Later, Grace’s medium with 10% fetal bovine serum, FBS, (Lonza) w

Later, Grace’s medium with 10% fetal bovine serum, FBS, (Lonza) was used for the isolation of other biovars. Since streptomycetes selleck chemicals growing in liquid medium form compact colonies, the following strategy was applied to isolate a pure culture: single colonies were transferred into individual wells of 24-well plate containing 500 μl fresh medium and were disrupted

by pipetting. After that, bacteria were incubated again until new micro-colonies appeared and the procedure was repeated three times. Finally, bacterial biomass was stored at −80°C with glycerol (15-20%) added to liquid medium. Bacterial isolates were named with the first three letters of the host species name, plus the running number for the host specimen according to our internal collection, and a number referring to the replicate isolate (e.g. alb539-2 refers to isolate 2 of the Philanthus albopilosus specimen no. 539). DNA extraction, PCR amplification, and identification of isolates Bacteria grown in appropriate liquid medium were collected in 1.5 ml tubes by centrifugation at 5000 × g for 1 min at room temperature and washed twice with sterile PBS (137 mM NaCl; 2.7 mM KCl; 10 mM

Na2HPO4; 2 mM KH2PO4). The bacterial Ibrutinib datasheet biomass was lysed as described elsewhere [39]: briefly, the biomass was resuspended in 500 μl TE25S buffer (25 mM Tris (pH 8.0), 25 mM EDTA (pH 8.0), 0.3 M sucrose) with lysozyme (2 mg/ml) and incubated at 37°C for 1 h. Afterwards, 50 μl proteinase K (20 mg/ml) and 30 μl SDS (10%) were added, mixed and the samples were incubated at 55°C with agitation for 20 min. 100–200 μl Protein Precipitation Solution Idelalisib cell line (Qiagen) was added to the transparent lysate, which was then thoroughly mixed and centrifuged at >16,000 × g for 10 min at 4°C to sediment proteins. The supernatant was transferred into a fresh tube, and an equal volume (i.e. 600–700 μl) of isopropanol was

added; the solution was thoroughly mixed and the tube was incubated at −20°C for ≥30 min, followed by centrifugation at ≥16,000 × g for 10 min to sediment DNA. The DNA pellet was then washed twice with 500 μl EtOH (70%), air-dried, and resuspended in EB buffer. Bacterial 16S rRNA gene fragments were amplified with the primers fD1 (5’-AGAGTTTGATCCTGGCTCAG-3’) and rP2 (5’-ACGGCTACCTTGTTACGACTT-3’); gyrase subunit A (gyrA) gene fragments were amplified with gyrA-5F (5’-AACCTGCTGGCCTTCCAG-3’) and gyrA-5R (5’-AACGCCCATGGTGTCACG-3’); gyrase subunit B (gyrB) gene fragments were amplified with primers gyrB-F1 (5’-GAGGTCGTGCTGACCGTGCTGCA-3’) and gyrB-R3 (5’-SAGCTTGACCGAGATGATCG-3’) [28].

PCR products were purified with QIAquick PCR Purification Kit (Qi

PCR products were purified with QIAquick PCR Purification Kit (Qiagen) and sequenced with primers fD1, rP2 and R1087 (5’-CTCGTTGCGGCACTTAACCC-3’), gyrA-5F, and gyrB-F1, respectively. Sequencing was done in the Department of Entomology at the Max Planck Institute for Chemical Ecology (Jena, Germany) or commercially by SEQLAB Sequence Laboratories (Göttingen, Germany).

Bacterial sequences were deposited in the GenBank database under following accession numbers: KM035545 – KM035652 (16S rRNA genes), KM035653 – KM035673 (gyrA genes) and KM035674 – KM035755 (gyrB genes). Diversity of bacterial strains in individual beewolf antennae Bacterial micro-colonies selleck compound were isolated from individual antennae of two different Philanthus multimaculatus and one Philanthus psyche

female with serial dilution in 24-well plates with liquid medium SB203580 mouse as described above. Individual micro-colonies were carefully transferred by pipette into 96-well PCR plates with 100 μl PCR lysis solution A without proteinase K (67 mM Tris–HCl (pH 8.8); 16.6 mM (NH4)2SO4; 6.7 mM MgCl2; 6.7 μM EDTA (pH 8.0); 1.7 μM SDS; 5 mM β-mercaptoethanol) [40]; samples were heated at 95°C for 5 min to destroy bacterial cells. Afterwards, gyrB gene fragments were amplified, purified and sequenced as described above. Obtained sequences were aligned and manually curated using Geneious software version 6.0.5 (Biomatters Ltd., http://​www.​geneious.​com/​). Clomifene Phylogenetic analysis 16S rRNA, gyrA and gyrB gene sequences of isolated symbionts were aligned with those obtained from field-collected beewolves

as well as representative outgroup sequences of free-living Streptomyces and other actinomycete strains (Additional file 4: Table S4). Alignments of individual genes were concatenated for phylogenetic analyses. Approximately-maximum-likelihood trees were reconstructed with FastTree 2.1 using the GTR model, with local support values estimated by the Shimodaira-Hasegawa test based on 1,000 resamplings without re-optimizing the branch lengths for the resampled alignments [41]. Bayesian inferences were run with MrBayes 3.1.2 [42–44], with the different genes defined as separate partitions in the concatenated alignment. The searches were conducted under the GTR + I + G model, with 4,000,000 generations per analysis. Trees were sampled every 1,000 generations. We confirmed that the standard deviation of split frequencies was consistently lower than 0.01, and a “burnin” of 25% was used, i.e. the first 25% of the sampled trees were discarded. We computed a 50% majority rule consensus tree with posterior probability values for every node.

PubMedCrossRef 27 Marraffini LA: Impact of CRISPR immunity on th

PubMedCrossRef 27. Marraffini LA: Impact of CRISPR immunity on the emergence of bacterial pathogens. Future Microbiol 2012, 5:693–695.CrossRef 28. Karginov FV, Hannon GJ: The CRISPR system: small RNA-guided defence in bacteria and archaea. Mol Cell 2010, 37:7–19.PubMedCrossRef 29. Rezzonico F, Smits TH, Duffy B: Diversity, evolution, and functionality of clustered regularly interspaced short palindromic repeat (CRISPR) selleck regions in the fire blight pathogen Erwinia amylovora. Appl Environ Microbiol 2011, 77:3819–3829.PubMedCrossRef 30. Barrangou R, Horvath P: CRISPR: new horizons in phage resistance and strain identification. Annu Rev Food Sci Technol 2012, 3:143–162.PubMedCrossRef 31. Brüggemann H, Lomholt HB, Tettelin H,

Kilian M: CRISPR/cas loci of type II Propionibacterium acnes confer immunity against acquisition of mobile

elements present in type I P. acnes. PLoS One 2012, 7:e34171.PubMedCrossRef 32. Rho M, Wu YW, Tang H, Doak TG, Ye Y: Diverse CRISPR evolving in human microbiomes. PLoS Genet 2012, 8:e1002441.PubMedCrossRef 33. Katoh K, Asimenos G, Toh H: Multiple alignment of DNA sequences with MAFFT. Methods Mol Biol 2009, 537:39–64.PubMedCrossRef 34. Crooks GE, Hon G, Chandonia JM, Brenner SE: WebLogo: a sequence logo generator. DAPT Genome Res 2004, 14:1188–1190.PubMedCrossRef 35. Makarova KS, Haft DH, Barrangou R, Brouns SJ, Charpentier E, Horvath P, Moineau S, Mojica FJ, Wolf YI, Yakunin AF, van der Oost J, Koonin EV: Evolution and classification of the CRISPR-Cas systems. Nat Rev Microbiol 2011, 9:467–477.PubMedCrossRef 36. Hofacker I: Vienna RNA secondary structure server. Nucleic Acids Res 2003, 31:3429–3431.PubMedCrossRef 37. Weinberger AD,

Sun CL, Pluciński MM, Denef VJ, Thomas BC, Horvath P, Barrangou R, Gilmore MS, Getz WM, Banfield JF: Persisting viral sequences shape microbial CRISPR-based immunity. PLoS Comput Biol 2012, 8:e1002475.PubMedCrossRef 38. Horvath P, Romero DA, Coûtè-Monvoisin AC, Richards M, Deveau H, Moineau S, Boyaval P, Fremaux C, Barrangou R: Diversity, activity, and evolution of CRISPR loci in Streptococcus thermophilus. J Bacteriol 2008, 190:1401–1412.PubMedCrossRef 39. Sapranauskas R, Gasiunas G, Fremaux C, Barrangou R, Horvath P, Siksnys V: The Streptococcus thermophilus CRISPR/Cas system provides immunity in Escherichia coli. Nucleic Acids Res 2011, mafosfamide 39:9275–9282.PubMedCrossRef 40. Semenova E, Jore MM, Datsenko KA, Semenova A, Westra ER, Wanner B, van der Oost J, Brouns SJ, Severinov K: Interference by clustered regularly interspaced short palindromic repeat (CRISPR) RNA is governed by a seed sequence. Proc Natl Acad Sci USA 2011, 108:10098–10103.PubMedCrossRef 41. Mojica FJ, Díez-Villaseñor C, García-Martínez J, Almendros C: Short motif sequences determine the targets of the prokaryotic CRISPR defence system. Microbiology 2009, 155:733–740.PubMedCrossRef 42. Swarts DC, Mosterd C, van Passel MW, Brouns SJ: CRISPR interference directs strand specific acquisition.

Figure 3 Current density-voltage ( J -

V ) characteristic

Figure 3 Current density-voltage ( J -

V ) characteristics of DSSCs based on PEDOT/FTO, TiO 2 -PEDOT:PSS/PEDOT:PSS/glass, STA-9090 and Pt/FTO CEs. Table 2 The performances of dye-sensitized solar cells with different CEs measured under an AM 1.5G illumination Counter electrode V oc (V) J sc (mA cm−2) FF η (%) PEDOT:PSS/FTO 0.72 11.63 0.43 3.64 TiO2-PEDOT:PSS/PEDOT:PSS/glass 0.73 12.45 0.51 4.67 Pt/FTO 0.75 10.54 0.63 5.11 Conclusions In summary, we utilize a facile wet method to fabricate a novel hierarchical Pt- and FTO-free CE for the dye-sensitized solar cell. It is found that the TiO2 doped PEDOT:PSS catalytic activity layer will dramatically affect the electrochemical properties of the final device. By adjusting the composition of TiO2, the properties of CE have been optimized preliminarily. Because of the large active area of TiO2 nanoparticles, the proposed composite CE shows excellent enhancement in the conductivity and the superior catalytic activity for the reduction of I3 − to I−. The conversion efficiency is increased

by 22% than that of the DSSC with PEDOT:PSS/FTO CE and is comparable to that of the DSSC with traditional Pt/FTO CE. After further optimization, the TiO2-PEDOT:PSS/PEDOT:PSS/glass CE can be more cost-effective, high efficient, and flexible to replace Pt and FTO CEs and more broadly used for future commercial applications. Acknowledgements We acknowledge the support partly from the National Natural Science Foundation of China (grant nos. 91333122, 51372082, 51172069, 50972032, 61204064, PLX3397 buy Paclitaxel and 51202067), the Ph.D. Programs Foundation of Ministry of Education of China (grant nos. 20110036110006, 20120036120006, and 20130036110012), and the Fundamental Research Funds for the Central Universities. References 1. O’Regan B, Grätzel M: A low-cost, high-efficiency

solar cell based on dye-sensitized colloidal TiO 2 films. Nature 1991, 353:737–740.CrossRef 2. Grätzel M: Photoelectrochemical cells. Nature 2001, 414:338–344.CrossRef 3. Xu HG, Zhang XY, Zhang CJ, Liu ZH, Zhou XH, Pang SP, Chen X, Dong SM, Zhang ZY, Zhang LX, Han PX, Wang XG, Cui GL: Nanostructured titanium nitride/PEDOT:PSS composite films as counter electrodes of dye-sensitized solar cells. ACS Appl Mater Interfaces 2012, 4:1087–1092.CrossRef 4. Song DD, Li MC, Bai F, Li YF, Jiang YJ, Jiang B: Silicon nanoparticles/PEDOT-PSS nanocomposite as an efficient counter electrode for dye-sensitized solar cells. Funct Mater Lett 2013,6(4):1350048.CrossRef 5. Li QH, Wu JH, Tang QW, Lan Z, Li PJ, Lim JM, Fan LQ: Application of microporous polyaniline counter electrode for dye-sensitized solar cells. Electrochem Commun 2008, 10:1299–1302.CrossRef 6. Bu CH, Tai QD, Liu YM, Guo SS, Zhao XZ: A transparent and stable polypyrrole counter electrode for dye-sensitized solar cell. J Power Sources 2013, 221:78–83.CrossRef 7. Lee KS, Lee HK, Wang DH, Park NG, Lee JY, Park OO, Park JH: Dye-sensitized solar cells with Pt- and TCO-free counter electrodes.

The oscillatory amplitude of ρ xx (B) was well fitted by the Shub

The oscillatory amplitude of ρ xx (B) was well fitted by the Shubnikov-de Haas (SdH) theory [21–23], with amplitude given by (1) where μ q represents the quantum mobility, D(B, T) = 2π 2 k B m * T/ℏeB sinh (2π 2 k B m * T/ℏeB), and C is a constant relevant to the value of ρ xx at B = 0 T. The observation of the SdH oscillations suggests the possible existence of a Fermi-liquid metal. It should be pointed out that the SdH theory is derived by considering Landau quantization

in the metallic regime without taking localization effects into account [24, 25]. By observing the T-dependent Hall slope, Cyclopamine order however, the importance of e-e interactions in the metallic regime can be demonstrated [26]. In addition, as reported in [27], with a long-range

scattering potential, SdH-type oscillations appear to Pritelivir nmr span from the insulating to the QH-like regime when the e-e interaction correction is weak. Recently, the significance of percolation has been revealed both experimentally [28] and theoretically [29, 30]. Therefore, to fully understand the direct I-QH transition, further studies on e-e interactions in the presence of background disorder are required. At low B, quantum corrections resulting from weak localization (WL) and e-e interactions determine the temperature and magnetic field dependences of the conductivity, and both can lead to insulating behavior. The contribution of e-e interactions can be extracted after the suppression of WL at B > B tr, where the transport magnetic field (B tr) is beta-catenin inhibitor given by with reduced Planck’s constant (ℏ), electron charge (e), diffusion constant (D), and transport relaxation time (τ). In systems with short-range potential fluctuations, the theory of e-e interactions is well established [31]. It is derived

based on the interference of electron waves that follow different paths, one that is scattered off an impurity and another that is scattered by the potential oscillations (Friedel oscillation) created by all remaining electrons. The underlying physics is strongly related to the return probability of a scattered electron. In the diffusion regime (k B Tτ/ℏ < < 1 with Boltzmann constant k B), e-e interactions contribute only to the longitudinal conductivity (σ xx) without modifying the Hall conductivity (σ xy). On the other hand, in the ballistic regime (k B Tτ/ℏ > > 1), e-e interactions contribute both to σ xx and σ xy, and effectively reduce to a renormalization of the transport mobility. However, the situation is different for long-range potential fluctuations, which are usually dominant in high-quality GaAs-based heterostructures in which the dopants are separated from the 2D electron gas by an undoped spacer.

Figure 5 Binding characteristics of Lsa33 and Lsa25 proteins to E

Figure 5 Binding characteristics of Lsa33 and Lsa25 proteins to ECM components. (A) Wells were coated with 1 μg of laminin, collagen type I, collagen type IV, cellular fibronectin, plasma fibronectin and the control

proteins gelatin and fetuin. One μg of the recombinant proteins Lsa33 and Lsa25 was added per well and the binding was measured by ELISA. In (A) the protein binding was detected by polyclonal antibodies against each protein, while in (B) protein binding was evaluated with monoclonal anti – polyhistidine serum. Data represent the mean ± the standard deviation from three independent experiments. For statistical analyses, the attachment of recombinant Vemurafenib chemical structure proteins to the ECM components was compared to its binding to gelatin by the two – tailed t test (*P < 0.05). (C) Dose - dependent binding experiments of recombinant proteins with laminin was performed by polyclonal antibodies against each protein; each point was performed check details in triplicate and expressed as the mean absorbance value at 492 nm ± standard error for each point. Gelatin was included

as a negative control. The dissociation constants (KD) are depicted and were calculated based on ELISA data for the recombinant proteins that reached equilibrium. (D) Immobilized laminin was treated with sodium metaperiodate (5 to 100 mM) for 15 min at 4°C in the dark. Mean absorbance values at 492 nm (± the standard deviations of three independent experiments) were compared to those obtained with untreated laminin (0 mM). Interaction of recombinant proteins to serum components Our group has recently reported that leptospires interact with PLG and that several proteins could act as PLG – receptors [17–19, 21]. Protein binding to complement regulators factor H and C4bp have also been shown [31, 32]. Therefore, we set out to evaluate whether the recombinant proteins Lsa33 and Lsa25 were capable of binding human PLG, factor H and C4bp in vitro. The components,

human PLG, factor H and C4bp and the control proteins, gelatin and fetuin, were individually immobilized onto 96 – wells plates followed by incubation with the recombinant leptospiral proteins. The results obtained using polyclonal antibodies against each protein to probe the reactions showed that Lsa33 and Lsa25 Edoxaban interact with C4bp while only Lsa33 appears to bind to PLG (Figure 6A). No reaction was observed with factor H and the control proteins (Figure 6A). Similar results were achieved when binding was performed using monoclonal anti – his tag antibodies (Figure 6B). Both data show that while Lsa33 protein depicted a statistically significant absorption value for the interaction with PLG, the Lsa25 appears to have only a weak or no adherence to this component. These data were further confirmed when the reaction between the recombinant proteins and PLG were assessed on a quantitative basis as illustrated in Figure 6C. Dose – dependent and saturable binding was observed when increasing concentrations (0 to 1.

4 6 1 ± 0 6 −0 7 ± 0 1 Photosynthesis measured at 217 μmol

4 6.1 ± 0.6 −0.7 ± 0.1 Photosynthesis measured at 217 μmol

photons m−2 s−1. Standard deviation based on biological triplicates anmol ml−1 min−1 106 cells−1 Fig. 3 C59 wnt Chlorophyll a content of photoheterotrophic versus phototrophic cells. Cells were grown in the presence (A) and absence (B) of acetate in various concentrations of iron and chlorophyll a abundance was determined by HPLC. Standard deviation based on biological triplicates Photosynthetic efficiency of photoheterotrophic versus phototrophic cells Photosynthesis was further assessed by determination of photosynthesis–irradiance curves. In the presence of acetate, the maximum photosynthetic rate (P max) was decreased with respect to decreased iron nutrition (Table 3). Conversely, P max was increased in phototrophically grown severely iron-limited cells (0.1-μM Fe). On the other hand, the relative quantum efficiency of oxygen evolution (α) was decreased in response to decreased iron concentration in both photoheterotrophically and phototrophically grown cells; although, in phototrophic cells the decrease in α is not seen until severe iron limitation (0.1-μM Fe), concomitant with the increase in P max. The increase in P max of phototrophic cells at 0.1-μM Fe results in an increase in the light saturation index

(E k , defined as P max/α). Table 3 Photosynthetic parameters Fe (μM) Acetate CO2 P max a αb E k (P max/α) c P max a αb E k (P max/α) c 0.1 1.7 ± 0.1 0.014 ± 0.001 130 ± 6 3.6 ± 0.2 0.013 ± 0.004 300 ± 140 Akt inhibitor 0.2 1.9 ± 0.0 0.014 ± 0.002 140 ± 21 2.8 ± 0.2 0.021 ± 0.001 140 ± 16 1 2.3 ± 0.2 0.024 ± 0.002 95 ± 1 2.5 ± 0.2 0.022 ± 0.003 120 ± 11 20 2.7 ± 0.4 0.023 ± 0.002 120 ± 25 2.7 ± 0.2 0.022 ± 0.002 120 ± 7 Standard deviation based on biological triplicates anmol O2 (nmol Chl a)−1 min−1 b(nmol O2 [nmol Chl a]−1 min−1)/(μmol photons m−2 s−1) cμmol photons m−2 s−1 Maximum quantum efficiency of PSII in photoheterotrophic versus phototrophic cells F v /F m

is relatively constant, but decreased in stressed cells (Björkman and Demmig ASK1 1987), such as those under iron limitation (Morales et al. 1990, 2000). This parameter, which assesses the maximum quantum efficiency of PSII photochemistry, was lower in iron-limited relative to iron-replete Chlamydomonas cells in the presence of acetate, but remained high in iron-limited cells growing phototrophically, indicating maintenance of light reactions and photochemistry (Table 4). Non-photochemical quenching (NPQ) was likewise decreased in iron-limited acetate-grown cells, but remained high (and perhaps slightly increased) in iron-limited cells without acetate (Fig. 4). This suggests that increased NPQ contributes to the ability of phototrophic iron-limited cells to maintain photosynthesis. At a biochemical level, the abundance of photoprotective xanthophyll cycle pigments was increased in photoheterotrophic iron-limited cells when compared to phototrophic iron-limited cells (Fig. 5).

The immunity proteins coded by the usp gene operon have

The immunity proteins coded by the usp gene operon have AZD6244 price a characteristic two-histidine region which appears to enable the inactivation of the Usp DNase activity [10]. However, Usp-encoding strains that do not have all three orfU immunity protein genes have been described. All three immunity proteins are thus not essential for the protection of the Usp producers, although Usp is lethal when it is expressed alone in E. coli. It has been postulated that none of the three proteins is exclusively required for Usp protein synthesis [6]. As protection of the Usp-producing bacterial

cell might be provided by a mechanism that is different from that of the colicins, we have investigated the E. coli Usp-associated immunity protein Imu3, previously designated

OrfU3. Our study indicates that Imu3 has protective non specific DNA-binding abilities that could have possible biotechnological potential. Results and discussion Isolation of Immunity protein 3 (Imu3) with Ni-NTA affinity chromatography provided protein fractions with appropriate purity; (Figure  1A). DNA binding ability was not affected by the presence or absence of the his-tag, as both precipitated linear DNA (Additional file 1: Figure S1). The theoretical and actual mass (11.497 kDa) of the purified Imu3 differed by 1.5 Da (measured by ESI + and Q-Tof; Waters-Micromass, United Kingdom, data not shown), indicating that Imu3 is not post-translationally modified. Parret and DeMot [5] previously Ergoloid described an approximately 45% sequence identity of the C-terminal region of the Usp protein with known nuclease colicins, such as colicins E7 and E9. Although it has been shown that colicin E7 and its immunity

protein form a high-affinity complex [11], we were not able to confirm the formation of a high affinity complex between Usp and any of the three smaller proteins encoded downstream of the usp gene (data not shown) which were previously proposed to protect the Usp-producing cell against its endonucleolytic activity [5]. Nevertheless, our results showed that Imu3 protects isolated DNA from digestion by the nuclease colicin E7, indicating a nonspecific protection mechanism that is distinct from that of the colicin immunity proteins (Figure  2). Figure 1 Purified Imu3 protein. (A) SDS PAGE gel of Imu3 isolated using Ni-NTA agarose affinity chromatography, M: PageRuler Prestained Protein Ladder (Fermentas). (B) Superimposed chromatograms of Imu3 protein monomers (darker line) (HPLC, size-exlusion) with absorption values at 280 nm normalised. LexA protein self-cleavage products were used as standards (lighter line). Figure 2 Imu3 protection against colicin E7 DNase activity.

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of the smallest Apicomplexan genome from the human pathogen Babesia microti . Nucleic Acids Res 2012,40(18):9102–9114.PubMedCentralPubMedCrossRef 66. Huang B, Troese MJ, Ye S, Sims JT, Galloway NL, Borjesson DL, Carlyon JA: Anaplasma phagocytophilum APH_1387 is expressed throughout bacterial intracellular development and localizes to the pathogen-occupied vacuolar membrane. Infect Immun 2010,78(5):1864–1873.PubMedCentralPubMedCrossRef 67. Coumou J, van der Poll T, Speelman P, Hovius JW: Tired of Lyme borreliosis. Lyme borreliosis in the Netherlands. Neth J Med 2011,69(3):101–111.PubMed 68. Stanek G, Fingerle V, Hunfeld see more KP, Jaulhac B, Kaiser R, Krause A, Kristoferitsch W, O’Connell S, Ornstein K, Strle F, et al.: Lyme borreliosis: clinical case definitions for diagnosis and management in Europe. Clin Microbiol Infect 2011,17(1):69–79.PubMedCrossRef 69. Adams DA, Gallagher KM, Jajosky RA, Kriseman J, Sharp P, Anderson WJ, Aranas AE, Mayes M, Wodajo MS, Onweh DH, Abellera JP: Reports of nationally notifiable infectious diseases—United States, 2011. MMWR Morb Mortal Wkly Rep 2013,60(53):1–117.PubMed 70. Schnittger L, Rodriguez AE, Florin-Christensen

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Thereby, quadrats with high observed species richness acquire few

Thereby, quadrats with high observed species richness acquire fewer additional species from interpolation while quadrats with a low number of observed species could acquire a larger fraction

of additional species—if the unadjusted interpolation results predict additional species. We accepted overestimating species richness in some quadrats, knowing that vast areas of the Neotropics are under-sampled (Prance et al. 2000; Ruokolainen et al. 2002; Tobler et al. 2007). Although detailed maps of botanical sampling effort are available for some areas within the Neotropics (e.g. for Amazonia by Schulman et al. 2007), Sorafenib they are not available everywhere and therefore not used in the present work. Also, the procedure to adjust for sampling effort proposed here has the advantage of only requiring information inherent in the available point-to-grid data. Species richness Areas of elevated levels of species richness are the result of multiple overlapping species ranges. Most species occupy small ranges (Fig. 2a). Weighting of the species ranges (Eq. 3) demonstrates that the range sizes increase when applying our interpolation approach (Fig. 2f), but with a lower skewness and a lower maximum number of species compared

to a medium interpolation distance of five quadrats (Fig. 2c), thus avoiding overestimation of ranges of widespread species. The ‘smoothed’ increase of the range sizes due to the interpolation approach is reflected in the species richness maps (Fig. 3b, c). Whereas the inclusion of 340 more species (Fig. 3a) showed no major differences to the point-to-grid

species richness map presented in Morawetz and Raedig (2007), considerable distinctions are evident in both maps of species richness (Fig. 3b, c). For the weighted interpolation, these differences are plotted in Fig. 4. For all centers of diversity as well as for the unassigned quadrats, interpolated species richness is above the equity line. Edoxaban The different effect of interpolation on the species richness according to diversity center is particularly revealing for Amazonia. Even for small distances, the interpolation of species ranges here is consistently high. Comparison of maps 3b and 3c reveals the effect of adjusting species richness for sampling effort: the range of species richness is reduced, whereas the peaks of species richness found in Fig. 3b are retained in Fig. 3c. This effect is also apparent in the lower mean and standard deviation values for the centers of adjusted species richness, and in their closer range (Table 1). The Andean species richness center (Fig. 3c, polygon 2) shows the lowest standard deviation relative to the mean values (Table 1), suggesting more equal species richness and sampling effort of these Andean quadrats. The most obvious difference is that the Amazonian species richness center is by far the largest.