated Phos-tag complex (Nakanishi et al., 2007). Prior to LC-MS/MS analysis, the protein bands of interest on the blotting membranes were cut out and digested with 10−7 M lysyl endopeptidase (Wako) for 1 h at 37 °C. Peptides

produced by protease digestion were separated by a 0–40% linear acetonitrile gradient for 60 min, followed by analyses with a Waters UPLC Xevo Qtof. Obtained data were processed with ProteinLynx Global Server 2.4 and searched against Alveolata protein entries in the Uniprot Knowledgebase (UniprotKB) Dasatinib nmr and Alveolata protein records in Entrez. When cells of C. cucullus cultured for 1–2 days were stimulated to encyst by

overpopulation in the presence of 0.1 mM Ca2+, the phosphorylation level of several proteins was enhanced within 1 h after onset of encystment induction (Fig. 1a, ‘P-tag’ right lane). In this case, the integrated optical density between in two lanes of CBB-stained blots (Fig. 1a, ‘CBB’) determined by NIH Image analysis was almost equivalent, indicating that an elevation of the Phos-tag signal (Fig. 1a, ‘P-tag’) should not be attributed to a difference in amounts of proteins between the lanes, but rather to a real enhancement of the protein phosphorylation level. In most of these cases, the increased phosphorylation 5-FU level was maintained for at least 12 h (data not shown). In the preparation of the protein samples shown in Fig. 1a, the sample buffer for SDS-PAGE did not contain inhibitors of proteases or phosphatases. Therefore, the phosphorylated proteins detected in this assay may have contained protein fragments digested by endogenous proteases, and some of the proteins may have been dephosphorylated by endogenous phosphatases during sample preparation. Therefore, as shown in Fig. 1b, the phosphorylation assay of the encystment-induced Colpoda proteins was re-examined in the presence of protease and phosphatase inhibitors (PPI).

Identical restriction patterns were detected for all these 16 phages, in spite of their host range difference. Phage Bf7 was selected for further investigations, because of its outstanding ability for infection. On the basis of the electron microscopic studies, the morphology of phage Bf7 seems to be similar to some other bacteriophages infecting the members of the genus Pseudomonas.

We assigned phage Bf7 to the family Podoviridae based on its icosahedral phage head with a diameter of about 60 nm, the short tail (Fig. 1), and the size of the dsDNA genome (Ackermann, 2001). The phages of this family infect enteric and related Gram-negative bacteria (Van Regenmortel et al., 2000). Well-known pseudomonad-infecting members of the family are phages, for example gh-1, φPLS27, φPLS743, Pssy9220 (Van Regenmortel et al., 2000), φGP100 (Keel et al., 2002), φIBB-PF7A (Sillankorva et al., 2011), http://www.selleckchem.com/products/Rapamycin.html and BVPaP-3 (Ahiwale et al., 2012). Phage

find more Bf7 forms clear plaques (1–3 mm in diameter) after 18 h incubation at 20 °C on P. tolaasii 2342T. This property depends mostly on the temperature. At 5, 10, 20, and 25 °C clear plaques are formed after 18–48 h incubation, but no plaques are generated at 30 and 35 °C. This phenomenon is similar to the plaque forming characteristics of phage φGP100 (Keel et al., 2002). Based on these observations, we performed our further experiments at 20 or 25 °C. The single-step growth experiments have revealed that the Bf7 bacteriophage had a latent period of about 140 min. This latent period is nearly similar to phage φGP100 infecting P. fluorescens CHA0 (Keel et al., 2002) belonging to the Podoviridae family. The calculated burst

size was 237 PFU per infected cell before at 20 °C, MOI of 0.06, taken into account the latent period, the eclipse and the rise periods (Fig. 2). Comparing this value with those of other pseudomonad-infecting members of the Podoviridae family, it can be concluded that it is higher than the average. On the basis of single-step growth, Bf7 bacteriophage has a latent period of 140 min and relatively high burst size. These phenomena could be good indicators of an effective biocontrol agent, because it could infect large number of target bacteria in the same time, so there is less chance for the development of resistant strains. Moreover, the phage was resistant to chloroform treatment for at least one month. The genome of Bf7 bacteriophage proved to be dsDNA, 40 058 bp in size, including direct terminal repeats (DTRs) of 417 bp. The length of the DTRs was confirmed by direct sequencing with outward-directed primers, leading to stop of the reactions at both ends of the genome. G+C content of the Bf7 genome was 58.4% (GenBank accession number: JN991020.) Analysis of the genome sequence revealed the presence of 46 ORFs, most of them had an ATG start codon (43), but there were 2 with GTG and one with TTG start codon (Table 4).