The interaction between insects and plants takes myriad forms in the generation of spectacular diversity. In this association a species host range is fundamental and often measured using an estimate of phylogenetic concordance between species. Pollinating fig wasps display extreme host species specificity, but the intraspecific variation in empirical accounts of host affiliation has previously been underestimated. In this investigation, lineage delimitation and codiversification tests are used to generate and discuss hypotheses elucidating on pollinating fig wasp associations with Ficus.
Statistical parsimony and AMOVA revealed deep divergences at the COI locus within several pollinating fig wasp species that persist on the same host Ficus species. Changes in branching patterns estimated using the generalized mixed Yule coalescent test indicated lineage duplication on the same Ficus species. Conversely, Elisabethiella and Alfonsiella fig wasp species are able to reproduce on multiple, but closely related host fig species. Tree reconciliation tests indicate significant codiversification as well as significant incongruence between fig wasp and Ficus phylogenies.
The findings demonstrate more relaxed pollinating fig wasp host specificity than previously appreciated. Evolutionarily conservative host associations have been tempered by horizontal transfer and lineage duplication among closely related Ficus species. Independent and asynchronistic diversification of pollinating fig wasps is best explained by a combination of both sympatric and allopatric models of speciation. Pollinator host preference constraints permit reproduction on closely related Ficus species, but uncertainty of the frequency and duration of these associations requires better resolution.
Several lines of theory have been proposed to account for the enormous diversity of phytophagous insects. Diversification conceivably ensues by ecological opportunity and adaptation to the exploitation of previously unattainable resources [1, 2]; by restricted gene flow through allopatric means [3, 4]; and disruptive selection and sympatric speciation [5, 6]. In order to discern among potential mechanisms driving speciation, both historical pattern and ecological scale processes are important to consider [7–10]. Comparative phylogenetic approaches that test congruence between host and associate populations can contribute to greater resolution in unravelling ecological scale processes [11–14]. Here we interpret the codiversification between Ficus host species and populations of a group of African fig wasp pollinator species.
No single adaptive hypothesis is yet to explain the conditions determining the origin, maintenance, and extinction of insect-plant mutualisms  and the overwhelming amount of literature surrounding the field has led to periodic reassessment of central concepts [2, 16, 17]. Pollination mutualisms are iconic examples of highly specialized interactions [18–20]. Fig trees (Moraceae: Ficus) are singularly dependent on pollination by fig wasps (Chalcidoidea: Agaonidae) that in turn are dependent on the fig 'fruit' for reproduction [18, 21]. Pollinating fig wasps show remarkable conservatism in host Ficus preference having concordant and ancient associations in the vicinity of 60 Myr [22, 23]. Correlated evolution between figs and fig wasps is supported by numerous examples [18, 24]. These include respiratory adaptation to fluid-filled figs  and head and mouthpart adaptation to the receptacle ostiole . The presence of close phylogenetic correspondence within the mutualism has been presented in previous work [22, 27–30] and strongly supports the hypothesis of a historically conservative interaction. The causal mechanisms supporting the maintenance of extremely conserved interactions remain unclear.
The estimation of the strength of these evolutionarily conserved interactions has come about through testing phylogenetic congruence at the species level and above. The presence of strict-sense cospeciation between one pollinator species and one Ficus species remain an outstanding case in insect-plant interactions, albeit greater scrutiny over the last decade has revealed species complexes and multiple pollinator species present on single host species [8, 13, 31]. From an empirical point of view, tests of phylogenetic correspondence that seek to estimate the level of 'cospeciation'  can in theory indicate up to four codiversification scenarios . Thus, the taxonomic scale at which tests of congruence are conducted can influence interpretation of the cospeciation pattern in uncovering process. Analyses of congruence among distantly related taxonomic subdivisions in poorly sampled clades can bias interpretation in favour of 'cospeciation' . Jackson and colleagues  recently demonstrated that the level of resolution of genetic variation can bias the interpretation of host species associations. Earlier work on the fig-fig wasp mutualism [26, 28] tended to be conducted at the level of genus and species. Fig wasp host fidelity is perhaps most strong at the genus/host section level [8, 22, 23, 26, 34]. Hypotheses explaining discordance in phylogenetic congruence implicate independent speciation, extinction, lineage duplication of either host or associate as well as horizontal transfer between host species.
Recent work has highlighted uncertainties concerning taxonomic classification and difficulty in understanding the phylogenetic associations between African pollinator genera and Ficus section Galoglychia . In this investigation, we test how independent diversification of agaonid pollinators is from host species of Galoglychia. Global measures of fit and reconciliation approaches are used to test phylogenetic congruence between pollinator fig wasp populations and Ficus species. Fig wasp within-species genetic variation at the cytochrome oxsidase one (COI) locus is assessed using statistical parsimony, AMOVA, and diversification modelling. Alternative pollinator topologies inferred with mitochondrial (mtDNA) and nuclear gene sequences are explored using likelihood ratio tests. The results were optimised over a set of hypotheses and are best characterised by pollinator populations reproducing on multiple Ficus species.
Our Bayesian and parsimony bootstrap consensus inferences based on EF-1α, COI, and Cytb produced the same topology (Additional file 1). The Markov chain reached apparent stationarity after 2.0 × 106 generations and the last 10.0 × 106 generations of 20.0 × 106 generations were used to generate the pollinator and Ficus consensus phylogenies. Posterior probabilities (PP) for all genera were well resolved and well supported (PP = 100). Bootstrap support for the genus Elisabethiella was poor (47) as was the support for the bifurcations of genera especially for the Courtella/Alfonsiella node (PP = 83; bootstrap = 62). This topology was in general agreement with a phylogeny of Machado and colleagues  that was based on COI showing Courtella and Elisabethiella as sister-clades. However, the majority of other cases support a topology where Elisabethiella is sister-clade to Alfonsiella though topology outside this relationship is inconsistent  and pollinator body length compatibility with syconium size measurements supports this hypothesis as well . Therefore, we used the harmonic means of four separate Bayesian runs to generate Bayes Factors  that compared constrained topologies of each case. The results were inconclusive (BF ~ 0.07) of one topology over the other so we used both in the reconciliation tests. We elected to use the Elisabethiella and Alfonsiella sister-genera inference for congruence testing as most studies support this topology and pilot analyses showed negligible differences between the two.
The Ficus phylogenetic inferences (Additional file 2) generated by Bayesian and parsimony approaches had the same topologies and are in general agreement with previous work . The subsections Chlamydodorae, Platyphylla, and Caulocarpae form well-supported clades in the Bayesian inference (posterior probabilities > 99). The former two subsections show bootstrap support of 97 & 73 respectively and more equivocal support (52) for the Caulocarpae clade. Subsections Urostigma and Sycomorus were constrained to outgroup taxa. Singleton representatives of subsections Crassicostae and Galoglychia are consistent with the topology of previous work . Due to the placement of F. ilicina (Sonder) Miq., the resulting polyphyly of Chlamydodorae necessitates future revision.
The GMYC likelihood test was robust to both molecular clock and relaxed clock assumptions and identified 40 'entities' or clades from a total of 65 terminal taxa, which are inferred to include coalescing lineages below the level of species (Figure 1). A total of 16 clusters represent clades that comprise potential below-species divergence at the COI locus. The inferred shift at which species diversification changes to within-lineage processes was significant (P = 0.000). The threshold time (-0.3040073) was converted to a relative time scale. The likelihood of the null model was less than the alternative model (Ln null model = 79.27529; Ln GMYC model = 96.37995) indicating a switch between-species branching according to a Yule pure-birth model and neutral coalescence. Within 5 of these 'coalescing' clades are host associations with more than one Ficus species. Elisabethiella stuckenbergi (Grandi), E. glumosae Wiebes, Alfonsiella Binghami Wiebes, and Courtella bekiliensis (Wiebes) show species level divergences within each of these 'species groups': (i) E. socotrensis (Mayr) was collected from F. natalensis natalensis Hochst. and F. burkei (Miq.) Miq.; (ii) E. stuckenbergi (between 3 clades indicating coalescence) from F. burkei, F. natalensis, and F. lingua lingua De Wild. & T. Durand, F. petersii Warb.; and (iii) A. binghami (between 2 clades showing coalescence, and a species-level lineage) from F. craterostoma Mildbr. & Burret, F. stuhlmannii Warb., F. petersii, and F. natalensis. The log-lineages through time plot (Additional file 3) shows a sharp increase in lineage accumulation that represents the inferred shift between speciation and coalescent processes.
Tanglegram of pollinator lineages and host Ficus species. Strict molecular clock (COI) fig wasp and (ETS & ITS) Ficus ultrametric phylogenies were generated using BEAST. Coloured lines connect wasp specimens with the fig species they were collected from (each connecting line is coloured according to fig wasp genus). A relative time scale is given. The nodes used to calibrate the divergence timing estimates are indicated by large open circles. Horizontal open bars represent the 95% upper and lower posterior density intervals around the mean node age. Black vertical bars on the right indicate Ficus subsections. The red vertical line indicates the threshold at which the Yule pure-birth model is inferred to have switched to a neutral coalescent model. Grey boxes indicate clades falling within the coalescent region.
The statistical parsimony analysis partitioned the COI sequence data into a series of 14 haplotype networks possessing two or more samples and 20 singleton haplotypes (Additional file 4). Of the Agaonidae pollinator networks, at least 4 groups (species) were collected from more than 2 Ficus species. Differentiation among species was significant (P < 0.000) at F ST = 0.78. Analysis of molecular variance showed significant levels of differentiation for all species (a 'species' compared to all other 'species' groups) except for within the A. binghami group (group specific F ST = 0.55) that was much lower than the those of other groups that ranged from F ST = 0.77 to 0.86 (Table 1). Species-specific fixation indices ranged from F ST = 0.55 to F ST = 0.86 (Table 2) and were largely in the vicinity of the latter. Both GMYC and statistical parsimony tests divide E. stuckenbergi into at least three putative species clades with substantial differentiation between each. Statistical parsimony tended to be more sensitive to differentiation among putative species lineages within A. binghami and E. socotrensis. Species delimitation that relies on single loci is not conclusive and has to be treated with caution. However, divergence at the COI locus provides credible evidence of negligible differentiation among pollinator lineages sampled from different Ficus species and is important to the interpretation of host species associations and co-diversification.
Fixation indices (F ST ) among each fig wasp species at the COI locus.
Species-specific fixation indices (F ST ) at the COI locus.
A comparison between 42 terminal pollinator taxa with 26 Ficus species resulted in non-significant congruence using ParaFit and under particular event cost schemes in TreeFitter tests. The application ParaFit tests a global null hypothesis that the association between the 'host' and 'parasite' phylogenies is independent. The global test of independence was insignificant (P = 0.18). Tests on individual associations indicate that the inferred lack of cophylogeny was a result of only 4 of the 42 links being significant (P < 0.05) in their contribution to the global test statistic. Instances of host switching relative to codivergence events were not particularly sensitive to cost assignment modulation and indicated preponderance to more switching than codivergence (Table 3). When event costs were set to default values (0,0,1,2) or where switching was similarly down-weighted (0,1,1,1), instances of codivergence (13-14; P < 0.05 and 9-13; P < 0.05 respectively) by comparison to host-switching (20-23; P < 0.05 and 23-29; P < 0.05) were significantly less (P < 0.001) where the total cost of random trees was greater than observed associations. These results are perhaps realistic given undisputed species-level phylogenetic congruence shown in the literature (i.e. pollinator fig wasp host-switching is less likely than codivergence). Significant instances of duplication events of pollinator lineages (5-7; P < 0.05) were indicated by the analysis using default cost assignments.
Event-based analysis of host associations between fig wasp lineages and Ficus species implemented in TreeFitter. Event costs weighting schemes (left column) are ordered as follows: codivergence, duplication, sorting, and switching. Redundant exemplars of the same fig wasp and host-species association were removed for the purposes of the test. * Observed event significantly more than randomised trees P < 0.05. ** Observed events significantly less than randomised trees P < 0.05. *** Total event costs of observed trees were significantly less than the randomised trees P < 0.05.
The general agreement between reconciliation tests of alternative topologies (not shown) indicates switching events exert a considerable influence, but does not alter the approximate numbers co-divergence events. Phylogenetic uncertainty inherent in these types of analyses, acts to obscure accuracy in tree topology and divergence estimates. The results show that the tests of congruence are robust to changes in topology. Incomplete representation of Ficus population genetic variation might bias the frequency of codivergence events. However, the results provide a reasonable indication of switching and duplication in terms of Ficus species. Overall, these results show that pollinators are highly constrained to Ficus subsections and switching between these higher taxonomic scales is rare and likely represents very ancient events (Figure 1). For instance, host associations of Allotriozoon heterandromorphum Grandi and F. lutea Vahl, E. enriquesi (Grandi) and F. ilicina, and an undescribed Elisabethiella species and F. usambarensis Warb. Divergence timing estimates of both pollinator and Ficus phylogenies show a high incidence of overlap among the 95% upper and lower posterior density intervals around the mean node ages. Regardless of this uncertainty, host-switching events of single pollinator species appears to occur between Ficus lineages that share a common ancestor that predates the origin of the pollinator populations. The age of the split between subsections Sycomorus/Urostigma and the subsections of section Galoglychia is inferred as relatively ancient. Inconsistency between the performance of COI and ETS/ITS fragments and sampling effects likely explains this discrepancy in the reconstruction of deep nodes.
Species of Elisabethiella pollinating fig wasps reproduce on at least three Ficus subsections. Long branches tend to be associated with the most extreme instances of phylogenetic incongruence implying ancient host shifts have shaped contemporary relationships. Although reconciliation tests indicated significant phylogenetic correspondence, host-switching patterns among pollinator lineages inferred to be below species level were more prolific than codivergence patterns. The discordance between the pollinator and Ficus phylogenies was in part due to restricted transfer of Elisabethiella and Alfonsiella populations amongst closely related host species in subsection Chlamydodorae. Reconciliation analyses also detected duplication patterns that can be partly explained by fig wasp differentiation via host switching, or in allopatry on the same host species, followed by secondary contact with sibling lineages. Repeated evidence of divergent putative species-lineages within what is considered the same 'good morphological species' of pollinator is consistent with a time lag between speciation of pollinators and that of hosts. Together these findings suggest that diversification in pollinating fig wasps is driven by preferential host-use amongst closely related species in conjunction with ecological factors contributing to speciation.
Phylogenetic reconciliation analyses (Table 3) show significant evidence of independence between Ficus and pollinator divergence due to either 'duplication' or 'host switching.' Both these types of patterns largely preclude synchronistic wasp-host diversification hypotheses. However, the phylogenetic discordance reported here might also be a function of the dissimilar taxonomic scales used to test associations between wasps and hosts. For instance, breakdown in phylogenetic concordance patterns has been argued to be partly the result of genetic introgression and hybridization across different fig species [8, 38, 40]. The Ficus phylogeny (Additional file 2) considered only species and not intraspecific variation. It is difficult to determine the distribution of host traits that are actually targeted by pollinators and how closely correlated these traits are to host phylogeny. Pollinator transfer between different Ficus species should increase the tendency for post-speciation introgression between host lineages  and presumably influence the evolution of fig wasp and host fig phenotypes. It would appear by the results presented in this study that the intensity of this type of genetic exchange should be predomiantly confined among closely related host species. Therefore, underestimation of intraspecific genetic variation of hosts might result in overestimation of phylogenetic incongruence at the species level. However, multiple host species associations with a single pollinator species and associations of Elisabethiella with several host subsections (Figure 2) indicates horizontal transfer between different Ficus species does play some role in the evolution of wasp and host fig phenotypes.
Higher level phylogenetic congruence. Broader taxonomic scale depiction of pruned phylogenies showing host associations between fig wasp genera and Ficus subsections. Note that the genus Elisabethiella has a wider host subsection range than the other genera sampled. Host switching has occurred between relatively divergent clades that comprises at least two different subsections. *Requires revision or poorly resolved.
Generally, host-plant switches can occur between distantly [41–45] or more closely related host lineages [20, 46, 47]. Arguably, pollinator fig wasps occupy the most extreme end of host specificity observed in insect-plant associations. Horizontal transfer by pollinator wasps between host lineages appears to be the most important contributor to the phylogenetic incongruence shown by this study (Table 3). The patterns of incongruence can be interpreted in a number of ways. The empirical evidence of 'host switching' derived from the reconciliation simulations does not discriminate between the strength of fidelity between natal and novel host species or the duration of associations with different host lineages. Thus, intensity of pollinator specialisation for a given host species and the magnitude of gene flow between pollinator populations sharing different host species are difficult to decipher [17, 48]. The COI haplotype tree shows E. socotrensis, E. stuckenbergi, and A. binghami all possess low-level (below species) genetic divergence among lineages on different host species (Figure 1). Alfonsiella binghami populations are able to reproduce on F. stuhlmannii, F. natalensis, F. burkei, F. petersii, and F. craterostoma (Figure 1). However, this does not imply reproductive isolation among populations on different the host species. It is uncertain whether the pollinator populations that use alternative host species for reproduction are evolutionarily viable and become good species rather than go extinct. Conversely, Elisabethiella glumosae, E. socotrensis, E. stuckenbergi, and A. binghami show considerable divergence at the COI locus between lineages on the same host species. As these incipient/putative species of Elisabethiella and Alfonsiella belong to lineages that are mostly monophyletic and not polyphyletic, the haplotype tree (Figure 1) likely shows orthologous alleles that have evolved from a common ancestor . One exception was the paraphyletic relationship Elisabethiella glumsosae has with another lineage of this 'morphological species' and might reflect phylogenetic uncertainty or a morphologically similar but different species.
Together, phylogenetic inference, statistical parsimony, and reconciliation analyses strongly imply that horizontal transfer among host species and lineage duplication are responsible for much of the phylogenetic incongruence. Previous work [8, 13] has shown evidence of both lineage duplication and host switching while more recent studies have debated the role extinction and duplication play in explaining phylogenetic incongruence [30, 38]. The results presented in this study do not refute any of these arguments, especially as extinction scenarios are difficult to substantiate. Assuming wasp and host fig extinction was negligible, more pollinator species should have evolved in relation to host species given duplication or host switching, and this is consistent with other studies [8, 13, 31]. Pollinator lineage 'duplication' was the least frequent of the significant tree reconciliation scenarios (Table 1). Eventual extinction by competitive exclusion of one pollinator species that shares a host with another has been used to explain phylogenetic incongruence . Phylogenetic relatedness has been shown to be a good predictor of competitive exclusion . However, this hypothesis relies on saturation of available resources or niche and does not seem applicable to Ficus. Resource utilisation of ephemeral and unpredictable host species by fig wasps in general, occurs at low levels . Pollinator recruitment does not occur for all available Ficus at any given period and should relax selection based on competitive exclusion. Duplication is characteristic of insect populations being isolated from one another while the host is not and does not imply that a duplicated lineage has to arise on the same host species nor in allopatry . Host switching among different Ficus species in sympatry might be responsible for pollinator lineage duplication via secondary colonisation of an ancestral host species that still supports a closely related pollinator lineage . Differentiation in allopatry between populations supported by the same host species followed by secondary contact would result in the same incongruence pattern. Presumably, duplication results from either transfers among closely related sympatric host species, or in allopatry on the same host species, or a combination of both. The contribution of any one of these processes should depend on limitations set by ecological, physiological, and morphological specialisation [54, 55].
It is difficult to speculate accurately about the contribution of extinction to the observed phylogenetic incongruence shown in this or any other study. The relatively inconsequential number and non-significance of sorting events (pollinator extinction independent of the host) detected in this study (Table 3) might be a consequence of low extinction probability overall . Local extinctions have been caused by drought where up to 17 pollinator species became locally extinct . This could ultimately lead to the local extinction of the corresponding figure. However, co-extinction of pollinator species might not be as influential when host species are broadly distributed over large regions such as Southern Africa. Local extinction of pollinators might not be a threat due to relatively long-lived fig species and re-colonization from other source populations . The relative lack of pollinator lineage loss from host lineages might be disguised by a predominance of co-extinction, but would not be detected without fossil evidence [59–61]. Low levels of local endemnicity of Ficus hosts sampled by this work suggest extinction might not play a substantial role in the low levels of phylogenetic correspondence.
Evidence of phylogenetic congruence is indicative of restricted switching among closely related hosts. Host species of Elisabethiella and Alfonsiella species are largely sympatric over their host distributions . Host switching per se could lead to sympatric speciation via disruptive selection. For sympatric speciation to operate, selection must overcome gene flow where different host species are geographically arranged such that insect dispersal does not limit migration between them . Fig wasp dispersing capability is believed to be substantial . Trade-offs between alternative host-species that reduce fitness on one host and increase fitness on another is necessary for disruptive selection and sympatric speciation . However, the best-known example of disruptive selection is inconclusive and might have involved divergence in allopatry preceding secondary contact [6, 65, 66]. Furthermore, it is difficult to test hypotheses that reject allopatry . Selection imposed by ecological interactions might also induce speciation. Host adaptation to different habitat types in close proximity suggests ecologically driven speciation . Habitat selection and physiological tolerance limitations of pollinator populations to different environmental variables  might support ecologically-driven speciation  in both pollinator and Ficus lineages. Host transfers among closely related Ficus species should be linked to life history constraints or preferential targeting of traits possessed by closely related phenotypes. Fig wasp host-use behaviour has been shown to tightly adhere to specific volatile cues emitted by their host species [70–72]. This suggests that fig wasps track plant secondary chemistry where colonisation of a novel host occurs between chemically similar lineages, assuming convergence of chemical traits is absent . Such behaviour is characterized by delayed associate speciation and has been described as host tracking [74, 75]. Host tracking of gall wasps that specialise on oaks  appears to be at least partially conserved due to galling life history constraints. Rare and periodic switches were shown to occur between more distantly related host section lineages and this is similar to the patterns of host-use by pollinating fig wasps. Limitations set by galling behaviour of pollinating fig wasps might have consequences for the breadth of host phenotypes that can be used for reproduction and between which pollen is transferred.
Significant levels of concordant and independent divergence between pollinating fig wasp lineages and Ficus species revealed several processes important to the evolution of the mutualism. This study supports recurrent backcrossing between closely related Ficus species and very rarely between distantly related host lineages. Evidence showing pollinator lineage duplication could have arisen both in sypmatry by switching among different host species, or allopatrically on the same host species, followed by secondary contact between sibling lineages. Unrealized within-species genetic variation of both fig wasps and Ficus need to be accounted for to accurately identify causal links between host lineage conservatism and speciation. Analyses using single pollinator species-representatives might result in misleading interpretation of host associations and obscure patterns relevant to speciation processes. This work shows that the propensity for host transfer among closely related Ficus species is variably independent of host speciation and permits ecological infuences on the speciation of pollinating fig wasps.
Most African Ficus are recognised in section Galoglychia [77, 78] with fewer in sections Sycomorus and Urostigma that are most strongly represented in the Indo-Australasian region . A large majority of African Ficus are monoecious, containing male and female flowers within the same syconium. Multiple specimens of pollinating fig wasp species (Agaonidae) from the genera Elisabethiella Grandi, Alfonsiella Waterston, and Courtella Kieffer were most abundant in our collections and served as the focal group in this investigation. All known available and compatible sequence data was incorporated into the study. No DNA material was available for the genera Blastophaga and Kradibia that each comprises single species. Other less well-represented taxa were used in the analysis to improve species delimitation tests and provide a broader taxonomic context. These comprised species from the genera Allotriozoon Grandi, Nigeriella Wiebes, Ceratosolen Mayr, and Platyscapa Motschoulsky. Together, the sample included 24 of the 71 described Afrotropical agaonid species. Both pollinator and Ficus outgroup constraints were based on previous inferences ; however, taxonomic distinctions among Ficus [9, 38] and pollinator  species remain controversial. All pollinator sequences (Additional file 5) were sequenced de novo from recent tissue collections with voucher specimens deposited at the Iziko South African Museum. Ficus sequences were sourced from GenBank ([22, 80]; Additional file 6).
Phylogenetic inferences of pollinator and Ficus sequence data were used for relative divergence timing estimation and for tests of congruence between them. To generate phylogenetic inferences among the wasps, fragments of mtDNA cytochrome oxidase one (COI ~ 620 bp), cytochrome b (Cytb ~ 380 bp), and nuclear DNA (nDNA) elongation factor-one alpha F2 copy (EF-1α ~ 500 bp) were sequenced. A comprehensive explanation of DNA extraction, PCR, and alignment protocols is given in . The species delimitation test requires an (mtDNA) ultrametric tree. Substitution rates of COI mtDNA tend to be higher than in nuclear protein coding genes and therefore more prone to saturation bias that impedes deep node resolution. In order to implement reasonable prior tree topology constraints for ultrametric pollinator tree inference and for co-phylogenetic analyses, we used species for which multiple gene fragments including nDNA were available to infer a phylogeny using Bayesian and parsimony approaches. Sequence data of up to 767 bp's of a ribosomal internal transcribed spacer (ITS) and up to 479 bp of external transcribed spacer (ETS) were used to infer Ficus species phylogenies under Bayesian and parsimony methods. Analyses presented in this work assume the Ficus species phylogeny is fully resolved and does not consider population-level genetic variation influence on host associations. Species-level appraisal of host lineages does provide evidence of comparative genetic distances for instances of departures from one-to-one species specificity. Sequences of ITS and ETS were amplified using the protocol outlined in previous work .
A Bayesian approach implemented in MrBayes 3.1.1  was used to partition the COI, Cytb, and EF-1α data into gene fragments and also codon positions. The Ficus sequence data was partitioned into ITS and ETS for the Bayesian phylogenetic analyses. A general time reversible DNA substitution model was used with gamma distributed (+G) rates with a proportion of invariant sites (+I). Posterior probabilities and mean branch lengths were derived from 15000 post-burnin trees sampled every 1000 trees from generations 5 to 20 million. Four separate Bayesian reconstructions were run to verify consistency of phylogenetic outcomes. The consensus trees were derived from post-burnin generations of Markov chains that had reached apparent stationarity. Convergence was assessed using the MCMC Tracer Analysis Tool v.1.4.1  by plotting the log likelihoods to assess the point in the chain where stable values were reached and with the standard deviation of split frequencies of all runs. Parsimony bootstrap analysis implemented using PAUP version 4.0b10  was used to assess the robustness of the Bayesian consensus phylogeny. The parsimony bootstrap consensus trees were derived from a search consisting of 500 bootstrap replicates using a full heuristic search. To calculate bootstrap support, we used branch-swapping by stepwise addition on best trees only, 100 random additional sequences holding 10 trees at each step, and the TBR search algorithm.
To distinguish population-level processes from species diversification we used the generalised mixed Yule coalescent (GMYC) likelihood test . The mixed Yule coalescent analysis approach has been shown to be relatively robust to sampling effects, marker use, the numbers of markers used . The test uses models that describe ultrametric phylogenetic trees that have either within-population coalescent branching or species branching signatures. The GMYC test assumes between-species branching according to a Yule pure-birth model  and within-species neutral coalescent model . A likelihood test of the mixed model estimates the shift from speciation to within-population branching. Although the threshold at which this split occurs might not reflect true diversification processes across all lineages in large trees , the approach has been used with high rates of success on preliminary species delimitation of diverse groups including insects . The test is intended to be complementary to traditional morpho-taxonomic approaches, but useful for detecting the presence of within-species diversification. The GMYC test was implemented using the 'R'  package SPLITS (available from: http://R-Forge.R-project.org). An ultrametric tree reconstruction was generated using a strict molecular clock (the GMYC test is based on the strict clock assumption) implemented in BEAST v.1.4.8  with gamma distributed invariant sites, GTR substitution prior, empirically estimated base pair frequencies, and unlinked 1st plus 2nd positions, and 3rd codon position. We also used an ultrametric tree based on a relaxed clock using the same priors as above. To compare branching time estimates between fig wasps and figs, we reconstructed a molecular clock Ficus tree under the same priors, but with linked codon positions. We set topology priors that constrained the clade of each fig wasp genera and the monophyletic subsections as inferred by our Bayesian consensus. Sequence data comprising approximately 680 bp's of a COI mtDNA fragment was used in GMYC tests of the fig wasps and 1345 bp's of the ITS and ETS regions for the Ficus analysis. All available sequence data for Elisabethiella and Alfonsiella were used in the analysis as GMYC test performance is optimised using larger sample numbers comprising non-zero branch length information . We generated a log-lineages through time plot to visualize the 'coalescent region' or shift between species and population branching patterns. Outgroup constraints were the same as for the phylogenetic inferences above. We also constrained the topology of fig wasp genera in the BEAST analysis and verified the generic relationships with the abridged multiple gene fragment phylogenetic inference. A tanglegram between the two BEAST molecular clock inferences was constructed by hand to illustrate fig wasp associations with Ficus species.
Statistical parsimony implemented using TCS v. 1.8  was used to partition the COI sequence data into independent networks connected by non-homoplasious mutations. Statistical parsimony estimates the maximum number of single substitutions among haplotypes (referred to as the connection limit) preceded by the connection of haplotypes into a network differing by increasing numbers of single site changes . All taxa within a given genus were analysed using statistical parsimony and the resulting networks assumed to be putative genetic species [95, 96] for AMOVA below. A 95% connection limit probability was used, where gaps were treated as missing with no connection limit step priors. F ST coefficients (at the 0.05 significance level) were estimated using AMOVA as implemented in Arlequin v. 3.1.  by permutating groups (species) of haplotypes among networks to assess fixation indices among the putative species. The F ST coefficient is the proportion of the total genetic variance within a subpopulation (S) relative to the total genetic variance (T) and ranges from 0 to 1. A high F ST implies substantial differentiation among groups and was expected under the hypothesis of statistical parsimony networks representing putative species. The P-value of each test is the proportion of permutations resulting in an F ST value larger or equal to the observed proportions. We used 10,000 permutations to estimate F ST and P-values and a gamma distribution of 0.5 that was determined using Modeltest v.3.0 .
A statistical test of congruence implemented in ParaFit  and a tree reconciling method implemented in TreeFitter  were used to investigate the magnitude of phylogenetic correspondence between the fig wasp COI haplotype tree and the Ficus species tree. These methods were selected on the basis of being able to accommodate the unequal numbers of terminal taxa between host and parasite tree, and to gauge both a global indication of congruence and the relative contribution of host switching (parasite horizontal transfer to another host lineage), duplication (parasite speciation without host speciation), sorting (parasite loss from host lineage) and codivergence (co-cladogenesis of parasite and host) patterns to tree correspondence. Both methods assume that the trees are fully resolved without polytomies. ParaFit was used to test a global null hypothesis of independent host-parasite associations revealed by two phylogenetic trees. This approach is not directed at reconstructing a putative history among hosts and parasites. The associations between the phylogenies are randomized and tested. Patristic distance matrices were calculated for both fig wasp (COI) and Ficus (ETS and ITS) sequence data using PAUP and then converted to principal coordinates using the R package ade-4 . A matrix explaining the associations between hosts and parasites is permutated at random and used in conjunction with the other two matrices to produce a test statistic. We used 9999 permutations to calculate the test statistic. By comparison, TreeFitter is an event-based parsimony method that is useful for exploring the relationships between gene trees (parasite tree) and species trees (host tree). In the absence of reliable species delimitation, Ricklefs and colleagues  treated each haplotype as a separate entity. Inclusion of 'associate' lineages below species (e.g. entities) can result in reflecting excessive duplication (speciation within host lineages). In order to minimize this effect, all sequence data of replicate host-pollinator associations were removed for both congruence analyses. The number of codiversification, sorting, duplicating, and switching events was tested against random datasets drawn by a heuristic search of tree space generated by the Markov process. TreeFitter calculates the event frequencies according to the minimum cost solutions for various runs where the host tree was permutated. Default cost-assignments were used (co-divergence equal to 0, duplication equal to 0, lineage sorting set at 1, and switches a cost of 2; i.e. 0,0,1,2) in addition to separate analyses involving equal and down weighting of costs of each event in different tests (1,1,1,1; 0,1,1,1; 1,0,1,1; 1,1,0,1; 1,1,1,0).
This work was supported by the Claude Leon Foundation and START secretariat and by a South African National Research Foundation grant GUN 61497. We thank Jenny Underhill for assisting with specimen collection and bench work, Krystal Tolley for assistance with specimen collection, and anonymous reviewers for their recommendations. This material is partially supported financially by the National Research Foundation, Ref. no. IFR2009090800013 and the South African National Biodiversity Institute (SANBI).
The conception and design of the analysis was developed by MJM. Bench work, sequence alignments, data analyses, interpretation of results, and manuscript drafting were conducted by MJM. Morphological appraisal of specimens, intellectual contributions, and interpretation of analyses were given by SvN. Field collections were contributed to by SvN and MJM. All authors have read and approved the final manuscript.


text:
« Filton's unsung heroes honoured at awards ceremony
Conygre House to be offered for sale by South Glos Council »
Construction of new Airbus office block set to start next month
Posted on Tuesday 24th July 2012 at 11:06 am by SH (Editor)
Work on the construction of a massive new office building for aircraft manufacturer Airbus is due to start at its Filton site next month following the appointment of a building contractor.
Miller Construction has been selected to deliver two projects totalling £37 million at the new Airbus Aerospace Park.
The contracts include a £10 million project comprising the refurbishment and alteration of Pegasus House, together with the partial demolition and refurbishment of Old Filton House and construction of a new 25,000sqm office building, totalling £27 million.
Chris Webster, Chief Executive, Miller Construction, said:
"We have been working closely with Airbus and their consultants over the past six months and are looking forward to progressing together to deliver the new office building and fully refurbished Pegasus House."
"We have a proven track record in the delivery of award winning accommodation and we are confident these projects will both meet and exceed Airbus' requirements and expectations."
"These contract wins are in line with our strategy to develop our presence in one of our key developing markets, science and technology, providing us with the opportunity to widen our expertise and grow our reputation as a cross-market leader."
Pegasus House, an existing five-storey, 1936, Grade 2 listed art-deco office building will undergo a complete refurbishment and alteration, and Old Filton House, a Grade 2 listed mid-eighteenth century, three storey country house, will be partially demolished and refurbished, in order to return the building to its former glory. The works also include substantial structural alterations, new mechanical and electrical (M&E) installation and an upgrade of the current thermal installation. Construction works commenced in May and are due for completion next summer.
Work on the office building is due to commence in August and will see the delivery of a new four-storey, naturally ventilated office building incorporating four atria and a roof level plant room. Once complete in autumn 2013, the building will provide office accommodation for 2,500 staff with changing facilities for up to 600 cyclists, an energy centre, and 900 car parking spaces.
Source: Press release from Miller Construction
Tags: Airbus, Airbus Aerospace Park, Pegasus House
This entry was posted on Tuesday 24th July 2012 at 11:06 am	and is filed under Business. You can follow any responses to this entry through the RSS 2.0 feed. Both comments and pings are currently closed.
2 Responses to "Construction of new Airbus office block set to start next month"
Wednesday 25th July 2012 at 10:19 am
Great news cant wait to see it all when its built
Adam101 Says:
Sunday 29th July 2012 at 8:16 pm
Exciting news, been waiting for years for Pegasus House to receive its much deserved restoration, I have studied the plans on the Council website and must say Airbus cannot be faltered for the plan of preservation of some of the distinctive parts to the building, such as the huge stain glass window. Hopefully there will be some kind of opening ceremony and tour once the restoration has been completed.

redpajama_set_name: RedPajamaCommonCrawl


>gnl|bcl2dbcds|H9GL49_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Anolis carolinensis GN=BMF (length=669 residues).
>gnl|bcl2dbcds|A0A2K5E1W9_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Aotus nancymaae GN=BMF (length=390 residues).
>gnl|bcl2dbcds|A0A2K5E1W9_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Aotus nancymaae GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A3B1K1X5_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Astyanax mexicanus GN=BMF (length=468 residues).
>gnl|bcl2dbcds|Q05KI3_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Bos taurus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|Q05KI3_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Bos taurus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|F7HPK0_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Callithrix jacchus GN=BMF (length=390 residues).
>gnl|bcl2dbcds|F7HPK0_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Callithrix jacchus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|J9PB65_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Canis lupus familiaris GN=BMF (length=612 residues).
>gnl|bcl2dbcds|A0A286XXB0_BMF-03 Bcl2 modifying factor gene OS=[Eukaryota] Cavia porcellus GN=BMF (length=570 residues).
>gnl|bcl2dbcds|A0A286XXB0_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Cavia porcellus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A286XXB0_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Cavia porcellus GN=BMF (length=711 residues).
>gnl|bcl2dbcds|A0A2K5MJY6_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Cercocebus atys GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2K5MJY6_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Cercocebus atys GN=BMF (length=390 residues).
>gnl|bcl2dbcds|A0A0D9R4R5_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Chlorocebus sabaeus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|Q0GKC7_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Danio rerio GN=BMF (length=429 residues).
>gnl|bcl2dbcds|M3WRS0_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Felis catus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|M3WRS0_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Felis catus GN=BMF (length=390 residues).
>gnl|bcl2dbcds|U3JS06_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Ficedula albicollis GN=BMF (length=582 residues).
>gnl|bcl2dbcds|G3QIN5_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Gorilla gorilla gorilla GN=BMF (length=555 residues).
>gnl|bcl2dbcds|G3QIN5_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Gorilla gorilla gorilla GN=BMF (length=390 residues).
>gnl|bcl2dbcds|W5UAU7_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Ictalurus punctatus GN=BMF (length=459 residues).
>gnl|bcl2dbcds|I3MDS9_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Ictidomys tridecemlineatus GN=BMF (length=570 residues).
>gnl|bcl2dbcds|I3MDS9_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Ictidomys tridecemlineatus GN=BMF (length=777 residues).
>gnl|bcl2dbcds|W5N4P7_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Lepisosteus oculatus GN=BMF (length=462 residues).
>gnl|bcl2dbcds|G3T7Z4_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Loxodonta africana GN=BMF (length=807 residues).
>gnl|bcl2dbcds|G7PAW6_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Macaca fascicularis GN=BMF (length=555 residues).
>gnl|bcl2dbcds|G7PAW6_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Macaca fascicularis GN=BMF (length=390 residues).
>gnl|bcl2dbcds|F7CM09_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Macaca mulatta GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2K5Z4C3_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Mandrillus leucophaeus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2K5Z4C3_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Mandrillus leucophaeus GN=BMF (length=390 residues).
>gnl|bcl2dbcds|G1NHG8_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Meleagris gallopavo GN=BMF (length=582 residues).
>gnl|bcl2dbcds|F6X3N6_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Monodelphis domestica GN=BMF (length=555 residues).
>gnl|bcl2dbcds|M3YAD5_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Mustela putorius furo GN=BMF (length=555 residues).
>gnl|bcl2dbcds|G1PAU1_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Myotis lucifugus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2I3HD83_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Nomascus leucogenys GN=BMF (length=390 residues).
>gnl|bcl2dbcds|I3K2D6_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Oreochromis niloticus GN=BMF (length=492 residues).
>gnl|bcl2dbcds|G1SR62_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Oryctolagus cuniculus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A3B3I0V9_BMF-04 Bcl2 modifying factor gene OS=[Eukaryota] Oryzias latipes GN=BMF (length=510 residues).
>gnl|bcl2dbcds|A0A3B3I0V9_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Oryzias latipes GN=BMF (length=492 residues).
>gnl|bcl2dbcds|A0A3B3I0V9_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Oryzias latipes GN=BMF (length=492 residues).
>gnl|bcl2dbcds|A0A3B3I0V9_BMF-03 Bcl2 modifying factor gene OS=[Eukaryota] Oryzias latipes GN=BMF (length=492 residues).
>gnl|bcl2dbcds|A0A3B3CGR9_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Oryzias melastigma GN=BMF (length=492 residues).
>gnl|bcl2dbcds|H0WYH6_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Otolemur garnettii GN=BMF (length=549 residues).
>gnl|bcl2dbcds|W5QFV1_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Ovis aries GN=BMF (length=567 residues).
>gnl|bcl2dbcds|A0A2R9BS98_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Pan paniscus GN=BMF (length=390 residues).
>gnl|bcl2dbcds|A0A2R9BS98_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Pan paniscus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2J8QDD5_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Pan troglodytes GN=BMF (length=390 residues).
>gnl|bcl2dbcds|A0A2J8QDD5_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Pan troglodytes GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A096NTE9_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Papio anubis GN=BMF (length=591 residues).
>gnl|bcl2dbcds|A0A096NTE9_BMF-03 Bcl2 modifying factor gene OS=[Eukaryota] Papio anubis GN=BMF (length=390 residues).
>gnl|bcl2dbcds|A0A096NTE9_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Papio anubis GN=BMF (length=621 residues).
>gnl|bcl2dbcds|A0A3B3RH63_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Paramormyrops kingsleyae GN=BMF (length=441 residues).
>gnl|bcl2dbcds|K7FRX2_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Pelodiscus sinensis GN=BMF (length=582 residues).
>gnl|bcl2dbcds|A0A3B4B6Y3_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Periophthalmus magnuspinnatus GN=BMF (length=363 residues).
>gnl|bcl2dbcds|A0A096LRV0_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Poecilia formosa GN=BMF (length=483 residues).
>gnl|bcl2dbcds|A0A3B3WU80_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Poecilia mexicana GN=BMF (length=483 residues).
>gnl|bcl2dbcds|A0A2J8T301_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Pongo abelii GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2K6FFR1_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Propithecus coquereli GN=BMF (length=390 residues).
>gnl|bcl2dbcds|A0A2K6FFR1_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Propithecus coquereli GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A3B4ETH0_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Pundamilia nyererei GN=BMF (length=492 residues).
>gnl|bcl2dbcds|A0A3B4CNJ8_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Pygocentrus nattereri GN=BMF (length=468 residues).
>gnl|bcl2dbcds|Q8K589_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Rattus norvegicus GN=BMF (length=558 residues).
>gnl|bcl2dbcds|A0A2K6L933_BMF-03 Bcl2 modifying factor gene OS=[Eukaryota] Rhinopithecus bieti GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2K6L933_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Rhinopithecus bieti GN=BMF (length=615 residues).
>gnl|bcl2dbcds|A0A2K6L933_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Rhinopithecus bieti GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2K6RAW8_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Rhinopithecus roxellana GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2K6RAW8_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Rhinopithecus roxellana GN=BMF (length=372 residues).
>gnl|bcl2dbcds|A0A2K6TW86_BMF-03 Bcl2 modifying factor gene OS=[Eukaryota] Saimiri boliviensis boliviensis GN=BMF (length=390 residues).
>gnl|bcl2dbcds|A0A2K6TW86_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Saimiri boliviensis boliviensis GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2K6TW86_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Saimiri boliviensis boliviensis GN=BMF (length=555 residues).
>gnl|bcl2dbcds|G3WDQ2_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Sarcophilus harrisii GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A0A2U9CJH3_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Scophthalmus maximus GN=BMF (length=654 residues).
>gnl|bcl2dbcds|A0A3B4U5H8_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Seriola dumerili GN=BMF (length=492 residues).
>gnl|bcl2dbcds|A0A3B4UNJ0_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Seriola dumerili GN=BMF (length=420 residues).
>gnl|bcl2dbcds|A0A3B4WM88_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Seriola lalandi dorsalis GN=BMF (length=492 residues).
>gnl|bcl2dbcds|A0A3B5B8F6_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Stegastes partitus GN=BMF (length=495 residues).
>gnl|bcl2dbcds|A0A287AIU8_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Sus scrofa GN=BMF (length=552 residues).
>gnl|bcl2dbcds|A0A287AIU8_BMF-03 Bcl2 modifying factor gene OS=[Eukaryota] Sus scrofa GN=BMF (length=552 residues).
>gnl|bcl2dbcds|A0A287AIU8_BMF-04 Bcl2 modifying factor gene OS=[Eukaryota] Sus scrofa GN=BMF (length=552 residues).
>gnl|bcl2dbcds|A0A287AIU8_BMF-05 Bcl2 modifying factor gene OS=[Eukaryota] Sus scrofa GN=BMF (length=474 residues).
>gnl|bcl2dbcds|A0A287AIU8_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Sus scrofa GN=BMF (length=552 residues).
>gnl|bcl2dbcds|H0ZMX5_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Taeniopygia guttata GN=BMF (length=582 residues).
>gnl|bcl2dbcds|A0A3B5KWG6_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Takifugu rubripes GN=BMF (length=444 residues).
>gnl|bcl2dbcds|A0A2U4BC98_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Tursiops truncatus GN=BMF (length=555 residues).
>gnl|bcl2dbcds|A3KND0_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Xenopus laevis GN=BMF (length=354 residues).
>gnl|bcl2dbcds|F6TGJ4_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Xenopus tropicalis GN=BMF (length=546 residues).
>gnl|bcl2dbcds|A0A3B5QK08_BMF-02 Bcl2 modifying factor gene OS=[Eukaryota] Xiphophorus maculatus GN=BMF (length=483 residues).
>gnl|bcl2dbcds|A0A3B5QK08_BMF-01 Bcl2 modifying factor gene OS=[Eukaryota] Xiphophorus maculatus GN=BMF (length=483 residues).


text:
Home Breaking News 'Trinkets' Confirms Season 2 With a Short Teaser Trailer
'Trinkets' Confirms Season 2 With a Short Teaser Trailer
The 30 second teaser video announcing season two of Trinkets assures fans of the popular, binge-worthy series that their questions will be answered. Unfortunately, it also confirms the second season will be the series' final season.
Netflix announced Trinkets' story will conclude with season two. New showrunner Sarah Goldfinger (Charmed) will take over to guide the final season.
The first season debuted on June 14, 2019 and consisted of 10 30-minute episodes. Season two will also be made up of 10 episodes, per Netflix. Expect to see the second and final season arrive in 2020.
Trinkets is based on the young adult novel by Kirsten "Kiwi" Smith and has Smith involved as an executive producer. Shelley Zimmerman, Brin Lukens, and Rebecca Glasho also executive produce along with writers Amy Andelson and Emily Meyer. The drama's produced by AwesomenessTV.
The series stars Brianna Hildebrand (Deadpool, The Exorcist), Kiana Madeira (The Flash), Quintessa Swindell (Euphoria), Odiseas Georgiadis (The Perfect Date), Henry Zaga (13 Reasons Why), and Brandon Butler (13 Reasons Why). Larry Sullivan (Big Little Lies), October Moore (Baskets), Larisa Oleynik (Mad Men), Joy Bryant (Parenthood), Linden Ashby (Teen Wolf), and Kat Cunning (The Deuce) also star in the drama.
Quintessa Swindell, Brianna Hildebrand, and Kiana Madeira star in 'Trinkets' (Photo by Allyson Riggs / Netflix)
The Plot:
"When three teenage girls from different corners of the high school cafeteria find themselves in the same mandated Shoplifter's Anonymous meeting, an unlikely friendship forms. Elodie (Hildebrand) — the grieving misfit, Moe (Madeira) — the mysterious outsider, and Tabitha (Swindell) — the imperfect picture of perfection, will find strength in each other as they negotiate the complicated dilemma of trying to fit in while longing to break out."
joy bryant
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Maryhill invites all Wine Club members to attend an exclusive evening opportunity to preview our newest tasting room prior to the public grand opening. RSVP required. Mingle with fellow club members and enjoy complimentary tastings of our award-winning wines and small bites in the Club Room along with a live music performance by The Juleps (5-8pm). You must be an active Maryhill Wine Club member to attend, no additional guests.


text:
Abortion doctor signals she'll sue Indiana AG over comments
Updated: 10:22 PM EDT Jul 19, 2022
Darron Cummings
Indiana AG Todd Rokita
SOURCE: Darron Cummings
An Indianapolis doctor who performed an abortion on a 10-year-old rape victim from Ohio took the first step Tuesday toward suing Indiana's attorney general for defamation.Dr. Caitlin Bernard, an Indianapolis obstetrician-gynecologist who gave the girl a medication-induced abortion on June 30, filed a tort claim notice over what she says are false statements that Attorney General Todd Rokita has made about her and her work.Bernard received widespread attention after she gave an interview to the Indianapolis Star about the child, who traveled to Indiana from Ohio for the abortion. A so-called fetal heartbeat law took effect in Ohio last month after the Supreme Court overturned Roe v. Wade. Such laws ban abortions from the time a fetus' cardiac activity can be detected, which is typically around the sixth week of pregnancy.A 27-year-old man was charged last week in Columbus, Ohio, with raping the girl, confirming the existence of a case that was initially met with skepticism by some news outlets and Republican politicians. Bernard's lawyer, Kathleen DeLaney, filed the "tort claim notice" against Rokita just days after she sent a cease and desist letter. The claim starts a 90-day period for the state to settle it. If it's not settled, Bernard could file a lawsuit. The claim didn't say how much money Bernard is seeking, noting that "the harm is ongoing.""Mr. Rokita's false and misleading statements about alleged misconduct by Dr. Bernard in her profession constitute defamation," the claim reads. "The statements have been and continue to be published by or on behalf of Mr. Rokita and the Office of the Attorney General."After the news of the 10-year-old's abortion broke, Rokita told Fox News that he would investigate whether Bernard violated child abuse notification or abortion reporting laws. He also said his office would look into whether anything Bernard said to the Indianapolis Star about the girl's case violated federal medical privacy laws. Rokita offered no specific allegations of wrongdoing.Records obtained by The Associated Press and local news outlets show that Bernard submitted her report about the girl's abortion on July 2, which is within Indiana's required three-day reporting period for an abortion performed on a girl younger than 16.A spokesperson for Rokita said Bernard's claim is "baseless" and attempts to "distract from the important work of the office.""Attorney General Rokita and the Office of Attorney General are leaders in the pro-life movement," spokesperson Kelly Stevenson said via email. "His historic work has further distinguished Indiana as a protector of unborn life and women."The Indiana Democratic Party criticized Rokita for the impact the lawsuit would have on taxpayers."Hoosier taxpayers will now have to pay for Attorney General Todd Rokita's national smear campaign against a doctor who helped a 10-year-old rape survivor attain a legal and safe abortion," party spokesperson Drew Anderson wrote in an email.
INDIANAPOLIS —
An Indianapolis doctor who performed an abortion on a 10-year-old rape victim from Ohio took the first step Tuesday toward suing Indiana's attorney general for defamation.
Dr. Caitlin Bernard, an Indianapolis obstetrician-gynecologist who gave the girl a medication-induced abortion on June 30, filed a tort claim notice over what she says are false statements that Attorney General Todd Rokita has made about her and her work.
Bernard received widespread attention after she gave an interview to the Indianapolis Star about the child, who traveled to Indiana from Ohio for the abortion. A so-called fetal heartbeat law took effect in Ohio last month after the Supreme Court overturned Roe v. Wade. Such laws ban abortions from the time a fetus' cardiac activity can be detected, which is typically around the sixth week of pregnancy.
A 27-year-old man was charged last week in Columbus, Ohio, with raping the girl, confirming the existence of a case that was initially met with skepticism by some news outlets and Republican politicians.
Bernard's lawyer, Kathleen DeLaney, filed the "tort claim notice" against Rokita just days after she sent a cease and desist letter. The claim starts a 90-day period for the state to settle it. If it's not settled, Bernard could file a lawsuit. The claim didn't say how much money Bernard is seeking, noting that "the harm is ongoing."
"Mr. Rokita's false and misleading statements about alleged misconduct by Dr. Bernard in her profession constitute defamation," the claim reads. "The statements have been and continue to be published by or on behalf of Mr. Rokita and the Office of the Attorney General."
After the news of the 10-year-old's abortion broke, Rokita told Fox News that he would investigate whether Bernard violated child abuse notification or abortion reporting laws. He also said his office would look into whether anything Bernard said to the Indianapolis Star about the girl's case violated federal medical privacy laws. Rokita offered no specific allegations of wrongdoing.
Records obtained by The Associated Press and local news outlets show that Bernard submitted her report about the girl's abortion on July 2, which is within Indiana's required three-day reporting period for an abortion performed on a girl younger than 16.
A spokesperson for Rokita said Bernard's claim is "baseless" and attempts to "distract from the important work of the office."
"Attorney General Rokita and the Office of Attorney General are leaders in the pro-life movement," spokesperson Kelly Stevenson said via email. "His historic work has further distinguished Indiana as a protector of unborn life and women."
The Indiana Democratic Party criticized Rokita for the impact the lawsuit would have on taxpayers.
"Hoosier taxpayers will now have to pay for Attorney General Todd Rokita's national smear campaign against a doctor who helped a 10-year-old rape survivor attain a legal and safe abortion," party spokesperson Drew Anderson wrote in an email.


Fly Veil made with elastic material and quality cotton, it adapts perfectly to the head horse and provides a elegant result.
Fly veil by Veredus made with elastic material and cotton that adapts perfectly to the shape of the ears. Design created with professional rider Scott Brash. Machine washable 30º.


Cryptocurrency is getting hotter by the second. Though Bitcoin is the big name in the cryptocurrency game, there are many other crypto players claiming to serve a unique and underserved purpose in the market.
I realize for a cryptocurrency to have succeed like Bitcoin, it needs to have network effects. In other words, regular users should be passionate about the coin in order to tell their friends about it. And if the code of the coin is open source, developers should care enough about it to make new contributions.
If these two requirements fall into place, a coin has a greater chance of achieving astronomical success. I'm always on the lookout for the "next Bitcoin" and am always curious which coin is being talked about more and more. I realized the simplest (read 'easiest') way for me to determine this would be to monitor Google Trends.
Google Trends gives you an idea of how much hype is behind a coin. The first place many newcomers come to learn about a coin is the Google search bar. Knowing this, I built a web application that investors or those interested in cryptos can use to view any cryptocoin and monitor its Google trends over the past six months.
Thus, CryptoTrend was born. I built the app using Vue.js. This is just a start and only a small piece of the network effects puzzle, but nonetheless it's easy to visualize and reason about.
We favor React and Angular at Rangle, but when I was chatting with our Director of Software Delivery, Alamelu Radhakrishnan, she encouraged me to explore my interest and try building something on Vue as it's a framework we may want to incorporate into future projects.
CryptoTrend leverages data from two APIs: Google Trends and Coin Market Cap. This allows the application to present the price fluctuations/patterns of individual cryptocoins in comparison to their search trends. Cryptocoins can be sorted by market cap, 1 price change, 24-hour price change and 7-day price change.
By clicking "Google Trends" you can go into that specific cryptocurrency to view its trend history over the past six months and basically visualize the public interest in this coin. You can also see the usual metrics of 1 hour, 24 hour and 7-day price change.
I found my experience building an application with Vue.js to be a relatively natural progression and I appreciated the opportunity to explore another form of front end development.
Although we're not using Vue.js at Rangle at the moment, I think it's important for developers like myself to experiment with new technologies, libraries and frameworks. It's one of the best ways to develop your skills. Plus, you never know when a client is going to want to try something new.
For a closer look at CryptoTrend, check out cryptotrend.io.


text:
Q: StreamWriter.Writeline throws "Not enough storage is available to process this command." IO exception I have an application which will copy 1000 files from One folder to another. After each file is copied we will write the Copy Success/Failure information into a separate file. At some time, during writing this copy information in a file, StreamWriter,WriteLine throws the following exception"
10/28/2014 12.21.02.068 Message : Not enough storage is available to process this command.
Filename : Copying file C:\Program Files\XYZ\SampleFile.xml to C:\Program
Files\ABC\SampleFile.xml.xml | Inner Exception :  | Type : System.IO.IOException | Source : mscorlib

Not enough storage is available to process this command.

at System.IO.__Error.WinIOError(Int32 errorCode, String maybeFullPath)
at System.IO.__ConsoleStream.Write(Byte[] buffer, Int32 offset, Int32 count)
at System.IO.StreamWriter.Flush(Boolean flushStream, Boolean flushEncoder)
at System.IO.StreamWriter.Write(Char[] buffer, Int32 index, Int32 count)
   at System.IO.TextWriter.WriteLine(String value)
   at System.IO.TextWriter.SyncTextWriter.WriteLine(String value)
   at System.Console.WriteLine(String value)
   at VersionActivator.VersionActivationController.LogMessage(String message)
mscorlib : Not enough storage is available to process this command.**

But this application works fine when the PC initialized in safe mode.
Can any one please help me?

A: It is developed in C#.
My code is
private void CopyAll(DirectoryInfo source, DirectoryInfo target)
      {
          try
          {
              if (!source.Exists)
              {
                  LogMessage(string.Format("{0} does not exist!", source.FullName));
                  return;
              }
              // Check if the target directory exists, if not, create it.
              if (Directory.Exists(target.FullName) == false)
              {
                  Directory.CreateDirectory(target.FullName);
              }

              // Copy each file into it's new directory.
              foreach (FileInfo fi in source.GetFiles())
              {
                  if (fi.Exists)
                  {
                      string destFileName = Path.Combine(target.FullName, fi.Name);
                      // If the file is readonly, make it non-readonly
                      if (File.Exists(destFileName))
                      {
                          File.SetAttributes(destFileName, FileAttributes.Normal); // IMS 2793051209
                      }
                      LogMessage(string.Format("Copying file {0} to {1}", fi.FullName, destFileName));
                      fi.CopyTo(destFileName, true);
                  }
              }
          }
          catch (Exception ex)
          {
              System.Diagnostics.Trace.WriteLine(ex.ToString());
              throw;
          }
      }

public void LogMessage(string message)
      {
          try
          {
              Console.WriteLine(message);
              if (m_LogWriter != null)
              {
                  m_LogWriter.WriteLine(
                     string.Format(@"{0} {1}"
                     , DateTime.Now.ToString("MM/dd/yyyy HH.mm.ss.fff")
                     , message)
                     );
              }
          }
          catch (Exception ex)
          {
              WriteException(ex, message);
              throw;
          }
      }

And the m_LogWriter will be initialized at Windows_load event.
if (m_LogWriter != null)
            return;

         DirectoryInfo logFolder = new DirectoryInfo(versionActivatorConfigObj.Folders.LogFolder.Path);

         try
         {
            if(!(logFolder.Exists))
            {
               logFolder.Create();
            }
            logFolder.Refresh();
            if(logFolder.Exists)
            {
               string logFileName = string.Format(@"MyCopyLog{0}.txt", DateTime.Now.ToString("M-d-yyyy_h-m-s"));
               m_LogWriter = new StreamWriter(Path.Combine(logFolder.FullName, logFileName), true);
               m_LogWriter.AutoFlush = true;
            }
         }

redpajama_set_name: RedPajamaStackExchange


text:
Home / Government / Commuter rail dispute threatens project engineering
Commuter rail dispute threatens project engineering
By: Joe Yovino	June 17, 2010 3:11 pm
(Map courtesy of the Southeast Wisconsin RTA)
Local supporters of the Kenosha-Racine-Milwaukee commuter rail project are divided as Milwaukee County officials try to delay project engineering.
The Southeast Wisconsin Regional Transit Authority within a week will apply for federal approval to begin engineering the estimated $232.7 million rail project. But Milwaukee County RTA representatives are asking federal officials to deny the application.
"Why are you going to waste money for engineering if you don't have an operating source?" said Milwaukee County Supervisor Michael Mayo, an RTA member opposing the application.
Mayo and Milwaukee County Board Chairman Lee Holloway last month were the only two members of the RTA board to oppose asking the Federal Transit Administration to approve the KRM project engineering.
They opposed it because the Wisconsin Legislature has not yet passed a law that would let the RTA raise a half-cent sales tax to pay for local public transit, including the Milwaukee County bus system.
Mayo said the KRM should not move forward until the county's budget-strapped bus system is supported by a local sales tax. The sales tax also likely would pay for the KRM's operating costs, he said, and prevent the KRM rail line from competing with the local bus system for transit operating money.
"I don't want to really submarine them," he said of the KRM supporters. "But I want them to put it on hold."
Karl Ostby, RTA chairman, said Milwaukee County's opposition is not necessarily the death of the project because the fight is over support for buses rather than whether the KRM is worthy. Federal officials are likely to take that into consideration, he said.
"I think, naturally, most people would say our best application is if you have unanimity from the total board," he said. "Hopefully, they would look a little bit further and see the reason."
The federal approval for the KRM application would let the RTA use up to $6 million set aside by Congress to pay for 80 percent of the engineering costs, said Ken Yunker, executive director of the Southeastern Wisconsin Regional Planning Commission. It also would make the project eligible to apply for federal construction money, he said.
Yunker said a letter from Mayo and Holloway asking the FTA to reject the application will be included in the information sent to the federal agency. He said he has no idea how the letter will affect the RTA's chances of gaining federal approval.
Milwaukee County's opposition is unlikely to hamper the RTA's overall effort to improve regional transit, including buses, Ostby said.
Mayo said Milwaukee County will not get state action on the sales tax until after the elections this November. Until then, he said, he wants the KRM on ice.
"Milwaukee County could have done their own RTA," Mayo said. "We really did not have to be married to the SERTA board, and that is really bothering me right now.
"But we decided to bring in Racine County and Kenosha County. We decided to be a team player."
Kenosha-Racine-Milwaukee commuter rail KRM Lee Holloway Michael Mayo Ostby Southeastern Wisconsin Regional Planning Commission Yunker 3:11 pm Thu, June 17, 2010 The Daily Reporter - WI Construction News & Bids
Tagged with: Kenosha-Racine-Milwaukee commuter rail KRM Lee Holloway Michael Mayo Ostby Southeastern Wisconsin Regional Planning Commission Yunker
Previous: Milwaukee bid preference cuts city contractor
Next: Wisconsin unemployment drops to lowest level in 14 months
Jesse Hagen
Michael Mayo & Lee Holloway are now presenting their roadmap to delay. Why move forward when you can do nothing at all?
Dave Wagner
Referencing the last line in the article quoting Chmn. Holloway as saying that Milw Co wanted to be a "team player", I think his action in opposing this application is anything but being a team player.
I think the County Board Chairs and County Executives involved need to show some leadership on this issue if they think that regional mass transit is a good idea. One county holding the three county project funding hostage is unseemly at best and counterproductive for sure.
As a Milwaukee County resident, it is these types of actions that cause me to wonder about whether hope and change are anything other than convenient rhetoric to get elected.


text:
import mentorCreationTmpl from './mentor-create-view.html!text';

function createMentoringInitialData(){
    console.log("createMentoringInitial data came here");
}
createMentoringInitialData.$inject = [];

function createMentoringController(){
    console.log("mentoring controller reaches here");
}
createMentoringController.$inject = [];

export{
    createMentoringInitialData,
    createMentoringController,
    mentorCreationTmpl
};

redpajama_set_name: RedPajamaGithub


Benches within the Jean Sweeney Park are available for sponsorship and memorial plaques are available. Jean Sweeney Open Space Park Fund is managing the bench project for ARPD.
We would like to thank the Alameda Community Fund for starting off the bench project with the sponsorship of two benches: One of each style.
Maria Young Erik R. Lee - Martial Arts Instructor, Father, Friend "The More You Sweat, the Better You Get"
Nancy Hird Partial donation toward bench honoring Jean Sweeney. Donations may be made toward this bench.
Ivana Krajcinovic & Andrew Slivka In memory of Natalie Silvka. Text to be determined.
Charles & Linda Spencer (Jr.) In memory of Jean Sweeney - Text to be determined.
Antonette G. Vella For Jaime Garlit (1927-2000) & Mary Garlit With Love From Their Children & Grandchildren with whom they enjoyed so many of Alameda's Parks.


Until the end of 2018, ATA carnets were only accepted in China for trade fairs and exhibitions. The Chinese government have now announced they will accept all categories, to include commercial samples and professional equipment.
Changes are being formally ratified by the ICC and represent a boost for those wishing to export into a growing economy. Improved trade facilities with a nation of 1.4 billion people is good news.
Wider ATA carnet support will also aid film, or TV producers and any business wishing to demonstrate their products, or equipment.
Procedure is well understood at major ports and airports. A number of provincial centres for carnet acceptance are also being opened, a sign of China's wish to increase carnet use.
At present, validity is restricted to 6 months, although an extension is normally possible. An application needs to be made before expiry, working with your ATA carnet supplier on this makes sense.
Where you are taking carnet items as hand luggage, customs clearance is rarely an issue, although you then need to visit the nearest China Chamber of International Commerce (CCOIC) office within three working days.
For shipped goods on a carnet, electronic data registration is normally required. As with meeting other regulations, your freight forwarder should assist.
Through our own management and our specialist ATA carnet company, we can provide all the assistance you require. When travelling to China, using professional support is a useful idea.
China are making an effort to build trade and welcome business visitors. They are still a country where being administratively correct helps.
Lists of goods on carnets needs to be clear and accurate, along with labelling of items. Regulatory procedures for categories of goods needing extra clearance should be followed.
Don't let that put you off, simply a case of being systematic, to visit an incredibly diverse country full of opportunity.


The 5' diameter Drone Launch Pad from Hoodman provides a takeoff and landing surface for your DJI Inspire, Yuneec Typhoon, or similar-sized RC heli. The pad prevents dust and debris being kicked up by the downdraft, and features an 8 lb weighted perimeter to avoid being blown away. A bright orange coloration makes the Launch Pad easy to locate and serves to alert others in the vicinity that a drone is in use.


Kelly Kandra Hughes is conducting a basic skating class at the town rink on Saturday morning. Children under 10 should wear a bike helmet. Kelly competed in high school for the Hershey, Pa., Figure Skating Club.
We will announce any necessary change here.


text:
TunnelTECH
Critical analysis of FRS durability study Mar 2015
Pierre Rossi, Research Director, fiber reinforced concrete consultant, Paris, France
In critical analysis of a paper about the durability of fiber-reinforced shotcrete (FRS) in an aggressive environment, consulting engineer and research director Pierre Rossi suggests that the study parameters are flawed, the criteria for the study are not applicable for the concrete reinforced structures in an 'in-service' state, and that providing evidence that metal corrodes more than synthetic products is not a real or new scientific advancement.
The durability of different fiber-reinforced concretes used in shotcrete-type applications is the topic of a paper entitled Durability performance of fiber-reinforced shotcrete in aggressive environments, written by J P Kaufmann and presented and published in the Proceedings of the WTC Congress of Iguassu Falls, Brazil, in 2014.
More precisely, the comparison targets the effects of cracking of these materials when they are thrust into different environments – ranging from slightly aggressive to very aggressive. Five fiber-reinforced concretes are studied; one consisting of metal fibers, the remaining four of different synthetic fibers; and five increasingly aggressive environments were studied.
It must be specified that the matrix of all fiber-reinforced concretes studied is the same and representative of the usual matrix in the intended type of industrial application.
Set-up of test samples used in the durability study for exposure to liquid solutions (top), coverage (center) and different square panels for mechanical testing and prisms for microscopy (bottom).
As a general opinion, the author evidently wishes his article to be considered as a scientific one. The beginning of the article – including the description of chosen materials, the production of tested samples, and the tests carried out – is relatively precise and supports the idea that it could be considered as a scientific article on this type of subject. However the subsequent parts do not keep standards with the beginning.
In fact, when the author begins to deal with the very heart of the subject matter, details become less specific. Different types of aggressive environments are selected without substantiating the choice, either by explaining the situations in which these environments are found or by citing examples of tunnels in which such environments have led to serious problems.
Further, there is an assumption that all square slabs consisting of different fiber-reinforced concretes are pre-cracked before undergoing chemical aggression, and that the crack openings initially created are between 0.5 and 1mm at a certain place on the slabs (the same for all of them); and further that they reach "around" 2mm at the center of the slabs, without specifying which slab has a 0.5mm crack opening and which has a 1mm crack opening. Such imprecision is not permissible considering the importance these crack openings have with respect to corrosion of fibers, particularly metal ones.
There are also some basic scientific weaknesses in the study in that the author does not specify the number of slabs and prisms tested per type of fiber-reinforced concrete and per type of aggressive environment.
It would seem to be a matter of:
one cracked slab per type of fiber-reinforced concrete
one prismatic sample pre-cracked to 1.2mm, plus
one prismatic sample pre-cracked to 0.5mm per type of fiber-reinforced concrete and per type of environment.
How can one draw such strong conclusions when so few results are available, especially in a supposedly scientific article that concerns concrete and the problems of durability and within the framework of a problem where dispersion of reinforcement is important?
The principal weaknesses of this work concern the outline of the problem that the author has chosen to study. Focusing on the cracking-durability relationship of fiber-reinforced concretes only makes sense in a situation where the structure is in a service state. Generally, when in a service state situation crack openings should not exceed 100 for a very aggressive environment, to 200 or 300µm in a slightly aggressive environment. This is far from the 2mm crack openings studied by the author.
The author should have known that when a crack opening is less than 300µm in fiber-reinforced concrete, it presents a very tortuous and, at times, discontinuous path, which makes the circulation of aggressive agents more difficult.
When the crack opening is less than 300µm, self-healing mechanisms can occur and the corrosion products (in the case of metal fibers) can be deposited in the interior of the cracks.
These two physical mechanisms consequently obstruct the cracks and therefore prevent circulation of aggressive ions.
The author, in a laudable attempt at scientific objectivity, should have cited other researchers and previous studies which show that metal fiber-reinforced concretes do not present problems, except, in certain cases, where soiling poses an aesthetic concern because of the above-mentioned physical phenomena.
In conclusion, it must be emphasized that, in a situation where the fiber-reinforced shotcrete plays a mechanical role in the tunnel (recovery of forces exerted by the ground), as well as a sealing role (for example, preventing water infiltration), it is easier to realize this by using metal fibers than synthetic fibers – for two reasons.
First, the Young's modulus of synthetic fiber is much lower than that of metal fiber, even when the synthetic fiber in question is macro-synthetic.
Secondly, the crack openings in synthetic fiber-reinforced concrete are significantly larger than those in metal fiber reinforced concrete, for the same service load.
A crack crossed by a synthetic fiber is significantly more likely to increase in size over the course of time than a crack crossed by a metal fiber when kept under constant load over time. This is due to the fact that synthetic fibers have more of a tendency to creep than metal fibers.
For these various considerations, the article in question cannot be considered objective, rigorous, or truly scientific. It presents results that do not represent real or pertinent situations for fiber-reinforced concrete usage. Providing evidence that metal corrodes more than synthetic products is neither a real nor a new scientific advancement.
Review of FRC brittle failure considerations – TunnelTalk, March 2015
fiber-reinforced concrete for precast segments – TunnelTalk, May 2014
Critique of synthetic-fiber FRS paper – TunnelTalk, September 2014
Response to macro-synthetic fiber critique – TunnelTalk, November 2014
20 years of fiber concrete linings in the UK – TunnelTalk, Oct 2010