Synanceia Verrucosa Research Articles

Diel activity patterns and habitat preferences of stonefish (Synanceia spp.) in captivity

Synanceia verrucosa (reef stonefish) and Synanceia horrida (estuarine stonefish) are known as the world’s most venomous fishes. However, little is known about their habitat preference and activity patterns. This study aimed to determine the habitat preferences and diel activity patterns of stonefish. The movements of four adults of each species that were offered complex and simple habitats in captivity were recorded by filming during day and night periods. Generally, both species exhibited similar behaviour, though S. verrucosa travelled further per movement than S. horrida. Collectively, stonefish spent 94% of their time associated with complex habitat and moved significantly less frequently on complex (1.6 movements per hour) relative to simple habitat (17.3 movements per hour). Stonefish were least active around sunset in the late afternoon and evening, which may relate to an active feeding period during the morning or paradoxically to a higher probability of ambush feeding success near sunset. These findings have enhanced our current understanding of the ecology of these species.

A Chromosome-Level Genome Assembly of the Reef Stonefish (Synanceia verrucosa) Provides Novel Insights into Stonustoxin (sntx) Genes.

Reef stonefish (Synanceia verrucosa) is one of the most venomous fishes, but its biomedical study has been restricted to molecular cloning and purification of its toxins, instead of high-throughput genetic research on related toxin genes. In this study, we constructed a chromosome-level haplotypic genome assembly for the reef stonefish. The genome was assembled into 24 pseudo-chromosomes, and the length totaled 689.74 Mb, reaching a contig N50 of 11.97 Mb and containing 97.8% of complete BUSCOs. A total of 24,050 protein-coding genes were annotated, of which metalloproteinases, C-type lectins, and stonustoxins (sntx) were the most abundant putative toxin genes. Multitissue transcriptomic and venom proteomic data showed that sntx genes, especially those clustered within a 50-kb region on the chromosome 2, had higher transcription levels than other types of toxins as well as those sntx genes scatteringly distributed on other chromosomes. Further comparative genomic analysis predicted an expansion of sntx-like genes in the Percomorpha lineage including nonvenomous fishes, but Scorpaenoidei species experienced extra independent sntx duplication events, marking the clear-cut origin of authentic toxic stonustoxins. In summary, this high-quality genome assembly and related comparative analysis of toxin genes highlight valuable genetic differences for potential involvement in the evolution of venoms among Scorpaeniformes fishes.

Polypharmacological Cell-Penetrating Peptides from Venomous Marine Animals Based on Immunomodulating, Antimicrobial, and Anticancer Properties.

Complex pathological diseases, such as cancer, infection, and Alzheimer's, need to be targeted by multipronged curative. Various omics technologies, with a high rate of data generation, demand artificial intelligence to translate these data into druggable targets. In this study, 82 marine venomous animal species were retrieved, and 3505 cryptic cell-penetrating peptides (CPPs) were identified in their toxins. A total of 279 safe peptides were further analyzed for antimicrobial, anticancer, and immunomodulatory characteristics. Protease-resistant CPPs with endosomal-escape ability in Hydrophis hardwickii, nuclear-localizing peptides in Scorpaena plumieri, and mitochondrial-targeting peptides from Synanceia horrida were suitable for compartmental drug delivery. A broad-spectrum S. horrida-derived antimicrobial peptide with a high binding-affinity to bacterial membranes was an antigen-presenting cell (APC) stimulator that primes cytokine release and naïve T-cell maturation simultaneously. While antibiofilm and wound-healing peptides were detected in Synanceia verrucosa, APC epitopes as universal adjuvants for antiviral vaccination were in Pterois volitans and Conus monile. Conus pennaceus-derived anticancer peptides showed antiangiogenic and IL-2-inducing properties with moderate BBB-permeation and were defined to be a tumor-homing peptide (THP) with the ability to inhibit programmed death ligand-1 (PDL-1). Isoforms of RGD-containing peptides with innate antiangiogenic characteristics were in Conus tessulatus for tumor targeting. Inhibitors of neuropilin-1 in C. pennaceus are proposed for imaging probes or therapeutic delivery. A Conus betulinus cryptic peptide, with BBB-permeation, mitochondrial-targeting, and antioxidant capacity, was a stimulator of anti-inflammatory cytokines and non-inducer of proinflammation proposed for Alzheimer's. Conclusively, we have considered the dynamic interaction of cells, their microenvironment, and proportional-orchestrating-host- immune pathways by multi-target-directed CPPs resembling single-molecule polypharmacology. This strategy might fill the therapeutic gap in complex resistant disorders and increase the candidates' clinical-translation chance.

Inter-species variation in stonefish (Synanceia spp.) ichthyocrinotoxins; an ecological perspective

Although stonefish (Synanceia spp.) are well-known to harbour a highly noxious defensive venom in their dorsal spines, very little is known about the composition and ecological function of the ichthyocrinotoxins that they secrete onto their epidermis. This study profiled reef (Synanceia verrucosa) and estuarine (Synanceia horrida) stonefish ichthyocrinotoxins via electrophoresis, liquid chromatography, and mass spectrometry to visualise and compare the composition of these toxins between the two species. Stonefish ichthyocrinotoxins were found to be multifarious concoctions that exhibited subtle differences between reef and estuarine species. We speculate that these variations and similarities are driven by the different and similar ecology of these fish species. Further research into the activity of the toxins components is now required to better understand their ecological role.

First record of stonefish metabolism: baseline respiration and spontaneous activity of tropical marine Synanceia verrucosa

ABSTRACT For the first time oxygen consumption and spontaneous activity of 16 individuals of stonefish Synanceia verrucosa were determined simultaneously. The experiments were conducted in an intermittent-flow respirometer at 25°C, and salinity (PSU 32). Continuous measurements of fish activity and oxygen consumption were combined in order to calculate routine (RMR) and standard metabolic rate (SMR) and at the same time quantify spontaneous activity. The relationship between RMR and activity was found to be highly significant (P < 0.001). Relative SMR values varied between 24.0 and 54.4 mg O2 h−1 kg wet mass−1 for large (> 500 g) and small (102–394 g) specimens, respectively. Spontaneous activity rates ranged between 4–6 and 40–75 movements h−1 (mean), for large and small specimens, respectively. Absolute SMR for a standard mass of 100 g was calculated to be 3.58 mg O2 h−1, using a wet mass vs. oxygen consumption relationship of SMR [mg O2 h−1] = 0.0643*WM [g] 0.873. The investigated stonefish is so far the most motionless fish found in coral reef/mangrove areas, with activity levels much lower than closely related scorpionfish (Scorpaenidae), and scorpaenids from lower latitudes (e.g. sculpins). SMR of fish in this study was extremely low in comparison with other marine tropical species.

Getting stoned: Characterisation of the coagulotoxic and neurotoxic effects of reef stonefish (Synanceia verrucosa) venom

The reef stonefish (Synanceia verrucosa) is a venomous fish which causes excruciatingly painful envenomations. While some research on the pathophysiology and functions of the venom have been conducted, there are still some gaps in the understanding of the venom effects due to the extreme lability of fish venom toxins and the lack of available testing platforms. Here we set out to assess new functions of the venom whilst also attempting to address some unclear pathophysiological effects from previous literature. Utilising a biolayer interferometry assay, our results highlight that the venom binds to the orthosteric site of the α-1 nicotinic acetylcholine receptor as well as the domain IV of voltage-gated Ca2+ (CaV1.2) channel mimotopes. Both these results add some clarity to the previously ambiguous literature. We further assessed the coagulotoxic effects of the venom using thromboelastography and Stago STA-R Max coagulation analyser assays. We reveal that the venom produced anticoagulant activity and significantly delayed time until clot formation of recalcified human plasma which is likely through the degradation of phospholipids. There was a difference between fresh and lyophilised venom activity toward the nicotinic acetylcholine receptor mimotopes and coagulation assays, whilst no difference was observed in the activity toward the domain IV of CaV1.2 mimotopes. This research adds further insights into the neglected area of fish venom whilst also highlighting the extreme labile nature of fish venom toxins.

Bioinformatics Research Through Image Processing of Histopathological Response to Stonefish Venom

The present study utilizes coastal and environmental engineering to investigate the histopathological effects of Synanceia verrucosa venom on Albino BALB/c mice. S. verrucosa is the most hazardous venomous marine fish that belong to the family Synanceiidae, generally known as the "Reef Stonefish". Crude venom was collected from venom glands of the dorsal spines of stonefish samples taken from the Jordanian coastline of the Gulf of Aqaba, Red Sea. The mice were given intramuscular injections of the venom. Consequently, the research evaluated the acute toxicity and influence on selected serum biomarker enzymes, as well as possible histological alterations of the soleus skeletal muscles. The mice 24 h LD50 was 0.107 µg toxin/kg mouse body weight. After the treatment using venom sublethal dose, the serum biomarkers, including Lactate dehydrogenase (LDH) and Alanine aminotransferase (ALT), were significantly improved (P≤0.05). In addition, Lipid Peroxidation (LPO) contents were significantly increased (P≤0.05) after venom treatment. Moreover, we combined routine medical procedures and artificial intelligence-assisted image analysis for a rapid qualitative and quantitative diagnosis of stonefish injury, based on the histophotography of mice tissue samples during the observation period (1, 2, and 3 hours respectively). The novelty of our method is that we could detect severe and mild damage with an accuracy of 93% and 91%, respectively. The most histological abnormalities in muscles were the great variety in diameters, content, and widespread among randomly distributed muscle fibres. In addition, loss of the tissue's striated appearance was noticed in toxin-treated groups compared with the control group. Consequently, our findings indicate the Stonefish's harmful influences that may endanger human life and highlight the need for appropriate measures to be considered. This, in turn, can ensure beach safety in the Gulf of Aqaba.

The complete mitochondrial genome sequence and phylogenetic analysis for Inimicus japonicus (Scorpaeniformes: Synanceiidae)

The complete mitochondrial genome (mitogenome) of the Inimicus japonicus was analyzed by next-generation sequencing in this study. The mitogenome is 16,978 base pairs (bp) long and codes for 22 transfer RNA (tRNA) genes, 13 protein-coding genes (PCGs), 2 ribosomal RNA genes (rRNA), and 1 non-coding control region. The overall nucleotide composition of the mitogenome is: 29.61% for A, 29.16% for T, 25.26% for C, and 15.97% for G. Twenty-two tRNAs range from 67 to 74 bp in length, and 2 rRNA (12S and 16S) were 953 and 1,687 bp long, respectively. Phylogenetic analysis by neighbor-joining (NJ) method indicated that I. japonicus showed considerable genetic similarity (82%), and had a closer relationship in the phylogenetic tree to Synanceia verrucosa.

Clinical characteristics of stonefish “Oni‐daruma‐okoze” envenomation in Japan

AimStonefish, known in Japan as “Oni‐daruma‐okoze” (Synanceia verrucosa Block et Schneider), inhabit the waters surrounding the Okinawa/Amami districts. By far, habubites have been limited to these areas mainly due to climatic conditions. Due to changes promoted by global warming and increasing tourism, concerns regarding the potential for stonefish envenomation as well as jellyfish stings on the main island of Japan are increasing. However, few studies of stonefish envenomation have been reported in Japan. The current study aimed to comprehensively examine for the first time the clinical characteristics of stonefish envenomation in Japan.MethodsWe administered retrospective questionnaire surveys between January 2013 and December 2017 to determine patient characteristics, treatment, and clinical outcomes of stonefish envenomation. We compared patient characteristics between tourists and non‐tourists.ResultsOver the 5‐year study period, 15 patients were identified from four hospitals. More than 70% of stonefish envenomation cases occurred in the northern part of Okinawa Main Island. All patients recovered well, including four patients with severe stonefish envenomation. In two severe cases, the length of hospitalization was more than 1 week (8–9 days). Antivenom was not given. There were no significant differences between tourists and non‐tourists with regard to baseline characteristics.ConclusionsDuring a 5‐year period, 15 cases of stonefish envenomation occurred in Japan. The patients recovered well without treatment with antivenom.

Stonefish Synanceia verrucosa Bloch & Schneider, 1801 (Actinopterygii: Synanceiidae): the first record in the Syrian coast and the fourth in the Mediterranean

Fish species introduction into the Mediterranean Sea is constantly increasing, and this is what worries biologists especially after the arrival of poisonous species . In this paper, Synanceia verrucosa is recorded for the first time in the Syrian marine waters, filling the gap of its distributions between north and south of the eastern Mediterranean.

The association of economically important fish with mangroves in Maumere Bay, Indonesia

Mangroves are known to play a role in fisheries production, but specifically, however the associations between economically important fish species and mangroves with varied characteristics are not yet widely known. The aim of this study was to analyse the relationship (association) between economically important fish species and mangroves with varied characteristics in Maumere Bay, Indonesia. The research used spatial and non-spatial methods; the research on economically important fish species used an experimental fishing sampling method. Correspondence analysis was applied to answer the question why certain economically important fish species were associated with certain mangrove characteristics. The mangroves at the study locations with characteristically dominant species Bruguiera parviflora, Avicennia alba, Bruguiera gymnorrhiza, Ceriops tagal, with a moderate value of the Shannon diversity index (1< H’ ≤3), and moderate value of the Simpson dominance index (0.5< C ≤0.75), associated with the economically important fish species Psammoperca waigiensis, Synanceia verrucosa and Pomadasys maculatus. The locations of mangrove with high mangrove cover (≥ 80%), Avicennia marina as dominant species, large area (100 ha) and high density (≥ 3000 trees / ha), Rhizophora mucronata as dominant species, and high value of Pielou’s evenness index (0.6< e ≤1.0) were associated with the economically important fish species Upeneus mollucensis, Lethrinus lentjan, and Lutjanus bitaeniatus. The mangrove locations with medium cover (50% - 75%), moderate values of Simpson dominance index (0.5< C ≤0.75) but high mangrove belt width (≥ 400 m) were associated with the economically important fish species Siganus javus, Stegostoma fasciatum, Mugil cephalus, Carangoides malabaricus and Chanos chanos.

Sequence and phylogenetic analysis of the complete mitochondrial genome for the grey stingfish Minous monodactylus (Scorpaeniformes: Synanceiidae)

The mitochondrial genome sequence of Minous monodactylus (16,459 bp) was determined by the MiSeq platform for the first time in the genus Minous. It encoded the 37 genes (13 protein-coding genes, 22 tRNAs, and two ribosomal RNAs) and two conserved noncoding regions; control region (D-loop) and the origin of light strand (OL). Twenty-eight genes were encoded on the heavy (H) strand, while eight tRNAs and ND6 genes were on the light (L) strand. According to the phylogenetic analysis with other complete mitogenomes from order Scorpaeniformes, M. monodactylus was clustered with Synanceia verrucosa as family Synanceiidae with 81% sequence identity.

Biliary Tract-Infecting Myxosporeans from Estuarine and Reef Stonefish (Scorpaeniformes: Synanceiidae) Off Eastern Australia, with Descriptions of Sphaeromyxa horrida n. sp. and Myxidium lapipiscis n. sp. (Myxosporea: Bivalvulida).

Two new species of myxosporeans are described from the gallbladders of estuarine stonefish, Synanceia horrida, and reef stonefish, Synanceia verrucosa, from localities off Cairns, in tropical north Queensland and in Moreton Bay in southern Queensland, Australia. Sphaeromyxa horrida n. sp. can be distinguished from congeners in the morphologically distinct "balbianii" species group within Sphaeromyxa on the basis of morphometric differences in length and width of mature spores, length and width of polar capsules, and unique small-subunit (SSU) ribosomal (rDNA) sequence composition relative to other taxa. Replicate SSU rDNA sequences generated from Sph. horrida n. sp. collected from Sy. horrida and Sy. verrucosa in tropical north Queensland and from Sy. horrida in Moreton Bay were identical, suggesting that this species is widely distributed along the east coast of Australia. Myxidium lapipiscis n. sp. can be distinguished from the majority of described Myxidium species on the basis of its relatively small mature spore size (6.1-7.9 μm long × 3.1-3.9 μm wide), and its unique SSU rDNA sequence. Specimens putatively identified as M. lapipiscis n. sp. were found in Sy. horrida from both tropical north Queensland and Moreton Bay, suggesting that this taxon is also widely distributed along the east coast of Australia. However, no molecular data were available for the specimens from tropical north Queensland for comparative genetic analyses. Bayesian inference and maximum likelihood analysis of the SSU rDNA sequences for these 2 new species revealed that Sph. horrida n. sp. formed a strongly supported clade with Sphaeromyxa zaharoni Diamant, Whipps, and Kent, 2004, which was described from the scorpaeniform, Pterois miles, from the Red Sea. This is the first report of myxozoans infecting stonefish (Synanceiidae).

A Case Report on the Survivability of Marine Luminous Bacteria Vibrio campbellii STF1 under Starvation Conditions

Recently luminous Vibrio campbellii has been recognised as a cosmopolitan pathogen in seafood and aquaculture industry. The significance of this study is to present the findings derived from longer-term starvation experiment on intense luminescence producing bacteria Vibrio campbellii strain STF1. This strain was isolated from surface swab of Synanceia verrucosa (stone fish) and subjected to the starvation-survival procedure. The viability rate of this culture was determined following spread plate method by counting viable cells in starter inoculum and in the same starvation culture. It was observed that 90 to 95% of luminous bacteria could remain viable for over 4 years under nutrient deficiency condition. This study also determines the effectiveness of the preservation of luminous bacteria in sterile sweater.

Biochemical and histopathological effects of the stonefish (Synanceia verrucosa) venom in rats

The Reef Stonefish (Synanceia verrucosa) is one of the most dangerous venomous fish known, and has caused occasional human fatalities. The present study was designed to examine some of the pathological effects of the venom from this fish in Sprague Dawley rats. Crude venom was extracted from venom glands of the dorsal spines of stonefish specimens collected from coral reefs in the Gulf of Aqaba (in the northeastern branch of the Red Sea). The rats were given intramuscular injections of the venom and acute toxicity and effect on selected serum marker enzymes as well as normal architecture of vital organs were evaluated. The rat 24 h LD50 was 38 μg/kg body weight. The serum biochemical markers; alanine transaminase (ALT), lactate dehydrogenase (LDH) and creatine kinase (CK) increased after 6 h of administration of a sub lethal dose of the venom and remained significantly raised at 24 h. Amylase levels also significantly increased after venom injection. The venom caused histological damage manifested as an interstitial hemorrhage, inflammatory cell infiltration, and necrosis. The demonstrated rises in the levels of different critical biochemical parameters in the serum may have led to the observed abnormal morphological changes in these organs. These results may account for some of the clinical manifestations observed in victims of stonefish envenomation. Thus, the presented data provide further in vivo evidence of the stonefish toxic effects that may threaten human life and call for the need for special measures to be considered.

Purification of a novel lectin from the dorsal spines of the stonefish, Synanceia verrucosa

Purification of a novel lectin from the dorsal spines of the stonefish, Synanceia verrucosa

New Mediterranean Biodiversity Records (October 2015)

The Collective Article “New Mediterranean Biodiversity Records” of the Mediterranean Marine Science journal offers the means to publish biodiversity records in the Mediterranean Sea. The current article is divided per countries, listed according to a Mediterranean west-east geographic position. New biodiversity data are reported for 7 different countries, although one species hereby reported from Malta is overall new for the entire Mediterranean basin, and is presumably present also in Israel and Lebanon (see below in Malta). Italy: the rare native fish Gobius kolombatovici is first reported from the Ionian Sea, whilst the alien jellyfish Rhopilema nomadica and the alien fish Oplegnathus fasciatus are first reported from the entire country. The presence of O. fasciatus from Trieste is concomitantly the first for the entire Adriatic Sea. Finally, the alien bivalve Arcuatula senhousia is hereby first reported from Campania (Tyrrhenian Sea). Tunisia: a bloom of the alien crab Portunus segnis is first reported from the Gulf of Gabes, from where it was considered as casual. Malta: the alien flatworm Maritigrella fuscopunctata is first recorded from the Mediterranean Sea on the basis of 25 specimens. At the same time, web researches held possible unpublished records from Israel and Lebanon. The alien crab P. segnis, already mentioned above, is first formally reported from Malta based on specimens collected in 1972. Concomitantly, the presence of Callinectes sapidus in Maltese waters is excluded since based on misidentifications. Greece: the Atlantic northern brown shrimp Penaeus atzecus, previously known from the Ionian Sea from sporadic records only, is now well established in Greek and international Ionian waters. The alien sea urchin Diadema setosum is reported from the second time from Greece, and its first record date from the country is backdated to 2010 in Rhodes Island. The alien lionfish Pterois miles is first reported from Greece and concomitantly from the entire Aegean Sea. Turkey: the alien rhodophyte Antithamnion hubbsii is first recorded from Turkey and the entire eastern Mediterranean. New distributional data are also offered for the native fishes Alectis alexandrina and Heptranchias perlo. In particular, the former record is constituted by a juvenile of 21.38 mm total length, whilst the latter by a mature male. Cyprus: the rare native cephalopod Macrotritopus defilippi, and the alien crab Atergatis roseus, sea slug Plocamopherus ocellatus and fish Cheilodipterus novemstriatus are first recorded from the entire country. Lebanon: the alien crabs Actaea savignii and Matuta victor, as well as the alien fish Synanceia verrucosa, are first recorded from the entire country. In addition, the first Mediterranean record of A. savignii is backdated to 2006, whilst the high number of M. victor specimens observed in Lebanon first suggest its establishment in the basin. The Atlantic fishes Paranthias furcifer and Seriola fasciata, and the circumtropical Rachycentron canadum, are also first reported from the country. P. furcifer record backdate its presence in the Mediterranean to 2007, whilst S. fasciata records backdate its presence in the eastern Mediterranean to 2005. Finally, two of these latter species have been recently ascribed to alien species, but all the three species may better fit the cryptogenic category, if not a new one.

Proteomic Identification of Stonefish Synanceja verrucosa Venom

Three venom toxins, neoverrucotoxin (neoVTX) α-subunit and β-subunit as well as verrucotoxin (VTX) β-subunit, were identified in the stonefish Synanceja verrucosa by SDS-PAGE, Native-PAGE and two-dimensional electrophoresis (2-DE) coupled with Matrix Assisted Laser Desorption Ionization-Quadrupole-Time-of-Flight (MALDI-Q-TOF). The venom estimated by Native-PAGE were 471, 358, 260 and 166 kDa. The predominate protein bands of crude venom were 84 and 75 kDa by SDS-PAGE. The crude venom protein fell in the region with pI values of 7-9 and molecular weights of 75-90 kDa by 2-DE. Peptide mass fingerprints (PMF) and MS/MS ions originated from MALDI-Q-TOF were used to identify the protein. Our results showed that the complete components of neoverrucotoxin (neoVTX) α-subunit and β-subunit as well as verrucotoxin (VTX) β-subunit were identified from SDS-PAGE and 2-DE patterns. Native-PAGE did not yield protein identifications but revealed the presence of protein complexes.

The complete mitochondrial genome of the Synanceia verrucosa (Scorpaeniformes: Synanceiidae)

The complete mitochondrial genome of the Synanceia verrucosa has been sequenced. The mitochondrial genome is 16,506 bp in length, containing 13 protein-coding genes, two ribosomal RNA genes, 22 transfer RNA genes, and one control region. The gene order and the composition of S. verrucosa mitochondrial genome were similar to that of most other vertebrates. The overall nucleotides base composition of the heavy strand is A (31.01%), G (15.06%), C (25.60%), and T (28.34%). With the exception of the NADH dehydrogenase subunit 6 (ND6) and eight tRNA genes, all other mitochondrial genes are encoded on the heavy strand. The tRNA-Ser2 gene lacked DHC arm and could not fold into a typical clover-leaf secondary structure. Seen from the phylogenetic tree, a stonefish (S. verrucosa), two lionfishes, and eight rockfishes from the same order (Scorpaeniformes) clustered into one branch.

Toxin gene determination and evolution in scorpaenoid fish

In this study, we determine the toxin genes from both cDNA and genomic DNA of four scorpaenoid fish and reconstruct their evolutionary relationship. The deduced protein sequences of the two toxin subunits in Sebastapistes strongia, Scorpaenopsis oxycephala, and Sebastiscus marmoratus are about 700 amino acid, similar to the sizes of the stonefish (Synanceia horrida, and Synanceia verrucosa) and lionfish (Pterois antennata and Pterois volitans) toxins previously published. The intron positions are highly conserved among these species, which indicate the applicability of gene finding by using genomic DNA template. The phylogenetic analysis shows that the two toxin subunits were duplicated prior to the speciation of Scorpaenoidei. The precedence of the gene duplication over speciation indicates that the toxin genes may be common to the whole family of Scorpaeniform. Furthermore, one additional toxin gene has been determined in the genomic DNA of Dendrochirus zebra. The phylogenetic analysis suggests that an additional gene duplication occurred before the speciation of the lionfish (Pteroinae) and a pseudogene may be generally present in the lineage of lionfish.