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You say Conchaphila, I say Lurida: molecular evidence for restricting the Olympia oyster (Ostrea lurida Carpenter 1864) to temperate western North America.
KEY WORDS: Olympia oyster, Ostrea conchaphila, Ostrea lurida, Ostreidae, systematics, phylogeny, cryptic species
Many marine taxa are known to exhibit a high degree of phenotypic plasticity that challenges systematic biologists and creates problems for conservation efforts. Knowlton (1993) described the abundance of cryptic marine species, which are characterized as being almost impossible to distinguish based on morphology alone. Species definitions have been the subject of many debates (Donoghue 1985) and, in recent decades, even the most popular species concepts have been criticized, such as the biological species concept (Mishler & Donoghue 1982, Donoghue 1985, Cracraft 1987, Kluge 1990) and the phylogenetic species concept (Avise 2004). Regardless of one's species definition, the use of molecular markers has enabled scientists to identify previously cryptic species and to resolve taxonomic inconsistencies (Knowlton 1993, Simison & Lindberg 1999, Dawson & Jacobs 2001, Crummett & Eernisse 2007, Shilts et al. 2007, Hewson 2008).
Recent efforts to restore populations of the Olympia oyster ([double dagger]), Ostrea conchaphila, some involving transplants of oysters between estuaries separated by large distances, have been conducted despite lingering uncertainty about the oyster's true identity. The taxonomic problem is whether Ostrea lurida Carpenter 1864, the name once widely used for the Olympia oyster, should be considered a junior synonym of O. conchaphila Carpenter 1857. This synonym was proposed by Harry (1985) without explanation but is presumed to be based on morphological examination of museum specimens combined with the notion that the species was more widely ranging than previously thought. Prior to Harry's study, O. lurida and O. conchaphila were thought to be separate species, with the range of O. conchaphila from Mazatlan, Mexico to Panama (Dall 1914) and the range of O. lurida from Sitka, AK, USA to Cabo San Lucas, Baja California Sur, Mexico. Neither species was thought to exist in the contiguous coastline between the two species' ranges, including all of the Gulf of California, Mexico (Dall 1914). An exception (although not addressing the Gulf of California) was Hertlein's (1959) discussion of a possible transition zone between these species based on intermediate-appearing oysters from San Diego, CA and northern Baja California, Mexico, which would necessitate a considerable northern extension of the range of O. conchaphila. After Harry (1985) proposed this was all a single species, some (e.g., Baker 1995) have questioned the synonymy whereas others have indiscriminately used one name or the other, mostly in reference to oysters from Washington or British Columbia.
A separate issue is Harry's (1985) revival of Ostreola Monterosato 1884 (type species Ostrea stentina [Payraudeau, 1826]), which he also extended to include O. conchaphila. Subsequent morphological (Coan et al. 2000) and molecular (Kirkendale et al. 2004, Lapegue et al. 2006, Shilts et al. 2007) studies have not supported Ostreola as distinct from Ostrea. In particular, Shilts et al. (2007) have called for the dissolution of Ostreola and reassignment to Ostrea.
Until the present study, no one has tested Hertlein's (1959) predicted zone of overlap in southern California and along the Baja California peninsula, and this information would be especially useful considering ongoing native oyster restoration efforts. For example, government agencies such as NOAA and nongovernmental organizations such as The Nature Conservancy are interested in expanding restoration projects to locations south of San Francisco Bay, CA (Dick Vander Schaaf, The Nature Conservancy, pers. comm.) to include Hertlein's (1959) zone of overlap.
Given the high phenotypic plasticity of these oysters, the use of molecular markers applied in a geographic context could potentially be useful for resolving whether the native oyster is one or more species. Parallel studies with oysters in different ocean basins (Lam & Morton 2006, Lapegue et al. 2006, Shilts et al. 2007, Wang & Guo 2008) have already demonstrated the usefulness of molecular markers. Here, for the first time, we have tested Harry's (1985) synonymy of the two species with DNA sequence comparisons, followed by morphological comparisons that were posthoc with respect to groupings identified by our molecular analyses. We analyzed two mitochondrial DNA (mtDNA) markers, 16S ribosomal DNA (16S) and cytochrome oxidase subunit III (CO3), for oysters collected at multiple localities between Willapa Bay, WA (the type locality of O. lurida) and Mazatlan, Mexico (the type locality for O. conchaphila). Our sampling sites also included seven localities between southern California, USA and the Pacific coast of Baja California, Mexico to test for the presence of Hertlein's (1959) predicted zone of overlap.
Specimen Acquisition
All samples collected whose sequences were used in the analysis, along with localities and sample numbers, are listed in Table 1. Samples from Willapa Bay, WA were collected and identified as the Olympia oyster by Russell Rogers (WA Department of Fish and Wildlife) and were given to us as soft tissue only. One ingroup sequence O. "conchaphila" from Barkley Sound, B.C., Canada, and all outgroups used in the analyses were downloaded from GenBank (Table 2). Some of the names listed in Table 2 reflect recent taxonomic changes, such as favoring Ostrea over Ostreola for those species commonly assigned to the latter (see Introduction). When collected in the field, all oysters were opened and then immediately preserved in 95% ethanol. We randomly selected a subset of samples collected from each site for analysis, and these were each given unique voucher labels. Because samples from Willapa Bay, WA were soft tissue only, the entire sample was extracted.
DNA Extraction and "Touchdown" Polymerase Chain Reaction (PCR)
For the DNA extraction and purification, we used DNeasy tissue kits from Qiagen (Valencia, CA) and followed the manufacturers' recommendations. For the tissue, approximately 25 mg of the adductor muscle was dissected and digested in a Proteinase-K and lysis buffer bath overnight at 56[degrees]C. Double-stranded products of partial 16S were amplified via PCR with universal animal primers (Palumbi 1996): 16Sar: 5'--CGC CTG TTT ATC AAA AAC AT--3', 16Sbr: 5'--GCC GGT CTG AAC TCA GAT CAC GT--3'. Double-stranded products of CO3 were amplified via PCR with primers modified for oysters (P. Baker unpublished): CO3F: 5'--AAA AGT TCA AAG CGG TCT TA--3', CO3R: 5'--AGC TAA CAT ACG AAC AAG GC--3'.
A HotStarTaq kit from Qiagen was used to prepare 50 [micro]L reactions for amplification of 16S and CO3. Each reaction included 4.0 [micro]L of 25 mM Mg[Cl.sub.2] (CO3 only), 5 [micro]L of 10 x buffer, 5 [micro]L of dNTPs (10 mM), 2 [micro]L of primer (1 [micro]L forward/1 [micro]L reverse, 10 [micro]M), 1 [micro]L genomic DNA template, 0.25 [micro]L HotStarTaq polymerase, and deionized sterile water to bring the reaction volume to 50 [micro]L per reaction. Reactions were carried out via a GeneAmp PCR System 2400 (Perkin-Elmer) thermocycler. HotStarTaq activation and initial denaturation temperatures were 95[degrees]C for 15 min (a step specific to the HotStarTaq) and then 94[degrees]C for 1 min thereafter (denaturation only). For 16S, initial annealing temperatures of 53[degrees]C for eight cycles of 1 min each (decreased by 1[degrees]C each cycle) were used; final annealing temperatures of 48[degrees]C for 25 cycles of 1 min each were used and an extension temperature of 72[degrees]C was used for 1 min per cycle and for the final extension period of 10 min, followed by a hold at 10[degrees]C until samples were removed to storage at 5[degrees]C. For CO3, initial/final annealing temperatures of 50/45[degrees]C for 30 cycles of 1 min each were used and an extension temperature of 72[degrees]C was used for 1 min per cycle and for the final extension period of 10 min. PCR product was visualized with agarose-gel electrophoresis (1.5%), ethidium bromide staining, and UV illumination to confirm successful amplification of the mtDNA region of interest.
DNA Sequencing Alignment and Analyses
PCR product cleanup and sequencing were performed commercially at the Duke University Institute for Genome Sciences and Policy DNA sequencing facility (Duke IGSP, Durham, NC). Each sequence contig was assembled from a pair of sequences from opposite strands using CodonCode Aligner software (CodonCode Corp, Dedham, MA). All new sequences were submitted to GenBank and have been assigned the accession numbers FJ768501-FJ768589 (16S) and FJ768590-FJ768673 (CO3).
All sequences were aligned by eye with the use of MANIA software (D. L. Swofford & D. J. Eernisse, unpublished). There was very little sequence length variation, and so alignment was generally not problematic. The DNA sequence alignments of 16S and CO3 were 502 and 417 sites, respectively. Heuristic searches were performed for the combined, and separate, data set, or sets, using maximum parsimony (MP) and maximum likelihood (ML) optimality criteria. Altogether, 161 specimens were in our combined data set, including 111 for 16S and 86 for CO3. 16S sampling emphasized the contrast between Ostrea conchaphila and O. lurida and tested for Hertlein's (1959) proposed region of overlap whereas CO3 sequencing emphasized tests of phylogeographic structure within O. lurida. The larger 16S data set included 22 sequences downloaded from GenBank, whereas only two of these (Crassostrea virginica and C. gigas) were available for inclusion in our CO3 data set. The other 84 CO3 sequences were either Ostrea "lurida" (n = 24 from 8 widely separated localities) or O. conchaphila (n = 7 from 2 localities in close proximity). To combine the two data sets, 50 ingroup specimens with CO3 sequences but without 16S sequences were deleted from the analysis (i.e., the combined analysis included the same 111 taxa as in the 16S-only analysis). This avoided confounding tree searching by having a "missing data" set of taxa from one data set that shared no characters with a similar set from the other.
For the MP criterion, we used the PAUP* 4.0b10 software package (Swofford 2003). All gaps were treated as missing data to avoid treatment of adjacent gaps as separate characters. The two-part search strategy used by Kelly and Eernisse (2008) was used to improve the efficiency of searching, specifically to avoid having searches become unnecessarily trapped in nonoptimal tree islands. The first part involved 1,000 replicate searches with only 10 trees held per replicate, keeping all minimum length trees found. Then the second part involved swapping on all minimum-length trees found with the trees-held restriction removed, starting with trees already in memory and with maxtrees set to 1,000. Branch robustness was estimated by 1,000 bootstrap replicate searches, each with 10 replicate random stepwise addition sequence heuristic searches.
For the ML criterion, we used genetic algorithm for rapid likelihood inference (GARLI) v. 0.951 (Zwickl 2006). Because this program searches for trees with a fast and accurate genetic algorithm whose speed is not substantially influenced by the number of taxa, it can handle much larger data sets than are conventionally analyzed with the ML criterion. The version of GARLI used has automatic model selection, which is optimized for the particular data set during analysis. (A newly released v. 0.96 of the program adds additional model assignment options that were not previously available.) Ten separate GARLI searches were performed and the tree selected was the one with the most optimal (closest to zero) final likelihood score. Branch robustness was estimated with 100 bootstrap replicate searches using GARLI. To check whether these results agreed with a more conventional ML search, we performed such a search with PAUP* with the number of taxa greatly reduced to 29 taxa to allow the feasible completion of searches. For this analysis, we used FINDMODEL (Tao et al. 2005) to identify an appropriate model for each data set. Based on this selection, we performed a heuristic search for the combined data set using the general time-reversible plus gamma (GTR + [GAMMA]) model with optimality criterion set to minimum evolution and tree-bisection-reconnection (TBR) branch swapping algorithm.
Corrected pairwise differences within and between species were calculated using Arlequin version 3.11 (Excoffier et al. 2005) with 1,000 permutations, transitions and transversions equally weighted, and acceptable level of missing data set to 5 percent. Corrected pairwise differences were used to estimate intra and interspecific percent sequence divergence by dividing the mean pairwise differences between species by the number of base pairs used in the pairwise analysis.
Morphological Description
To verify the identities of the samples collected from and around Mazatlan, several morphological characters previously identified as potentially diagnostic within the Ostreidae (Keen 1958, Torigoe 1981, Harry 1985, Castillo Rodriguez & Garcia-Cubas 1986, Quayle 1988, Coan et al. 2000) were considered. The morphologies of the samples from Mexico were examined in a "blind" manner, with no a priori knowledge of the molecular outcomes. Characters were examined in selected specimens from mainland Mexico from which DNA was extracted (Table 3). Post-hoe examinations of nine specimens (Table 3) from Hertlein's (1959) zone of overlap (Southern California, USA and Baja California, Mexico) were also completed and compared with the blind-examined specimens in an effort to validate findings from the molecular analyses. Also examined was the shell of a syntype specimen of Ostrea lurida from the Redpath Museum at McGill University, Montreal, Canada. The specimen, RMM 125, is presumed to be part of the type lot described by Philip Carpenter. The only reason that we have avoided designating this specimen the lectotype for O. lurida is our lack of knowledge of other syntype specimens. The syntypes of Ostrea conchaphila are housed at The Natural History Museum in London, England. Images of one of these specimens selected by Kathie Way, as a candidate for lectotype designation, were obtained, but too late to include in this manuscript.
Shell and soft-tissue characters used in the morphological examinations are described individually later. For illustrations of some of these characters see Harry (1985).
Chomata Sweep and Chomata Shape
Chomata are small (often under 1 mm), repeating, tubercule or denticle-like shell features on the internal shell margin on either side of the hinge. Pits (catachomata) lie in the left, or attached valve, and matching denticles (anachomata) lie in the right valve (Torigoe 1981). Chomata shape may be rounded (less than twice as long as wide) or elongate (more than twice as long as wide).
Chomata extend (sweep) in a series from the hinge area around the margin towards the ventral side. Chomata can be a useful character to distinguish the genus Crassostrea from Ostrea, as Crassostrea are believed to lack this character (Harry 1985). In some taxa (e.g., Saccostrea) the chomata extend all the way to the ventral margin (100% sweep), but, in other taxa, the chomata extend only a fraction of that distance.
Plicae (plications) are matching folds in both shell valves that are expressed in a zigzag manner, particularly near the ventral shell margin, so that the valves close precisely. Individual plica-like folds can be mimicked by the shell conforming to substratum topology.
Shell Color
Shell color, both internal and external, is sometimes listed as a character for oysters, especially for Crassostrea (e.g., Keen 1958, Quayle 1988, Coan et al. 2000), where the color is typically described as chalky white, but these same sources admit its high variability. Furthermore, shell color can be altered or eliminated by weathering and preservation techniques (Patrick Baker, pers. obs.). Descriptions of Ostrea conchaphila (and lurida) often note an olive-green interior color (e.g., Keen & McLean 1971, Hertlein 1959).
Other Shell Characters
External shell sculpture, in the form of foliations or spines, has been described for some taxa (Torigoe 1981). Secondary mantle retractors occur in the genus Saccostrea and appear on the shell as a series of small oval muscle scars paralleling the pallial line (Torigoe 1981).
Soft Tissue Characters
Torigoe (1981) and Harry (1985) used mantle tentacles and the morphology of anal papillae as diagnostic characters although Coan et al. (2000) questioned their value and, likewise, Baker (pers. obs.) observed that preservation techniques might change their appearance. Therefore, characters were reduced to polymorphism of tentacles on the inner two mantle folds (those with the most distinct tentacles), and the presence or absence of an external anal papilla.
Molecular Analyses
Trees that were optimal for the maximum parsimony (MP) and maximum likelihood (ML) optimality criteria, for the combined and separate 16S and CO3 datasets, all resulted in similar topologies: a northern group (O. "lurida" from British Columbia to Baja California) and a southern group (O. "conchaphila" from EdPn and EdPs in Sinaloa, combined as EdP hereafter) are reciprocally monophyletic with respect to all other included taxa. Variation among the first 1,000 MP trees found is adequately represented by one of these and was selected arbitrarily (Fig. 1). The sampling scheme for 16S (n = 111) and CO3 (n = 84) data sets differed considerably in the localities sampled (see Methods and Table 1). Only 36 of the 86 taxa with CO3 data were included in our combined analysis of 111 total taxa (Fig. 1 and Fig. 2). Analysis of the separate CO3 data set with either only these 36 included taxa or else with the complete 86-taxon CO3 data set produced results entirely consistent with analysis of the 16S data set and the combined set. For example, for our MP analysis, the nodes for each focal species or for both sister species together in the CO3-only data set are supported with high (>95%) bootstrap support. These results agree with the 16S-only and combined sets, though the latter produced somewhat lower bootstrap support (e.g., Fig. 1). The MP and ML bootstrap support values mostly agreed with each other, but with somewhat lower support for nodes with ML. Results of a PAUP * search using the ML criterion, with a necessarily reduced set of taxa, were consistent with those from the more extensive GARLI analysis (Fig. 2). This implies that the particular topological results emphasized here were independent of the optimality criterion (MP or ML) or ML approach (PAUP* or GARLI) used. The best MP and ML trees did differ somewhat in the topology of the most proximal outgroups for O. conchaphila + O. lurida (Fig. 1 and Fig. 2, respectively). In our best ML tree, a greater proportion of the total branch length is found in the separation of our ingroup and the most proximal outgroups (Ostreinae + Lophinae) from our more distal outgroups from Saccostrea and Crassostrea (both in Crassostreinae). Although Ostreinae is paraphyletic to Lophinae in our best trees (as indicated by the quotation marks around Ostreinae in Fig. 2A), there is no strong bootstrap support for such nesting of Lophinae within Ostreinae, so it is possible that further resolution will support their reciprocal monophyly.
The bootstrap support supporting the monophyly of O. lurida and O. conchaphila are each high for both MP and ML (Figs. 1 and 2B). However, the joining of these as sister species is more equivocal. Our best trees for MP and ML resolves them as sister taxa but with low bootstrap support. The bootstrap proportions supporting the node joining O. conchaphila + O. lurida is higher for MP (69%) than for ML (43%). The only alternative grouping observed for MP and ML with >10% bootstrap support had O. lurida as sister taxon of all other Ostreinae + Lophinae species, including O. conchaphila. This arrangement was supported in 11% of the MP bootstrap replicates and 35% of the ML replicates.
The average percent sequence divergence between paired combinations of 16S sequences for O. conchaphila versus O. lurida was 2.06%. Samples with 16S only from Mazatlan (Maz) and Bahia de Kino (BdK) did not group with either Ostrea species. All Maz samples and one BdK sample grouped with Saccostrea spp., whereas the other BdK sample grouped with Crassostrea spp. (almost certainly C. gigas, assuming the sequence from GenBank is correctly identified as from this species). Independent morphological examinations only partially supported their assignment to these genera as described later. We used morphology to identify one of the specimens (see later) as S. palmula (Carpenter, 1857), and this was later resolved as Saccostrea with DNA. Because this is the only recognized species of Saccostrea from the Panamic region of the eastern Pacific (P. Valentich-Scott, pets. comm.), we have provisionally assigned all of the specimens resolved with DNA as Saccostrea to this species.
Separate analysis of the complete CO3 data set (as well as especially our MP combined data set analysis) supported a phylogeographic separation within O. lurida. Ignoring autapomorphic substitutions in six of the 84 total ingroup taxa, the entire CO3 data set reduces to the variable sites summarized in Figure 3 and the corresponding haplotype network in Figure 4. In particular, sites for either side of the southern end of Vancouver Island (B.C., Canada) were identical to each other but separated from Willapa Bay (southwestern Washington, USA). All other O. lurida sites from Oregon, CA, and Baja California, Mexico were identical to the Willapa Bay site, ignoring autapomorphies. The southern and northern haplotype groups within O. lurida are equally separated from the much more distant O. conchaphila, with each having an identical nucleotide state in common with O. conchaphila, relative to the other at two sites. Because two highly divergent CO3 sequences for Crassostrea spp. were the only available outgroups for our CO3 data set, these did not provide a useful rooting for inferring whether these alternative matching states are plesiomorphic or derived. Because of this lack of proximal outgroups and because O. conchaphila is relatively far from O. lurida, the rooting depicted in Figure 4 for O. conchaphila midway between the Vancouver Island and more southern haplotype group is arbitrary.
The phylogeographic contrast discovered for CO3 was not seen in the three 16S sequences obtained from the Vancouver Island populations versus the other 108 more southern sequences. This is probably best explained by the generally slower substitution rate of the 16S gene region.
Morphological Examinations
Independent Morphological Identification of Mainland Mexico Samples
The findings from the independent morphological examinations of samples from Mexico were in agreement with the molecular results in most cases (Table 3). All samples from two adjacent sites within Ensenada del Pabellon, Sinaloa, Mexico were tentatively identified as Ostrea conchaphila based on the presence of a rounded or slightly elongate chomata that extended <50% along the internal shell margin on either side of the hinge (Table 3); this was in agreement with the molecular results. In contrast, one sample from Mazatlan (Maz 0503) was identified as Saccostrea palmula based on the presence of chomata all the way around the shell, a purple interior margin and the absence of anal papilla and secondary mantle retractor scars. The identifications of the other two samples from Mazatlan could not be positively resolved using morphological characters. All three Mazatlan samples grouped with Saccostrea sp. in the molecular results. From Bahia de Kino, one specimen (BdK 0701) was identified as Crassostrea sp. based on the absence of chomata and the thick, chalky shell, and this finding was congruent with the molecular results. Despite the presence of chomata in the other BdK sample examined (BdK0702), this sample was also tentatively identified as Crassostrea sp. based on the thick, chalky shell (Table 3), however, this sample grouped with Saccostrea sp. in the molecular findings.
Post-Hoc Morphological Examination of Samples from Hertlein's Zone of Overlap
Our post-hoc morphological examinations were not able to resolve any consistent differences between the putative northern O. lurida and the southern O. conchaphila (Table 3). For each character there was at least one sample that differed from the mode (e.g., chomata shape, Table 3). Morphological distinctions were also not apparent from the examination of a syntype specimen of Ostrea lurida.
Specific observations from the independent and post-hoc examinations are reported later for each character.
Chomata Sweep and Chomata Shape
Chomata were present in all specimens except BdK 0701 (Table 3). In some specimens, this chomata shape was intermediate and/or variable, designated as round-elongate in Table 3. In none of the specimens examined, however, was the presence or absence of chomata ambiguous, at least under low-power magnification.
Figure 3. Summary of observed CO3 variation by three haplotype groups. Variation was consistent with dividing localities into southernmost, northernmost, and all other localities in between, as listed in the CO3 column of Table 1. Oco-Southern (S) includes both Ostrea conchaphila sites, EdPn and EdPs; Olu-Northern (N) includes both Vancouver Island sites for O. lurida, LH and AI. Olu-All Other (A) includes all other O. lurida localities. Sample sizes for three observed haplotypes: S = 7; N = 7; A = 70. This includes three individuals (two S and one A) that differed from the other normal haplotype patterns by only a single unique base pair difference, but none of this variation occurred at sites a-m. The similarity "Pattern" tallies to 12 for A + N ("1") and two each for A + S and N + S ("2" and "3"). No assumption was made to root the similarity matching patterns because the only available outgroups (Crassostrea spp.) were highly divergent from these three haplotypes. Alignment sites summarized above were as follows, relative to the complete mitochondrial genome for Crassostrea gigas, AF177226: a = 190; b 210; c = 231; d = 258; e = 309; f = 366; g = 395; h = 399; i = 435; j = 453; k = 459; I = 495; m = 501; n = 519; o = 558; and P = 582. Of these, only site 'g' represents a nonsynonymous substitution. CO3Site abcdefghijklmnop Oco-Southern TGTCACATGTTTGTAA Olu-Northern AACTGTGTAGTCACGC Olu-Allother AACTGTACGGCCACGC Pattern 1111112321311111
Plicae were scored as absent if they did not appear to arise independently of underlying substratum topology. In some specimens, plicae were numerous and regular, but in others they were irregular or few in number (Table 3).
Shell Color
Internal shell color was ambiguous in all samples except for Maz 0503 in which it was deep purple; internal shell color is not included in Table 3. External shell color, where it differed unambiguously from white or off-white, was recorded in Table 3.
Other Shell Characters
External shell sculpture in the form of foliations or spines was not present in any of the specimens examined in this study. Secondary mantle retractor scars were present in specimen Maz 0503, which helped in diagnosing it as Saccostrea. Both samples from BdK had relatively thick shells throughout (>2 mm) and were chalky in texture, which is a common description to Crassostrea shells.
Soft Tissue Characters
For some of the samples, characters such as anal papillae and mantle tentacles could not be defined precisely because of tissue damage, poor preservation, and possible environmental plasticity. The presence/absence of an anal papilla and the relative size of mantle tentacles on the two inner folds were variable within suspect taxa.
The combined and separate analyses of the 16S and CO3 sequence data sets do not support Harry's synonymy of Ostrea lurida with Ostrea conchaphila, but instead support the original classifications by Carpenter (1857, 1864), who treated them as separate oyster species. Our evidence thus provisionally supports the reinstatement of Ostrea lurida as the Olympia oyster. From the analysis of the 16S data set, samples from the type locality of O. lurida, Willapa Bay, WA, along with seven other locations in Southern California and Baja California are unambiguously supported as a monophyletic grouping, within which we noted only minor variation. Likewise, 16S sequences of oysters from two adjacent locations within Ensenada del Pabellon (EdP in Fig. 1), Sinaloa, Mexico, near the type locality of O. conchaphila, group as a well-supported phylogenetic lineage. Relative to the most proximal available outgroups, specifically various Ostrea spp. listed in Table 2, O. lurida and O. conchaphila are weakly supported as sister species. However, Keen and McLean (1971) note the presence of at least 3 other putative Ostrea species in the Gulf of California, for which molecular data are unavailable, but who might serve as sister taxa to either focal species in this study.
Bootstrap support for the sister relationship between the focal species was higher for MP (68%) than for ML (43%). For either MP or ML, the highest and lowest bootstrap support proportions were observed for the CO3-only and 16S-only data sets, respectively, but even the latter was reasonably well supported in the case of MP. As detailed earlier in our results, alternative grouping with the next highest bootstrap proportion for both MP (11%) and ML (35%) placed O. lurida as basal within the "Ostreinae" + Lophinae monophyletic grouping. Still, the best trees for either optimality criterion places O. conchaphila and O. lurida as sister species. The lack of bootstrap support could be caused by the considerable branch length divergence of the involved nodes in their separation from the next available outgroups (Saccostrea and Crassostrea spp.), or the presently sparse intraspecific sampling for our other "Ostreinae" + Lophinae outgroups. Alternatively, the low bootstrap support might reflect the phylogenetic possibility that these native North American oyster species might not be sister species. For example, we have not included any extant or extinct Ostrea spp. from the northwestern Pacific and perhaps one or more of these could have a closer relationship to the similarly cooler water O. lurida.
Interesting phylogeographic structure was observed within O. lurida for the CO3 data set and this coincided with the geographic separation between Willapa Bay, WA and the southern part of Vancouver Island, B.C., Canada. A break in this vicinity is unexpected. For marine organisms, most have found little evidence of a biogeographic break in this area (e.g., Kelly & Eernisse 2007), although some have found a break further north (e.g., Hart et al. 2005) or coinciding with the exposed coast versus inland passage discontinuity between British Columbia and Alaska (e.g., Lindstrom et al. 1997). It is possible that the Vancouver Island localities are populated with oysters that are relicts of Pleistocene glacial refugia, which have been inferred for areas north of these Vancouver Island localities. Specifically, glacial refugia during the late Wisconsin glaciation have been supported for the Queen Charlotte Islands, northern British Columbia (Hetherington et al. 2003) and in the Alexander Archipelago of southeastern Alaska (Carrara et al. 2007). At least one terrestrial species, the lodgepole pine, might have relicts in glacial refugia on the outer coast of Vancouver Island (Godbout et al. 2008). However, the present evidence for phylogeographic separation needs further study with more localities and more gene regions before such historical hypotheses can be accurately inferred.
Because of their extremely plastic morphologies, oysters present difficult challenges for identifications based solely on shell characteristics in the field and such identifications can be especially challenging or even impossible, even after careful examination of internal anatomy and shell structure in the laboratory. For example, the two samples from Bahia de Kino were identified as Ostrea conchaphila in the field, and were reidentified as Crassostrea sp. based on shell morphology in the laboratory. The subsequent molecular analyses grouped only one of the two samples as nearly identical to C. gigas with 100% MP bootstrap support, the other grouped with our other specimens presumed to be Saccostrea palmula (Fig. 1, Table 2). The need, in some cases, to rely on molecular analyses to positively identify specimens underscores the challenges associated with resolving the systematics of this troublesome group. Regarding the morphological characters examined, the presence/absence of chomata was the only unambiguously diagnostic character and it was only informative at a generic level, not between species of Ostrea.
Aside from the presence/absence of chomata, no other shell or soft tissue characters were 100% congruent with molecular data. Only one of the three Saccostrea samples examined had unambiguously elongate chomata all the way around the shell margin. Whereas all Ostrea samples examined had chomata, some were rounded whereas others were slightly elongate. Specimens identified as O. lurida from molecular study (Tables 1 & 3) were nearly as likely to have external shell color or shell plications as not. Soft tissue characters were more consistent, but for each character, at least one specimen, even in this small sample size, differed from the mode. The potential value of anal papillae was further limited by tissue damage. The variability we observed is consistent with comments by Keen (1958) and Coan et al. (2000) on Ostreidae morphology; Coan et al. (2000) emphasize the need for molecular data for more reliable identification. In all, there was considerable intraspecific and interpopulation variation that confounded any attempt to separate the genetically distinct lineages of O. lurida from O. conchaphila by morphology alone.
For the combined analyses of the 16S and CO3 datasets, O. conchaphila and O. lurida were well supported as distinct taxa (Figs. 1-2). Unfortunately, we were unable to find any examples of a calibrated molecular clock for either 16S or CO3 for other bivalves, but an approximate comparison is available for a molecular clock calibrated for 16S in Pacific porcelain crabs (Stillman & Reeb 2001), based on this, the range of estimated pairwise divergence times for O. conchaphila versus O. lurida is about 1.5-3.9 mya. The evidence for the reciprocal monophyly of O. conchaphila and O. lurida is indicative of a significant absence of gene flow between the two groups. However, again, we found no diagnostic morphological character corresponding to either the northern or southern groups (Table 3). Although we have contrasted oysters from near the type locality of O. conchaphila with oysters from the type locality of O. lurida, much further north in Washington, we have not carefully studied the range of morphological variation across our O. lurida localities with enough detail to determine whether there might be diagnostic shell or anatomical traits to separate O. lurida from O. conchaphila. Moreover, we have only collected O. conchaphila from one pair of nearby sites near Mazatlan. Clearly, further morphological study, including more intensive sampling throughout the range of O. conchaphila, is needed.
Based on our molecular evidence, we only found O. lurida within Hertlein's (1959) putative zone of overlap that is supposed to exist between these two species. Samples from the seven localities in southern California and northern Baja California (Table 1) all were effectively indistinguishable from O. lurida at the type locality, Willapa Bay, WA, and this was true for both mtDNA gene regions. In the future, sampling from other locations is warranted to determine the existence, if any, of a zone of overlap, and any such zone is predicted to occur further south than where we sampled, perhaps on the outer coast of Baja California Sur. A transition zone might also exist within the Gulf of California, however, previously published reports imply that neither species occurs within the Gulf of California (e.g., Keen & McLean 1971). Still, this could be because of lack of study, because molecular data on Gulf of California oyster species are mostly lacking. Thus, the only stretch of ostensibly inhabited coastline that was not sampled in this study extends from locations south of Bahia San Quintin, Baja California to the previously reported southern range endpoint for Ostrea lurida at Cabo San Lucas, Baja California Sur. Recent extensive intertidal searches in Cabo San Lucas failed to locate any oysters (Polson & Zacherl, this issue). The only mention of the existence of a zone of overlap was by Hertlein (1959), who also described three separate morphotypes found in southern California. Considering the high degree of phenotypic plasticity in oyster shell morphology, a zone of overlap more likely never was present in southern California and northern Baja California. Instead, Hertlein (1959) might have documented evidence of the high degree of phenotypic plasticity exhibited by O. lurida.
To bridge the 2 most popular species concepts, the biological species concept and the phylogenetic species concept, the concordance principle (CP) introduced by Avise & Ball (1990), recognizes a species based on the phylogenetic concordance of multiple independent molecular markers. Moreover, CP calls for reproductive isolation but is based on intrinsic rather than extrinsic (i.e., geographic) forces. With this study, we have provided molecular evidence across two nonindependent (i.e., tightly linked) mtDNA markers in a geographic context. Our phylogenetic estimates support O. lurida and O. conchaphila as two distinct taxa, with data still lacking for any transitional localities, should they exist. Finding a zone of overlap (sympatry) within which there is little evidence for ongoing hybridization would have provided stronger support that O. lurida and O. conchaphila are separate species and not simply extremes of a latitudinal cline. Future studies would benefit from sampling variable nuclear markers to test for such hybridization. Recently Wang and Guo (2008) found ITS length variation across multiple species of Crassostrea but unfortunately no variation was detected across the Ostrea spp. compared.
The provisional decision to again regard these clades as separate species here, reviving Carpenter's Ostrea lurida and rejecting Harry's (1985) synonymy, is based on the evidence from this study for historical separation of the oysters from the Panamic versus the cooler northeastern Pacific. Whereas no one has yet demonstrated diagnostic shell or anatomical characters, this situation is in no small part caused by their notorious plasticity, and having the present phylogenetic estimate will help define where morphological contrasts with a genetic basis are likely to be found. With roughly 1.5-3.9 my separation, our mitochondrial markers were successful in distinguishing these species. The Olympia oyster becomes a sister species whose southern range limit has yet to be carefully explored along the Baja California Peninsula, and much less is yet known about O. conchaphila. In addition to molecular distance, this pair of oyster species is separated both by geography and contrasting climates. Although other Ostrea spp. and other locations in Mexico need to be sampled, evidence to date implies a sibling species pair with one in the subtropical Panamic marine biogeographical province and the other extending along the cooler Californian and Oregonian temperate provinces to the north. Our preliminary evidence for further phylogeographic structure within O. lurida is potential evidence that such historical processes might be ongoing. Thus, calling for the reinstatement of Ostrea lurida as it was originally described and once widely accepted will help focus research on the remaining mysteries of the Olympia oyster throughout its range.
The authors thank Dr. Michel Hendrickx and Jose "Pepe" Salgado (Universidad Nacional Autonoma de Mexico, Instituto de Ciencias del Mar y Limnologia, Mexico) for collecting samples from Sonora and Sinaloa, Mexico and for all their help with our sampling in Mazatlan. The authors also thank Russell Rogers (Washington Department of Fish and Wildlife) for the Willapa Bay samples and Dr. Anthony Ricciardi from the Redpath Museum, Canada for lending us a syntype of Ostrea lurida. Paul Valentic-Scott (Santa Barbara Museum of Natural History) helped with acquisition of our vouchers into that museum and provided valuable information on Panamic Ostreidae. The authors also appreciate the help of Ms. Lisa Bukovnik at Duke University's IGSP for her help with sequencing our samples. Financial support was provided by grants to D. Zacherl from the CSU Special Fund for Research, Scholarship, and Creative Activity and from NSF-OCE # 0351860, and grants to M. Poison from the Department of Biological Science and the Dr. and Mrs. Donald B. Bright Environmental Scholarship. D. Eernisse acknowledges sabbatical support by CSUF and a fellowship from the National Evolutionary Synthesis Center (NESCent) at Duke University, NSF-EF#0423641.
Avise, J.C. 2004. Molecular markers, natural history, and evolution. Second edition. Sinauer Associates, Inc. Publishers. Sunderland, Massachusetts. 684 pp.
Avise, J. C. & R. M. Ball. 1990. Principles of genealogical concordance in species concepts and biological taxonomy. Oxford Surv. Evol. Biol. 7:45-67.
Baker, P. 1995. Review of ecology and fishery of the Olympia oyster, Ostrea lurida, with annotated bibliography. J. Shellfish Res. 14:501-518.
Carrara, P. E., T. A. Ager & J. F. Baichtal. 2007. Possible refugia in the Alexander Archipelago of southeastern Alaska during the late Wisconsin glaciation. Can. J. Earth Sci. 44:229-244.
Castillo Rodriguez, Z. G. & A. Garcia-Cubas. 1986. Taxonomia y anatomia comparada de las ostras en las costas de Mexico. An. Inst. Cienc. Mar Linmol. Univ. Nac. Aulon. Mex. 13:249-314.
Clement, M., D. Posada & K. Crandall. 2000. TCS: a computer program to estimate gene genealogies. Mol. Ecol. 9:1657-1660.
Coan, E.V., P. Valentich-Scott & & F.R. Bernard. 2000. Bivalve seashells of western North America: marine bivalve mollusks from Arctic Alaska to Baja California. Natural History Monographs No. 2, Santa Barbara Museum of Natural History, Santa Barbara, California, USA. 764 pp.
CodonCode Corporation. Software for DNA sequencing, www.codoncode. com.
Cracraft, J. 1987. Species concepts and the ontology of evolution. Biol. Philos. 2:329-346.
Crummett, L. T. & D. J. Eernisse. 2007. Genetic evidence for the cryptic species pair, Loilia digitalis, and Lottia austrodigitalis and microhabitat partitioning in sympatry. Mar. Biol. 152:1-13.
Dall, W. H. 1914. Notes on west American oysters. Nautilus 28:1-3.
Dawson, N. D. & D. K. Jacobs. 2001. Molecular evidence for cryptic species of Aurelia aurita (Cnidaria, Scyphozoa). Biol. Bull. 200:92-96.
Donoghue, M. J. 1985. A critique of the biological species concept and recommendations for a phylogenetic alternative. Bryologist 88:172-181.
Excoffier, L., G. Laval & S. Schneider. 2005. Arlequin ver. 3.0: an integrated software package for population genetics data analysis. Evol. Bioinform. Online 1:47-50.
Godbout, J., A. Fazekas, C. H. Newton, F. C. Yeh & J. Bousquet. 2008. Glacial vicariance in the Pacific Northwest: evidence from a lodgepole pine mitochondrial DNA minisatellite for multiple genetically distinct and widely separated refugia. Mol. Ecol. 17:2463-2475.
Harry, H. 1985. Synopsis of the supraspecific classification of living oysters (Bivalvia: Gryphaeidae and Ostreidae). Veliger 28:121-158.
Hart, M. W., J. Sunday & C. C. Keever. 2005. Dispersal and climate history in sea star population genetic structure. Integr. Comp. Biol. 45:1009. [abstract].
Hetherington, R., J. Vaughn Barrie, R. G. B. Reid, R. MacLeod, D. J. Smith, T. S. James & R. Kung. 2003. Late Pleistocene coastal paleogeography of the Queen Charlotte Islands, British Columbia, Canada, and its implications for terrestrial biogeography and early postglacial human occupation. Can. J. Earth Sci. 40:1755-1766.
Hertlein, L. 1959. Notes on California oysters. Veliger 2:5-10.
Hewson, W.E. 2008. Biogeography of Lottia strigatella (Carpenter 1864) and a description of a cryptic species. Master thesis. California State University, Fullerton, CA. 82 pp.
Keen, A. M. 1958. Sea shells of tropical west America. Stanford, California: Stanford. 624 pp.
Keen, A. M. & J. H. McLean. 1971. Sea shells of tropical west America. Second edition. Stanford, California: Stanford University Press. 1,064 pp.
Kelly, R. P. & D. J. Eernisse. 2007. Southern hospitality: A latitudinal gradient in gene flow in the marine environment. Evolution hit. J. Org. Evolution 61:700-707.
Kelly, R. P. & D. J. Eernisse. 2008. Reconstructing a radiation: The chiton genus Mopalia in the north Pacific. Invertebr. Syst. 22:1-12.
Kirkendale, L., T. Lee, P. Baker & D.O. Foighil. 2004. Oysters of the Conch Republic (Florida Keys): a molecular phylogenetic study of Parahyotissa megintyi, Teskeyostrea weberi and Ostreola equestris. Malacologia 46:309-326.
Kluge, A. G. 1990. Species as historical individuals. Biol. Philos. 5:417-431.
Knowlton, N. 1993. Sibling species in the sea. Atom. Rev. Ecol. Syst. 24:189-216.
Lam, K. & B. Morton. 2006. Morphological and mitochondrial-DNA analysis of the Indo-West Pacific rock oysters (Ostreidae: Saccostrea species). J. Molluscan Stud. 72:235-245.
Lapegue, S., I. B. Salah, F. M. Batista, S. Heurtebise, L. Neifar & P. Boudry. 2006. Phylogeographic study of the dwarf oyster, Ostreola stentina, from Morocco, Portugal and Tunisia: evidence of a geographic disjunction with the closely related taxa, Ostrea aupouria and Ostreola equestris. Mar. Biol. 150:103-110.
Lindstrom, S. C., J. L. Olsen & W. T. Stare. 1997. Postglacial recolonization and the biogeography of Palmaria mollis (Rhodophyta) along the Northeast Pacific coast. Can. J. Zool. 75:1887-1896.
Mishler, B. D. & M. J. Donoghue. 1982. Species concepts: a case for pluralism. Syst. Zool. 31:491-503.
Palumbi, S. 1996. Nucleic Acids II: the polymerase chain reaction. In: D. M. Hillis, C. Mortiz & B. K. Mable. editors. Molecular Systematics, 2nd edition. Sunderland, MA: Sinauer and Assoc. Inc. pp. 205-246.
Quayle, D.B. 1988. Pacific oyster culture in British Columbia. Canadian Bulletin of Fisheries and Aquatic Sciences 218. Department of Fisheries and Oceans, Ottawa, Ontario, Canada. 192 pp.
Shilts. H. M., M. S. Pascual & D.O. Foighil. 2007. Systematic, taxonomic and biogeographic relationship of Argentine flat oysters. Mol. Phylogenet. Evol. 44:467-473.
Simison, W. B. & D. R. Lindberg. 1999. Morphological and molecular resolution of a putative cryptic species complex: a case study of Notoacmea fascicularis (Menke, 1851) (Gastropoda: Patellogastropoda). J. Molluscan Stud. 65:99-109.
Stillman. J. H. & C. A. Reeb. 2001. Molecular phylogeny of eastern Pacific porcelain crabs, genera Petrolisthes and Pachycheles based on mtDNA 16S rDNA sequence: phylogeographic and systematic implications. Mol. Phylogenet. Evol. 19:236-245.
Swofford, D. L. 2003. PAUP*. Phylogenetic Analysis Using Parsimony (* and Other Methods). Version 4.0b10. Sinauer Associates, Sunderland, Massachusetts.
Tao, N., W. J. Bruno, W. Abfalterer, B. M. E. Moret, T. Leitner & C. Kuiken. 2005. FINDMODEL: a tool to select the best-fit model of nucleotide substitution, http://hcv.lanl.gov/content/hcv-db/findmodel/findmodel.html.
Torigoe, K. 1981. Oysters in Japan. J. Sci. Hiroshima Univ., B. 1. 29:291-481.
Wang, Y. P. & X. M. Guo. 2008. ITS length polymorphism in oysters and its use in species identification. J. Shellfish Res. 27:489-493.
Zwickl, D.J. 2006. Genetic algorithm approaches for the phylogenetic analysis of large biological sequence datasets under the maximum likelihood criterion. PhD thesis. University of Texas, Austin. 125 pp.
(1) Department of Biological Science, California State University Fullerton, P.O. Box 6850, Fullerton, California 92834; (2) Department of Fisheries and Aquatic Sciences, University of Florida, 7922 NW 71st Street, Gainesville, Florida 32653
* Corresponding author. E-mail: firstname.lastname@example.com
([dagger]) The taxonomy of the Olympia oyster has been in dispute since Harry (1985) proposed synonymy of Ostrea lurida Carpenter 1864 and Ostrea conchaphila Carpenter 1857. Poison et al. 2009 provide molecular evidence that the Olympia oyster refers to the nominal species, Ostrea lurida Carpenter 1864. In view of their genetic data, and for consistency, the original taxon, Ostrea lurida, is used throughout this volume to refer to the Olympia oyster, which is distributed from approximately Baja California (Mexico) to southeast Alaska.
([double dagger]) The common name as sanctioned by the American Fisheries Society is Olympia oyster, though other names are commonly used including, "native oyster," "California oyster," "Yaquina oyster."
TABLE 1. Sampling localities for sequenced western North American oysters with the number of samples sequenced for each gene. Names are as determined in this study. 16S sampling emphasized the contrast between Ostrea conchaphila and O. lurida and tested for Hertlein's (1959) proposed region of overlap whereas C03 sequencing emphasized tests of phylogeographic structure within O. lurida. Species Locality (Code) Ostrea lurida Ladysmith Harbor, Vancouver Id.. B.C. (LH) Ostrea lurida Ahmah Island, Vancouver Id., B.C. (AI) Ostrea lurida Willapa Bay, WA (WB) Ostrea lurida Yaquina Bay, OR (YB) Ostrea lurida Coos Bay, OR (CB) Osirea lurida Humboldt Bay, CA (HB) Ostrea lurida Tomales Bay West, CA (TB) Ostrea lurida Drakes Estero, CA (DE) Ostrea lurida Pt, San Quentin, San Francisco Bay, CA (SF) Ostrea lurida Elkhorn Slough, Monterey Co., CA (ES) Ostrea lurida Mugu Lagoon. CA (ML) Ostrea lurida Alamitos Bay, CA (AL) Ostrea lurida Newport Bay, CA (NB) Ostrea lurida Aqua Hedionda, CA (AH) Ostrea lurida Batiquitos Lagoon, CA (BL) Ostrea lurida Scripps Pier, La Jolla, CA (LJ) Ostrea lurida Mission Bay, CA (MB) Ostrea lurida San Diego Bay, CA (SD) Ostrea lurida Bahia San Quintin, Baja California (SQ) Ostrea conchaphila Ensenada del Pabellon (N), Sinaloa (EdPn) Ostrea conchaphila Ensenada del Pabellon (S), Sinaloa (EdPs) Saccostrea spp. Urias Lagoon, Mazatlan, Sinaloa (Maz) Saccostrea sp. Bahia de Kino, Sonora (BdK) Crassostrea gigas Bahia de Kino, Sonora (BdK) Species GPS Coordinates Ostrea lurida N 48[degrees]59.321', W 123[degrees]48.241' Ostrea lurida N 48[degrees]56.911', W 125[degrees]5.239' Ostrea lurida N 46[degrees]29.916', W 124[degrees]01.766' Ostrea lurida N 44[degrees]36.807', W 124[degrees]4.799' Ostrea lurida N 43[degrees]19.313', W 124[degrees]23.156' Osirea lurida N 40[degrees]45.225', W 124[degrees]12.92' Ostrea lurida N 38[degrees]13.763', W 122[degrees]58.556' Ostrea lurida N 38[degrees]0.302', W 123[degrees]0.307' Ostrea lurida N 37[degrees]56.508', W 122[degrees]30.315' Ostrea lurida N 36[degrees]48.631', W 121[degrees]46.996' Ostrea lurida N 34[degrees]6.104', W 119[degrees]5.882' Ostrea lurida N 33[degrees]44.264', W 118[degrees]7.114' Ostrea lurida N 33[degrees]37.173', W 117[degrees]53.542' Ostrea lurida N 33[degrees]08.579', W 117[degrees]19.370' Ostrea lurida N 33[degrees]08.579', W 117[degrees]19.370' Ostrea lurida N 32[degrees]52.087', W 117[degrees]15.307' Ostrea lurida N 32[degrees]47.673', W 117'13.468' Ostrea lurida N 32[degrees]42.505', W 117[degrees]10.260' Ostrea lurida N 30[degrees]27.858', W 115'57.834' Ostrea conchaphila N 24[degrees]30.063', W 107[degrees]41.245' Ostrea conchaphila N 24[degrees]28.843', W 107[degrees]33.396' Saccostrea spp. N 23[degrees]12.33', W 106[degrees]24.315' Saccostrea sp. N 28[degrees]45.55', W 111[degrees]54.8' Crassostrea gigas N 28[degrees]45.55', W 111[degrees]54.8' Species 16S Seqs CO3 Seqs Ostrea lurida 2 4 Ostrea lurida 1 3 Ostrea lurida 21 5 Ostrea lurida 0 5 Ostrea lurida 0 5 Osirea lurida 0 5 Ostrea lurida 0 5 Ostrea lurida 0 4 Ostrea lurida 0 4 Ostrea lurida 0 3 Ostrea lurida 0 4 Ostrea lurida 5 5 Ostrea lurida 4 5 Ostrea lurida 4 3 Ostrea lurida 4 5 Ostrea lurida 5 1 Ostrea lurida 4 5 Ostrea lurida 4 3 Ostrea lurida 4 3 Ostrea conchaphila 2 2 Ostrea conchaphila 7 5 Saccostrea spp. 24 0 Saccostrea sp. 1 0 Crassostrea gigas 1 0 TABLE 2. GenBank sequences used in analyses (16S except as noted). Species Locality in GenBank Accession Ostrcct lurida * Barkley Sound, BC. AF052071 Canada Outgroups Ostrea algoensis Port Alfred, AF052062 South Africa Ostrea angasi St. Helen. Tasmania. AF052063 Australia Ostrea auporia Hauraki Gulf, AF052064 New Zealand Ostrea chilensis Hauraki Gulf, AF052065 New Zealand Ostrea denselamellosa South Korean AF052067 hatchery stock Ostrea edidis France DQ280032 Ostrea equestris Big Pine AF052074 Key, FL, USA Ostrea puelchana San Matias AF052073 Gulf, Argentina Ostrea spreta Argentina DQ640402 Ostrea weberi Florida, USA AY376601 Alectryonella plicatula Pohnpei, Micronesia AF052072 Cryrptostrea permollis Florida panhandle, USA AF052075 Dendostrea folium Aitutaki, Cook AF052069 Islands Dendostrea frons Carrie Bow Cay, AF052070 Belize Lopha cristagalli Guam AF052066 Saccostrea commiercialis Moreton Island, AF353100 Queensland, Australia Saccostrea cuccullata Australia AF458901 Crassostrea ariakensis China AY160757 Crassostrea rhizophorae Brazil AJ312938 Crassostrea gigas Bangor, U.K. (cultured) AJ553903 Crassostrea gigas(CO3) None listed AF177226 (native to northwestern Pacific) Crassostrea virginica None listed AF092285 (native to northwestern Atlantic) Crassostrea virginica Delaware Bay. USA AY905542 (CO3) * This ingroup sequence from GenBank. O. "conchaphila," was reidentified in the current study as O. lurida. TABLE 3. Morphological characters examined among a subset of samples used in the molecular analyses (see Table 1 and Figure 1 for locality codes in the first column). ID Molecular Specimen ID Field * ID Laboratory ** ([section]) BdK 0701 O. conchaphila Crassostrea sp. (a) Crassostrea sp. BdK 0702 O. conchaphila Crassostrea sp. (a) Saccostrea sp. Maz 0503 O. conchaphila Saccostrea sp. (a) Saccostrea sp. Maz 0510 O. conchaphila Unresolved (a) Saccostrea sp. Maz 0512 O. conchaphila Unresolved (a) Saccostrea sp. EdPn 0702 O. conchaphila O. conchaphila (a) O. conchaphila EdPs 0701 O. conchaphila O. conchaphila (a) O. conchaphila EdPs 0703 O. conchaphila O. conchaphila (a) O. conchaphila EdPs 0704 O. conchaphila O. conchaphila (a) O. conchaphila SD 0601 O. conchaphila O. conchaphila (a) O. lurida SD 0603 O. conchaphila O. conchaphila (a) O. lurida AH 0505 O. conchaphila O. conchaphila (a) O. lurida NB 0601 O. conchaphila O. conchaphila (a) O. lurida NB 0602 O. conchaphila O. conchaphila (a) O. lurida SQ 0504 O. conchaphila O. conchaphila (a) O. lurida SQ 0505 O. conchaphila O. conchaphila (a) O. lurida LJ 0503 O. conchaphila O. conchaphila (a) O. lurida LJ 0505 O. conchaphila O. conchaphila (a) O. lurida Chomata Chomata Specimen Sweep Shape Plicae BdK 0701 None Absent No BdK 0702 <50% Round Yes Maz 0503 100% Elongate Yes Maz 0510 <50% Round Yes Maz 0512 <50% Round Yes EdPn 0702 <50% Round-elongate Yes EdPs 0701 <50% Round-elongate Yes EdPs 0703 <50% Round-elongate Yes EdPs 0704 <50% Round-elongate Yes SD 0601 <50% Round Yes SD 0603 <50% Round Yes AH 0505 <50% Round Yes NB 0601 <50% Round No NB 0602 <50% Round No SQ 0504 <50% Round Yes SQ 0505 <50% Round Yes LJ 0503 <50% Round No LJ 0505 <50% Round-elongate No External Mantle Anal Specimen Color Tentacles Papilla BdK 0701 No Monomorphic NA BdK 0702 No Polymorphic Internal Maz 0503 Yes Monomorphic None Maz 0510 No Monomorphic None Maz 0512 No Polymorphic Simple EdPn 0702 Yes Polymorphic Simple EdPs 0701 No Monomorphic None EdPs 0703 Yes Polymorphic Simple EdPs 0704 Yes Polymorphic Simple SD 0601 Yes Polymorphic NA SD 0603 Yes Polymorphic Simple AH 0505 Yes Polymorphic NA NB 0601 No Polymorphic Simple NB 0602 No Polymorphic Simple SQ 0504 Yes Polymorphic NA SQ 0505 Yes Polymorphic Simple LJ 0503 No monomorphic NA LJ 0505 No polymorphic NA * Taxon ID based upon field identification using external morphological characters. ** Taxon ID based upon both (a) pre and (b) posthoc laboratory examination of internal and external morphological characters. ([section]) Taxon ID after molecular results; Saccostrea sp. is likely S. palmula, the only Panamic member of this genus.
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IFC Midnight takes Zack Parker's "PROXY"
Zack Parker's psychothriller look at grief and obsession, PROXY, has been picked up for North American distribution out of the Toronto International Film Festival.Read more »
Zack Parker's psychothriller look at grief and obsession, PROXY, has been picked up for North American distribution out of the Toronto International Film Festival.Read more »
INSIDIOUS: CHAPTER 2 is a surprising film. Less the slow build scare-a-thon of the first (and even James Wan's summer hit THE CONJURING), this film is an aggressive whirlwind that's no longer approaching The Further with trepidation, but charging through and searching for its weirdest bits. Of course, in what's become something of a recurring motif in the work of Wan and writer Leigh Whannell, it's presided over by opposing forces, and guided by sage elderly characters. In a conversation that's largely focused on death and my budding theory that Wan & Whannell find the elderly to be last vestiges of superstition, the writer tells FANGORIA of the film, their undervalued DEAD SILENCE and his preoccupation with our eventual end.Read more »
The fantastic, strange and unsettling HERE COMES THE DEVIL, from COLD SWEAT and PENUMBRA director Adrián García Bogliano, was picked up by Magnolia Pictures/Magnet Releasing out of Fantastic Fest 2012. With little word since, it's exciting to see the film finally has a release date.Read more »
Open Road Films, who released last year's fantastic THE GREY and the not-so-fantastic SILENT HILL: REVELATIONS has picked up the latest from Eli Roth, his first directorial effort in six years.Read more »
It seems crude to put it in such general, comparative terms, but I do suspect that DARK TOUCH will be the better rendition of "Carrie" we see this year. It's an unapologetic, dark Ireland-set film that re-envisions fundamentals of that teenaged, oncoming womanhood tale (telekinetic girl lashing out) and warps it to a more unsettling place. While hope is held for Kimberly Peirce's handling of the Stephen King novel, will the re-adaptation have the interest in being so stark and prodding as DARK TOUCH?Read more »
"Bad things happen everywhere," so says a character in Randy Moore's acclaimed, controversial ESCAPE FROM TOMORROW. Everywhere extends to the so-called Happiest Place on Earth where the manufactured magic of Disneyland—complete with iconic characters and rides—and the family-friendly company's recognizable title cards are utilized to frame a surreal, horrifying trip in the first trailer for the film.Read more »
This is the most anticipated television show of the season.Read more »
Music video affords so many possibilities. Most are performance-based, or utilize some sort of short form storytelling in reference to the song, but often the best are stream-of-consciousness images fueled by the fever of music and free from worrying about any bit of structure. Veteran director Diane Martel (of this year's massively popular "Blurred Lines" video for Robin Thicke and much more you'll recognize) has helmed the latest from Scottish band Franz Ferdinand and steeped it very much in shamelessly bloody madness. Titled, "Evil Eye," the video goes for a shot-on-video look and features dismemberment, necrophilia, a seeming Fulci nod, poolside instrument playing and a strange face attached to a dude's giant gut. Fun stuff!Read more »
Philadelphia-based The Awesome Fest is living the dream. As in, it's directors and organizers are capping off summer with a six film all nighter this Friday the 13th, and planning summer camp activities and encouraging BYOB and tents to boot.Read more »
Currently playing TIFF as part of the Vanguard Section, SAPI is the first horror film from Filipino filmmaker Brillante Ma Mendoza. A film about television producers hoping to document actual spiritual possession, SAPI looks to capture something rough.Read more »
Having long planned to pit his occult detective Harry D'Amour against possibly his most iconic creation Pinhead, the great Clive Barker has confirmed completion of that novel, THE SCARLET GOSPELS.Read more »
As we await word on OCULUS, the second film from ABSENTIA director Mike Flanagan, out of TIFF, his next nightmare-inducer has been revealed.Read more »

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Adequate, sturdy. Exactly what we expected from description
it was great! I have used it many times. Its great of old people or handicapped people.
As I am getting older, it is harder for the old body to twist and bend the right way anymore, and I needed something to help get my shoes on. This telescopic shoe horn fit the bill perfectly, it is not only adjustable, but is made of pretty firm material. I am very happy with it and would highly recommend it.
Works great. Very "tele-scoping" but easy to make rigid and fixed. Plastic with some aluminum, but quite sturdy. Be prepared for a very long, large box to arrive. And one is thinking, what is that? Hmmmm, I did not order golf clubs. Inside, lots of those plastic air pocket things for protection, and one's new shoe horn.
I gave this to my husband as a gift. He is pleased with the length of the shoe horn, but finds there is too much flexibility. He seems to have difficulty using it because of this lack of rigidity. The spoon is not big enough and the spring is not strong enough to easily assist in putting on his shoes. As for appearance, it looks very nice - just the kind of thing you'd like to have on display in your home entrance
Usually leaves our warehouse in 1-2 business days. *
Items must be returned in new or unused condition and contain all original materials included with the shipment. More details
FINAL SALE EXCLUSION: Items marked as FINAL SALE are not returnable unless the problem you experience is the result of our error.
Our standard shipping method is ground shipping. We also offer expedited shipping for most items. Your final shipping cost and estimated delivery timeframe will be available at checkout.
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* For your safety, some orders go through our loss prevention department. If we select your order for this process, we will immediately notify you that there may be up to a 3-day delay on your order.
** Most Oversize orders are delivered within 1-4 weeks. Some orders may take 6 weeks to be delivered.


text: The Streets Of Paris Silk Scarf by Amanda Jane's £44$83.94€57.93$73.66 In stock FREE mainland UK delivery - Order now and receive this item on or before Tuesday 2 December FREE mainland UK deliveryEstimated delivery: 2-5 days International delivery available colour story choicecolour story choice (required)Grey with blue tipsPink with blue tipsBlue with pink tipsGrey with purple tipscolour story choice (required)Grey with blue tipsPink with blue tipsBlue with pink tipsGrey with purple tipscolour story choice (required)Grey with blue tipsPink with blue tipsBlue with pink tipsGrey with purple tipscolour story choice (required)Grey with blue tipsPink with blue tipsBlue with pink tipsGrey with purple tips quantity by Amanda Jane's Location: Cumbria, GB read seller's profile see seller's complete range more items from this seller The Streets Of Paris Silk Scarf £44$83.94€57.93$73.66 Paris Streets Scene Les Petite Silk Scarf £28$53.42€36.86$46.88 Paris Catacombs Scene Silk Scarf £38$72.50€50.03$63.62 Paris Stamps With Love Silk Scarf £38$72.50€50.03$63.62 Paris Carousel Horse Silk Scarf £38$72.50€50.03$63.62 Paris Penny Farthing Silk Scarf £38$72.50€50.03$63.62 view all you may also like PRODUCT DETAILS CUSTOMER REVIEWS DELIVERY RETURNS Made in Britain gift wrapping available ask seller a question A beautifully handmade 100% silk scarf that is hand dyed, stitch and then screen printed with a vintage style Paris Street Scene! Made with such delicate tones of colour these scarves can go with almost anything on anyone, even complimenting some skins tones that are normally challenging. Suitable for both the ladies and gentleman to wear most of the year around as the 100% silk keeps you cosy but still allows your skin to breathe! No scarf ever comes out the same so you know you are getting something truely original and special! Each scarf is presented in a luxury gift box and presentation card as complimentary. All materials sourced within the UK and handmade in England! This design of scarf is available in four colours stories: Grey with blue tips. Pink with blue tips. Grey with purple tips. Each scarf is dipped dye with a main colour in the middle then dipped dyed at either end with another colour, blending as they meet. These colour choices can be seen in the photos that are in the order of above. Blue with pink tips. made from: This 100% Silk scarf should only be hand washed in lukewarm water. When ironing always start from a low heat setting to test on a small corner of the scarf, then adjust accordingly. dimensions: Approx. H150 X W40cm product code: 219353 1 customer review Mon 23 Dec 2013 No comment supplied. read about our product rating system ratings guide trusted independent customer feedbackforum excellent bad good poor This product is dispatched to you by Amanda Jane's. MAINLAND UK order now and receive this item on or before price Free Delivery Tuesday 2 December FREEFREEFREEFREE NON-MAINLAND UK estimated delivery price Highlands & Islands of Scotland 2-5 daysfrom dispatch £2.75$5.24€3.62$4.60 Northern Ireland 2-5 daysfrom dispatch £2.75$5.24€3.62$4.60 Other UK Islands 2-5 daysfrom dispatch £4.95$9.44€6.51$8.28 INTERNATIONAL DELIVERY estimated delivery price Western Europe 2-5 daysfrom dispatch £9.95$18.98€13.10$16.65 Rest of Europe 5-10 daysfrom dispatch £9.95$18.98€13.10$16.65 USA & Canada 5-10 daysfrom dispatch £17.95$34.24€23.63$30.05 Far East & Australasia 5-10 daysfrom dispatch £17.95$34.24€23.63$30.05 Rest of the world 5-14 daysfrom dispatch £17.95$34.24€23.63$30.05 You have 28 days, from receipt of cancellable goods, to notify the seller if you wish to cancel your order or exchange an item. Please note: goods that are personalised, bespoke or made-to-order to your specific requirements, perishable products and personal items sold with a hygiene seal (cosmetics, underwear) in instances where the seal is broken are non-refundable, unless faulty. Read more about our returns policy.Should you choose to return or exchange your order you will need to deliver the item(s) to the UK, where this seller is based. Amanda Jane's's return policy


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- Northern Illinois to get $8.3 million for state construction projects
- Tree-lighting festival kicks off holiday season in Machesney Park
- Roscoe Boy Scout Troop's tree stand at new location
- Tips for selecting safe toys for kids this holiday season
- Prayer service for World AIDS Day Nov. 30
- Food Bank joins national #GivingTuesday movement
- Lee Hamilton: What lies ahead for Congress
- Rockford Public Schools faces $8.8 deficit, board OKs flat tax, HR chief
- Literary Hook: A holiday tradition: 'This Thanksgiving, Remember'
- Cold snap does not negate global warming
Madison geek rockers bring interactive show to Freeport Friday, Aug. 13
By Jim Hagerty
Staff Writer
Billed as the KISS of geek rock and the Van Halen for nerds, Madison's Sunspot will bring its Major Arcana tour to Freeport's Jumpin' Joe's this Friday, Aug. 13.
Unlike a traditional rock show, Major Arcana includes videos, crowd interaction and quirky storyline, marking a rock opera that leans toward a tech-driven Rocky Horror Picture Show.
"It's like watching a movie with a band in front of it," vocalist and bassist Mike Huberty said.
The story follows the main character, Joe, an IT geek away from home for the first time after college. Missing his girlfriend, Joe immerses himself in personal relationships over the Internet, most notably with a web psychic. The songs, Huberty said, come from the emotional center of what's going on in the minds of each character.
"Sunspot brings the swagger of the stadium to the pocket-protected masses," Huberty said. "We've been on the road for several weeks bringing our show to the East Coast, South and Midwest. Our Freeport stop is the second-to-last night of the tour."
A three-piece act, Sunspot debuted the first half of Major Arcana at the SXSW festival in Austin, Texas, last year. It has since been developed into a 90-minute production.
Show starts at 9 p.m. Admission is free. Jumpin' Joe's is at 1100 W. Galena Ave. in Freeport. The club can be reached at (815) 235-6145. More information about Sunspot is at sunspotmusic.com.
From the Aug. 11-17, 2010 issue


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Donate glasses and change someone's life. If you own glasses you are no longer wearing and want them out of your junk drawer please bring them to Maplewood where they will be donated to the Lion's Club. It costs the Lion's Club less than $0.08 to provide a pair of recycled eyeglasses and change someone's life.
For children, clear vision means a better education, healthier development and a better quality of life. For adults, it means greater employment opportunity and economic strength. For seniors it means less dependence on others. Thank you.
On November 1st the winter dress code is implemented. Students are no longer able to wear shorts, flip flops, sandals, etc.
November 4. LICE-DOC-(2)-(1).pdf Lice-letter-in-Spanish.pdf
During these next months of chilly weather, you may feel inclined to drop off or pick up your student directly in front of the building. Unfortunately, our parking area at Maplewood is not ideal, and we must continue to follow the drop off and pick up routines by pulling as far forward to the West side of the building in order for this to run as smoothly as possible. Students can safely walk down the sidewalk back to the front doors of the building. In the afternoon during pick up, please do not park along the yellow curb when you enter the driveway of Maplewood. There is not enough space for two cars to pass each other when a car is parked along that curb line, which creates a hazard for us all. If possible, pull into a parking spot in the lot on the East side of the building and have your student meet you there, or consider coming 5-10 minutes later to avoid the heavy congestion that occurs right at 2:45pm. Any help you can provide in alleviating the headaches in our parking lot will be appreciated by all. The school district is working on a plan to try and redo the lot, but we all know that takes time and money. Until then, let's all do our part to keep our kids safe!
Thank you.
Officer Cook
PSLO Maplewood Middle School
Information for Parents and Guardians about:
English Language Arts standards
Curriculum & Instruction
Click Here
Maplewood referral slip
Click here to be redirected to our on-line forms
(anticipated absences, travel consent forms, etc).
Professional Learning Community
PBIS Corner
Cool Tools
Tardy Table
Hero in the Hall
Organization 2013
Goal Setting #1
Goal Setting #2
Maplewood Technology Club (MTC)
Scholarships and Contest
Welcome to Maplewood Middle School, Home of the Blue Jays. The middle school years are an exciting time in students' lives and our school reflects that in our learning activities, support programs and co-curricular activities.
Maplewood Middle School provides an array of academic courses like science, math, integrated language arts, and social studies, as well as, fine arts, world languages, and introductory courses to vocational education. Students are given the opportunity to expand their K-5 world language experience in Japanese, German or Spanish or try a new language altogether. Pride rallies are held approximately every two months to honor many students who have been involved in sports, school-sponsored contests, or just to create school spirit among the staff and students.
Maplewood Middle School is a great place to learn. We have a supportive community, involved parents, talented and committed staff, and the best students on the planet that surround us. We invite you to browse our website to find out what is happening at school.
Guidance Corner
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Our World History 5 Facts
(SRI, SAM, READ180 only available on district machines)


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Photos and Description of this 1 bedroom holiday apartment in Lamporecchio - POGGIO ALLA CAVALLA
Holiday Apartments POGGIO ALLA CAVALLA in the heart of Tuscany
POGGIO ALLA CAVALLA FARMHOLIDAYS is made up of 5 Apartments, and is surrounded by a large garden, where there is a swimming pool with a large panoramic terrace and a private car park.
THE APARTMENTS: ORANGE, BLUE, YELLOW, AND GREEN have the same surface sq m 40/45 with a doublebedroom, each of them can accommodate up to 4 people.
THE MASTER APARTMENT has sq m 110 with two doublebedrooms, can accommodate up to 6 people.
THE PRIVILEGED GEOGRAPHICAL LOCATION of the tourist farm makes it possible to go on convenient day trips to tourism and culture centres and Tuscan cities of art: Firenze, Siena, San Gimignano, Volterrra, Pisa, Lucca and the sea of Versilia, Pistoia, Arezzo e Cortona.
THE SWIMMING POOL with springboard (10x6 m - open from 24.04 to 04.10) equipped with a gazebo, shower, deck chairs, sun beds, parasols and tables.
Set on a splendid terrace solarium, where you can enjoy a wonderful view of the surrounding countryside.
Details for holiday apartment BLUE APARTMENT, 1 Bedroom, Sleeps 4, 1 Bathroom
- Suitability:
- Events Allowed
- Long-term Renters Welcome
- children welcome
- pets considered
- smoking allowed
- wheelchair inaccessible
About Massimo Camelli
The owner Massimo Camelli, follows the reception directly of the guests to the farmhouse and looks during their stay to make enrich their holiday with pleasant sensations, thanks to the beauty of the landscape and genuine hospitality.
The organic farm is run by owner, who directly follows the various phases of growing and producing wine and olive oil according to organic farming methods.
Massimo Camelli purchased this apartment in 2000
Why Massimo Camelli chose Lamporecchio
We recommend the 'Poggio alla Cavalla' Farmholiday, because is located in a quiet and well maintained place
Guests can spend a holiday rich in pleasurable sensations, thanks to the beautiful scenery and hospitality.
What makes this apartment unique
POGGIO ALLA CAVALLA farmholidays, is situated in Lamporecchio (Pistoia) in the heart of Tuscany, near Florence, between Montecatini Terme and Vinci, the birth town of Leonardo.
The privileged geographical location of the farmholidays makes it possible to go on convenient day trips to tourism and culture centres and Tuscan cities of art: Firenze, Siena, San Gimignano, Volterrra, Pisa, Pistoia, Arezzo e Cortona, Lucca and the sandy beaches of Versilia.


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Posted by: BHMACIN on Apr-28-13 10:49 AM (EST)
I've always wondered about that. I've noticed that in most boats. I've gotta think it's just an illusion though.
Gedi Convertible Helmet
2-3 Canoe/Kayak Trailer
Deck Rigging Gear
The Kayak Wing
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Water Tower Campus Move-in and Block Party
Loyola University is excited to welcome back students to the Water Tower Campus. There are many activities planned to get students safely moved in and acclimated to campus life.
We will be welcoming 400 students as they move into Raymond C. Baumhart Hall at 26 E. Pearson. Please take note of the following days and times during which the City has provided a permit to obstruct meters on the north side of Pearson Street. This will help us accommodate the families and students that will be unloading cars and moving trucks.
Move-in activities have moved to Pearson Street from Chestnut in previous years due to the construction taking place on Chestnut. The north side of Pearson between State and Wabash will be used for safe unloading of students and their vehicles for access to Baumhart Hall. As always, the Campus Safety officers will be on hand to direct traffic. Vehicles are given a 15-minute unloading window and are quickly directed to nearby parking garages.
The table below reflects days and times activities will take place.
Sunday, August 18:
1-26 E Pearson (northside only)
1-26 E Pearson (northside only)
Block Party:
The annual Fall Block Party is scheduled for Thursday, August 29, from 11:00 a.m. – 2:00 p.m. The event welcomes students, staff and faculty to the new school year. Loyola is permitted to close Pearson Street from State Street to Wabash Avenue and Wabash from Chicago Avenue to Chestnut Street from 6:00 a.m. until 4:00 p.m. Please plan accordingly.
I would like to thank you sincerely for your patience throughout the move-in and block party processes each year. If you should have any questions, comments or concerns during move-in, please feel free to speak to any staff member on site or call me directly at 773-508-7453 or at email@example.com.
More Featured Stories
Ignatian Heritage
Sobrino: We must speak the truth, defend the poorRethink what your responsibility is to the truth. That was the message Fr. Jon Sobrino, S.J., wanted to pass on Thursday night to an auditorium packed with Loyola students, faculty, staff, and others in the Chicago community.
Students shine at data conferenceTwo Quinlan teams, composed mostly of undergraduate students, took home two out of three awards at one of the nation's largest data analytics and technology conferences. "Our teams presented flawlessly," said Professor Nenad Jukić, who served as the coach.
Online exhibit commemorates martyrsThe History Department of Loyola University Chicago commemorates the 25th anniversary of the Jesuit martyrs of El Salvador through the launch of the public history project, "The Ellacuría Tapes: A Martyr at Loyola."
Hear Archbishop Cupich on Relevant RadioHear newly installed Archbishop Blase Cupich discuss his first week in Chicago on Relevant Radio, hosted by Fr. Greg Sakowicz of St. Mary Parish in Evanston and Wayne Magdziarz, Vice President and Chief of Staff for the President's Office at Loyola.
Ramblers name new athletics directorSteve Watson was named the new athletics director at Loyola on November 12. Watson, a former Division 1 student-athlete who played professional basketball overseas, comes to Loyola from St. Bonaventure University. "(Loyola) is a great school, in a great city, with great people," Watson said.
Student spotlight
Senior named 2014 Student LaureateLoyola senior David Lewis is a busy man. He's a member of numerous student groups and is also pursuing a double major, along with two minors. On November 1, Lewis added one more item to his growing resume: Loyola's 2014 Lincoln Academy of Illinois Student Laureate.
In the news
Dean Wheeler makes "most influential" listDean Darrell Wheeler, PhD, of the School of Social Work was recently named one of the 30 most influential social workers alive today. Selection was based on merit, scholastic study, and political activism.
In the news
Finding a solution to Chicago's food desertsQuinlan hosted a special event this fall on campus, bringing together leaders from the governmental, nonprofit, and corporate sectors to come up with solutions to food deserts in Chicago
Staff spotlight
Her mission: Help fellow veteransAnita Lumpkin, who served for 10 years in the Army Reserves before getting her master's degree in higher education administration, brings a unique background to her post as coordinator of Loyola's new office of Military Veteran Student Services.
He helps children, families live with spina bifidaLoyola psychology professor Grayson Holmbeck has been studying children with spina bifida for more than 20 years. In that time, he says: "We've learned a lot about what their problems and issues are, what we can do to help them, and more importantly, what they're capable of."
Adult Learners
FASTRACK your college degreeStarting in 2015, Loyola will offer several FASTRACK degree programs for adult learners at its Cuneo Mansion & Gardens in Vernon Hills. Courses will be on alternating Saturdays with an online component—perfect for anyone looking to balance work, life, and school.
Professor profile
Jukić named Faculty Member of the YearQuinlan Professor Nenad Jukić was named Loyola's Faculty Member of the Year on September 14 as part of the University's Faculty Convocation. This latest award caps off a string of impressive accolades for Jukić, who also was named Quinlan's Outstanding Undergraduate Teacher of the Year.
Helping others
Students receive prestigious Schweitzer FellowshipsFour Loyola graduate students were recently selected for the prestigious Albert Schweitzer Fellowship program and will spend the next year working on healthcare-related projects to help underserved communities in Chicago.
Phi Beta Kappa chapter puts University in elite companyLoyola is one of just 283 universities to have a Phi Beta Kappa chapter, a claim that only about 10 percent of the nation's colleges can make.
Loyola is No. 4 on list of greenest collegesLoyola is ranked No. 4 on the Sierra Club's 2014 list of the greenest colleges in America. The annual rankings are designed to spotlight universities that are deeply committed to environmental responsibility.
In the news
IC named one of the 'coolest' libraries in U.S.Loyola's Information Commons joins an elite group of peers on Business Insider's list of the "coolest" college libraries in the country.
Institute focuses on sustainable livingThe Institute of Environmental Sustainability combines academics and research with agriculture and community living—all in one facility.
Damen Center
Damen Center takes campus life to next levelThe Damen Center was designed from top to bottom with students in mind, making it the center of social life on Loyola's Lake Shore Campus.


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Council Committee Patch Plastic/Gauze Back. USED but in very good condition.
Explorer Crew Leader Patch on Blue Square. USED but in fair condition.
Journalist Lapel Pin. Excellent condition.
Organizer Lapel Pin. Excellent condition.
Neighborhood Commissioner Lapel Pin. Excellent condition.
District Commissioner Colar Brass. Two posts. Excellent condition.
National Staff Colar Brass. Excellent condition.
Patrol Leader Patch 1940s. Tan Square Material. Mint condition.
Junior Assistant Scoutmaster Patch 1950s Type 1. Mint condition.
District Commissioner Patch 1940s Fine Twill. USED but in excellent condition.
Employee Patch Computer Design Border. Mint condition.
Den Mother Pin. Mint condition.
Assistant Scoutmaster Lapel Pin. Horizontal Safety Pin. Mint condition.
Boatswain's Mate Patch Navy Blue on White. Plain back. Mint condition.
Crew Leader Patch Navy Blue on White. Plain back. Mint condition.
Crew Leader Patch Blue on White. Plain back. Mint condition.
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Best Schools in Central East Austin
Blackshear Elementary School
Public • Grades PK-5
236 students • 14 student/teacher
Kealing Middle School
Public • Grades 6-8
1215 students • 14 student/teacher
Garza Independence High School
Public • Grades 9-12
289 students • 13 student/teacher
Texas Empowerment Academy
Charter • Grades 5-12
121 students • 19 student/teacher
Which schools are assigned to my home?
About Central East Austin
Central East Austin contains 4 primary and secondary schools. One of the highest rated schools is Blackshear Elementary School, which has a rating of 9. The school has 236 students. Public schools in Central East Austin belong to Austin Independent School District or Texas Empowerment Academy. University of Texas Elementary Charter School is close and scores higher than Austin Independent School District. Nearby University of Texas University Charter School scores lower, with a rating of 2.
School Districts Near Central East Austin
- University of Texas Elementary Charter School
- University of Texas University Charter School
- Texas School for the Deaf
- The East Austin College Prep Academy
- Texas School for the Blind & Visually Impaired
Disclaimer: School attendance zone boundaries are supplied by Maponics and are subject to change. Check with the applicable school district prior to making a decision based on these boundaries.


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Britt Hayes
Britt Hayes is a writer, movie lover, and sensible sweater enthusiast living in Austin, Texas. She watches too much TV and never eats her vegetables. Her mom is worried about her.
With 'Godzilla' stomping into theaters next month, Legendary and Warner Bros. are helping us get better acquainted with director Gareth Edwards' vision for the monster via some interesting new featurettes that take you behind the scenes of this exciting new film. Where else are you going to see Bryan Cranston do his best impression of Godzilla's iconic roar?
August seriously cannot get here soon enough, and while we've still got plenty of summer blockbusters to get through before we get to James Gunn and Marvel's 'Guardians of the Galaxy,' we've got two more new pics from the upcoming movie to help us bide the time. Both images put the focus on Chris Pratt's Star-Lord, the leader of the rag-tag group of unlikely heroes.
Well that didn't take long -- 'Jem and the Holograms' has announced the four young actresses that will play the leading roles of Jem and her Hologram bandmates in the upcoming adaptation of the hit '80s cartoon, directed by 'G.I. Joe: Retaliation' helmer Jon M. Chu. And although Chu and the production team previously mentioned that casting was open to anyone -- even unknowns -- the four stars are all established young actresses.
Legendary debuted the first trailer (and poster!) for their upcoming horror flick 'As Above, So Below,' which takes found footage below ground and deep into the heart of the creepy catacombs of Paris. And while found footage has certainly been wearing out its welcome, the idea of a horror movie taking place in the catacombs is enough to pique our interest.
'Jurassic Park 4' (now officially titled 'Jurassic World') only just began filming in Hawaii, but life finds a way and Universal is already planning sequels to the upcoming, uh, sequel. (Please read previous sentence in your best Jeff Goldblum voice.) The studio has some pretty big plans for the upcoming film, and those plans involve setting the stage to continue the franchise, if they can.
Not to be outdone by 'Jem and the Holograms,' Barbie will also be getting her own live-action movie -- but of course homegirl has to one-up the competition, so she's getting a franchise because dream homes and matching pink cars aren't cheap, you know.
While we'll likely never see Spider-Man fighting side by side on the big screen with the Avengers, you can see all your favorite heroes teaming up in a different arena with Marvel Universe LIVE!, a new traveling production set to hit the road this summer. And now you can go behind the scenes with this new featurette on the upcoming production, which unites all your favorite Marvel heroes and villains for one spectacular show.
Hot on the heels of Mark Wahlberg receiving his achievement award at the 2014 MTV Movie Awards last night, Paramount unveiled the first official TV spot for 'Transformers 4,' ushering in a new era of 'Transformers' ... with Dinobots! We've also got a few new pics featuring Wahlberg striking some serious action poses.
Sony has plans for two more 'Amazing Spider-Man' movies, as well as spin-offs for villains, including Spidey's greatest foes: the Sinister Six. And while they are the most villainous of the villains, the producers say we can expect a slightly different take on their story when it comes time for their tale to be told in the 'Sinister Six' film, which is being helmed by director Drew Goddard.
We've seen tons of pictures of Dwayne Johnson as the legendary hero from director Brett Ratner's 'Hercules,' but now you can see the action star in, well, action! In this new clip from the upcoming film, see Johnson's Hercules in his fabled lion's hood (which does look pretty badass, we gotta say) preparing to take on a whole army of bad guys.
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Golden Tower Apartments
Official apartment prices and pictures. Check Availability!
107 S Spring St Ofc, Princeton, IN 47670
See Princeton Apartments near Golden Tower Apartments…
Vail Estates Apartments (more info)
Princeton, IN 47670
You just found your new home!
1 to 2 bedroom apartments from $625 to $715
Call: (866) 930-2135check availability
North Park Apartments (more info)
Evansville, IN 47710
Happiness isn't far away at all.
1 to 2 bedroom apartments from $530 to $683
Call: (812) 618-1628check availability
Arbors at Eastland Apartments (more info)
Evansville, IN 47715
A higher standard of living.
1 to 4 bedroom apartments from $581 to $845
Call: (866) 466-5963check availability
Brumfield Place Apartments (more info)
Princeton, IN 47670
Call: (812) 386-8200
Rainwood Apartments (more info)
Fort Branch, IN 47648
Call: (812) 753-3434
Sycamore Apartments (more info)
Petersburg, IN 47567
Call: (812) 354-2885
Ginger Hills Apartment (more info)
Winslow, IN 47598
Call: (812) 789-2067
Vincennes Niblack Apartments (more info)
Vincennes, IN 47591
Call: (812) 882-4968
Highland Hills Apartments (more info)
Evansville, IN 47710
Call: (812) 425-0629
Vincennes Park Apartments (more info)
Vincennes, IN 47591
Call: (812) 886-6339
See Official Princeton Apartments Prices and Pictures. Check Availability!
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I love you guys. You have a great selection with tons of pictures and updated rents. Thanks - I found the perfect place.
— Jasmine W.
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Official Living Guide
Did You Know?
The average Princeton resident moves 34 days after starting their Princeton rental search.
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Got questions? We're here to help. Get answers to all your apartment related questions…


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Product DetailsDimensions: 33w x 33h x 33d, Seat 15h, Arm 20.5h
Specifications: Materials: natural cherry, walnut, santos palisander, ebony or white ash; leather upholstery; chair has a 5-star black or white enameled base (white ash only) with polished aluminum trim; ottoman has a 4-star black or white enameled base (white ash only) with polished aluminum trim; urethane 6" foam padding; stainless steel glides.Shipping: These items are made to order and we estimate they will ship in 6 - 8 weeks, though that is ultimately up to Herman Miller. Because these pieces are custom-built, returns will not be accepted unless the product is flawed. We will charge your credit card 2 weeks after your order is placed, at which time ALL sales are final. Shipments to "lower 48" US states only.


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a graphical study…
exquisite work Richard
Excellent work
Great lines and tones!
And not a snail hole in sight! Nice work Richard
Fantastic lines and contrast Rick!! :)
Love the broken lines and compo…you sure have the eye…beautiful!
wow, very well processed! :)
Excellent work.
Wellllll.. how good is this & ALL in your Gallery! You works are all a fav to me.
Love the hosta. Your capture so perfect in B&W.
Absolutely stunning Richard


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Amanda's Blog
They let me carve ice with a CHAINSAW!
by Amanda
posted Feb 11 2013 12:57PM
Phew. I had quite the eventful weekend. Thursday I was sick. Like, can't breathe, feel like my head is going to explode sick. I got progressively better, but I still have a nasty cough today. Thankful for no flu though!
Friday I had plans to meet my mom and a few friends in Connecticut to see Bon Jovi's dress rehearsal concert for his world tour. The wicked storm that left 3-4 feet of snow in New England threatened to ruin that for us, but thankfull we arrived before the storm, and wound up camping out at Mohegan Sun for an extra day to wait out the weather and travel ban. Lucky for us, Bon Jovi ended up cancelling his show on Saturday night and instead played a free show for those who were stranded at the hotel because of the storm. HOW AWESOME! They put on a GREAT show and their new music actually brought my mom to tears. Some really great stuff.
I left CT at 4am on Sunday morning (they lifted the travel ban) to get back to PA in time to compete in the Celeb Charity Ice Sculpting Competition at CRYSTAL CABIN FEVER in Lakeville. Oh my gosh . . . I HAD SO MUCH FUN. I didn't actually think they were going to let me a.) hold a chainsaw b.) turn it on or c.) USE IT TO CARVE ICE, but they did and I had the best time. Tom Clark from WNEP was there as well as Tim Kelchner from WBRE. We were all competing for a $1,000 donation to our favorite local charity and I was there in support of Candy's Place. We were each paired up with a professional ice sculptor and had 2 hours to create something awesome (dino themed) from our block of ice. Neil was my partner and he was awesome. He explained how everything works, how it should look, and how to use all of the tools. He actually let me do a lot of the work and I think I found a new hobby - I had so much fun and check this out . . . WE WON!!!!!!
If you haven't checked out the ice displays at Crystal Cabin Fever, definitely get there soon as a winter activity. It was SO much fun!
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