Genetic differentiation reflects geological history in the Azorean land snail,Leptaxis azorica

The land snail Leptaxis azorica, endemic on the Azores, was subjected to an electrophoretic (allozymes) and morphometric (genital tract) analysis. Genetic distances suggest the presence of four distinct lineages and are compatible with colonisation proceeding from the eastern, older islands (Santa Maria and S?o Miguel) to the west (Flores and Corvo). On S?o Miguel, genetic and morphometric differentiation is concordant with the separate colonisation of two islands that gave rise to the current island 50,000 years ago. The maximum time available for differentiation in isolation (0.55 million years) suggests a high rate of allozyme change between the two lineages on S?o Miguel. This may be related to population isolation and bottlenecks caused by human and volcanic activity on S?o Miguel in relatively recent times. This is more prominent in the eastern region where populations are also characterised by reduced genetic variation (loss of alleles and heterozygosity) compared to populations elsewhere.     

The genus Dryopteris in the Canary Islands and Azores and the description of two new tetraploid species

From studies in the field, botanic garden and herbarium we concluded that the following four species of Dryopteris grow on the Canary Islands: D. aemula (n = 41) so far found only on Gomera; D. pseudomas (n = 82, apogamous) similarly recorded only for Gomera; D. oligodonta (n = 41) the most common representative of the genus in the Islands is possibly an old Canary endemic related to some African species; D. guanchica a new tetraploid species ( n = 82) hitherto confused with D. austriaca (D. dilatata). On four islands of the Azores the following were found: D. aemula (n = 41); D. azorica (n = 41); D. pseudomas (n = 82, apogamous); and D. crispifolia a new tetraploid species ( n = 82) which is very distinct when living but in the dried pressed state is difficult to differentiate from D. austriaca and D. guanchica.     

Mitochondrial DNA phylogenetic analysis of the genus Laparocerus (Coleoptera, Curculionidae, Entiminae). I. The Madeiran clade

Laparocerus are plant‐chewing flightless weevils distributed on oceanic islands in Macaronesia, with a single species in Morocco. The genus has a complicated taxonomic history with several subgenera described. To assist in a taxonomical revision of the group, a molecular study was undertaken. In this first contribution, the species from the Azores and Madeira archipelagos are studied together with representatives of subgenera from the Canary Islands and the single known continental species (46 OTUs). Phylogenetic analyses are based on sequence data from mitochondrial cytochrome oxidase II (COII) and the ribosomal 16S ribosomal RNA (16S rRNA) genes (combined data set 1023 bp), with additional data from the nuclear elongation factor 1α (EF‐1α) for some selected species. Maximum likelihood (ML) and Bayesian analyses show that all Madeiran species are monophyletic and form a monophyletic group with the Afro‐Canarian samples. Species of the genus Lichenophagus appear within the Madeiran and Canarian Laparocerus clades, but separated in accordance with their respective island origin. Thus, Lichenophagus is here restricted to Madeiran species and proposed as subgenus (status novo) of Laparocerus. Conversely, the Laparocerus subgenus Drouetius from the Azores is revealed to be a separate and distant outgroup, in agreement with its morphological distinctiveness, deserving an independent genus status. Internal relationships within the Madeiran clade are discussed and compared with morphologically defined species groups. The Madeiran monotypic subgenus Cyphoscelis is not supported by the genetic data and synonymized (nov. syn.) with Laparocerus. Subgenera Laparocerus and Atlantis prove to be polyphyletic. Consequently a restriction to monophyletic and morphologically congruent clades is proposed. A cryptic species vicariant of Laparocerus morio was detected and recognized as L. chaoensis, status novo. Other cases of discrepancy between the genetic results and the traditional taxonomy are discussed in detail in the light of mitochondrial introgression, incomplete lineage sorting or poor taxonomy hypotheses.     

Variation in the genitalia of the land snail Heterostoma paupercula (Lowe, 1831) (Helicidae) in Madeira

The small helicid snail Heterostoma paupercula occurs in the Azores and Madeira. Previously regarded as a single species, it was split into two genera on the basis of variation in shell morphology and genital anatomy: Heterostoma having a toothed shell and being hemiphallic, while Steenbergia had a toothless shell and was euphallic. This division was first questioned when toothed shells {Heterostoma) in the Azores were found not to be hemiphallic. In this study the genitalia of 361 specimens from islands in the Madeira archipelago were examined. Four components of the genitalia were measured (penis/epiphallus, flagellum, bursa stalk and bursa) for 158 individuals. Hemiphally was found to be rare, occurring in only a proportion of the populations at two localities and it did not correspond with the expected shell morphology–the shells were untoothed (= Steenbergia) , which is the opposite of that previously described.
Variation in the genitalia was geographically based: Madeira and its nearest neighbour islands forming one group and the Porto Santo islands another. There is no evidence to suggest that the hemiphallic animals are more than an intraspecific form. It is suggested that the variation between the two island groups is reflective of allopatric differentiation. Without für ther evidence of divergence, H. paupercula should be regarded as the sole species throughout the range and the genus Steenbergia should be suppressed.     

Of mice and the ‘Age of Discovery’: the complex history of colonization of the Azorean archipelago by the house mouse (Mus musculus) as revealed by mitochondrial DNA variation

Humans have introduced many species onto remote oceanic islands. The house mouse (Mus musculus) is a human commensal and has consequently been transported to oceanic islands around the globe as an accidental stowaway. The history of these introductions can tell us not only about the mice themselves but also about the people that transported them. Following a phylogeographic approach, we used mitochondrial D‐loop sequence variation (within an 849‐ to 864‐bp fragment) to study house mouse colonization of the Azores. A total of 239 sequences were obtained from all nine islands, and interpretation was helped by previously published Iberian sequences and 66 newly generated Spanish sequences. A Bayesian analysis revealed presence in the Azores of most of the D‐loop clades previously described in the domesticus subspecies of the house mouse, suggesting a complex colonization history of the archipelago as a whole from multiple geographical origins, but much less heterogeneity (often single colonization?) within islands. The expected historical link with mainland Portugal was reflected in the pattern of D‐loop variation of some of the islands but not all. A more unexpected association with a distant North European source area was also detected in three islands, possibly reflecting human contact with the Azores prior to the 15th century discovery by Portuguese mariners. Widening the scope to colonization of the Macaronesian islands as a whole, human linkages between the Azores, Madeira, the Canaries, Portugal and Spain were revealed through the sharing of mouse sequences between these areas. From these and other data, we suggest mouse studies may help resolve historical uncertainties relating to the ‘Age of Discovery’.     

On the taxonomy of recent and fossil species of Laurus (Lauraceae)

A detailed comparison of the vegetative and reproductive structures in Laurus nobilis and L. azorica is presented. Most of the characters used by authors to differentiate the two species show a marked degree of overlap. The only characters for which absolute differences could be demonstrated were the leaf tomentum and chromosome number. The fossil record is discussed. None of the taxa referred to Laurus can be accepted as representatives of the genus. However, an examination made of leaf-remains known as Laurophyllum abchasicum established that these belong to the genus Laurus.     

Diversification of the forest beetle genus Tarphius on the Canary Islands, and the evolutionary origins of island endemics

The flightless beetle genus Tarphius Erichson (Coleoptera: Colydiidae) is a distinctive element of the beetle fauna of the Canary Islands with 29 species distributed across the five western islands. The majority of Tarphius species are rare and intimately associated with the monteverde forest and only two species occur on more than one island. In this study we investigate the phylogeography of the Canary Island Tarphius, and their relationship to Tarphius from the more northerly archipelagos of Madeira and the Azores using maximum parsimony and Bayesian inference analysis of mitochondrial cytochrome oxidase I and II sequence data. We use geological datings for the Canary Islands, Azores, and Madeira to calibrate specific nodes of the tree for the estimation of divergence times using a penalized likelihood method. Data suggest that the Canary Island species assemblage is of some antiquity, however, much of this species diversity is relatively recent in origin. The phylogenetic relationships of species inhabiting the younger islands of El Hierro and La Palma indicate that colonization events between islands have probably been a significant factor in the evolutionary history of the Canary Island species assemblage. A comparison of molecular phylogenetic studies of arthropods on the Canary Islands suggests that, in the evolution of the arthropod species community of an island, the origin of endemic species is initially the result of colonizing lineages differentiating from their source populations. However, as an island matures a greater proportion of endemic species originate from intra-island speciation.     

Drivers of diversity in Macaronesian spiders and the role of species extinctions

Aim To identify the biogeographical factors underlying spider species richness in the Macaronesian region and assess the importance of species extinctions in shaping the current diversity. Location The European archipelagos of Macaronesia with an emphasis on the Azores and Canary Islands. Methods Seven variables were tested as predictors of single‐island endemics (SIE), archipelago endemics and indigenous spider species richness in the Azores, Canary Islands and Macaronesia as a whole: island area; geological age; maximum elevation; distance from mainland; distance from the closest island; distance from an older island; and natural forest area remaining per island – a measure of deforestation (the latter only in the Azores). Different mathematical formulations of the general dynamic model of oceanic island biogeography (GDM) were also tested. Results Island area and the proportion of remaining natural forest were the best predictors of species richness in the Azores. In the Canary Islands, area alone did not explain the richness of spiders. However, a hump‐shaped relationship between richness and time was apparent in these islands. The island richness in Macaronesia was correlated with island area, geological age, maximum elevation and distance to mainland. Main conclusions In Macaronesia as a whole, area, island age, the large distance that separates the Azores from the mainland, and the recent disappearance of native habitats with subsequent unrecorded extinctions seem to be the most probable explanations for the current observed richness. In the Canary Islands, the GDM model is strongly supported by many genera that radiated early, reached a peak at intermediate island ages, and have gone extinct on older, eroded islands. In the Azores, the unrecorded extinctions of many species in the oldest, most disturbed islands seem to be one of the main drivers of the current richness patterns. Spiders, the most important terrestrial predators on these islands, may be acting as early indicators for the future disappearance of other insular taxa.     

Isozyme differentiation in the endemic genusArgyranthemum (Asteraceae: Anthemideae) in the Macaronesian Islands

The genusArgyranthemum (Asteraceae: Anthemideae) comprises 38 taxa and is restricted to the archipelagos of the Canaries, Selvagens, Madeira, and Desertas in the Macaronesian biogeographic region. An electrophoretic study, including 17 enzyme loci and at least one population of each of the described taxa, was carried out. High identity (low distance) values between taxa (mean of 0.893) were obtained despite the old age of the islands, their close proximity to the African continent, and the fact thatArgyranthemum is the most species-rich and variable genus in Macaronesia. These results suggest that the genus is monophyletic and that it has evolved very rapidly in these islands. There is little correspondence between taxonomy and neighbor-joining analysis based on Roger's genetic distances, but in several instances populations from the same islands cluster together despite being from different species or even different sections. It is suggested that repeated genetic bottlenecks associated with the founding of new populations during radiation of the genus resulted in lineage sorting of ancestral allozyme polymorphisms. Because every population has a high average identity with all other populations, lineage sorting could result in populations of different taxa being slightly more similar than populations of the same taxon. Gene flow between different species on the same island could account for some populations clustering by island of origin rather than taxonomic disposition. Average allozyme diversity within populations (0.098) is 50% higher than the mean total diversity for species endemic to oceanic islands.     

Hybridization and speciation in the genusDryopteris (Pteridophyta:Dryopteridaceae) on Pico Island in the Azores

The cytology of wild hybrids ofDryopteris from Pico, Azores has been examined. This has confirmed the parentage of the hybrids, and provided evidence to support the view that the tetraploid speciesD. crispifolia has evolved in the Azores from two diploid species,D. aemula andD. azorica, by hybridization and polyploidy.     

A molecular phylogenetic study of the genus Ribeiroia (Digenea): trematodes known to cause limb malformations in amphibians

Species of Ribeiroia (Trematoda: Psilostomidae) are known to cause severe limb malformations and elevated mortality in amphibians. However, little is known regarding the number of species in this genus or its relation to other taxa. Species of Ribeiroia have historically been differentiated by slight differences among their larval stages. To better understand the systematics and biogeography of this genus and their potential relevance to the distribution of malformed amphibians, specimens identified as Ribeiroia were collected across much of the known range, including samples from 5 states in the United States (8 sites) and 2 islands in the Caribbean (Puerto Rico and Guadeloupe). A cercaria from East Africa identified as Cercaria lileta (Fain, 1953), with attributes suggestive of Ribeiroia (possibly R. congolensis), was also examined. The intertranscribed spacer region 2 (ITS-2) of the ribosomal gene complex was sequenced and found to consist of 429 nucleotides (nt) for R. ondatrae (United States) and 427 nt for R. marini (Caribbean), with only 6 base differences noted between the 2 species. The ITS-2 region of C. lileta (429 nt) aligned closely with those of the 2 other Ribeiroia species in a phylogenetic analysis that included related trematode genera. This evidence suggests that a third Ribeiroia species exists in tropical Africa. Variation in ITS-2 within R. ondatrae was nonexistent among the 8 populations from North America. Our study further suggests that Ribeiroia spp. originally parasitized Biomphalaria sp., and that a host switch to a closely related snail, Helisoma sp., may have occurred in the lineage represented by R. ondatrae. However, relationships within the Echinostomatidae are not understood well enough to make any robust conclusions at this time.     

Environmental factors driving arbuscular mycorrhizal fungal communities associated with endemic woody plant Picconiaazorica on native forest of Azores

Arbuscular mycorrhizal fungi (AMF) play important key roles in the soil ecosystems as they link plants to the root-inaccessible part of soil. The aims of this study were to investigate which environmental factors influence the spatial and temporal structuring of AMF communities associated to Picconia azorica in two Azorean islands (Terceira and São Miguel islands), and investigate the seasonal variation in AMF communities between the two islands. Communities of AMF associated with P. azorica in native forest of two Azorean islands (Terceira and São Miguel) were characterised by spore morphology or molecular analysis. Forty-five AMF spore morphotypes were detected from the four fragments of P. azorica forest representing nine families of AMF. Acaulosporaceae (14) and Glomeraceae (9) were the most abundant families. AMF density and root colonisation varied significantly between islands and sampling sites. Root colonisation and spore density exhibited temporal patterns, which peaked in spring and were higher in Terceira than in São Miguel. The relative contribution of environmental factors showed that factors such as elevation, relative air humidity, soil pH, and soil available P, K, and Mg influenced AMF spore production and root colonisation. Different sporulation patterns exhibited by the members of the commonest families suggested different life strategies. Adaptation to a particular climatic and soil condition and host phenology may explain seasonal differences in sporulation patterns. Cohorts of AMF associated to P. azorica are shaped by regional processes including environmental filters such as soil properties and natural disturbance.     

Phylogeography and molecular phylogeny of Macaronesian island Tarphius (Coleoptera: Zopheridae): why are there so few species in the Azores?

Aim We used a phylogenetic framework to examine island colonization and predictions pertaining to differentiation within Macaronesian Tarphius (Insecta, Coleoptera, Zopheridae), and explain the paucity of endemics in the Azores compared with other Macaronesian archipelagos. Specifically, we test whether low diversity in the Azores could be due to recent colonization (phylogenetic lineage youth), cryptic speciation (distinct phylogenetic entities within species) or the young geological age of the archipelago. Location Macaronesian archipelagos (Azores, Madeira and the Canary Islands), northern Portugal and Morocco. Methods Phylogenetic analyses of mitochondrial and nuclear genes of Tarphius beetles of the Azores, other Macaronesian islands and neighbouring continental areas were used to investigate the origin of island biodiversity and to compare patterns of colonization and differentiation. A comparative nucleotide substitution rate test was used to select the appropriate substitution rate to infer clade divergence times. Results Madeiran and Canarian Tarphius species were found to be more closely related to each other, while Azorean taxa grouped separately. Azorean taxa showed concordance between species and phylogenetic clades, except for species that occur on multiple islands, which segregated by island of origin. Divergence time estimates revealed that Azorean Tarphius are an old group and that the most recent intra‐island speciation event on Santa Maria, the oldest island, occurred between 3.7 and 6.1 Ma. Main conclusions Our phylogenetic approach provides new evidence to understand the impoverishment of Azorean endemics: (1) Tarphius have had a long evolutionary history within the Azores, which does not support the hypothesis of fewer radiation events due to recent colonization; (2) the current taxonomy of Azorean Tarphius does not reflect common ancestry and cryptic speciation is responsible for the underestimation of endemics; (3) intra‐island differentiation in the Azores was found only in the oldest island, supporting the idea that young geological age of the archipelago limits the number of endemics; and (4) the lack of evidence for recent intra‐island diversification in Santa Maria could also explain the paucity of Azorean endemics. Phylogenetic reconstructions of other species‐rich taxa that occur on multiple Macaronesian archipelagos will reveal whether our conclusions are taxon specific, or of a more general nature.     

Helminths of the wild rabbit (Oryctolagus cuniculus) in Macaronesia

Two hundred and four rabbits from 8 Macaronesian islands (Pico, San Jorge, San Miguel, Terceira, and Flores from Azores Archipelago; Tenerife and Alegranza from Canary Islands; and Madeira from Madeira Archipelago) were examined for helminth parasites between 1995 and 2000. Three species of cestodes, Taenia pisiformis (larvae), Andrya cuniculi, and Mosgovoyia ctenoides, and 5 species of nematodes, Trichuris leporis, Graphidium strigosum, Trichostrongylus retortaeformis, Passalurus ambiguus, and Dermatoxys hispaniensis, were identified. Only 3 species (M. ctenoides, T. retortaeformis, and P. ambiguus) were regularly distributed over the 3 archipelagos. Taenia pisiformis was not collected in Madeira, nor was A. cuniculi in the Azores and G. strigosum in the Canary Islands. Trichuris leporis and D. hispaniensis were only found in Madeira. Significant differences in the general prevalence of the nematodes G. strigosum and T. retortaeformis were detected between Azores and Madeira. The prevalence of T. retortaeformis differs significantly between the Azores and the Canaries and that of P. ambiguus was higher in Madeira than in Azores and Canaries. The helminth richness found in the wild rabbit in these Macaronesian archipelagos was very low compared with the Palearctic helminth fauna of this host. The wild rabbit was introduced from the Iberian Peninsula into different Macaronesian islands. Helminths introduced with Oryctolagus cuniculus into these islands also are commonly found in Iberian wild rabbits, which are excellent colonizers, as demonstrated in this study.     

Spider biodiversity patterns and their conservation in the Azorean archipelago,with descriptions of new species

Abstract

In this contribution, we report on patterns of spider species diversity of the Azores, based on recently standardised sampling protocols in different habitats of this geologically young and isolated volcanic archipelago. A total of 122 species is investigated, including eight new species, eight new records for the Azorean islands and 61 previously known species, with 131 new records for individual islands. Biodiversity patterns are investigated, namely patterns of range size distribution for endemics and non‐endemics, habitat distribution patterns, island similarity in species composition and the estimation of species richness for the Azores. Newly described species are: Oonopidae ‐ Orchestina furcillata Wunderlich; Linyphiidae: Linyphiinae ‐ Porrhomma borgesi Wunderlich; Turinyphia cavernicola Wunderlich; Linyphiidae: Micronetinae ‐Agyneta depigmentata Wunderlich; Linyphiidae: Erigoninae ‐ Acorigone gen. nov. with its type species Acorigone zebraneus Wunderlich; Clubionidae ‐ Cheiracanthium floresense Wunderlich; Cheiracanthium jorgeense Wunderlich; Salticidae ‐ Neon acoreensis Wunderlich. Other major taxonomic changes are: Diplocentria acoreensis Wunderlich, 1992 (Linyphiidae) is transferred to Acorigone (comb. nov.), Leucognatha Wunderlich 1995 (Tetragnathidae) is not an endemic genus of the Azores but an African taxon and synonymous with Sancus Tullgren, 1910; Leucognatha acoreensis Wunderlich, 1992 is transferred to Sancus. Minicia picoensis Wunderlich, 1992 is a synonym of M. floresensis Wunderlich, 1992. For each species additional information is presented about its known distribution in the islands, its colonisation status, habitat occurrence and biogeographical origin.     

Systematics of the genus Capra inferred from mitochondrial DNA sequence data

Traditional classification in the genus Capra is based mainly on horn morphology. However, previous investigations based on allozyme data are not consistent with this classification. We thus reexamined the evolutionary history of the genus by analyzing mitochondrial DNA (mtDNA) sequence variation. We collected bone samples from museums or dead animals found in the field. Thirty-four individuals were successfully sequenced for a portion of the mtDNA cytochrome b gene and control region (500 bp in total). We obtained a star-like phylogeny supporting a rapid radiation of the genus. In accordance with traditional classification, mtDNA data support the presence of two clades in the Caucasus and the hypothesis of a domestication event in the Fertile Crescent. However, in conflict with morphology, we found that C. aegagrus and C. ibex are polyphyletic species, and we propose a new scenario for Capra immigration into Europe.     

Tracking macroalgae introductions in North Atlantic oceanic islands

The Azores archipelago was selected as a case study since there are few studies on macroalgae introduction in oceanic islands. While at a global scale, around 3 % of macroalgae are considered non-indigenous; in the remote oceanic islands of the Azores, over 6 % of the marine algal flora is non-indigenous. The taxa distribution pattern of non-indigenous species in the Azores is significantly different from the distribution pattern in the globe. The most representative group was Rhodophyta species, being 84 % of the total non-indigenous macroalgae, mainly introduced via maritime traffic. This study highlights the vulnerability of remote islands to the introduction of macroalgae and the need to develop further studies on other archipelagos to understand whether the observed vulnerability is generally characteristic of oceanic islands. The development of local monitoring and mitigation programs and the necessity of regulatory and preventive measures for the maritime traffic vector are strongly suggested.     

Genetic structure of house mouse (Mus musculus Linnaeus 1758) populations in the Atlantic archipelago of the Azores: colonization and dispersal

We analyzed the genetic structure and relationships of house mouse (Mus musculus) populations in the remote Atlantic archipelago of the Azores using nuclear sequences and microsatellites. We typed Btk and Zfy2 to confirm that the subspecies Mus musculus domesticus was the predominant genome in the archipelago. Nineteen microsatellite loci (one per autosome) were typed in a total of 380 individuals from all nine Azorean islands, the neighbouring Madeiran archipelago (Madeira and Porto Santo islands), and mainland Portugal. Levels of heterozygosity were high on the islands, arguing against population bottlenecking. The Azorean house mouse populations were differentiated from the Portuguese and Madeiran populations and no evidence of recent migration between the three was obtained. Within the Azores, the Eastern, Western, and Central island groups tended to act as separate genetic units for house mice, with some exceptions. In particular, there was evidence of recent migration events among islands of the Central island group, whose populations were relatively undifferentiated. Santa Maria had genetically distinctive mice, which may relate to its colonization history. © 2013 The Linnean Society of London     

Ranking protected areas in the Azores using standardised sampling of soil epigean arthropods

Nineteen areas in seven of the nine Azorean islands were evaluated for species diversity and rarity based on soil epigean arthropods. Fifteen out of the 19 study areas are managed as Natural Forest Reserves and the remaining four were included due to their importance as indigenous forest cover. Four of the 19 areas are not included in the European Conservation network, NATURA 2000. Two sampling replicates were run per study area, and a total of 191 species were collected; 43 of those species (23%) are endemic to the archipelago and 12 have yet to be described. To produce an unbiased multiple-criteria index (importance value for conservation, IV-C) incorporating diversity and rarity based indices, an iterative partial multiple regression analysis was performed. In addition, an irreplaceability index and the complementarity method (using both optimisation and heuristic methods) were used for priority-reserves analyses. It was concluded that at least one well-managed reserve per island is absolutely necessary to have a good fraction of the endemic arthropods preserved. We found that for presence/absence data the suboptimal complementarity algorithm provides solutions as good as the optimal algorithm. For abundance data, optimal solutions indicate that most reserves are needed if we want that at least 50% of endemic arthropod populations are represented in a minimum set of reserves. Consistently, two of the four areas not included in the NATURA 2000 framework were considered of high priority, indicating that vascular plants and bird species used to determine NATURA 2000 sites are not good surrogates of arthropod diversity in the Azores. The most irreplaceable reserves are those located in older islands, which indicates that geological history plays an important role in explaining faunal diversity of arthropods in the Azores. Based both on the uniqueness of species composition and high species richness, conservation efforts should be focused on the unmanaged Pico Alto region in the archipelago’s oldest island, Santa Maria.