First records of Hippocampus algiricus in the Canary Islands (north‐east Atlantic Ocean) with an observation of hybridization with Hippocampus hippocampus

Morphometric and genetic analyses confirmed the first records of the West African seahorse Hippocampus algiricus at Gran Canaria Island (north‐east Atlantic Ocean), and also the first evidence of interspecific hybridization in seahorses. These results provide additional data on the distribution of H. algiricus that may help to establish future conservation strategies, and uncover a new potential sympatric scenario between H. algiricus and Hippocampus hippocampus.     

Novel microsatellite loci in the threatened European long-snouted seahorse (<Emphasis Type="Italic">Hippocampus guttulatus</Emphasis>) for genetic diversity and parentage analysis

The long-snouted seahorse Hippocampus guttulatus is one of the two European seahorse species. We describe the isolation of the first 12 microsatellite loci in this threatened species. These new markers were tested in non-invasive samples of 32 seahorses from NW Spain. The number of alleles ranged from 2 to 15 (mean: 6.3) and expected heterozygosity from 0.031 to 0.912 (mean: 0.500). All loci conformed to Hardy–Weinberg expectations and no genotypic disequilibrium was observed between any pair of loci. The theoretical exclusion probabilities for this set of loci, when no parental information exists or when one parent is known, were 0.973 and 0.998, respectively. This study indicates the usefulness of these novel loci for population analysis and kinship studies in Hippocampus guttulatus. Their potential application is extended to the other European seahorse species, since all loci were successfully cross-amplified in H. hippocampus.     

Seahorse (Hippocampinae) population fluctuations in the Ria Formosa Lagoon,south Portugal

Comparisons of three sets of surveys in the Ria Formosa Lagoon, Portugal, over a 13 year period (2001–2002, 2008–2009 and 2010–2013) revealed significant population fluctuations in at least one of the two seahorse (Hippocampinae) species living there, and that those fluctuations were potentially associated with habitat changes in the lagoon. After a significant decline between the first two survey periods (2001–2002 v. 2008–2009), long‐snouted seahorse Hippocampus guttulatus populations increased significantly between 2008–2009 surveys and new 2010–2013 surveys. There were no significant differences in H. guttulatus populations between the 2001–2002 and 2010–2013 surveys. In contrast, there were no significant differences in short‐snouted seahorse Hippocampus hippocampus densities among the 16 sites surveyed throughout the three sampling periods, although the ability to detect any change was hampered by the low densities of this species in all time periods. Fluctuations in H. guttulatus densities were positively correlated with the percentage of holdfast coverage, but with none of the other environmental variables tested. These results highlight the importance of holdfast availability in maintaining stable seahorse populations. While population fluctuations are certainly more promising than a consistent downward decline, such extreme fluctuations observed for seahorses in the Ria Formosa Lagoon could still leave these two species vulnerable to any additional stressors, particularly during low density periods.     

Serial monogamy in the European long-snouted seahorse Hippocampus guttulatus

Seahorses (Hippocampus spp.) are non-sex-role-reversed members of the Syngnathidae family that provide extensive brood care. Previous studies of seahorses have revealed monogamy within a single brood, but their longer term mating system had not been comprehensively evaluated. The parental contribution to 29 wild-born broods of Hippocampus guttulatus, sampled from six Portuguese populations with differing seahorse densities and sex ratios, was assessed using microsatellite DNA markers. To assess the longer term genetic mating system of this species parentage was determined in eleven broods sampled from a captive population over two breeding seasons. Genetic data suggest that this socially polygamous seahorse is serially monogamous across breeding seasons, i.e. monogamous within a season but may switch mates between seasons, and that differing population densities and sex ratios do not affect the mating system.     

Field studies of seahorse population density,structure and habitat use in a semi-closed north-eastern Mediterranean marine area (Stratoni,North Aegean Sea)

The present study was carried out in the marine area of Stratoni, Greece, where two seahorse species are present (Hippocampus hippocampus and Hippocampus guttulatus). Two surveys were conducted (September 2016 and May 2019) to gather information regarding seahorse species' abundance, distribution and habitat characteristics. Four different seahorse natural and artificial habitat types were identified. The results revealed that the presence of H. hippocampus was relatively high, especially at sites with artificial structures, whereas the presence of H. guttulatus was rare. Data collected can provide baseline information for future population assessments.     

Identifying key environmental variables of two seahorse species (<Emphasis Type="Italic">Hippocampus guttulatus</Emphasis> and <Emphasis Type="Italic">Hippocampus hippocampus</Emphasis>) in the Ria Formosa lagoon,South Portugal

Recent findings reported a significant decrease in abundance of two seahorse species (Hippocampus guttulatus and H. hippocampus) in the Ria Formosa lagoon (South Portugal) and no direct causes have been, so far, clearly identified. This study aimed to describe fluctuations in the local seahorse populations through monthly surveys over a course of a year, in order to identify some of the potential drivers behind the seasonal fluctuations. A total of six sites were chosen based on their habitat characteristics. The highest H. guttulatus abundances were recorded at sites with higher holdfast availability and depth ranging from 3 to 6 m, while H. hippocampus were observed at highest numbers in sites with lower holdfast availability and patchy distribution. In most sites, seahorse density decreased during the summer months (from May to August) and increased from August to December. Holdfast use changed across the surveyed sites, according to the respective habitat characteristics. This study identified environmental variables that influenced the abundance of seahorse population, i.e., holdfast availability, depth and temperature in the Ria Formosa lagoon, underlining the importance of monitoring populations over a course of no less than a year in order to avoid bias due to seasonal fluctuations. Identifying critical habitats will provide valuable information for local authorities in order to implement protective measures towards seahorse conservation.     

An assessment of West African seahorses in fisheries catch and trade

This study provides the first assessment of a heavily traded West African seahorse species, Hippocampus algiricus, and the first information on short‐snouted seahorse Hippocampus hippocampus biology in Africa. A total of 219 seahorses were sampled from fisher catch in Senegal and The Gambia, with estimated height at reproductive activity for H. algiricus (161 mm) larger than mean ± s.d . catch height (150 ± 31 mm). Catch composition, height at reproductive activity and potential biases in fishery retention are discussed with regard to the current Convention on International Trade of Endangered Species (CITES) guidelines.     

Life history and ecology of the elusive European short‐snouted seahorse Hippocampus hippocampus

To improve the understanding of the life history and ecology of one of Europe's most elusive fishes, the short‐snouted seahorse Hippocampus hippocampus, data from wild populations in a shallow coastal lagoon in southern Portugal were analysed. The data were collected from 17 tagged seahorses on a focal‐study grid as well as from >350 seahorses encountered during underwater visual surveys and a fishery‐independent study using beach seines. These populations of settled juveniles and adults had a mean population density of 0·009 m^?2. During the study period (2000–2004), reproduction peaked in July and August. Juveniles recruited to the lagoon at c. 66 mm standard length (L_S) and 0·5 years of age and established small home ranges (0·8 to 18·2 m²). First reproduction was estimated at 100 mm and 1 year of age. Based on a fitted von Bertalanffy model, H. hippocampus grew quickly (growth coefficient K = 0·93) to a maximum theoretical size L_∞ = 150 mm and have a maximum lifespan of c. 3·2 years. Courtship behaviours were consistent with the maintenance of pair bonds and males brooded multiple batches of young per year. Estimated annual reproductive output averaged 871 young (±632). Together these analyses provide the first life‐history parameters for this species and indicate that H. hippocampus bears characteristics of opportunist and intermediate strategists. Such populations are predicted to exhibit large fluctuations in abundance, making them vulnerable to extended periods of poor recruitment.     

Characterization of microsatellite loci in Brachyderes rugatus,the Canary Islands pine weevil (Coleoptera: Curculionidae)

The Canary Island pine weevil Brachyderes rugatus (Wollaston) consists of four allopatric subspecies that are thought to have arisen from several historic colonization events within the archipelago. We have isolated and optimized seven microsatellite loci from Brachyderes rugatus calvus from Gran Canaria. Six of these loci are polymorphic within B. rugatus (11–22 alleles per locus; heterozygosity between 0.43 and 0.84). There is no evidence for heterozygote deficit within populations or for linkage disequilibrium between pairs of loci. These molecular markers are likely to prove useful tools for quantifying the genetic variability of bottlenecked island populations.     

Microsatellite loci in the stable fly,Stomoxys niger niger (Diptera: Muscidae) on La Réunion Island

Six polymorphic microsatellite markers have been isolated from a microsatellite‐enriched DNA library from the stable fly, Stomoxys niger niger. These loci exhibited five to 10 alleles per locus and an expected heterozygosity ranging from 0.57 to 0.81 in the studied populations from La Réunion Island (Indian Ocean). They should therefore be useful for the study of genetic diversity and population structure of S. niger. In addition, cross‐species amplification of four of these six loci in the closely allied species Stomoxys calcitrans produced interpretable results, two of which would be useful for population biology studies.     

Complex population genetic structure in the endemic Canary Island pine revealed using chloroplast microsatellite markers

The Canary archipelago, located on the northwestern Atlantic coast of Africa, is comprised of seven islands aligned from east to west, plus seven minor islets. All the islands were formed by volcanic eruptions and their geological history is well documented providing a historical framework to study colonization events. The Canary Island pine (Pinus canariensis C. Sm.), nowadays restricted to the westernmost Canary Islands (Gran Canaria, Tenerife, La Gomera, La Palma and El Hierro), is considered an old (Lower Cretaceous) relic from an ancient Mediterranean evolutionary centre. Twenty seven chloroplast haplotypes were found in Canary Island pine but only one of them was common to all populations. The distribution of haplotypic variation in P. canariensis suggested the colonization of western Canary Islands from a single continental source located close to the Mediterranean Basin. Present-day populations of Canary Island pine retain levels of genetic diversity equivalent to those found in Mediterranean continental pine species, Pinus pinaster and Pinus halepensis. A hierarchical analysis of variance (AMOVA) showed high differentiation among populations within islands (approximately 19%) but no differentiation among islands. Simple differentiation models such as isolation by distance or stepping-stone colonization from older to younger islands were rejected based on product-moment correlations between pairwise genetic distances and both geographic distances and population-age divergences. However, the distribution of cpSSR diversity within the islands of Tenerife and Gran Canaria pointed towards the importance of the role played by regional Pliocene and Quaternary volcanic activity and long-distance gene flow in shaping the population genetic structure of the Canary Island pine. Therefore, conservation strategies at the population level are strongly recommended for this species.Communicated by D.B. NealeA. Gómez and S.C. González-Martínez as joint authors     

Is filament clipping an effective tool for tissue sampling in <Emphasis Type="Italic">Hippocampus guttulatus</Emphasis>?

Examining genetic diversity, population structure, and geographic distribution has become an important part in the conservation strategies of endangered populations. However, these studies rely on tissue samples collection for DNA analysis which may be problematic for species with a sensitive conservation status. Partial fin-clipping has been employed for tissue collection, due to the increased popularity of DNA-based analysis for ecological and fisheries studies, however, fin clipping can potentially affect behavior, swimming performance, predator avoidance and the ability to find and capture prey. This study aimed to test the effects of filament clipping using captive breed seahorses (Hippocampus guttulatus) as an alternative tool for tissue sample. Clipped filaments regrew on average 0.05?±?0.02 mm/day for 3 months and no mortality or disease was observed during the experiment. Filaments provided enough tissue for DNA analysis. This study provided valuable information regarding a new sampling technique that does not impair the seahorse locomotion. This methodology may be used in population’s genetic studies of other species that have skin filaments as a morphological trait.     

High-throughput sequencing-based analysis of bacterial communities associated with Barbour's seahorses (Hippocampus barbouri) from Surigao del Norte,Philippines

This study identified the bacterial community composition associated with Barbour's seahorses (Hippocampus barbouri). Seahorses and samples from the surrounding environment (sediment and water) were collected from Cantiasay Island in Surigao del Norte, Philippines. Genomic DNA was isolated from these samples, and the V1–V3 region of the 16S rRNA gene was amplified and sequenced on the Illumina MiSeq platform. There were 929 803 sequence reads corresponding to 859 operational taxonomic units (OTUs), which were obtained from the six genomic libraries. The results showed that the most abundant OTUs were affiliated to the phylum Proteobacteria, particularly those belonging to the genera Shewanella, Pseudomonas, Acinetobacter and Aeromonas, which were found to be dominant in seahorse gut samples. The only exception was for the skin of male seahorses, which was dominated by members belonging to the genus Bacillus (phylum Firmicutes). These findings on bacterial community structure and composition may shed light on therapeutic claims of Barbour's seahorses and help in the conservation of this species.     

Two new species of seahorses (Syngnathidae, Hippocampus) from the Middle Miocene (Sarmatian) Coprolitic Horizon in Tunjice Hills, Slovenia: The oldest fossil record of seahorses

Two new and the only known extinct seahorse species Hippocampus sarmaticus and Hippocampus slovenicus are described from the Middle Miocene beds (Lower Sarmatian) in Slovenia, representing the oldest known fossil record of seahorses. Hippocampus sarmaticus was most similar to the extant seahorse species Hippocampus trimaculatus, while H. slovenicus can be most easily compared to the extant pygmy seahorses H. bargibanti, H. denise, and H. colemani. These Sarmatian seahorses lived among seagrasses and macroalgae in the temperate shallow costal waters of the western part of the Central Paratethys Sea.     

Population dynamics of the long-snouted seahorse (Hippocampus guttulatus Cuvier, 1829) in the Mar Menor coastal lagoon

A 10-year monitoring program was developed to quantify the population dynamics of the long-snouted seahorse population in the Mar Menor coastal lagoon. Based on 985 underwater visual censuses, we estimated the long-snouted seahorse (Hippocampus guttulatus Cuvier, 1829) population size in the Mar Menor lagoon and its reduction in size in the last decades, as well as the effect of eutrophication crises in 2016 and 2019 on the species. The annual recruitment for the 2013–2020 period was estimated by comparing the relative abundance of early seahorse life stages in the ichthyoplankton. The density ranged from 0.0458 specimens/m³ at the beginning of the sampling period to 0.0004 at the end, showing a statistically significant difference between the three analyzed periods (H_gl=2 = 14.0, p = 0.001). The long-snouted seahorse population from the Mar Menor lagoon exemplifies the impact of fishing activities and human pressure, especially euxinic episodes and habitat destruction. As a result of this, the Mar Menor population has decreased from several million specimens to a few thousand, in only three decades. This species showed considerable resilience, the seahorse population began to recover once fishing activity stopped. In contrast, the long-snouted seahorse showed high vulnerability to habitat loss and an episodic flooding event. Adult seahorses showed preferences for highly complex habitats, especially Caulerpa prolifera–Cymodocea nodosa mixed meadows and habitats of high complexity and anthropogenic origin, such as harbors, jetties, or breakwaters. In contrast, juvenile seahorses preferred monotonous seabeds with low complexity, such as the sandy beds that are characteristic of the Mar Menor lagoon littoral.     

Reduced genetic variation and strong genetic population structure in the freshwater killifish Valencia letourneuxi (Valenciidae) based on nuclear and mitochondrial markers

The genetic variation of the critically endangered Corfu killifish (Valencia letourneuxi), an endemic freshwater fish species of the western Balkans, was assessed for nine populations sampled in eight water systems in western continental Greece, the Peloponnese and the Ionian Island of Corfu, using mitochondrial and microsatellite markers. The analyses were based on data from three mtDNA regions (D‐loop, COI and 16S rRNA sequences) and 14 microsatellite loci. Samples from the congeneric species Valencia hispanica and the phylogenetically closely related species Aphanius fasciatus were also used in the study as outgroups. Both the mitochondrial and the microsatellite analyses revealed three distinct population groupings associated with the geographical distribution of the populations: one southern group occupying rivers draining to the Patraikos Gulf, the second one including the populations flowing into the Amvrakikos Gulf and the third, more northern group, including the other populations from rivers in Corfu Island and Epirus flowing into the Ionian Sea. Within these groupings there is limited genetic differentiation between populations; in addition, there is reduced intrapopulation genetic variation, evidenced by low heterozygosity values, number of alleles and haplotype diversity. In terms of taxonomic implications and appropriate management actions for conservation, our data suggest that the major population groups should be regarded at least as three distinct conservation units (CUs), with translocation and restocking actions to take place only within the geographical range of the CU concerned. © 2013 The Linnean Society of London, Biological Journal of the Linnean Society, 2014, 111 , 334–349.     

Population genetic structure of Sorex unguiculatus and Sorex caecutiens (Soricidae, Mammalia) in Hokkaido, based on microsatellite DNA polymorphism

We investigated the genetic structure of Sorex unguiculatus and Sorex caecutiens populations in Hokkaido, Japan, using hypervariable microsatellite DNA markers. We used five microsatellite loci to type 475 S. unguiculatus individuals from 20 localities on the Hokkaido mainland and four localities from each of four offshore islands (and 11 shrews from one locality in southern Sakhalin for a particular analysis). We used six microsatellite loci to type 240 S. caecutiens individuals from 13 localities on the Hokkaido mainland. Genetic variation was high in mainland populations of both species and low in the island populations of S. unguiculatus. Allelic richness and island size were positively correlated for S. unguiculatus, suggesting that genetic drift occurred on those islands due to small population size. In addition, four insular populations of S. unguiculatus were genetically differentiated from the mainland populations, although clear phylogeographic clustering was not confirmed among populations on the Hokkaido mainland for either S. unguiculatus or S. caecutiens. Heterozygosity excess was observed in more than half of the populations including the mainland populations of the two species, suggesting recent bottleneck events in these populations. Population dynamics of the shrews might be explained by a metapopulation scheme. According to autocorrelation analysis, the extent of non-random spatial genetic structure was approximately 100 km. Isolation by distance was observed in S. unguiculatus, but not in S. caecutiens although there is a positive trend. The lack of correlation for S. caecutiens might have been due to small sample size. Thus, no obvious differences in population genetic structure were found between the two species on the Hokkaido mainland in the present study, while previous investigations using mitochondrial DNA sequences inferred that these two species might have rather different biogeographic histories.     

Microsatellite loci development in endangered pamphagid grasshoppers endemic to the Canary Islands (Orthoptera)

The five recognized endemic pamphagid species (Orthoptera) of the Canary Islands have restricted and fragmented ranges due to habitat decline. Seven polymorphic microsatellite markers have been developed for Acrostira tamarani, and the performance of primer pairs amplifying these loci in related taxa has been tested. The number of alleles in A. tamarani samples taken from two distant localities in the island of Gran Canaria ranged from two to eight per locus. Observed heterozygosities were from 0.151 to 0.559. Up to four primer pairs amplified in related species with moderate heterozigosities (maximum of 0.687 and 10 alleles for Ata67 locus in Purpuraria erna). These markers could be useful tools to study the population structure and management of endemic threatened pamphagids of the Canary archipelago.     

First record of the Pacific seahorse Hippocampus ingens in Guadalupe Island,Mexico

This is the first record of the Pacific seahorse Hippocampus ingens at a northern oceanic island from the eastern Pacific Ocean. The photographic record of the juvenile female H. ingens was made in November 2015 during a cage diving trip at Guadalupe Island, Mexico. The presence of H. ingens in this area could be related to long distance dispersal mechanisms, as has been observed in other species of seahorses.     

Integrative taxonomy reveals Europe's rarest songbird species,the Gran Canaria blue chaffinch Fringilla polatzeki

The conservation of endangered taxa often critically depends on accurate taxonomic designations. The status of the Gran Canaria population of the blue chaffinch Fringilla teydea polatzeki has not been reevaluated since the early 1900s when this taxon was described as a subspecies and combined with the much more common Tenerife blue chaffinch F. t. teydea in a single species. We show that multiple diagnostic differences in plumage, songs, calls and morphometrics distinguish F. t. polatzeki from F. t. teydea. Preliminary playback experiments suggest that F. t. polatzeki is able to discriminate between songs of both taxa. Along with previously reported differences in mitochondrial DNA, these findings show that the blue chaffinches on Gran Canaria and Tenerife represent two distinctive species: F. polatzeki and F. teydea. Gran Canaria blue chaffinch is Europe's rarest passerine species and should be classified as critically endangered. Its long‐term survival in the wild currently depends on a very small (< 20 km²) area in southwest Gran Canaria. Reclassification of Gran Canaria blue chaffinch as a species increases the urgency of ongoing conservation efforts. Our study underscores the critical importance of taxonomic clarification of threatened taxa that are currently classified as ‘subspecies’.