A case of mistaken identity: skin filaments are unreliable for identifying Hippocampus guttulatus and Hippocampus hippocampus

The presence or absence of skin filaments is an unreliable character for identifying European seahorses. Growth of skin filaments on Hippocampus guttulatus coincides with maturation, while presence on Hippocampus hippocampus is highly variable. Multiple morphological traits including snout, head and trunk shape should be used for identification.     

Advances in tagging syngnathids, with the effects of dummy tags on behaviour of Hippocampus guttulatus

Artificial marking and tagging techniques have been used to study movement, population dynamics, behaviour, ecology, survival and growth of at least 25 syngnathid species. External necklace-style tags and injection of visible implant elastomer have been the most used techniques, uniquely identifying hundreds of individual syngnathids to study population dynamics, mortality, behaviour, ecology and growth in at least 13 and 12 species, respectively. Only two studies, both on larger syngnathid species, have tested the use of internal or electronic tags. This new case study reveals that dummy tags, weighing up to 6% of individual body mass, have minimal effect on normal ex situ behaviour of the long-snouted seahorse Hippocampus guttulatus, a smaller syngnathid. In paired aquarium trials, tags did not affect movement, holdfast use or general behavioural state, and only had a short-term effect (1 day) on vertical orientation. Tagged H. guttulatus gained more mass during the 5 day trials, a result which warrants further exploration but indicates that tags did not reduce feeding. This study shows promise for using electronic tagging to study H. guttulatus and similarly sized syngnathids in the wild.     

Male gonads morphology,spermatogenesis and sperm ultrastructure of the seahorse Hippocampus guttulatus (Syngnathidae)

Testes morphology, spermatogenetic process and mature sperm ultrastructure were analysed in Hippocampus guttulatus, using both light and transmission electron microscopy. Both testes were organized in a single large germinal compartment, with a central lumen. Spermatocysts only contained spermatogonia and primary spermatocytes. Inside the testis lumen, together with mature sperm, two types of large mono‐nucleate cells, flagellate and aflagellate, were present. Both types of cells were interpreted as developing germ cells precociously released inside the testis lumen, where their maturation was completed. According to the different morphological features of the nuclei, such as chromatin condensation degree, aspect of the nuclear fossa and others, the flagellate cells were unquestionably developing spermatids. On the contrary, the developmental stage of the aflagellate was more difficult to interpreted. They could be secondary spermatocytes or young spermatids. No dimorphic sperm were recognizable, the only sperm type observed have features typical of the intro‐sperm reports in other syngnathids species. They had a cylindrical head, a short midpiece, characterized by two mitochondrial rings housed inside a cytoplasmic collar, and a long flagellum. These and previous data about the same topic reported on other syngnathids species were compared and discussed.     

Dispersal, habitat differences, and comparative phylogeography of Southeast Asian seahorses (Syngnathidae: Hippocampus)

Four distinct phylogeographical patterns across Southeast Asia were observed for four species of seahorse (genus Hippocampus) with differing ecologies. For all species, genetic differentiation (based on cytochrome b sequence comparisons) was significantly associated with sample site (Phi(ST) = 0.190-0.810, P < 0.0001) and with geographical distance (Mantel's r = 0.37-0.59, P < 0.019). Geographic locations of genetic breaks were inconsistent across species in 7/10 comparisons, although some similarities across species were also observed. The two shallow-water species (Hippocampus barbouri and Hippocampus kuda) have colonized the Sunda Shelf to a lesser degree than the two deeper-water species (Hippocampus spinosissimus and Hippocampus trimaculatus). In all species the presence of geographically restricted haplotypes in the Philippines could indicate past population fragmentation and/or long-distance colonization. A nested clade analysis (NCA) revealed that long-distance colonization and/or fragmentation were likely the dominant forces that structure populations of the two shallow-water species, whereas range expansion and restricted dispersal with isolation by distance were proportionally more important in the history of the two deeper-water species. H. trimaculatus has the most widespread haplotypes [average clade distance (D(c)) of nonsingleton haplotypes = 1169 km], indicating potentially high dispersal capabilities, whereas H. barbouri has the least widespread haplotypes (average D(c) = 67 km) indicating potentially lower dispersal capabilities. Pleistocene separation of marine basins and postglacial flooding of the Sunda Shelf are extrinsic factors likely to have contributed to the phylogeographical structure observed, whereas differences among the species appear to reflect their individual ecologies.     

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.     

Conventional karyotype, nucleolar organizer regions and genome size in five Mediterranean species of Syngnathidae (Pisces, Syngnathiformes)

Conventional karyotypes, NOR-bearing chromosomes by means of silver impregnation and genome size were investigated in five Mediterranean species in three genera of the Syngnathidae. A karyotype of 48 subtelocentric-acrocentric chromosomes was found in the seahorse Hippocampus hippocampus (FN=48) while a diploid value of 44 occurred in H. guttulatus (2 sm-m+42 a; FN=46) and the pipefish Syngnathus abaster (44 a; FN=44) and S. typhle (44 a; FN=44). The pipefish Nerophis ophidion , possessing a diploid chromosomal set of 58 made up of 50 meta-submetacentric and eight subteloacrocentric elements (FN=108) and a genome size three to four times larger than those known to date, differs cytogenetically from all other Syngnathids studied so far. A single pair of active NOR-bearing chromosomes was found in both species of the genus Hippocampus while in Syngnathus and Nerophis species more than two silver positive chromosomes were found to be involved in nucleolus organization giving rise to NOR-bearing chromosome polymorphism. The possible evolutionary routes of quantitative and qualitative changes in chromosome and DNA are discussed. The resulting phylogenetic scheme is shown to coincide with that constructed from morphological characters.     

Activity rate of the seahorse Hippocampus reidi Ginsburg, 1933 (Syngnathidae)

This study aimed to compare activity levels of male and female Hippocampus reidi and to relate the level of activity of the males to their height and brood pouch circumference. Females appeared to be highly active, while pregnant males were less active and non-pregnant males presented more balance in time investment between active and inactive behavior. There was a positive correlation between height, brood pouch circumference, and inactive behavior.     

The dorsal fin engine of the seahorse (Hippocampus sp.)

The muscles, fin ray joints, and supporting structures underlying the dorsal fin are described for two seahorse species: Hippocampus zosterae and Hippocampus erectus. A fan-shaped array of cartilaginous bones, the pterigiophores, form the internal supporting structure of the dorsal fin. Each pterigiophore is composed of a proximal radial that extends from a vertebra to the dorsal side of the animal, where it fuses to a middle radial. The middle radials fuse with each other to form a dorsal ridge upon which sit the spheroidal distal radials. Each distal radial articulates with a fin ray on its dorsal side and is attached to the dorsal ridge on its ventral side by a material that has been histologically identified as elastic cartilage. Together these connections form a two-axis joint that permits elevation, depression, and inclination of the ray. Each fin ray is actuated by two bilateral pairs of muscles, an anterior pair of inclinators, and a posterior pair of depressors. The anteriormost fin ray is actuated by three bilateral pair of muscles, the inclinators, the depressors, and a pair of elevator muscles that are positioned anterior to the inclinators. Preliminary examinations of the ray joints of the pectoral and anal fins of adult H. zostera and the pectoral fins of newborn H. erectus revealed structures similar to that seen in the dorsal fins. To further explore the structure and function of the dorsal fin gross dissections and simple functional tests were performed on H. erectus and H. barbouri and behavioral observations were made of all three species plus Hippocampus kuda.     

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.     

三斑海马线粒体基因组核苷酸全序列结构与分析

通过PCR扩增并测序获得了三斑海马(Hippocampus trimaculatus)线粒体DNA(mt DNA)全序列。三斑海马线粒体基因组全序列长度为16 534 bp(Gen Bank登录号为KJ956892),编码37个基因,包括13个蛋白编码基因、22个t RNA基因和2个r RNA基因。非编码区域包括1个控制区(D-loop)及一个轻链复制起始区域。大部分基因由H-链编码,包括14个t RNA基因、2个r RNA基因、12个蛋白编码基因;只有ND6和8个t RNA基因是在L-链编码。预测的22个t RNA基因的二级结构均为典型的三叶草状。基因间隔一般1~14 bp不等。此外,还存在7处碱基重叠,其中,4处是鱼类和脊椎动物典型的基因重叠位点。总的碱基含量分别为,A 32.7%,C 23.4%,G 14.6%,T 29.3%,A+T含量为62.0%。其线粒体基因组序列的结构与脊椎动物的典型结构近似。邻接法和贝叶斯法构建的三斑海马系统进化树的拓扑结构相似,这与现有的三斑海马的系统演化地位一致。本研究为海马的进化研究以及保护工作提供了基础数据。     

Light–dark variations in plasma melatonin concentrations in the pot-bellied seahorse Hippocampus abdominalis Lesson, 1827

A diel plasma melatonin profile measured in adult pot-bellied seahorses Hippocampus abdominalis showed that this species produces elevated levels of melatonin during the scotophase, returning to basal levels during the photophase.     

Monthly population density and structure patterns of an endangered seahorse Hippocampus capensis: a comparison between natural and artificial habitats

This study investigated aspects of the population ecology of the endangered Knysna seahorse Hippocampus capensis within different habitat types. High densities of H. capensis were found within artificial Reno mattress habitat, within the Knysna Estuary, South Africa. Monthly surveys at three sites were conducted from October 2015 to August 2016 to compare population densities of H. capensis in this artificial habitat with natural eel grass Zostera capensis habitat. Hippocampus capensis densities varied significantly across all sites and highest population densities were consistently observed within the Reno mattress habitat. Hippocampus capensis were also found to be significantly larger within the Reno mattress habitat and pooled data showed that males were significantly larger than females. The overall sex ratio for all three sites was female biased, although this varied across seasons at two sites. The findings suggest that artificial Reno mattresses provide novel habitat for this endangered species and consideration should be given to the usefulness of these structures in future conservation actions.     

Population characteristics of two seahorses, Hippocampus coronatus and Hippocampus mohnikei, around seagrass beds in the southern coastal waters of Korea

Population characteristics of two seahorse species, Hippocampus coronatus and Hippocampus mohnikei, in seagrass beds in the southern coastal waters of Korea were determined based on field observations. In Zostera beds in the Yeosu area, there was a mean density of 2.9 seahorses per 1,000?m² for H. coronatus and 1.4 seahorses per 1,000?m² for H. mohnikei. The greatest numbers of young individuals of both species were observed in July, with numbers decreasing through November. The male:female sex ratio was 1:1.6 for H. mohnikei and 1:1.7 for H. coronatus. Breeding males of H. coronatus and H. mohnikei were observed from July to November and from July to September, respectively. Within the study site, the populations of the two seahorse species exhibited low density and patchy distributions, with variation in temporal abundance being significantly correlated with water temperature. The findings of this study provide the basis for future research on the population status of H. coronatus and H. mohnikei in seagrass beds.     

Fecundity, survival, and growth of the seahorse Hippocampus ingens (Pisces: Syngnathidae) under semi-controlled conditions

We studied fecundity, survival, and growth of the seahorse Hippocampus ingens under semi-controlled conditions. Three wild brood stock mature males of 14.8, 24.5, and 32.0 g released 1,598, 1,703, and 1,658 juveniles. Juvenile stocking densities of 12 were settled in 60-1 aquariums in groups of 1, 12, and 20 days old organisms. The rate of survival was 21.5, 61.9, and 59.0%, respectively, in 35 days. Juveniles were fed a mix diet of rotifers B. plicatilis and Artemia nauplii, then they were transferred to a cement tank of 100,000 1 at a density of 50/1,000 1 and fed with live adult Artemia for 60 days more. They grew from an average of 0.7, 1.5, and 2.7 to 4.5, 5.4, and 6.7 cm, respectively, in 95 days. The seawater temperature varied from 17 to 23 degrees C.     

Genetic monogamy despite social promiscuity in the pot-bellied seahorse (Hippocampus abdominalis)

Sexual selection theory predicts a positive correlation between relative parental investment and mate choice. In syngnathid fishes (seahorses and pipefish), males brood offspring in specialized brooding structures. While female-female mating competition has been demonstrated in some pipefishes, all seahorses (genus Hippocampus) studied to date have been found to have conventional sex roles with greater male-male competition for access to mates despite possessing the most complex brood structures in the family. Although multiple mating is common in pipefish, seahorses are again exceptional, exhibiting strict genetic monogamy. Both demographic and behavioural explanations have been offered to explain the lack of multiple mating in seahorse species, but these hypotheses have not yet been explicitly addressed. We investigated mating systems and brood parentage of the pot-bellied seahorse, Hippocampus abdominalis, a temperate-water species that is socially promiscuous with conventional sex roles in laboratory populations. We observed promiscuous courtship behaviour and sex-role reversal in high density, female-biased field populations of H. abdominalis. We hypothesize that sex roles are plastic in H. abdominalis, depending on local population density and sex ratio. Despite promiscuous courtship behaviour, all assayed male seahorses were genetically monogamous in both laboratory and wild populations. Physiological limitations associated with embryo incubation may explain the absence of multiple mating in seahorses and may have played an important role in the development of the unique reproductive behaviour typical in these species.     

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.     

Free radical and reactive oxygen species scavenging activities of the extracts from seahorse, Hippocampus kuda Bleeler

Seahorse, Hippocampus kuda (SH) a marine teleost fish, is well known not only for its special medicinal composition and used as one of the most famous and expensive materials of traditional Chinese medicine. It was extracted with water (SHW), methanol (SHM), and ethanol (SHE), respectively and evaluated by various antioxidant assays. The including reducing power, total antioxidant, DPPH radical scavenging, hydroxyl radical scavenging, superoxide anion radical scavenging, alkyl radical scavenging, and protective effect on DNA damage caused by hydroxyl radicals generated. Further, the ROS level was detected using a fluorescence probe, 2′,7′-dichlorofluorescin diacetate (DCFH-DA), which could be converted to highly fluorescent dichlorofluorescein (DCF) with the presence of intracellular ROS on mouse macrophages, RAW264.7 cell and inhibited myeloperoxidase (MPO) activity in human myeloid, HL60 cells, respectively. Those various antioxidant activities were compared to standard antioxidants such as α-tocopherol. Among SHM exhibited the highest antioxidant activity in linoleic acid system, effective reducing power, DPPH radical scavenging, hydroxyl radical scavenging, superoxide radical scavenging, alkyl radical scavenging, inhibitory intracellular ROS, and inhibited MPO activity. Furthermore, MTT assay showed no cytotoxicity on mouse macrophages cell (RAW264.7) and human cell lines (MRC-5, HL60, U937). This antioxidant property depends on concentration and increasing with increased amount of extracts. The results obtained in the present study indicated that the see horse (Hippocampus kuda Bleeker) is a potential source of natural antioxidant.     

Early life history of a seahorse, Hippocampus mohnikei, in Tokyo Bay, Japan

The early morphological development, seasonal and spatial occurrence patterns, and food habits of a seahorse, Hippocampus mohnikei, in offshore waters of Tokyo Bay, central Japan, were studied on the basis of 206 juvenile and young specimens (6.0–65.3 mm TL) collected between August 1995 and January 1999. All the specimens were collected within the period from May to January, inclusive, each year. In the least developed specimen (6.0 mm TL), the number of dorsal, anal, and pectoral fin rays had attained the adult complement, whereas the minute caudal fin, consisting of two rays, was present in juveniles of 6.0–26.4 mm TL. Hippocampus mohnikei≧35 mm TL, being larger than settlement size (ca. 30 mm TL), had very low gut fullness index values (GFI = 0, >70% of specimens), whereas those of 15–29 mm TL had higher values (GFI = 2–4, >80% of specimens). In addition, larger individuals selectively fed on larger planktonic animals (species of Brachyura), which occurred naturally at low densities, although smaller food items, such as Oithona davisae and Penilia avirostris, occurred abundantly, being consumed by smaller H. mohnikei individuals (15–34 mm TL). These results indicated that food availability for H. mohnikei in offshore waters of Tokyo Bay is significant for individuals larger than settlement size, because their food preference would shift from smaller food items to larger food items, which would be scarce in their environments. Received: January 12, 2001 / Revised: May 13, 2001 / Accepted: June 14, 2001     

Life history and population dynamics of an endangered seahorse (Hippocampus capensis) within an artificial habitat

Species-specific life-history information is critical for successful conservation, particularly in establishing an accurate baseline status. Obtaining such information is challenging for most species, but in particular for rare and threatened marine species. To facilitate future conservation of the endangered Knysna seahorse (Hippocampus capensis) this study aimed to determine important life-history information for this species. Visible implant fluorescent elastomer (VIFE) tags were used to mark 78 seahorses within a residential marina estate in the Knysna estuary, South Africa, in February 2018. Using a mark-resight approach, the size and movement patterns of the population and growth rate of seahorses were determined over a 14-month period. The closed population estimate for H. capensis, within Thesen Islands Marina, was estimated to be 134 (118–152 95% C.I. ) in February 2018 compared to only 72 (48–108 95% C.I. ) in February 2019. The species showed rapid initial growth with males and females having similar rates of growth based on the specialised von Bertalanffy growth function model. The importance of Reno mattresses as a habitat for H. capensis was confirmed based on the high abundance and site fidelity of the population, which emphasises the conservation potential of heavily modified environments for threatened seahorse species. The use of VIFE tags was deemed effective in studying this endangered seahorse and allowed the collection of important information for this species which can be used in future Red List assessments and conservation actions.     

The evolutionary history of seahorses (Syngnathidae: Hippocampus): molecular data suggest a West Pacific origin and two invasions of the Atlantic Ocean

Sequence data derived from four markers (the nuclear RP1 and Aldolase and the mitochondrial 16S rRNA and cytochrome b genes) were used to determine the phylogenetic relationships among 32 species belonging to the genus Hippocampus. There were marked differences in the rate of evolution among these gene fragments, with Aldolase evolving the slowest and the mtDNA cytochrome b gene the fastest. The RP1 gene recovered the highest number of nodes supported by >70% bootstrap values from parsimony analysis and >95% posterior probabilities from Bayesian inference. The combined analysis based on 2317 nucleotides resulted in the most robust phylogeny. A distinct phylogenetic split was identified between the pygmy seahorse, Hippocampus bargibanti, and a clade including all other species. Three species from the western Pacific Ocean included in our study, namely H. bargibanti, H. breviceps, and H. abdominalis occupy basal positions in the phylogeny. This and the high species richness in the region suggests that the genus evolved somewhere in the West Pacific. There is also fairly strong molecular support for the remaining species being subdivided into three main evolutionary lineages: two West Pacific clades and a clade of species present in both the Indo-Pacific and the Atlantic Ocean. The phylogeny obtained herein suggests at least two independent colonization events of the Atlantic Ocean, once before the closure of the Tethyan seaway, and once afterwards.