Acta Bot Mex 15:47–64 Sagástegui A (1995) Diversidad florística de Contumazá. Editorial Libertad, Trujillo Silva learn more RA, Santos AMM, Tabarelli M (2003) Riqueza e diversidade de plantas lenhosas em cinco unidades de paisagem da Caatinga. In: Leal IR, Tabarelli M, Silva JMC (eds) Ecologia e conservação da caatinga. Ed. Universitária da UFPE, Recife Svenson HK (1946) Vegetation of the coast of Ecuador and Peru and its relation to the Galápagos Islands. Am J Bot 33:394–498CrossRef The Nature Conservancy, Fundación Agua, EcoCiencia et al (2004) Portafolio de sitios prioritarios para la conservación dentro de la unidad de planificación ecorregional Pacífico Ecuatorial, Quito. http://​conserveonline.​org/​workspaces/​pe_​era.

Cited 17 Aug 2007 Ulloa Ulloa C, Neill DA (2005) Cinco años de adiciones a la flora del Ecuador. 1999–2004. Universidad Técnica Particular SAHA in vivo de Loja, Missouri Botanical Garden, FunBotanica, Loja, Ecuador

Ulloa Ulloa C, Zarucchi JL, León B (2004) Diez años de adiciones a la flora de Perú. Arnaldoa, ed. especial, Nov 2004 UNESCO-MAB (2002) Seville+5 Recommendations: Checklist for Action. http://​unesdoc.​unesco.​org/​images/​0012/​001266/​126629e.​pdf#xml=​http://​unesdoc.​unesco.​org/​ulis/​cgi-bin/​ulis.​pl?​database=​ged&​set=​44115CBA_​0_​13&​hits_​rec=​9&​hits_​lng=​eng. Cited 29 July 2009 Valencia R, Pitman NS, Léon-Yánez S (eds) (2000) Libro rojo de las plantas endémicas del Ecuador. Herbario QCA, Pontificia Universidad Católica del Ecuador, Montelukast Sodium Quito van der Werff H, Consiglio T (2004) Distribution and conservation significance of endemic species of flowering plants in Peru. Biodivers Conserv 13:699–713 Venegas PJ (2005) Herpetofauna

del bosque seco ecuatorial de Perú: taxonomía, ecología y biogeografía. Zonas Áridas 9:9–26 Weberbauer A (1945) El mundo vegetal de los Andes peruanos. Editorial Lume, Lima-Peru Wilson EO (1992) The diversity of life. Harvard University Press, Cambridge Wood JRI (2006) Inter-Andean dry valleys of Bolivia–floristic affinities and patterns of endemism: insights from Acanthaceae, PD173074 research buy Asclepiadaceae and Labiatae. In: Pennington RT, Lewis GP, Ratter JA (eds) Neotropical savannas and seasonally dry forests: plant diversity, biogeography and conservation. CRC Press, Florida”
“Erratum to: Biodivers Conserv DOI 10.1007/s10531-009-9680-9 The Author would like to add the following paragraph on page 4 after the sentence “…. Proper controls should consider animal behavior and spatial components such as pig home range size, movements, and plant distribution patterns. “Another potentially confounding factor is that large grazing and/or browsing ducks and geese where once common to the islands but are now extinct or greatly reduced in population size (Paxinos et al. 2002). One of these geese species was four times the size of a Canada goose (Branta canadensis) to which they were closely related.

Nanoscale Res Lett 2012, 7:241–248.CrossRef 16. Švorčík V, Siegel J, Šutta P, Mistrík J, Janíček P, Worsch Cell Cycle inhibitor P, Kolská Z: Annealing of gold nano-structures sputtered on glass substrate. Appl Phys A 2011, 102:605–611.CrossRef 17. Doron-Mor I, Barkay Z, Filip-Granit N, Vaskevisch A, Rubinstein I: Ultrathin gold island films on silanized glass. Morphology and optical properties. Chem Mater 2004, 16:3476–3483.CrossRef

18. Kan C, Zhu X, Wang GJ: Single-crystalline gold microplates: synthesis, characterization, and thermal stability. J Phys Chem B 2006, 110:4651–4656.CrossRef 19. Slepička P, Švorčík V, Šlouf M, Rybka V, Špirková M: Characterization of metal nanolayers sputtered on poly(ethyleneterephtalate). Optoelectron Adv Mater– Rapid Com 2008,2(Š): 153–160. 20. Hopfner U, Hehl H, Brehmer L: Preparation of ordered thin gold films. Appl Surf Sci 1999, 152:259–265.CrossRef

21. Roland T, Khalil A, Tanenbaum A, Berguiga L, Delichère P, Bonneviot L, Elezgaray J, Arneodo A, Argoul F: Revisiting the physical www.selleckchem.com/products/th-302.html processes of vapodeposited thin gold films on chemically modified glass by atomic force and surface plasmon microscopies. Surf Sci 2009, 603:3307–3320.CrossRef 22. Zhou HS, Honma I, Komiyama H, Haus JW: Controlled synthesis and quantum-size effect in gold-coated nanoparticles. Phys Rev 1994, B 50:12052–12056. 23. Vogel N, Zieleniecki Fenbendazole J, Köper I: As flat as it gets: ultrasmooth surfaces from template-stripping procedures. Nanoscale 2012, 4:3820–3832.CrossRef selleck 24. McDonnell JM: Surface plasmon resonance: towards an understanding of the mechanisms

of biological molecular recognition. Curr Opin Chem Biol 2001, 5:572–577.CrossRef 25. Niu J, Shin YJ, Son J, Lee Y, Ahn JH, Yang H: Shifting of surface plasmon resonance due to electromagnetic coupling between graphene and Au nanoparticles. Opt Express 2012, 20:19690.CrossRef 26. Li Y, Liu X, Lin Z: Recent developments and applications of surface plasmon resonance biosensors for the detection of mycotoxins in foodstuffs. Food Chem 2012, 132:1549–1554.CrossRef 27. Zhang J, Liu Y, Ke Y, Yan H: Periodic gold nanoparticle arrays templated by self-assembled 2D DNA nanogrids on a surface. Nano Lett 2006, 6:248–251.CrossRef Competing interests The authors declare that they have no competing interests. Authors’ contributions AS carried out the sample preparation and participated on the AFM analysis and paper corrections. PS analyzed the surface morphology, evaluated the surface roughness and thickness, and designed the study. IK analyzed the electrical properties and carrier concentration of evaporated and annealed samples. PM and AM performed the RBS analysis. VŠ participated in the study coordination and paper correction. All authors read and approved the final manuscript.

This is also reflected in gill associated microbial communities of other oyster species that differ more strongly from the surrounding sea water than INCB018424 for example gut communities [18]. The numerical abundance of α-proteobacteria in open water could however partly been attributed to PCR bias by preferential amplification of sequences from this taxonomic group [61]. The dominant genus detected, was Sphingomonas which contains opportunistic species [62] and can also commonly be found in gill tissue of European plaice Pleuronectes platessa from the same region [38]. It was also abundant on freshly

prepared cod in Iceland [63], indicating that this genus can reach high numbers on living hosts but is quickly outcompeted after the host’s death. Dominance of a few closely related OTUs has been reported for other species of oysters. Zurel et al. [18] for example found that https://www.selleckchem.com/products/chir-98014.html between 59 – 79% of OTUs in Chama spp. oysters in the Red Sea and the Mediterranean

belonged to OTUs from the class Oceanospirialles closely related to the genera Spongiobacter or Endozoicomonas (Hahellaceae), which is known for symbiotic associations. While we also observed 47 OTUs from the Oceanospirialles, these were relatively rare (99 reads in total) and only a single OTU was affiliated to the family Hahellaceae. Similarly, we only found very few OTUs classified as Arcobacter spp. (13 OTUs, 16 reads), which represent a major and common component of Chilean oysters Tiostrea chilensis[60]. selleckchem This suggests that oyster microbiomes can have similar structures in terms of abundances but dominant taxa differ strongly between species, habitats and sampled tissues. Under certain environmental conditions gut communities of other Crassostrea species were found to be dominated by Mycoplasma[17], which also became dominant in some oysters after disturbance in our experiments (Figure 5A).

The natural dominance of Mycoplasma in oysters from much warmer habitats [17] may thus suggest that Mycoplasma represents a temperature sensitive part of oyster microbiota and may proliferate preferentially at higher temperatures. Host stress and abiotic disturbance both could have contributed to the major shift in microbial PLEKHB2 community structure (Figure 3). The direction and magnitude of the shift was dependent on the initial community composition, and although no significant differences were observed between oyster beds in ambient conditions there was some indication for oyster bed specific shifts (Figures 3 and 4). The strongest shifts occurred in the beds with initially high microbial diversity (OW and PK), manifested in a sharp decrease in microbial diversity. In the oyster bed with low diversity on the other hand we observed no significant change in bacterial diversity (Figure 2).

Table 1 Minimum

inhibitory concentrations (MICs) of antibiotics used in this study Antibiotics Drug class MICs againstOrientia a) MICs against mycoplasmasb) Lincomycin Lincosamide No available data 0.25–2 μg/mL Ciprofloxacin New Quinolone 6.25–25 μg/mL 0.125–2 μg/ml Gentamicin Aminoglycoside No available TEW-7197 solubility dmso datac) 2.5–500 μg/mL Kanamicin Aminoglycoside No available data 2.5–500 μg/mL Minocycline Tetracycline 0.024–0.195 μg/mL 0.016–32 μg/mL MICs were obtained from previous reports. a) from [8] and b) from [5–7]. c) Gentamycin was not effective against Orinetia tsutsugamushi in a mouse model [25]. Our result of the direct sequencing showed that Ikeda and Kuroki strains of O. tsutsugamushi were contaminated with Mycoplasma hominis and M. orale respectively. M. hominis and M. orale are 10 to 30% of contaminants of cell cultures (Table 2) [11]. Previous reports showed that M. fermentas, M. hyorhinis, M. arginini and Acholeplasma laidlawii are the most common contaminants selleck products as well as M. hominis

and M. orale. More than 90% of the contaminants were caused by these six mycoplasmas [11, 12]. The TaqMan PCR and the nested PCR can detect not only all the 6 most common contaminants also some other mycoplasmas. These facts suggested that the detection methods were very reliable Suplatast tosilate to monitor mycoplasmas-contaminations in this study. Table 2 Major mycoplasmas, and their detection and sequencing methods in this study Species   PCR for detection PCR for Sequencingd)       Frequency of contaminationa) tufgene (TaqMan PCR)b) 16S-23S ribosomal RNA intergenic region (nested PCR)c) Match of new PCR primers Strains Sequence ID Most common contaminant

species             Mycoplasma fermentans 10%-20% + + Match human B cell lymphoma contaminants, 16054780 AY838558 Mycoplasma hyorhinis 10%-40% + + Match HUB-1 NC_014448.1 Mycoplasma orale 20%-30% + + Partial Match ATCC 23714D gi|315440428 Mycoplasma arginini 20%-30% No Data + Partial Match G230 gi|290575476 Acholeplasma laidlawii 5%-20% + + Match PG-8A CP000896 Mycoplasma hominis 10%-20% + + Match ATCC 23114 G418 M57675 Other species             Mycoplasma arthritidis No Data + No Data Match 158L3-1 NC_011025.1 Mycoplasma bovis No Data + No Data Match PG45 NC_014760.1 Mycoplasma buccale No Data + No Data No data – - Mycoplasma faucium No Data + No Data No data – - Mycoplasma gallisepticum No Data + No Data Match PG31 X16462 Mycoplasma genitalium No Data + + Match ATCC33530 X16463 Mycoplasma hyopneumoniae No Data + No Data Match 7448 NC_007332.1 Mycoplasma penetrans No Data + No Data Match HF-2 NC_004432.

Co-purification of DNA from these extractions were preformed Lazertinib chemical structure from the separated organic layer, using a DNeasy® Blood & Tissue Kit according to protocols for total bacterial DNA extractions (Qiagen, Valencia, CA). Purified DNA were kept in 1x Tris-EDTA Buffer and concentrations were measured spectrophotometerically at a ratio of 260/280 nm (Nanodrop 1000, Wilmington, DE). DNA at concentrations of 40–50 ng/μl in 50 μl of water was provided for sequencing. High throughput sequencing was conducted using 454 ®pyrosequencing technology (Roche Laboratories, Branford,

CT) at Research and Testing Laboratories, LLC (Lubbock, TX). Duplicate samples of RNA, collected from triplicate animals from each sex for each experimental condition were prepared for quantitative Real Time- PCR (qRT-PCR). High- Osimertinib Capacity® cDNA Reverse Transcription kit was used (ABI, Foster City, CA). For RNA samples with concentrations below 60 ng/μl a High® Capacity RNA-to-cDNA Master Mix kit was used for cDNA synthesis (ABI; Foster City, CA). cDNA were analyzed using SYBR green probes for genes of interest for Open® Array platform (Life Technologies Inc.; Carlsbad, CA). Probes for all genes were selected from array panels and customized for our study- 9 plates were used in the analysis. Assays were performed by The University of Texas, Southwestern at Dallas. Analysis of data was

conducted using Open® Array Real Time qPCR Analysis Software Version 1.0.4. Each cDNA sample was analyzed in duplicate,

from triplicate animals and both sexes. qRT-PCR analysis of MAP concentrations from tissues The template DNA used for construction of GS-9973 mw standards was extracted from MAP culture. (-)-p-Bromotetramisole Oxalate Briefly, 10 ml of the MAP culture was pelleted using centrifugation (Marathon 2100R, Thermo-Fisher Scientific, Houston, TX) at 5000 × g for 15 minutes. The cells were washed twice with HPLC-grade water (Ricca Chemical Company; Arlington, TX) and again suspended in new HPLC-grade water. DNA was extracted by heating 50 μl of cell suspension in PCR tubes (VWR Int, Westchester PA) at 99°C for 15 minutes in Gene Amp PCR system 2700 Thermocycler (Applied Biosystems, Foster City, CA). The heated sample was centrifuged to pellet the cell debris and the supernatant was used as template for successive experiments. The primers used for this assay amplifies a 163 bp region of the IS-Mav region in the MAP genome. Various primer pairs were tested before selecting the ISMav2 primers [3, 4, 41–43]. By using plasmids with the 163 bp fragment DNA insertion as standards, serial dilutions were tested to develop a standard curve and then enumerate the number of MAP cells in the experimental samples by plotting the Ct values on the curve. This was confirmed using the melting curve analysis of the PCR product which showed only one peak for ISMav2; thus the amplicon was very specific for MAP.

Sem Sci and Tech 2010, 25:024003.CrossRef

8. Nassiopoulou AG, Grigoropoulos S, Gogolides E, Papadimitriou D: Visible luminescence from one- and two-dimensional silicon structures produced by conventional lithographic and reactive ion etching techniques. Appl Phys Let 1995, 66:1114.CrossRef 9. Dresselhaus MS, Lin YM, Oded R, Black MR, Kong J, Dresselhaus GN: Springer Handbook of Nanotechnology. Edited by: Bhushan B. Berlin: Springer; 2010:99. 10. Peng K, Fang H, Hu J, Wu Y, Zhu J, Yan Y, Lee S: Metal-particle-induced, highly localized site-specific etching of Si and formation of single-crystalline Si nanowires in aqueous fluoride solution. Chemistry (Weinheim an der Bergstrasse, Germany) 2006, 12:7942–7947.CrossRef 11. Hochbaum AI, Gargas D, Hwang YJ, Yang P: Single crystalline mesoporous silicon nanowires. GSK126 Nano Lett 2009, 9:3550–3554.CrossRef 12. Zhong X, Qu Y, Lin YC, Liao L, Duan X: Unveiling the BYL719 manufacturer formation pathway of single crystalline porous silicon nanowires. ACS Appl Mater Interfaces 2011, 3:261–270.CrossRef 13. Qu Y, Liao L, Li Y, Zhang H, Huang Y, Duan X: Electrically conductive and optically active porous silicon nanowires. Nano Lett 2009, 9:4539–4543.CrossRef 14. Lin L, Guo S, Sun X, Feng J, Wang Y: Synthesis and photoluminescence properties of porous silicon nanowire arrays. Nano Res Lett 2010, 5:1822–1828.CrossRef 15. Voigt F, Sivakov V, Gerliz V, Bauer GH, Hoffmann

B, Radnoczi GZ, Pecz B, Christiansen S: Photoluminescence of samples produced by electroless wet this website chemical etching: between silicon nanowires and porous structures. Phys Status Solidi A 2011, 208:893–899.CrossRef 16. Chen H, Zou R, Chen H, Wang N, Sun Y, Tian Q, Wu J, Chen Z, Hu J: Lightly doped single crystalline porous Si nanowires with improved optical and electrical properties. J Mater Chemistry 2011, 21:801.CrossRef 17. He H, Liu C, Sun L, Ye Z: Temperature-dependent photoluminescence properties of porous silicon nanowire arrays. Appl Phys Let 2011, 99:23106.CrossRef 18. Artoni P, Irrera A, Iacona F, Pecora EF, Franzo G, Priolo F: Temperature dependence and aging effects on silicon nanowires photoluminescence. Opt Express 2012, 20:1483–1490.CrossRef 19. To

WK, Tsang CH, Li HH, Huang Z: Fabrication of n-type mesoporous silicon nanowires by one-step etching. Nano Lett 2011, 11:5252–5258.CrossRef 20. TCL Nassiopoulou AG, Gianneta V, Katsogridakis C: Si nanowires by a single-step metal-assisted chemical etching process on lithographically defined areas: formation kinetics. Nano Res Lett 2011, 6:597.CrossRef 21. Sailor MJ: Porous Silicon in Practice: Preparation, Characterization, and Applications. Weinheim: Wiley-VCH; 2012. 22. Salcedo WJ, Fernandez FJR, Galeazzo E: Structural characterization of photoluminescent porous silicon with FTIR spectroscopy. Brazilian J Phys 1997, 27:158–161. 23. Canham LT: Silicon quantum wire array fabrication by electrochemical and chemical dissolution of wafers. Appl Phys Let 1990, 57:1046.

Since filamentation was not responsible for the death of the macrophages incubated with the ISRIB environmental strains, maybe other virulence factors could account for these observations. Secretion of hydrolytic enzymes such as aspartic proteinases and phospholipases have been associated with C.albicans virulence [14, 16, 26, 27] and also with C. parapsilosis virulence [15, 18, 28–31]. Eighty percent of the tested C. parapsilosis strains were found to have high proteinase activity, being the majority blood isolates. To our knowledge, no other study compared Sap production in clinical and environmental C. parapsilosis

isolates, but Dagdeviren et al. [32] observed a higher production of acid proteinase among C. parapsilosis blood isolates compared to non-blood isolates. From the eight C. orthopsilosis tested only 25% were Sap producers, whereas selleck products none of the C. metapsilosis was. This is in accordance with Lin et al. [33], who also reported differences in proteinase activity within the three major groups of C. parapsilosis. No correlation was observed between hydrolytic enzymes secretion and environmental or clinical isolates,

or with cell damage (p > 0.05). Macrophage activation induces releasing of several key mediators, including proinflammatory cytokines such as TNF-α, which are important for protecting the host against disseminated candidiasis [34–36]. The amount of TNF-α produced by macrophages infected with C. parapsilosis isolates from bloodcultures was significantly higher than the amount produced by macrophages infected with environmental isolates, indicating that clinical isolates induce a higher pro-inflammatory https://www.selleckchem.com/products/Vorinostat-saha.html response than environmental strains. The fact that a high macrophage cell lysis occurred in the co-incubations with the environmental strains could also account for these results. In contrast, Orsi Casein kinase 1 et al. [23] reported little or no TNF-α production in the co-incubations of strains of the C. parapsilosis complex with microglial cells. This

discrepancy may result from the fact that the 6-hour incubation time used in their study was insufficient to trigger cell response. Our results showed a positive correlation between filamentation and TNF-α release (p = 0.0119) for C. parapsilosis. Candida orthopsilosis strains induced TNF-α levels similar to the clinical isolates, whereas C. metapsilosis isolates induced the production of lower amounts, which is in agreement with Gácser et al. [19] who showed that C. metapsilosis appears as the less virulent of the three species of the C. parapsilosis complex. Nevertheless, recent literature indicates that C. metapsilosis can be retrospectively identified at a frequency similar to C. orthopsilosis and from virtually all body sites [37, 38]. In addition, a meta-genomic study has found C. metapsilosis sequences in the oral cavity of healthy carriers, suggesting the possibility of oral commensalism for this species [39].

[15] the cytotoxic activity and IFN-γ production by CTLs are independent PD-1/PD-L1 Inhibitor 3 cost functions which may follow different regulatory pathways. In fact, not all CD8+ T cells function as “”killer”" cells. Indeed, during the acute phase of a CD8+ T-cell response, IFN-γ production, cytotoxicity, and proliferation appeared as independently regulated in cancer and infections [15, 33, 34]. The simultaneous determination of the different functions exerted by T cells can

offer a valuable tool for ex vivo analysis of the immune response against cancer as well as infections, but also in assessing autoimmune diseases as well as to identify correlates of immune protection exploitable for therapeutic strategies based on vaccine development. The assay we developed is based on a dual-colour LysiSpot

method aimed at measuring the extent of the recognition of tumour cells by CTLs, as elicited in a rat model harbouring a colorectal tumour induced by the DHD-K12 cell line. In this assay the simultaneous determination of the different functions APR-246 mouse exerted by T cells can offer a valuable tool for ex vivo analysis of the immune response against cancer as well as furnish a base to evaluate the number and function of lytic effector cell. DHD-K12 cells naturally express a tumour-associated antigen that induces specific cytotoxic responses in immune competent syngeneic animals [16, 17]. The synthetic nonapeptide antigen, CSH-275, was previously used in a IPI-549 cell line vaccination protocol and gave proof of the induction of an antitumour activity as elicited by

the vaccination [17]. By the ELISPOT assay illustrated in Figure 1 we have further demonstrated the specific recognition of this nonapeptide, epitope constitutionally express in DHD-K12 during cells In the present study, the DHD-K12 cell line was transiently transfected, using a pCMV-LacZ vector containing the nuclear-targeted β-gal coding region. This method permits to easily “”mark”" [35] the tumour cell line. We chose to use the plasmid DNA- Lipofectamine complex to introduce a gene expressing a marker protein because this methodology with non-viral vectors, either plasmids or siRNAs, efficiently transfects human colon cancer cells [36–39] as well primary neurons. In the latter, optimized protocols gives transfection efficiencies of 20-30%, a great improvement compared with less than 3% previously reported [40]. Non-viral vectors have been receiving increasing attention, since they are safer and cheaper, and can be produced easily in large quantities. A recent study comparatively examined a panel of non-viral gene transfer systems in several cells of different origins, including human colorectal carcinoma, and in human primary cells [41]. In this work, the authors evaluated the requirements for successful transfection and the potential for optimization of transfection efficiency.

8%) for cc32, 49/66 (74.2%) for cc162, 15/18 (83.3%) for cc41/44 while the highest value was found among the cc269 isolates (32/33; 97%). Figure 3 Contribution of each antigen to selleck compound coverage in relation to clonal complex. The numbers indicate the percentage of isolates predicted to be covered by each individual antigen. Isolates were defined as covered if they expressed PorA VR2 4 or had a MATS relative potency greater than the positive bactericidal threshold (PBT) for fHbp, NHBA, or NadA. The lowest fHbp contribution was found among the cc162 (24/66 36.3%) while higher contributions were

found among cc41/44 (12/18; 66.7%), cc269 (26/33; 78.8%) and cc32 isolates (16/16; 100%). PorA contribution to coverage in relation to clonal complexes revealed that PorA 1.4 was found mainly among the cc41/44 (9/18; 50%) while low PorA contribution was found for cc162 (2/66; 3%) and no PorA contribution to coverage was found for cc269 and cc32 strains. In contrast, Peptide 17 cost buy XAV-939 NadA contribution to coverage was low among cc41/44 isolates (1/18; 5.6%),

while it was not found in other clonal complexes (Figure  3). The recent licensure of the 4CMenB vaccine in Europe may promote recommendations for its use by national immunization technical advisory groups. Data on strain coverage are therefore crucial for decision making. This study provides the first such data on the potential coverage of Greek MenB isolates by 4CMenB. The relevance of this study is related to the high incidence, in Greece, of cc162, which is rare in Europe. cc162 has been described to be present both in disease-associated and in carrier isolates in Greece, with a high degree of heterogeneity among the isolates [35, 36]. When compared with killing of MenB strains in the hSBA, MATS-PBT was shown to provide a conservative prediction of strain coverage, especially in older age groups (children, adolescents, and adults) [37]. Notably, the MATS from assay was not designed to assess synergistic killing effects for strains having multiple MATS relative

potencies for different antigens slightly below their positive bactericidal thresholds. Using this conservative predictor, the 4CMenB vaccine is expected to provide good strain coverage globally (89.2%) among the tested isolates (148 strains isolated from cases of IMD during 1999–2010) and in particular for the most prevalent ccs, which include cc162 and cc269 predicted to be covered at 86.4% and 97%, respectively. The components of the 4CMenB vaccine contributed to MATS-PBT predicted strain coverage singularly (for a total of 44.6% of strains covered by one antigen) or in combination each other (44.6% covered by two or more antigens). A key antigen contributing to the coverage of Greek isolates was NHBA, predicted to cover the 78.4% of isolates. The greater contribution of NHBA to coverage with respect to the other antigens was evident for three out of the four most frequent MLST genotypes in Greece, cc162, cc41/44 and cc269.

5–2.5/42–72 BKD584 3 2.1–4.1/48–95 BKD694 3 1.9–2.9/19–27 BKD1023 2 1–2/37–71 BKD1506 3 5–8/34–56 BKD1850 2 4–5/43–55 BKD1935 3 click here 1–3/52–154 BKD2126 8 5.3–11.3/37–79 BKD2770 4 2.8–4.8/26–44 BKD3038 2 2.6–4.6/46–81 *locus name reflects its position within genome of R. salmoninarum reference isolate ATCC33209T (Accession number NC_010168). Loci in italics represent a minimum combined loci required to sufficiently recognized 17 R. salmoninarum haplotypes with the HGDI of value 0.81. The allelic selleck products diversity ranged from two (BKD 92, 396, 494, 526, 1023, 1850 and 3038) to eight different alleles (BKD 2126) per locus. The largest observed variation in allele size was

found in locus BKD2126 which varied between five to eleven repeats (Table 1). The VNTR MG-132 molecular weight typing system has a discriminatory power value of 0.81 and seventeen different haplotypes of R. salmoninarum were distinguished using 16 combined polymorphic VNTRs (Table 1, Table 2). A VNTR typing system relying on only six combined loci (BKD23, BKD305, BKD694, BKD1506, BKD1935, BKD2126) also sufficiently recognized 17 R. salmoninarum haplotypes, with the same discriminatory power value of

0.81. Table 2 Renibacterium salmoninarum isolates haplotype identified using multilocus tandem repeat sequencing Haplotype Isolate name Country of origin Host species Environment (wild/farmed fish) Data of isolation A MT1470, MT1511b, MT2119c, MT2622c Scotland RT FW, SW (F) 1994–2002 B MT452a, MT839, MT1351, MT1363, MT1880, MT2979, MT3277a, MT3314, MT3315b, MT3402, N4245, N6642, N6552d, N6553d, N6694, N6765, N6863e, N6864e Scotland, Norway AS, RT FW, SW (F) 1988–2009 C MT2943, MT3320 Scotland

AS SW (F) 2005–2008 D MT3482, MT3483 Scotland AS, RT SW (F) 2009 E N3769, N6695 Norway AS, RT FW, SW (F) 1997–2008 F N5298 Norway AS SW (F) 2005 G MT3106, MT3479, TERV Scotland AS, RT FW, SW (F) 2006–2009 H MT861 Scotland AS FW (F) 1990 I MT1262 Scotland AS FW (F) 1992 J ATCC33209 N. America Chinook salmon SW (F) 1974 K MT3313 Scotland RT FW (F) 2008 L MT444 Scotland AS SW (F) 1988 M N5223 Norway AS SW (F) 2005 N N6975 Norway AS SW (F) 2009 O NCIMB1116 Scotland AS FW (W) 1960 P NCIMB1114 Scotland AS FW (W) 1960 Q N7443 Norway Bcl-w AS FW (W) 1985 a,b,c,d,erepresent R. salmoninarum isolates from different disease outbreaks occurring on the same aquaculture site. RT – rainbow trout, AS – Atlantic salmon, FW – freshwater, SW – seawater, FA – farmed fish, W – wild wish. Phylogenetic relationships among R. salmoninarum isolates inferred from VNTRs The phylogenetic relationships among the R. salmoninarum strains inferred from 16 polymorphic VNTRs are illustrated in Figure 1. Two distinct groups comprising haplotypes A-L (group 1) and M-Q (group 2) were supported with a high bootstrap value (92%). Group 1 comprised R. salmoninarum from both Atlantic salmon and rainbow trout farmed in Scotland and Norway, recovered over a period of more than 40 years. This group also includes the type strain of R.