Deep intra‐island divergence of a montane forest endemic: phylogeography of the Puerto Rican frog Eleutherodactylus portoricensis (Anura: Eleutherodactylidae)

Aim Hypotheses proposed for lineage diversification of tropical montane species have rarely been tested within oceanic islands. Our goal was to understand how basin barriers and Pleistocene climatic fluctuations shaped the distribution of diversity in Eleutherodactylus portoricensis (Eleutherodactylidae), a frog endemic to the montane rain forests of Puerto Rico. Location The north‐eastern (Luquillo) and south‐eastern (Cayey) mountains of Puerto Rico. Methods We generated mitochondrial DNA (mtDNA) control region sequences (c. 565 bp) from 144 individuals of E. portoricensis representing 16 localities, and sequenced 646 bp of cytochrome b and 596 bp of nuclear DNA (nDNA) rhodopsin exon and intron 1 from a subset of individuals. We conducted a phylogenetic analysis on the mtDNA sequence data and explored population substructure with maximum parsimony networks, a spatial analysis of molecular variance, and pairwise F_ST analysis. Coalescent simulations were performed to test alternative models of population divergence in response to late Pleistocene interglacial periods. Historical demography was assessed through coalescent analyses and Bayesian skyline plots. Results We found: (1) two highly divergent groups associated with the disjunct Luquillo and Cayey Mountains, respectively; (2) a shallow mtDNA genetic discontinuity across the La Plata Basin within the Cayey Mountains; (3) phylogeographic congruence between nDNA and mtDNA markers; (4) divergence dates for both mtDNA and nDNA pre‐dating the Holocene interglacial (c. 10 ka), and nDNA suggesting divergence in the penultimate interglacial (c. 245 ka); and (5) historical demographic stability in both lineages. Main conclusions The low‐elevation Caguas Basin is a long‐term barrier to gene flow between the two montane frog populations. Measures of genetic diversity for mtDNA were similar in both lineages, but lower nDNA diversity in the Luquillo Mountains lineage suggests infrequent dispersal between the two mountain ranges and colonization by a low‐diversity founder population. Population divergence began prior to the Holocene interglacial. Stable population sizes over time indicate a lack of demonstrable demographic response to climatic changes during the last glacial period. This study highlights the importance of topographic complexity in promoting within‐island vicariant speciation in the Greater Antilles, and indicates long‐term persistence and lineage diversification despite late Pleistocene climatic oscillations.     

Metazoan parasites of sticklebacks on Sable Island, Northwest Atlantic Ocean: biogeographic considerations

Three species of sticklebacks ( Apeltes quadracus, Gasterosteus aculeatus , and Pungitius pungitius ( n = 236) were collected from five ponds on Sable Island. The nematodes Pseudoterranova decipiens, Contracaecum sp., Paracuaria adunca , and Cosmucephalus obvelatus , and the cestode Diphyllobothrium ditremum parasitized three-spined sticklebacks ( G. aculeatus ) and four-spined sticklebacks ( A. quadracus ) inhabiting four brackish water ponds. All the parasites except P. decipiens infected nine-spined sticklebacks ( P. pungitius ) from a freshwater pond. In addition, the cestode Schistocephalus pungitii , the copepod Thersitina gasterostei , and the monogenean Gyrodactylus canadensis occurred in nine-spined sticklebacks from the freshwater pond. The two cestodes, the copepod, and the sealworm, P. decipiens , were the most common parasites encountered. The remaining helminths were relatively rare. Most of the parasite species were larval forms which use gulls or seals as definitive hosts. These parasites probably colonized Sable Island with their definitive hosts, whereas only two species ( T. gasterostei and G. canadensis ) successfully colonized the island ponds with their fish hosts. The low parasite species richness encountered is attributed to the impoverished nature of the host fauna of Sable island, and the difficulty of colonization as a result of the island's isolation with respect to the mainland.     

Novel summary metrics for insular biotic assemblages based on taxonomy and phylogeny: Biogeographical,palaeogeographical and possible conservational applications

Using the land‐bound vertebrates on the marine islands as model organisms, two metrics are presented that permit quantitative and succinct synopses of the ‘evolutionary maturity’ of the hosted faunal assemblages. In turn, these reflect the geo‐physical settings and geological developments of the substrates. The assemblage lineage‐taxonomy spectrum (ALTS) is based on the constituent lineages’ taxonomic distinctiveness and diversity. Individual lineages within assemblages can in most cases be assigned to one of six categories, LT1?LT6: LT1 is a non‐endemic taxon, whereas LT6 comprises multiple endemic genera from a family that arose elsewhere. If required, the scheme can be expanded: LT9 is an endemic order. The data can then be combined to provide an assemblage spectrum, for example, 00:08:38:30:08:15[ 13 ]. Here, the first six values denote the number of lineages assigned to each category expressed as percentages of the overall total, with the sum of the processed lineages listed as the seventh (in brackets and bold). The ALTS metric highlights efficiently the key features of a marine island's biological assemblage. Notably, the contrast between spectra for suites on geologically and geo‐physically varied island types can be striking, for instance the squamate suite on the young, proximate orogenic margin island of Taiwan is coded 78:16:05:00:00:00[ 37 ] whereas the one on the distantly located, Late Eocene composite terrane island of New Caledonia is 00:11:00:11:33:44[ 9 ]. To overcome the subjectivity that is inherent in assigning supraspecific ranks, an alternative assemblage lineage‐age spectrum (ALAS) is also introduced that makes use of the binary logarithm values of the colonization times of the island lineages (0–2, 2–4, … , 32–64, >64 Ma). It is represented using a seven‐plus‐two‐number code, for instance Madagascar's squamates are 00:06:00:00:19:62:12[ 19 ( 16 )]; most colonizations took place in the Palaeogene (66–23 Ma); there are 19 lineages, but only 16 are presently age‐dated. In addition to marine‐island biogeography studies, the ALTS–ALAS spectrum approach is potentially useful for encapsulating biotas in other sorts of insular setting (e.g. lakes, mountain tops), and for evaluating palaeogeographical models. Furthermore, it may help emphasize the conservation value of an island's faunal assemblage.     

Evaluation of nutrients and related environmental factors for wolfberry (Lycium barbarum) fruits grown in the different areas of China

The wolfberry (Lycium barbarumL.) fruit has been used in traditional Chinese medicine and food, but fruits grown in various geographic areas are significantly different in quality and content of functional components. In this study, we tested the internal nutritional ingredients and related environmental factors using forty-two wolfberry fruits collected from seven wolfberry producing regions in China (Yinchuan and Zhongning in Ningxia, Jingyuan in Gansu, Nuomuhong Farm in Qinghai,Hangjinhouqi and Wulateqianqi in Inner Mongolia, and Jinghe in Xinjiang). The nutritional ingredients of the forty-two wolfberry fruits are dependent on the environment. Correlation analysis revealed that the amount of nutrients was highly related to each other. Based on the analysis ofratio of total sugar to betaine (sugar-alkali ratio), we have successfully established quality standards for the wolfberry based on the levels of betaine and sugar. The standards were as follows: high betaine low sugar (sugar-alkali ratio≤15.1), high betaine medium sugar (15<sugar-alkali ratio≤60), medium betaine medium sugar (60<sugar-alkali ratio≤121), medium betaine high sugar (121<sugar-alkali ratio≤165), and low betaine high sugar (sugar-alkali ratio>165). Zhongning of Ningxia is the geo-authentic producing region of the wolfberry, whereby the wolfberries are rich in main pharmacological ingredients Lycium barbarum polysaccharide (LBP) and three trace elements (iron, manganese and selenium) and medium sugar-alkali ratio. In addition, the correlation analysis showed that both the altitude and average temperature difference between day and night had positive effects on the wolfberry flavonoids content but the annual sunshine duration negatively affected the betaine contents of the wolfberry fruit.     

MV-mimicking micelles loaded with PEG-serine-ACP nanoparticles to achieve biomimetic intra/extra fibrillar mineralization of collagen in vitro

Since their discovery, matrix vesicles (MVs) containing minerals have received considerable attention for their role in the mineralization of bone, dentin and calcified cartilage. Additionally, MVs' association with collagen fibrils, which serve as the scaffold for calcification in the organic matrix, has been repeatedly highlighted. The primary purpose of the present study was to establish a MVs–mimicking model (PEG-S-ACP/micelle) in vitro for studying the exact mechanism of MVs-mediated extra/intra fibrillar mineralization of collagen in vivo. In this study, high-concentration serine was used to stabilize the amorphous calcium phosphate (S-ACP), which was subsequently mixed with polyethylene glycol (PEG) to form PEG-S-ACP nanoparticles. The nanoparticles were loaded in the polysorbate 80 micelle through a micelle self-assembly process in an aqueous environment. This MVs–mimicking model is referred to as the PEG-S-ACP/micelle model. By adjusting the pH and surface tension of the PEG-S-ACP/micelle, two forms of minerals (crystalline mineral nodules and ACP nanoparticles) were released to achieve the extrafibrillar and intrafibrillar mineralization, respectively. This in vitro mineralization process reproduced the mineral nodules mediating in vivo extrafibrillar mineralization and provided key insights into a possible mechanism of biomineralization by which in vivo intrafibrillar mineralization could be induced by ACP nanoparticles released from MVs. Also, the PEG-S-ACP/micelle model provides a promising methodology to prepare mineralized collagen scaffolds for repairing bone defects in bone tissue engineering.