, 2007) Cloning and the heterogeneous expression of crtI from Rb

, 2007). Cloning and the heterogeneous expression of crtI from Rba. azotoformans were performed to understand the product pattern of CrtI. A 1557 bp crtI gene was amplified via PCR from the Rba. azotoformans CGMCC 6086 genome with primers Ra-If and Ra-Ir (Table 1). A 518-amino acid protein was encoded with a predicted molecular

mass of 57.28 kDa. The crtI gene was inserted into pET22b and transformed into E. coli BL21 (DE3). The ratio of CrtI to total check details E. coli protein was approximately 7–10% after induction with IPTG. The subunit molecular mass of 57 kDa determined via SDS-PAGE (Fig. 3) was consistent with the predicted molecular mass. The product pattern of CrtI from

Rba. azotoformans was examined in vivo by co-transforming plasmid pET22b-I with plasmid pACYCDuet-EB into the E. coli BL21 (DE3). The transformant acquired a red color in LB culture after induction with IPTG. After cultivation for 5 h in LB medium with 0.5 mM IPTG, the recombinant E. coli produced three carotenoids (Fig. 4a) identified by molecular mass and absorption spectra as neurosporene, lycopene, and 3,4-didehydrolycopene DAPT (Fig. S3). Neurosporene has a relative molecular mass of 538.4 and three absorption maxima at 416, 440, and 469 nm. Lycopene has a relative molecular mass of 536.4 and three absorption maxima at 444, 472, and 504 nm. Meanwhile, 3,4-didehydrolycopene has a relative molecular mass of 534.4 and three absorption maxima at 469, 496, and 528 nm. After cultivation for 24 h, the relative Aldehyde dehydrogenase contents of neurosporene and lycopene in recombinant E. coli were approximately 23% and 75%, respectively, whereas 3,4-didehydrolycopene almost disappeared (Fig. 4b). This in vivo result showed that CrtI from Rba. azotoformans

CGMCC 6086 could produce three-step desaturated neurosporene and four-step desaturated lycopene as major products, together with small amounts of five-step desaturated 3,4-didehydrolycopene. The present study is the first to report that 3,4-didehydrolycopene could be produced by CrtI from Rhodobacter. CrtI would be a three-step phytoene desaturase in situ because carotenoids of the spheroidene series were synthesized in Rba. azotoformans CGMCC 6086. Therefore, the formation of lycopene and 3,4-didehydrolycopene in recombinant E. coli were probably due to neurosporene accumulation caused by the lack of hydroxyneurosporene synthase (CrtC) and CrtI kinetics. In a crtC deletion mutant of Rba. azotoformans CGMCC 6086 obtained via EMS and LiCl mutagenesis, carotenoid products contained approximately 90% neurosporene and 10% lycopene (data not shown). The kinetics could also affect product patterns of CrtI. CrtI from Rvi. gelatinosus and P.

coelicolor is more proteolytic than

S lividans (Kieser e

coelicolor is more proteolytic than

S. lividans (Kieser et al., 2000; Jayapal et al., 2007). When supernatants of the Δppm mutant IB31 carrying the cloned apa gene (in pBL1) were analyzed, the SAHA HDAC nmr Apa protein was still expressed and secreted, as evidenced by the presence of a clear band detected by the 6A3 monoclonal antibodies (Fig. 1b, lane 2), but it was not glycosylated, as indicated by the slightly lower mass observed and by the lack of reaction with ConA (Fig. 1c, lane 2). This result indicates that PpmSco is essential for glycosylation of M. tuberculosis Apa by S. coelicolor. To determine whether the S. coelicolor Δppm mutant IB31 could be complemented by M. tuberculosis Ppm (PpmMtu), the Rv2051c gene was amplified from M. tuberculosis H37Rv DNA and cloned under the control of the strong PtipA promoter (plasmid pBL10, Table 1); the S. coelicolor ppm gene (sco1423) and upstream flanking region were cloned www.selleckchem.com/products/FK-506-(Tacrolimus).html in pSET152 as a control (plasmid pBL13, Table 1). Phage φC31 was able to form plaques in the S. coelicolor Δppm mutant IB31 carrying either pBL10 or pBL13, encoding PpmMtu and PpmSco, respectively (Fig. 1a, plates 3 and 4; Table S2). In addition, introduction of these same plasmids into the S. coelicolor Δppm mutant

expressing Apa [IB31(pBL1)] restored glycosylation of this protein, as indicated by the presence of bands in Western blots detected with monoclonal antibodies (Fig. 1b, lanes 3 and 4), which showed restoration of ConA reactivity (Fig. 1c, lanes 3 and 4). To demonstrate activity of PpmMtu in S. coelicolor, an in vitro assay was carried out to detect labeling of the membrane polyprenyl phosphate by GDP-[14C]mannose in purified membrane fractions.

Streptomyces coelicolor harbors oxyclozanide a single C45 membrane polyprenol (Wehmeier et al., 2009; Fig. S1), and clear labeling of this molecule was observed in membranes of wild-type S. coelicolor (J1928) as indicated by a single-labeled band (Fig. 2, lane 1), but not in membranes of the Δppm mutant (IB31; Fig. 2, lane 2). Complementation was confirmed by this in vitro assay, because labeling of the membrane polyprenyl phosphate was restored when either pBL13 (PpmSco) or pBL10 (PpmMtu) was introduced into the Δppm mutant (Fig. 2, lanes 3 and 4, respectively), confirming that PpmMtu is functional when expressed in S. coelicolor. PpmMtu is a protein composed of two distinct domains. The N-terminal hydrophobic domain D1 (Met1-Tyr593) is responsible for lipoprotein N-acyltransferase (Lnt) activity, whereas the C-terminal domain D2 (Met594-Glu874) is the Ppm catalytic domain (Gurcha et al., 2002; Tschumi et al., 2009). We therefore decided to analyze whether the isolated D2 domain of PpmMtu was functional in S. coelicolor in the absence of the D1 domain. To do this, the portion of the Rv2051c gene encoding the D2 domain was cloned in pIJ6902 under control of the PtipA promoter (pBL11) and introduced into the S. coelicolor Δppm mutant IB31.

Thus, the present study does not provide evidence of a general on

Thus, the present study does not provide evidence of a general ongoing detrimental effect on BMD following the early period after HAART initiation of either of these two drug classes. The results may suggest that HAART or HAART-induced immunological changes cause a temporary imbalance between bone resorption and bone formation or that treatment abrogates HIV-induced accelerated

bone loss. MG-132 concentration Randomized studies to evaluate the influence of earlier treatment initiation on bone metabolism are warranted. Author contributions Conception and design: A. B. Hansen, N. Obel, H. Nielsen, C. Pedersen and J. Gerstoft. Collection of data: A. B. Hansen, N. Obel, H. Nielsen, C. Pedersen and J. Gerstoft. Analysis and interpretation of the data: A. B. Hansen and J. Gerstoft. Drafting of the article: A. B. Hansen. Critical revision of the article: A. B. Hansen, N. Obel, H. Nielsen, C. Pedersen and J. Gerstoft. Financial support This study was supported by an unconditional grant from Abbott, Denmark. Abbott was not involved in the design or conduct of the study, data collection, management, analysis or interpretation of data, preparation or approval of the manuscript, or the decision to submit for publication. “
“Highly active antiretroviral therapy (HAART) has transformed HIV

infection into a manageable chronic illness, yet AIDS mortality among ethnic minorities persists in the USA. HAART nonadherence is associated with increased PD0332991 datasheet HIV viral load, low CD4 cell count and racial disparities in HIV outcomes. While there is no universal consensus on how to improve medical adherence in HIV-positive populations, the community health worker (CHW) model is emerging as an effective strategy to overcome barriers to HAART adherence. Although utilized in international settings, there is little evidence regarding the effects of CHWs on HIV outcomes in the USA. We performed a comprehensive search from May 2010 to November

2010 to identify studies carried out in the USA that www.selleck.co.jp/products/abt-199.html utilized CHWs to improve HAART adherence and measured HIV viral loads and CD4 cell counts to assess intervention effects. Sixteen studies met the inclusion criteria and were reviewed for this article. All studies reported clinical HIV outcomes. Interventions that lasted at least 24 weeks, provided frequent contact with participants, and focused on medication management were associated with improved HAART adherence, as indicated by reduced HIV viral load and increased CD4 cell count. Compared with current standards of care, CHW programmes may offer a practical and cost-effective alternative to improve HAART adherence, which may lead to reduced HIV viral load and increased CD4 cell counts among HIV-positive populations in the USA. Highly active antiretroviral therapy (HAART) can transform HIV/AIDS from a fatal diagnosis to a manageable chronic illness [1,2].

However, assessing students’ dosing knowledge is not straightforw

However, assessing students’ dosing knowledge is not straightforward. Asking students to memorize specific dosing, which can lead to medication errors, particularly in special populations (e.g. paediatrics, geriatrics) may not be optimal. While it is necessary to ask students which reputable source they used to identify correct doses, these items may be better served in another course (i.e. drug information). Also, given that most students will practice in a general retail or hospital staff setting, looking up every dose is not efficient given they will be entering,

reviewing and verifying numerous orders and medications per hour. Therefore, dosing UK-371804 items which contain dosing ranges may be better suited in providing a balance for students in understanding if the

dose is within the correct range and to look up any medication that draws a red flag (e.g. dosing outside the range). Overall, the strengths of the study are multi-fold. This is the first study to specifically evaluate dosing items in pharmacy TP courses and the first to simultaneously evaluate format and content. It also provides additional data on Case-based items and their ability to discriminate compared to other formats. Finally, the study also provides an KU-60019 cost approach (i.e. Delphi technique) to minimize the bias when determining how to group items into categories. Given these strengths, this study also has limitations. First, it is unknown whether having items with a better difficulty or discrimination level translates into students’ increase in knowledge and application. While these indices should not be the sole measurements used to predict success outside the classroom, they are a starting point. This study suggests that Case-based and dosing items provide our students with greater difficulties. Based on other studies, these Histamine H2 receptor items may be better at assessing and predicting

students’ professional expertise and ranking their performance among their peers. These results are also based on students who may not be representative of other colleges of pharmacy. Nova Southeastern University College of Pharmacy has a very culturally diverse student body consisting of over 240 students per year, with more than 70% of the intake classifying themselves as minorities or from outside of the USA. Therefore, the diversity may explain the unexpected finding that format is more important in determining difficulty than content, indicating that non-native English speakers may struggle more with diverse formats. Lastly, despite having over 500 assessment items, our sample size was small, especially when deconstructed into elements (e.g. format, content). Collaborating with other colleges of pharmacy may assist in obtaining additional items written by other faculty members and answered by other pharmacy students. This will increase the sample size and address the heterogeneity of the data.

Our results showed the potential of wheat bran to produce laccase

Our results showed the potential of wheat bran to produce laccase by white-rot fungi, because high laccase activities were obtained. However, the use of different strains

for the laccase production clearly modifies the culture conditions, especially for the close-to-the-substrate hyphae. In this region, the hyphae density and the number of layers clearly depend on the strain. On the one hand, P. ostreatus PI3K targets presented the highest hypha density and the largest number of layers in the interface structure, showing a tendency for a hair-like growing morphology. This morphology could be linked to the low oxygen diffusion presented in the inner layers. On the other hand, C. unicolor presented the lowest hypha density and the smallest number of layers in the interface structure; thus, this fungus was expected to have

the best oxygen diffusion into its inner layers. However, there is no clear relationship between fungal morphology, hypha density and laccase production. Pleurotus ostreatus produced nearly 50% more laccase per gram of total dry matter than the one produced by C. unicolor. Trametes versicolor, which presented medium hypha density, produced the highest activity of laccase per gram of total dry matter, but T. pubescens, with similar growth morphology likely related to their common genus (Trametes), produced the lowest quantity of laccase per gram of total dry matter among all the strains studied. This indicates that, although all strains presented differences in the hypha size, clump size, morphology and the number of layers in the interface structure, NU7441 concentration the laccase production in terms of activity per gram of total dry matter cannot be related to these characteristics of the culture, but directly to the strain. This is in agreement with the review by Grimm et al. (2005), which stated that no simple relationship was found between morphology and productivity in filamentous fungi. This research was financed by the Spanish Ministry of Education

and Science (Project CTQ2007-66541). We thank Tau-protein kinase Dr A. Hatakka from the Department of Food and Environmental Sciences at the University of Helsinki (Helsinki, Finland) for the Trametes versicolor K120a2 (FBCC564), Cerrena unicolor T71 (FBCC744) and Pleurotus ostreatus DSM 11191 (FBCC375) fungal strains and Erika Winquist from the Aalto University School of Science and Technology (Aalto, Finland) for her help and support reading the manuscript. “
“Here, we describe mutants of Mycoplasma pulmonis that were obtained using a minitransposon, Tn4001TF1, which actively transposes but is then unable to undergo subsequent excision events. Using Tn4001TF1, we disrupted 39 genes previously thought to be essential for growth. Thus, the number of genes required for growth has been overestimated. This study also revealed evidence of gene duplications in M.

[1] Leptospirosis is now considered an emerging disease in travel

[1] Leptospirosis is now considered an emerging disease in travelers. On July 1, 2011, two Australian male tourists aged 25 and 26 years were admitted to the emergency department of the Careggi Hospital, Florence, Italy, reporting a 1-week history of fever with sudden onset of headache, myalgia, nausea, vomiting, and diarrhea. They had

no significant past medical or surgical history and they had recently traveled from Venice to Florence. At the time of admission they showed similar clinical signs and symptoms, mainly jaundice, conjunctival hyperemia, and muscle tenderness. Routine hematological and biochemical profiles were similar (Table 1). In both cases laboratory findings evidenced acute renal failure and hepatic impairment. Vital signs were normal. On auscultation, the heart sounds had no abnormalities CT99021 mouse and air entry was equal on both lungs with occasional scattered wheezing. Results of neurological examination were normal. Electrocardiogram, abdominal ultrasound, and X-ray of the chest revealed no abnormalities. Asked about recent exposure to animals, mud, or potentially contaminated freshwater sources, the young tourists mentioned they had settled at a campsite near Venice 2 weeks earlier; because of the heat, they had both immersed their feet in the waters of a Venice canal close to Rialto Bridge. One of them had also swum in it for less than a minute without

any protection for the conjunctiva.

No skin lesions or trauma were observed at the time of possible infection, nor any swallowing of the canal water. The common this website history of exposure to possible contaminated water, along with hepatic and renal impairment, suggested the diagnosis of leptospirosis. However, blood and urine specimens were collected for culture and polymerase chain reaction Baricitinib (PCR) was also evaluated. Serum samples were tested by the microscopic agglutination test (MAT). Intravenous ceftriaxone (2 g every 24 h) was empirically administered.[2] Adequate fluids and a diuretic infusion were also started. Both patients required daily hemodialysis for 5 days as a result of the severe renal injury. Blood and urine cultures had no growth. The PCR result was positive for leptospiral DNA in the urine of both patients. First collected serum sample results (approximately the tenth day of disease) were positive by MAT in both subjects with titers to serovars icterohaemorrhagiae of 1 : 1,600 and serovars copenhageni of 1 : 200 (serogroup icterohaemorrhagiae). Serovars icterohaemorrhagiae and copenhageni are commonly associated with rats as reservoir hosts.[3] Ten days later, the titer of 1 : 1,600 was confirmed in the first subject, while that of 1 : 200 of the other attained 1 : 3,200 (Table 1). The clinical picture progressively improved, restoring normal function of liver and kidney, and they were discharged after 2 weeks.

02) Conclusions  Home visits and telephone contacts conducted 6

02). Conclusions.  Home visits and telephone contacts conducted 6 monthly from birth are effective in reducing ECC prevalence by 24 months. “
“International Journal of Paediatric Dentistry 2011 Aim.  To investigate the root canal microbiota of primary teeth

with apical periodontitis and the in vivo antimicrobial effects of a calcium hydroxide/chlorhexidine paste used as root canal dressing. Design.  Baseline samples GDC0068 were collected from 30 root canals of primary teeth with apical periodontitis. Then, the root canals were filled with a calcium hydroxide paste containing 1% chlorhexidine for 14 days and the second bacteriologic samples were taken prior to root canal filling. Samples were submitted to microbiologic culture procedure to detect root canal bacteria and processed this website for checkerboard DNA–DNA hybridization. Results.  Baseline microbial culture revealed high prevalence and cfu number of anaerobic, black-pigmented bacteroides, Streptococcus, and aerobic microorganisms. Following root canal dressing, the overall number of cfu was dramatically

diminished compared to initial contamination (P <0.05), although prevalence did not change (P > 0.05). Of 35 probes used for checkerboard DNA–DNA hybridization, 31 (88.57%) were present at baseline, and following root canal dressing, the number of positive probes reduced to 13 (37.14%). Similarly, the number of bacterial cells diminished folowing application of calcium hydroxide/chlorhexidine root canal dressing (P = 0.006). Conclusion.  Apical periodontitis is caused by a polymicrobial infection, and a calcium hydroxide/chlorhexidine paste Adenosine is effective in reducing the number of bacteria inside root canals when applied as a root canal dressing. “
“International Journal

of Paediatric Dentistry 2012; 22: 116–124 Background.  Intracanal medication is important for endodontic treatment success as it eliminates microorganisms that persist after biomechanical preparation. Aim.  To evaluate the effect of two intracanal medications against Porphyromonas gingivalis and Enterococcus faecalis in the root canals of human primary teeth with necrotic pulp with and without furcal/periapical lesion, using quantitative real-time polymerase chain reaction (qRT-PCR). Design.  Thirty-two teeth with necrotic pulp were used. Twelve teeth did not present lesion, and 20 teeth presented radiographically visible furca/periapical lesion. Microbiological samples were collected after coronal access and biomechanical preparation. The teeth were medicated with calcium hydroxide pastes prepared with either polyethylene glycol or chlorhexidine. After 30 days, the medication was removed and a third collection was performed. Microbiological samples were processed using qRT-PCR. Data were analysed by Wilcoxon and Mann–Whitney tests (α = 0.05). Results.  There was no significant difference in the microbiota present in the primary teeth with and without furcal/periapical lesion.

coelicolor (Yang et al, 2006) The specificity for dCMP incorpor

coelicolor (Yang et al., 2006). The specificity for dCMP incorporation into pORF102 leaves the possibility that either the

see more first or the second nucleotide of the 3′-end of pAL1 (… GCAGG-3′) may serve as a template for the deoxynucleotidylation reaction. In this study, we identified the gene product of pAL1.102 as a protein that is associated with both termini of the linear Arthrobacter plasmid pAL1. The proposed TP – at least when fused to MBP to ensure solubility – was not capable of specifically recognizing telomeric pAL1 DNA in vitro. However, in an in vitro deoxynucleotidylation assay, the pORF102 protein specifically incorporated dCMP, complementary to the 3′-ends of pAL1. This is consistent with its presumed role as a protein primer in DNA replication. The financial support of the Deutsche Forschungsgemeinschaft is gratefully acknowledged (FE 383/11). We thank Prof. Dr R. Brandsch (University of Freiburg, Germany) for kindly providing the vector pART2, and Prof. Dr A. Steinbüchel (Münster) for access to the phosphoimager. We also thank Manuel Tomm for initial EMSA experiments, and Gabriele Niester and Almut Kappius for technical assistance. Table S1. Primers and ssDNA template

GSK J4 molecular weight used in this study. 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. “
“Many bacteria produce siderophores for sequestration of growth-essential iron. Analysis of the Salinispora genomes suggests that these

marine actinomycetes support multiple hydroxamate- and phenolate-type siderophore pathways. We isolated and characterized desferrioxamines (DFOs) B and E from all three recognized Salinispora species and linked their biosyntheses in S. tropica CNB-440 and S. arenicola CNS-205 to the des locus through PCR-directed mutagenesis. Gene inactivation of the predicted iron-chelator until biosynthetic loci sid2-4 did not abolish siderophore chemistry. Additionally, these pathways could not restore the native growth characteristics of the des mutants in iron-limited media, although differential iron-dependent regulation was observed for the yersiniabactin-like sid2 pathway. Consequently, this study indicates that DFOs are the primary siderophores in laboratory cultures of Salinispora. Siderophores are small molecules secreted by bacteria to sequester growth-essential ferric iron that is poorly soluble under neutral pH and aerobic conditions (Neilands, 1995). The structures of siderophores vary considerably and are often suited to the environmental niche of the producing bacterium. For example, amphiphilic siderophores possess hydrophobic fatty acid chains that enable them to remain associated with the cell membrane (Martinez et al., 2003) – an attribute particularly advantageous in pelagic marine environments where dilution occurs rapidly.

coelicolor (Yang et al, 2006) The specificity for dCMP incorpor

coelicolor (Yang et al., 2006). The specificity for dCMP incorporation into pORF102 leaves the possibility that either the

see more first or the second nucleotide of the 3′-end of pAL1 (… GCAGG-3′) may serve as a template for the deoxynucleotidylation reaction. In this study, we identified the gene product of pAL1.102 as a protein that is associated with both termini of the linear Arthrobacter plasmid pAL1. The proposed TP – at least when fused to MBP to ensure solubility – was not capable of specifically recognizing telomeric pAL1 DNA in vitro. However, in an in vitro deoxynucleotidylation assay, the pORF102 protein specifically incorporated dCMP, complementary to the 3′-ends of pAL1. This is consistent with its presumed role as a protein primer in DNA replication. The financial support of the Deutsche Forschungsgemeinschaft is gratefully acknowledged (FE 383/11). We thank Prof. Dr R. Brandsch (University of Freiburg, Germany) for kindly providing the vector pART2, and Prof. Dr A. Steinbüchel (Münster) for access to the phosphoimager. We also thank Manuel Tomm for initial EMSA experiments, and Gabriele Niester and Almut Kappius for technical assistance. Table S1. Primers and ssDNA template

MAPK inhibitor used in this study. 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. “
“Many bacteria produce siderophores for sequestration of growth-essential iron. Analysis of the Salinispora genomes suggests that these

marine actinomycetes support multiple hydroxamate- and phenolate-type siderophore pathways. We isolated and characterized desferrioxamines (DFOs) B and E from all three recognized Salinispora species and linked their biosyntheses in S. tropica CNB-440 and S. arenicola CNS-205 to the des locus through PCR-directed mutagenesis. Gene inactivation of the predicted iron-chelator ifenprodil biosynthetic loci sid2-4 did not abolish siderophore chemistry. Additionally, these pathways could not restore the native growth characteristics of the des mutants in iron-limited media, although differential iron-dependent regulation was observed for the yersiniabactin-like sid2 pathway. Consequently, this study indicates that DFOs are the primary siderophores in laboratory cultures of Salinispora. Siderophores are small molecules secreted by bacteria to sequester growth-essential ferric iron that is poorly soluble under neutral pH and aerobic conditions (Neilands, 1995). The structures of siderophores vary considerably and are often suited to the environmental niche of the producing bacterium. For example, amphiphilic siderophores possess hydrophobic fatty acid chains that enable them to remain associated with the cell membrane (Martinez et al., 2003) – an attribute particularly advantageous in pelagic marine environments where dilution occurs rapidly.

, 2009) For many other zoosporic pathogens, including P alni, P

, 2009). For many other zoosporic pathogens, including P. alni, P. kernoviae, and P. ramorum, such information is missing. The objective of this study was to examine the survival of P. alni, P. kernoviae, and P. ramorum in response to different levels of pH. Specifically, zoospores were tested over a range of pH from 3 to pH 11 in 10% Hoagland’s solution. Responses of these pathogens to

pH were determined by relative Epacadostat concentration survival rates measured as colony-forming units (CFU) and behaviors of zoospores measured as relative counts of swimming zoospores, encysted zoospores (cysts), and germinating zoospores. Ten percent Hoagland’s solution (HS) used for tests was prepared as follows. The stock HS solutions were made using Hoagland’s basal salt mixture (MP Bio, OH), pH-adjusted with NaOH or HCl, and then filter-sterilized. Precipitation

observed for stock solutions at pH 9 and 11 was removed through filtration. The pH solutions were used immediately or stored at 4 °C until used. Sterile distilled water (SDW) to be used for dilution was also pH-adjusted with NaOH or HCl. To obtain 10% HS solution with appropriate pH at 3, 5, 7, 9, or 11, a stock solution was diluted with SDW with the same pH. Solutions were tested for the total concentration of salts and dissolved individual ions by JR Peters Laboratory (Allentown, PA) (Supporting ROCK inhibitor Information, Table S1). Zoospore survival of P. alni, P. kernoviae, and P. ramorum isolates (Table 1) was assessed with colony-forming units of zoospores (CFU mL−1) at each test pH and exposure time and zoosporic behavior at each test pH up to 24 h after exposure. Stock zoospore suspensions were prepared through a liquid culture for 7 days followed by sporangia induction and zoospore

release as described previously (Kong et al., 2012). To determine CFU in response to pH, a volume of fresh zoospore stock was added to diluted HS in a 175-mL tissue culture container (Greiner Bio One, Monroe, NC) to make 100 mL of Farnesyltransferase 10% HS so that there were about 50 zoosporic colonies when 1 mL was placed in a 90-mm Petri dish. Each treatment included three replicate containers. Samples were taken immediately after the addition of the zoospore suspension stock solution (day 0) and after 1, 3, 5, 7, and 14 days incubation. At each time point, two 1-mL aliquots were taken from each container and spread onto two 90-mm Petri dishes containing PARP-V8 agar (Ferguson & Jeffers, 1999). Dishes were incubated at 20 °C in a growth chamber for 2–3 days and emerging colonies were counted. C. Brasier (P834) C. Brasier (P1590) S. Jeffers (4398) To examine the behavior of zoospores in each pH treatment, 1-mL samples were also taken from each treatment container as noted previously at the first time point (day 0) and placed in wells of a 24-well plate.