Widmer G: Meta-analysis of a polymorphic surface glycoprotein of

Widmer G: Meta-analysis of a polymorphic surface glycoprotein of the parasitic protozoa Cryptosporidium parvum and Cryptosporidium hominis . Epidemiol Infect 2009, 137:1800–1808.PubMedCrossRef 39. Altschul S, Gish W, PF-573228 order Miller W, Myers EW, Lipman DJ: Basic local alignment search tool. J Mol Biol 1990, 215:403–410.PubMed 40. Bouzid M, Heavens

D, Elwin K, Chalmers RM, Hadfield SJ, Hunter PR, Tyler KM: Whole genome amplification (WGA) for archiving and genotyping of clinical isolates of Cryptosporidium species. Parasitology 2010, 137:27–36.PubMedCrossRef 41. Elwin K, Chalmers RM, Roberts R, Guy EC, Casemore DP: Modification of a rapid method for the identification of gene-specific polymorphisms in Cryptosporidium parvum and its application to clinical and epidemiological investigations. Appl Environ Microbiol 2001, 67:5581–5584.PubMedCrossRef 42. Chalmers RM, Elwin K, Thomas AL, Guy EC, Mason B: Long-term Cryptosporidium typing reveals the aetiology and species-specific epidemiology of human cryptosporidiosis in England and Wales, 2000 to 2003. Euro Surveill 2009., 14: 43. Tanriverdi S, Arslan MO, Akiyoshi DE, Tzipori

S, Widmer G: Identification of genotypically mixed Cryptosporidium parvum populations in humans and calves. Mol MK-0457 in vivo Biochem Parasitol 2003, 130:13–22.PubMedCrossRef 44. Xiao L, Singh A, Limor J, Graczyk TK, Gradus S, Lal A: Molecular characterization ABT-263 price of Cryptosporidium oocysts in samples of raw surface water and wastewater. Appl Environ Microbiol 2001, 67:1097–1101.PubMedCrossRef 45. Mallon M, MacLeod A, Wastling J, Smith H, Reilly B, Tait A: Population structures and the role of genetic Quisqualic acid exchange in the zoonotic pathogen Cryptosporidium parvum . J Mol Evol 2003, 56:407–417.PubMedCrossRef 46. Alves M, Xiao L, Antunes F, Matos O: Distribution of Cryptosporidium subtypes in humans and domestic and wild ruminants in Portugal. Parasitol Res 2006, 99:287–292.PubMedCrossRef 47. Xiao L: Molecular epidemiology of cryptosporidiosis: an update. Exp Parasitol 2010, 124:80–89.PubMedCrossRef 48. Soba B, Logar J: Genetic classification

of Cryptosporidium isolates from humans and calves in Slovenia. Parasitology 2008, 135:1263–1270.PubMedCrossRef 49. Huang X, Madan A: CAP3: A DNA sequence assembly program. Genome Res 1999, 9:868–877.PubMedCrossRef 50. Tamura K, Dudley J, Nei M, Kumar S: MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol 2007, 24:1596–1599.PubMedCrossRef Authors’ contributions MB carried out the experimental testing of the predicted putative species-specific genes, sequence alignment and data analysis and drafted the manuscript. KMT conceived the study, provided technical guidance, coordinated the study and helped to draft the manuscript. RC performed the comparative genomic analysis. RMC participated in the design of the study and helped to draft the manuscript.

The bbk32 gene was amplified from B31 genomic DNA, however, PCR p

The bbk32 gene was amplified from B31 genomic DNA, however, PCR product was not detected in the N40D10/E9 strain. (B) Southern blot of EcoR1-digested genomic DNA of both strains (top) was hybridized with the probe prepared

using the bbk32 PCR product from B31. An approximately 1.8 kb size selleck inhibitor fragment was detected only in B31, as expected, but not in the N40D10/E9 genomic DNA containing lane. In another study, we compared two important, highly variable virulence factors of B. burgdorferi, OspC and DbpA. As expected, both of these ACP-196 manufacturer molecules are present in both spirochete strains but showed high sequence variation [29]. Therefore, irrespective of the phylogenetic grouping of these strains using RST and OspC categorization, the presence of known virulence factors in both strains suggests that B31 and N40D10/E9 could possibly exhibit similar levels of pathogenicity. Furthermore, although BBK32 is an adhesin [41], previous

studies showed that its absence results in a subtle infectivity defect, exhibiting disease attenuation only at low dose of infection [45, 102, 103]. Divergence of fibronectin-binding adhesin gene bbk32 in N40D10/E9 strain BBK32 could possibly https://www.selleckchem.com/products/Trichostatin-A.html contribute to the adherence-mediated tissue colonization in B31 as compared to N40D10/E9 strain but a negative PCR result is not sufficient to demonstrate this difference. Since sequence divergence at the priming sites may lead to unsuccessful PCR amplification, Southern hybridization was conducted to determine the presence of a homolog of bbk32 gene in the N40D10/E9 strain. Absence of a band in N40

even under low stringency conditions (data not shown) indicated that either bbk32 homolog in the N40D10/E9 strain was absent or had substantial Cyclic nucleotide phosphodiesterase DNA sequence divergence from that in the B31 strain (Figure 3B). Therefore, irrespective of the presence of BBK32, the two B. burgdorferi strains examined here (B31 and N40D10/E9) show similar levels of binding to most cells, indicating redundancy of function. However, BBK32 may contribute to the binding of Lyme spirochetes to specific cell line(s), such as Vero cells, and potentially to epithelial cells in vivo. B31 and N40D10/E9 showed remarkably different protein expression profiles Although known virulence factors are present in both B31 and N40D10/E9 strains (Figure 3A), they only represent the molecular profile of previously identified virulence factors and molecules associated with infectivity. Therefore, it would be erroneous to conclude that they represent the full repertoire of the virulence factors of B. burgdorferi that play important roles during pathogenesis in the mammalian host.

CTM transformation medium was used to induce competence and for t

CTM transformation medium was used to induce competence and for transformation, as described

previously [11]. The CSP concentration was 100 ng ml-1 and DNA concentration was 1 μg ml-1. The chromosomal source of DNA carrying mutated PBP alleles was the 9V derivative Spain23F-1 clone (strain URA1258) which carries the following mutations near or within the conserved motifs on the PBPs: Gln443Glu, Thr451Ala, Glu481Gly, Ser485Ala and Thr494Ala in PBP2B, Thr338Ala, Met343Thr, Ala346Ser, Ala347Ser, Leu364Phe, Ile371Thr, Arg384Gly, Leu546Val and Asn605Thr in PBP2X, and Thr371Ala, Glu388Asp, click here Pro432Thr, Asn546Gly, Thr574Asn, Ser575Thr, Gln576Gly, Phe577Tyr, Leu606Ile, Asn609Asp, Leu611Phe and Thr612Leu SB202190 price in PBP1A. Transformants were selected on plates containing 0.1 μg ml-1 and 0.5 μg ml-1 penicillin, and appropriate integration of PBP mutations was confirmed by nucleotide sequencing. Plates containing 2 μg ml-1 rifampicin and 10 μg ml-1 chloramphenicol were used to select rif-23

and Δstkp::cat transformants. All constructions were verified by PCR with the primers AZD3965 described in Table 2[6, 12]. Spontaneous mutation to penicillin in DNA free medium was < 10-9. Penicillin G was from Atral, Castanheira do Ribatejo, Portugal, and rifampicin was from Aventis Pharma. To assess StkP and PBPs conservation 50 strains were randomly selected among those isolated between 1994 and 2005 in various areas in Portugal; they included forty invasive isolates from blood and cerebrospinal fluid and ten colonizing isolates from the nasopharynx of asymptomatic carriers. Half of the isolates (n = 25) were non-susceptible to penicillin [minimal inhibitory concentration (MIC) > 0.1 μg ml-1]. These isolates were compared to the following reference strains: the highly for resistant serotype 9V strain URA1258, two susceptible and three non-susceptible strains provided by the ATCC and the unencapsulated strain R6 (Table 1). Table 1 Strains and plasmids used in the study Strain or plasmid Genotype or description Phenotypea Source or reference S. pneumoniae       R6 Non-capsulated

D39 derivative, susceptible reference strain; genome sequence available (R6) AtbS Laboratory stock ATCC BAA-334 Virulent reference strain, genome sequence available (TIGR4) AtbS ATCC ATCC 51916 Reference strain Tennessee 23F-4 PenR, EryR, ATCC ATCC 700670 Reference strain Spain 6B-2 PenR, CmR, TetR ATCC ATCC 700673 Reference strain Hungary 19A-6 PenR, EryR, CmR, TetR ATCC URA1258 Multiresistant strain closely related to Spain 23F-1 clone PenR, CmR, TetR [21] Cp1015 D39 derivate, str1; hexA SmR [31] Cp1016 D39 derivate, str1; hexA, rif23 RifR [31] Cp7000 Cp1015, stkP::cat SmR CmR This study Pen1 Cp1015, penA, and pbpX from URA1258 SmR PenR This study Pen2 Cp1015, penA, pbpX and pbp1A from URA1258 SmR PenR This study Pen1STK Cp1015Pen1, stkP::cat SmR PenR CmR This study Pen2STK Cp1015Pen2, stkP::cat SmR PenR CmR This study E.

Curr Med Chem 2011, 18:256–279 CrossRef 56 Kohanski MA, Dwyer DJ

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“Background Nowadays, organic polymers have replaced many traditional engineering materials because of their superior performance and low cost [1].

CrossRef 9 Jaeger J, Liebler-Teneorio E, Kirschvink N, Sachse K,

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J: Cross-reactivity between Coxiella burnetii and Chlamydiae. Folia Microbiol (Praha) 1999, 44:579–584.CrossRef 19. Berri M, Laroucau K, Rodolakis A: The detection of Coxiella burnetii from ovine genital swabs, milk and faecal samples by the use of a single touchdown polymerase chain reaction. Vet Microbiol 2000, 72:285–293.CrossRefPubMed 20. Laroucau C, Souriau A, Rodolakis A: Improved sensitivity of PCR for Chlamydophila using pmp genes. Vet Microbiol 2001, 82:155–64.CrossRefPubMed 21. DeGraves FJ, Gao D, Hehnen HR, Schlapp T, Kaltenboeck B: Quantitative detection of Chlamydia psittaci and C. pecorum by high-sensitivity real-time PCR reveals high prevalence of vaginal infection in cattle. J Clin Microbiol 2003, 41:1726–1729.CrossRefPubMed 22.

A number of flavivirus infections may lead to acute lethal haemor

A number of flavivirus infections may lead to acute lethal haemorrhagic fever or encephalitis in patients and are therefore of great global public health concern. Flaviviruses are enveloped viruses with a single-stranded, non-segmented positive RNA genome [2]. The approximate 11 kb long genome contains only one open reading frame encoding a single polyprotein, which is thereafter cleaved by cellular and viral proteases to form three structural and seven non-structural proteins (NS1, NS2a, NS2b, NS3, NS4a, NS4b, NS5). Recent studies also reported that a NS1′ viral protein, which is often

detected during infection, is the possible result of ribosomal frameshifting [3]. The NS3 protein has a pivotal function in flavivirus RNA replication SCH727965 concentration and viral protein maturation [4, 5]. It consists of two functional domains, protease and helicase in N-and C-terminus, respectively. NS5 protein is constituted by two distinct Pictilisib cost domains as well, namely an N-terminal MLN8237 cell line methyltransferase and

a C-terminal RNA-dependent RNA polymerase that are required for capping and synthesis of the viral RNA genome, respectively [6]. NS3 and NS5 proteins are the major enzymatic components of the viral replication complex, which promotes efficient viral replication in close association with cellular host factors [7]. Due to their numerous functions and their central role in the Thymidylate synthase virus life cycle, NS3 and NS5 have been designated as important drug targets [8, 9]. To identify host factors interacting with flavivirus NS3 and NS5 proteins, we have conducted a high-throughput yeast two-hybrid (Y2H) screen. Since the pioneer study published by Uetz et al. in 2006 on Herpes viruses interactome, the use of the high-throughput yeast two-hybrid (Y2H) technique to conduct genome-scale screens of virus-host protein interactions has led to major advances in our understanding of viral infections [10–13]. These results from the integrative system biology approaches highlighted the ability of viral proteins to interfere with intracellular pathways

to the benefit of viral replication. Indeed, viruses not only take advantage of such interactions for their replication or to escape host defense but also induce cellular interactome perturbations leading eventually to infection-related diseases. Recently, studies using genome-wide RNA interference screens in human or insect cells were able to provide the identification of numerous host cell factors potentially required to interfere with DENV or WNV infection [14]. Some of the targets identified are host (mammalian) or vector (insect) exclusive, others are common to both. This suggests that conservation of required factors between dipteran and human hosts is associated to flavivirus propagation [15].

Appl Phys Lett 2013, 102:183505 CrossRef 14 Long S, Perniola L,

Appl Phys Lett 2013, 102:183505.OICR-9429 concentration CrossRef 14. Long S, Perniola L, Cagli C, Buckley J, Lian X, Miranda E, Pan F, Liu M, Suñé J: Voltage and power-controlled regimes

in the progressive unipolar RESET transition of HfO 2 -based RRAM. Sci Rep 2013, 3:2929. 15. Long S, Lian X, Cagli C, Perniola L, Miranda E, Liu M, Suñé J: A model for the set statistics of RRAM inspired in the percolation model of oxide breakdown. IEEE Electron Device Lett 2010, 34:999.CrossRef 16. Park J, Biju KP, Jung S, Lee W, Lee J, Kim S, Park S, Shin J, Hwang H: Multibit operation of TiO x -based ReRAM by Schottky barrier height engineering. IEEE Electron Device Lett 2011, 32:476.CrossRef 17. Park WY, Kim GH, Seok JY, Kim KM, Song SJ, Lee MH, Hwang CS: A Pt/TiO 2 /Ti Schottky-type selection diode for alleviating

the sneak current in resistance switching memory arrays. Nanotechnology Target Selective Inhibitor Library order 2010, 21:195201.CrossRef 18. Kim DC, Seo S, Ahn SE, Suh DS, Lee MJ, Park BH, Yoo IK, Baek IG, Kim HJ, Yim EK, Lee JE, Park SO, Kim HS, Chung UI, Moon JT, Ryu BI: Electrical observations of filamentary conductions for the resistive memory switching in NiO films. Appl Phys Lett 2006, 88:202102.CrossRef 19. Ielmini D, Nardi F, Cagli C: Physical models of size-dependent nanofilament formation and rupture in NiO resistive switching memories. Nanotechnology 2011, 22:254022.CrossRef 20. Panda D, Huang CY, Tseng TY: Resistive switching characteristics of nickel silicide layer embedded HfO 2 film. Appl Phys Lett 2012, 100:112901.CrossRef 21. Long S, Cagli C, Ielmini D, Liu M, Suñé J: Reset statistics of NiO-based resistive switching memories. IEEE Electron Device Lett 2011, check details 32:1570.CrossRef 22. Chien WC, Chen YC, Lai EK, Yao YD, Lin P, Horng SF, Gong J, Chou TH, Lin HM, Chang Dimethyl sulfoxide MN, Shih YH, Hsieh KY, Liu R, Chih-Yuan L: Unipolar switching behaviors of RTO WO

x RRAM. IEEE Electron Device Lett 2010, 31:126.CrossRef 23. Kim S, Biju KP, Jo M, Jung S, Park J, Lee J, Lee W, Shin J, Park S, Hwang H: Effect of scaling WO x -based RRAMs on their resistive switching characteristics. IEEE Electron Device Lett 2011, 32:671.CrossRef 24. Peng HY, Li GP, Ye JY, Wei ZP, Zhang Z, Wang DD, Xing GZ, Wu T: Electrode dependence of resistive switching in Mn-doped ZnO: filamentary versus interfacial mechanisms. Appl Phys Lett 2010, 96:192113.CrossRef 25. Peng CN, Wang CW, Chan TC, Chang WY, Wang YC, Tsai HW, Wu WW, Chen LJ, Chueh YL: Resistive switching of Au/ZnO/Au resistive memory: an in situ observation of conductive bridge formation. Nanoscale Res Lett 2012, 7:1.CrossRef 26. Lin CY, Wu CY, Wu CYC-Y, Lee TC, Yang FL, Hu C, Tseng TY: Effect of top electrode material on resistive switching properties of ZrO 2 film memory devices. IEEE Electron Device Lett 2007, 28:366.CrossRef 27. Lin CC, Chang YP, Lin HB, Lin CH: Effect of non-lattice oxygen on ZrO 2 -based resistive switching memory. Nanoscale Research Lett 2012, 7:187.CrossRef 28.

Raman spectra (shown in Figure  2d) were performed to detect and

Raman spectra (shown in Figure  2d) were performed to detect and characterize the graphene layers in the surface of the glass wires with the different growth times (10 and 20 min). Both of the Raman spectra present typical characteristics of selleck screening library graphene layers: obvious D, G, and 2D bands at PRIMA-1MET nmr approximately 1,340, approximately 1,588 and approximately 2,700 cm-1. For the Raman spectrum of the sample grown for 10 min, The I 2D/I G intensity ratio is approximately 1.1, and the full width at half maximum (FWHM) of 2D band is approximately 45 cm-1, which represents one to two layers of graphene film [28]. For the Raman spectrum of the sample grown

for 20 min, the I 2D/I G intensity ratio is approximately 0.5, and the full width at half maximum (FWHM) of 2D band is approximately 55 cm-1, which represents three to five layers of graphene film [28]. Compared with the Raman spectrum of the monolayer graphene [1], the 2D band of the multilayer graphene is broader and can be fitted into multipeaks, which can be explained by the double-resonance

theory: the electronic band YH25448 in vitro structure and electron-phonon interactions change with the number of the graphene layers [29]. In particular, the observed small D band intensity as compared to the G band intensity indicated low levels of defects or local disorder in our MLG films. Figure 2 SEM images before and after deposition and Raman spectra of the 3D graphene/glass fibers. (a) (b) The SEM images before and after deposition of the graphene for 20 min on the glass fiber membrane surface. (c) SEM image of the 3D graphene/glass fibers with the different diameter. (d) Raman spectra of Tyrosine-protein kinase BLK the 3D graphene/glass fibers deposited for 10 and 20 min. It is possible to investigate the state of the graphene by transferring it to a small holey copper grid using TEM. The HR-TEM image (shown in Figure  3) was conducted on the sample to identify the number of the graphene layer. The edge-on image of graphene

in Figure  3 indicates the thickness of the prepared graphene is three to five layers and the measured intergraphene spacing is approximately 0.34 nm, which is consistent with the previous report [28]. In addition, the electron diffraction (ED) pattern on the multilayer graphene film (the inset of Figure  3) reveals a hexagonal pattern, confirming the threefold symmetry of the arrangement of carbon atoms in graphene. These TEM results show direct evidence that multilayer graphene film is directly fabricated on the glass fibers. Figure 3 HR-TEM image of the graphene layers deposited for 20 min. The inset shows the ED pattern of the multi-layer graphene film. In our experiment, the lower-temperature (600°C) growth is necessary due to the lower melting point of the glass fiber, which can be obtained by the revised CVD system with the two-heating reactor. The mechanism of synthesis of core-shell graphene/glass fiber structures by using such revised CVD system has been discussed here.

Authors’ contributions MMR, LFM, and JFB designed the experiment

Authors’ contributions MMR, LFM, and JFB designed the experiment and analyzed and discussed the results. MMR fabricated the NAA-based DBR and performed the optical characterization. All authors redacted and revised the manuscript. All authors read and approved the final manuscript.”
“Background There is a need to develop

rapid and biocompatible pH sensors to monitor changes in the wound-healing trajectory that are, for example, caused by bacterial infection or biofilm formation. Chronic wounds do not heal within 3 months, and are considered an important and costly medical issue in Dinaciclib ic50 the world’s aging societies, imposing considerable pain, reduced mobility and decreased quality of life on the sufferers [1]. During the lengthy healing process, the wound is invariably exposed to bacteria that can colonize the wound bed and form biofilms. This alters the wound metabolism and brings about Ilomastat mw a change of pH [2]. Several recent studies have demonstrated an oscillation of the pH between 5.4 and 9, during a bacteria infection in the wounds [2, 3].

Recently, significant DNA Damage inhibitor research efforts have been devoted to pH sensors for the detection of pH variation in wound fluid [1]. These are typically based on dyes [4, 5] or on inductive transducers [6] incorporated into wound dressings. For example, Trupp et al. have synthesized a series of hydroxyl-substituted azobenzene derivatives as indicator dyes for optically monitoring pH between 6 and 10 [4]. However, there are concerns over the biocompatibility of these dyes. Sridhar and Takahata have O-methylated flavonoid developed a micro-fabricated wireless pH monitor involving a pH-sensitive hydrogel intended to be imbedded

into a wound dressing to track pH wirelessly. The authors observed changes in moisture level in a wound dressing in the pH range 2 to 7 [6]. The cost of this device may be a limiting factor for reduction to practice. Simultaneously, materials with optical features such as the porous silicon (pSi) have been associated with pH-responsive polymers in order to detect variation of pH [7–9]. PSi is an attractive candidate to use as a sensor in contact with wound fluid because the material is highly biocompatible and well tolerated in vivo, even when implanted into the eye [10]. The material displays strong thin-film interference effects, which result in the appearance of Fabry-Pérot interference fringes [11]. In turn, multilayers of pSi of alternating high and low refractive indices result in a sharp photonic resonances [11]. Changes in the effective refractive index of pSi films cause a shift in the interference pattern or the position of the photonic resonance peak in multilayered pSi resonators, respectively [12–15]. Perelman et al. developed a pH sensor based on pSi modified with thermo- and pH-responsive hydrogel poly(N-isopropylacrylamide-co-acrylic acid).

The flhD/C DNA was detected as previously described Construction

The flhD/C DNA was detected as GW2580 mouse previously described. Construction of the null alleles of flhD, fliC, and flhA genes The flhD gene was isolated from pBYL2DC by digesting with BsmI, which cleaves at two sites in pBYL2DC and thereby conveniently deletes flhC from the operon. The resulting plasmid was designated pBYL2D. A kanamycin resistant gene from pACYC177 was isolated, made blunt-ended using a DNA-blunting kit (Takara Co., Tokyo, Japan), and inserted in the unique EcoRV site of the flhD gene. The resulting plasmid was designated pBYL2D-Kan. The pBYL2D-Kan

Selleck Nec-1s was re-isolated and linearized after HpaI and SspI restriction enzyme digestion, which deleted the ampicillin resistance gene and replication site of the plasmid. The linearized construct was transferred into H-rif-8-6, resulting

in the homologous replacement of the native flhD gene and generating a null allele. The DNA fragment of fliC was amplified by PCR from H-rif-8-6. After PCR amplification using two oligonucleotide primers (fliC-sen and fliC-anti), the partial fliC DNA fragment was purified, digested using AhdI and HindIII, and subcloned into plasmid pBR322 to generate the fliC plasmid. A kanamycin resistant gene from pACYC177 was isolated, made blunt-ended, and inserted into the unique SalI site of the fliC gene. The resulting plasmid was designated pfliC-Kan. The pfliC-Kan was linearized after AhdI and HindIII restriction enzyme digestion, which deleted the ampicillin resistance gene and replication site of the plasmid. The linearized construct HDAC inhibitor was transferred into H-rif-8-6 resulting in the homologous

replacement of the native fliC gene and generating a null allele. The DNA fragment of flhA was amplified by PCR from H-rif-8-6 using Molecular motor oligonucleotide primers flhA-sen and flhA-anti. The partial flhA DNA fragment was purified, digested using the restriction enzymes ClaI and EcoRI, and subcloned into plasmid pBR322 using T4 ligase to generate the flhA plasmid. A kanamycin resistant gene from pACYC177 was isolated, made blunt-ended, and inserted in the unique SalI site of the flhA gene. The resulting plasmid was designated pflhA-Kan. Computer analysis of sequence data The nucleotide sequence and the deduced amino-acid sequence of FlhD/C were compared using the BLAST and FASTA programs of the National Center for Biotechnology Information server. Sequence data were compiled by DNASIS-Mac software (Hitachi, Tokyo, Japan). RNA preparation and Northern hybridization Bacteriocin synthesis medium (BSM; 0.5% sucrose, 0.1% NH4Cl, 0.2% KH2PO4, and 0.02% MgSO4·7H2O [pH = 7.5]) was used to produce Carocin S1. Total RNA was extracted from cells (Pectobacterium carotovorum subsp. carotovorum harboring constructs) that were grown without drugs at 28°C. To determine the stability of H-rif-8-6, TH12-2, TH12-2/pBYL2C, KH17, and KH17/pBYL2D strains, culture samples (8 ml each; with rifampicin [0.