Response to Britz (2022) regarding the validity of the giant sunfish Mola alexandrini (Ranzani, 1834) (Teleostei: Molidae).

The small family of ocean sunfishes has a long and complicated taxonomic and nomenclatorial history dating back several centuries. Most recently, Britz (2022) questioned the validity of Mola alexandrini (Ranzani, 1839) as the valid name for Mola sp. A sensu Yoshita et al. (2009). Specifically, he questioned the authenticity of the purported Orthragoriscus alexandrini holotype rediscovered by Sawai et al. (2018) due to uncertainties regarding the original units of measurement, along with discrepancies between the original illustration and the purported holotype. He also questioned the reliability of the illustration to reflect the fresh morphology, and the adequacy of the species description (sans illustration and holotype) to link it to any of the currently recognized Mola species. In summary, Britz (2022) opined that M. alexandrini is a species inquirenda. Here we respond to Britzs concerns with the findings from an additional literature review. Firstly, the original species description appears to give measurements in Parisian units, yielding a discrepancy in length with the purported holotype within human error. However, due to various uncertainties, length does not appear to be a robust piece of evidence to neither confirm nor refute the specimens authenticity. Secondly, the morphological differences between the original illustration and the purported holotype were found to be within the expected level of accuracy for Mola illustrations at the time, by both Ranzani and others. Thirdly, a contemporaneous publication describes how Ranzani himself bought the specimen which became the Or. alexandrini holotype, suggesting he would likely have seen it in its fresh state, rendering it unlikely he would have produced a species description and illustration incongruent with this. Fourthly, re-examining the original species description suggests this provides adequate and sufficient information to link Or. alexandrini to Mola sp. A, even in the absence of a holotype. Finally, during this review, we came upon an even older paper describing Or. alexandrini, and we conclude that the currently valid name for Mola sp. A sensu Yoshita et al. (2009) is Mola alexandrini (Ranzani, 1834).

Water column structure influences long-distance latitudinal migration patterns and habitat use of bumphead sunfish Mola alexandrini in the Pacific Ocean.

Satellite-tracking of adult bumphead sunfish, Mola alexandrini, revealed long-distance latitudinal migration patterns covering thousands of kilometers. Horizontal and vertical movements of four bumphead sunfish off Taiwan were recorded with pop-up satellite archival tags in 2019-2020. Two individuals moved northward and traveled to Okinawa Island and Kyushu, Japan and two moved southwards; crossing the equator, to Papua New Guinea and New Caledonia. During daytime, bumphead sunfish descended below the thermocline and ascended to mixed layer depths (MLD) during nighttime. The N-S migrants, however, demonstrated different habitat utilization patterns. Instead of using prevailing currents, the northward movements of sunfish cohorts exhibited extensive use of mesoscale eddies. Fish in anticyclonic eddies usually occupied deeper habitats whereas those in cyclonic eddies used near-surface habitats. On northward excursions, fish spent most of their time in regions with high dissolved oxygen concentrations. Southward movement patterns were associated with major currents and thermal stratification of the water column. In highly stratified regions, fish stayed below the thermocline and frequently ascended to MLD during daytime either to warm muscles or repay oxygen debts. These results for bumphead sunfish present important insights into different habitat use patterns and the ability to undergo long-distance migrations over varying spatial-temporal scales and features.

Age and growth of grey triggerfish Balistes capriscus from trans-Atlantic populations.

Anthropogenic factors that negatively impact reef fishes can include changes in life-history patterns of fisheries-targeted species. Understanding these impacts on growth and population age structure is essential in the management of exploited populations of fishes. This is the first study to directly compare age and growth for a major fisheries species between east and west populations of a transatlantic reef fish. The main goal of this study was to document age and growth in grey triggerfish Balistes capriscus from coastal waters of Ghana in the Gulf of Guinea (GOG) and compare those with the previous growth studies from that region and with the western Atlantic population. A secondary objective of this study was to evaluate the use of otoliths to age triggerfish and to provide a preliminary comparison with spine-derived age estimates. The results obtained from this study provided an updated understanding of the growth and age structure of the eastern B. capriscus population in GOG. The authors documented that shifts in population attributes occurred for B. capriscus after its major decline in abundance. The differences in physical and biotic characteristics of the East and West Atlantic regions and the differences in collection methods of samples make direct comparisons of growth parameters difficult. Nonetheless, overall differences in maximum sizes and ages were apparent; the western Atlantic population had a larger maximum size and older maximum age. The authors also documented that sagittal otoliths can be used to provide age estimates for triggerfish species, and otoliths as an ageing structure had better between-reader precision compared to dorsal spines.

Sound production, hearing sensitivity, and in-depth study of the sound-producing muscles in the cowfish (Lactoria cornuta).

The ability to produce sounds has been reported in various Ostraciidae but not deeply studied. In some Ostracion species, two different sound-producing muscles allow these boxfishes to produce two different kinds of sounds in a sequence. This study investigates sound production in another Indo-Pacific species, the longhorn cowfish Lactoria cornuta that also possesses two pairs of sonic muscles associated with the swim bladder: extrinsic sonic muscles (ESMs) and intrinsic sonic muscles (ISMs). The cowfish produces two kinds of sounds called hums and clicks. Hums are made of trains of low amplitude pulses that last for long periods of time, suggesting that they are produced by fatigue-resistant muscles, whereas clicks correspond to shorter sounds with greater amplitude than the hums, suggesting that they result from more powerful contractions. Ultra-structural differences are found between extrinsic and intrinsic sonic muscles. According to features such as long sarcomeres, long I-bands, a high number of mitochondria, and a proliferation of sarcoplasmic reticulum (SR), ESMs would be able to produce fast, strong, and short contractions corresponding to clicks (the shortest sounds with the greatest amplitude). ISMs have the thinnest cells, the smallest number of myofilaments that have long I-bands, the highest volume of mitochondria, and well-developed SR supporting these muscles; these features should generate fast and prolonged contractions that could correspond to the hums that can be produced over long periods of time. A concluding figure shows clear comparisons of the different fibers that were studied in L. cornuta. This study also compared the call features of each sound with the cowfish's hearing ability and supports L. cornuta was more sensitive to frequencies ranging between at least 100 and 400 Hz with thresholds of 128-143 dB re 1 µPa over this range, meaning that they are sensitive to the frequencies produced by conspecifics.

Intentional partial beaching in a coral reef fish: a newly recorded hunting behaviour of titan triggerfish, Balistoides viridescens.

Coral reef fishes use a multitude of diverse feeding behaviours to increase their ability to successfully capture a wide range of prey. Here, this study reports a novel hunting behaviour in a coral reef fish, the titan triggerfish, Balistoides viridescens, where an individual was seen partially beaching itself while attempting to catch a Red Sea ghost crab, Ocypode saratan. This is the first report of this behaviour in the order Tetraodontiformes and represents an astonishing capability of this species to exploit food resources outside their typical assumed ecological niche.

Does conspicuousness scale linearly with colour distance? A test using reef fish.

To be effective, animal colour signals must attract attention-and therefore need to be conspicuous. To understand the signal function, it is useful to evaluate their conspicuousness to relevant viewers under various environmental conditions, including when visual scenes are cluttered by objects of varying colour. A widely used metric of colour difference (ΔS) is based on the receptor noise limited (RNL) model, which was originally proposed to determine when two similar colours appear different from one another, termed the discrimination threshold (or just noticeable difference). Estimates of the perceptual distances between colours that exceed this threshold-termed 'suprathreshold' colour differences-often assume that a colour's conspicuousness scales linearly with colour distance, and that this scale is independent of the direction in colour space. Currently, there is little behavioural evidence to support these assumptions. This study evaluated the relationship between ΔS and conspicuousness in suprathreshold colours using an Ishihara-style test with a coral reef fish, Rhinecanthus aculeatus. As our measure of conspicuousness, we tested whether fish, when presented with two colourful targets, preferred to peck at the one with a greater ΔS - from the average distractor colour. We found the relationship between ΔS and conspicuousness followed-- a sigmoidal function, with high ΔS colours perceived as equally conspicuous. We found that the relationship between ΔS and conspicuousness varied across colour space (i.e. for different hues). The sigmoidal detectability curve was little affected by colour variation in the background or when colour distance was calculated using a model that does not incorporate receptor noise. These results suggest that the RNL model may provide accurate estimates for perceptual distance for small suprathreshold distance colours, even in complex viewing environments, but must be used with caution with perceptual distances exceeding- -10 ΔS.

Mystery pufferfish create elaborate circular nests at mesophotic depths in Australia.

In 2011, the enigma of "mystery circles," small but complex underwater structures first observed by divers from southern Japan in 1995, was solved when a new species of pufferfish, white-spotted pufferfish (Torquigener albomaculosus Matsuura 2014), was identified as the responsible agent. To date these circles have been described only from Japan, where they are formed on a sandy seafloor in water depths less than 30 m. A survey of oil field infrastructure on the North West Shelf of Western Australia in 2018 using a remotely operated vehicle and hybrid autonomous underwater vehicle (HAUV) recorded a high-resolution video and bathymetric data of 21 circular formations with similar features to those described in Japan. The circles display dimensions and morphology like those described from Japan, but were observed in water depths between 129 and 137 m. The HAUV also recorded high-resolution photographs which captured a Torquigener sp. fish in the immediate vicinity of the circles. An additional circle and Torquigener sp. were observed in images collected by baited remote underwater stereo-video in a nearby location in 129 m depth. These circles are the first to be found in Australia. The pufferfish species responsible cannot be identified from images collected. Such a discovery not only generates intrigue and wonder among scientists and the general public but also provides an insight into the reproductive behaviour and evolution of pufferfish globally.

Aracaniform Swimming: A Proposed New Category of Swimming Mode in Bony Fishes (Teleostei: Tetraodontiformes: Aracanidae).

The deepwater boxfishes of the family Aracanidae are the phylogenetic sister group of the shallow-water, generally more tropical boxfishes of the family Ostraciidae. Both families are among the most derived groups of teleosts. All members of both families have armored bodies, the forward 70% of which are enclosed in rigid bony boxes (carapaces). There is substantial intragroup variation in both groups in body shapes, sizes, and ornamentation of the carapaces. Swimming-related morphology, swimming mode, biomechanics, kinematics, and hydrodynamics have been studied in detail in multiple species of the ostraciids. Ostraciids are all relatively high-performance median and paired fin swimmers. They are highly maneuverable. They swim rectilinearly with substantial dynamic stability and efficiency. Aracanids have not been previously studied in these respects. This article describes swimming-related aspects of morphology, swimming modes, biomechanics, and kinematics in two south Australian species (striped cowfish and ornate cowfish) that are possibly representative of the entire group. These species differ morphologically in many respects, both from each other and from ostraciids. There are differences in numbers, sizes, and placements of keels on carapaces. The most important differences from ostraciids are openings in the posterior edges of the carapaces behind the dorsal and anal fins. The bases of those fins in ostraciids are enclosed in bone. The openings in aracanids free the fins and tail to move. As a result, aracanids are body and caudal fin swimmers. Their overall swimming performances are less stable, efficient, and effective. We propose establishing a new category of swimming mode for bony fishes called "aracaniform swimming."

The paraneuronal gill neuroendocrine system in ocellated puffer fish Leiodon cutcutia.

Pseudobranchial neurosecretory system (PSNS) is the third Neuroendocrine (NE) system found in the gill region of fishes in close association with pseudobranch/carotid labyrinth/carotid gland and can suitably be placed under the category of "Diffused NE system (DNES)." The cells belonging to this system fall under the category of "Paraneurons," a concept proposed by Fujita and coworkers. It is found uniformly in all the catfish species and some other noncatfish group of teleosts as Atheriniformes, Channiformes, Perciformes, and Clupeiformes. The fishes, in which the PSNS is present, belong to different breathing habits. Most of these have the capacity to tolerate low O2 conditions. Leiodon cutcutia although not an air-breathing fish, is known to retain air in its stomach for varied periods when threatened. In an attempt to verify the veracity of this system in a fish of another peculiar breathing habit, ocellated puffer fish L. cutcutia (order Tetradontiformes) was investigated. The histological observations undertaken on L. cutcutia revealed the presence of a well-developed extrabranchial NE system. The findings are discussed in the light of the association of PSNS with chemosensory system and its evolution in fishes, especially in the view of the transition from aquatic to terrestrial life.

New tetrodotoxin analogs in Brazilian pufferfishes tissues and microbiome.

While tetrodotoxin (TTX) is commonly found in pufferfish tissues, it is unclear if bacterial symbionts isolated from pufferfish tissues can produce TTX. In this investigation, UPLC qTOF-MS/MS analysis of tissue extracts obtained from Sphoeroides spengleri and Canthigaster figuereidoi identified TTX in their composition, indicating their consumption is unsafe. UPLC qTOF-MS/MS analysis coupled with Molecular Networking indicated new TTX analogs (methyl-TTX, TTX-acetate, hydroxypropyl-TTX and glycerol-TTX). Bacterial extracts from sixteen strains revealed a compound with a [M+H]+ ion at m/z 320.1088, identical to TTX. However, TTX itself was not detected in these cultures by UPLC-MS/MS. Neurotoxicity of Vibrio A665 purified fraction 2 (with precursor [M+H]+ ion at m/z 320.1088) was significant in human neural stem cells (hNSCs), but the Nav blockage activity was not confirmed by the veratridine/ouabain essays, indicating a possible difference in the mechanism of action between the bacterium A665 purified fraction 2 and TTX. Vibrios symbionts of pufferfish point out involving in the production of TTX precursors.

Adaptative responses of myenteric neurons of Sphoeroides testudineus to environmental pollution.

Contamination in estuarine regions affects the local biota damaging the ecosystems and reaching humans. The gastrointestinal tract is a dynamic environment capable of obtaining nutrients and energy from food while it protects the host against harmful toxins and pathogens from the external environment. These functions are modulated by the enteric nervous system and changes in its structure can result in gastrointestinal disorders. The objective of this study was to evaluate if the environmental contaminants have effects on the myenteric neuronal plasticity of pufferfish Sphoeroides testudineus. Animals were collected in Barra do Una River, located at Jureia-Itatins Mosaic of Protected Areas (reference area - RA) and in the Santos Estuarine System (impacted area - IA). Morpho-quantitative analyses of the general and metabolically active myenteric neuronal populations of the proximal and distal intestine were made. Disarrangement was observed in the general organization of the myenteric plexus, with an expressive reduction of the neuronal groups (nodes) in the animals of IA. The vulnerability of the myenteric plexus was evidenced by a decrease in density and cellular profile of the general neuronal population, followed by an increase of the metabolism of the remaining neurons, which in turn was verified by a growth of the area of the cellular and nuclear profiles of the metabolically active neuronal population. Through these analyses, we concluded that animals inhabiting polluted regions present alterations in the myenteric neuronal plasticity, as a way of maintaining the functions of the gastrointestinal tract.

Muscle extract of Arothron immaculatus regulates the blood glucose level and the antioxidant system in high-fat diet and streptozotocin induced diabetic rats.

In the present study pufferfish, Arothron immaculatus muscle methanol extract (AIME) was used to evaluate the antidiabetic activity against the high-fat diet (HFD) in streptozotocin (STZ) induced diabetic rat models. Initially, the In vitro antioxidant activity of the different muscle extract was evaluated which showed that AIME has higher efficiency to scavenge the free radicals. The animal study results revealed that the AIME could decrease the blood glucose level after 14 days of oral treatment and recover the animal from the severe progression of the disease. The LC-ESI/MS analysis of AIME extract revealed the presence of compounds such as docosahexaenoic acid, adrenic acid, docosanol, codeine and metoprolol. Among these compounds, docosahexaenoic acid, adrenic acid and docosanol are reported for its antidiabetic studies. Hence, the muscle is recommended to consume by humans as natural food in order to overcome the development of diabetes. This is the first study on the muscle extract of marine pufferfish which is used as antidiabetic agent to treat the diabetes-induced in the animal model.

Functional morphology of endurance swimming performance and gait transition strategies in balistoid fishes.

Triggerfishes and filefishes (Balistoidea) use balistiform locomotion to power steady swimming with their dorsal and anal fins, and transition to a gait dominated by body and caudal fin (BCF) kinematics at high speeds. Fin and body shapes are predicted to be strong determinants of swimming performance and gait transitions. The goal of this study was to combine morphometrics and critical swimming tests to explore the relationships between fin and body shapes and swimming performance in a phylogenetic context in order to understand the evolution of balistiform swimming. Among 13 species of balistoid fishes, those with high aspect ratio fins tended to achieve higher critical swimming speeds than fishes with low aspect ratio fins. Species with long, large median fins and wide caudal peduncles used the balistiform gait alone for a larger percentage of their total critical swimming speed than fishes with short, small median fins and narrow caudal peduncles. Although analyses revealed overall positive relationships between median fin aspect ratios and gait transition speeds, fishes on both ends of the aspect ratio spectrum achieved higher swimming speeds using the balistiform gait alone than fishes with median fins of intermediate aspect ratios. Each species is specialized for taking advantage of one gait, with balistiform specialists possessing long, large median fins capable of the large power requirements of high-speed swimming using the median fins alone, while BCF specialists possess short, small median fins, ill-suited for powering high-speed balistiform locomotion, but narrow caudal peduncles capable of efficient caudal fin oscillations to power high-speed locomotion.

Life history, distribution and molecular phylogenetics of the Upward-Mouth Spikefish Atrophacanthus japonicus (Teleostei: Tetraodontiformes: Triacanthodidae).

Ninety-six juvenile specimens (37-54 mm standard length; LS ) of the rarely collected Upward-Mouth Spikefish Atrophacanthus japonicus (Triacanthodidae) were obtained from the stomachs of three Yellowfin Tuna Thunnus albacares collected off Guam in the Mariana Islands in the central Pacific Ocean. These specimens extend the range of A. japonicus eastward into Oceania. We review the systematic characters of the monotypic genus Atrophacanthus and present colour photographs of freshly collected specimens. The diet of the juvenile specimens of A. japonicus consisted of thecosome pteropods and foraminiferans. We present a range map of A. japonicus based on all known specimens and show that specimen size is related to whether specimens were collected in the pelagic zone or on the bottom. Our results support that, compared to all other Triacanthodidae, A. japonicus has an unusually extended pelagic larval and juvenile period, up to 54 mm LS , before settling to the bottom as adults. Lastly, we provide a multilocus phylogeny addressing the phylogenetic placement of Atrophacanthus based on eight of 11 triacanthodid genera and six genetic markers. Our results reveal that Atrophacanthus is the sister group of Macrorhamphosodes and they provide new insights about the evolutionary history of the family.

An Ishihara-style test of animal colour vision.

Colour vision mediates ecologically relevant tasks for many animals, such as mate choice, foraging and predator avoidance. However, our understanding of animal colour perception is largely derived from human psychophysics, and behavioural tests of non-human animals are required to understand how colour signals are perceived. Here, we introduce a novel test of colour vision in animals inspired by the Ishihara colour charts, which are widely used to identify human colour deficiencies. In our method, distractor dots have a fixed chromaticity (hue and saturation) but vary in luminance. Animals can be trained to find single target dots that differ from distractor dots in chromaticity. We provide MATLAB code for creating these stimuli, which can be modified for use with different animals. We demonstrate the success of this method with triggerfish, Rhinecanthus aculeatus, which quickly learnt to select target dots that differed from distractor dots, and highlight behavioural parameters that can be measured, including success of finding the target dot, time to detection and error rate. We calculated discrimination thresholds by testing whether target colours that were of increasing colour distances (ΔS) from distractor dots could be detected, and calculated discrimination thresholds in different directions of colour space. At least for some colours, thresholds indicated better discrimination than expected from the receptor noise limited (RNL) model assuming 5% Weber fraction for the long-wavelength cone. This methodology could be used with other animals to address questions such as luminance thresholds, sensory bias, effects of sensory noise, colour categorization and saliency.

Energetics and behavior of coral reef fishes during oscillatory swimming in a simulated wave surge.

Oxygen consumption rates were measured for coral reef fishes during swimming in a bidirectional, oscillatory pattern to simulate station-holding in wave-induced, shallow-water flows. For all species examined, increases in wave intensity, as simulated by increases in frequency and amplitude of oscillation, yielded increased metabolic rates and net costs of swimming (NCOS; swimming metabolic rate minus standard metabolic rate). Comparing species with different swimming modes, the caudal fin swimming Kuhlia spp. (Kuhliidae) and simultaneous pectoral-caudal fin swimming Amphiprion ocellaris (Pomacentridae) turned around to face the direction of swimming most of the time, whereas the median-paired fin (MPF) swimmers, the pectoral fin swimming Ctenochaetus strigosus (Acanthuridae) and dorsal-anal fin swimming Sufflamen bursa (Balistidae), more frequently swam in reverse for one half of the oscillation to avoid turning. Contrary to expectations, the body-caudal fin (BCF) swimming Kuhlia spp. had the lowest overall NCOS in the oscillatory swimming regime compared with the MPF swimmers. However, when examining the effect of increasing frequency of oscillation at similar average velocities, Kuhlia spp. showed a 24% increase in NCOS with a 50% increase in direction changes and accelerations. The two strict MPF swimmers had lower increases on average, suggestive of reduced added costs with increasing frequency of direction changes with this swimming mode. Further studies are needed on the costs of unsteady swimming to determine whether these differences can explain the observed prevalence of fishes using the MPF pectoral fin swimming mode in reef habitats exposed to high, wave-surge-induced water flows.

Anatomical characterization of the digestive system of the pufferfish (Chilomycterus spinosus spinosus)

Descriptive studies of the fish digestive system are fundamental because they provide information on the biology of the species. Thus, the objective of this study was to morphologically describe the digestive system of the pufferfish, Chilomycterus spinosus spinosus. For this, adult specimens of pufferfish (n = 10) of both sexes were used. The animals were fixed with 10% aqueous formaldehyde solution, dissected, analyzed descriptively and photographed. The results demonstrate that the pufferfish has a morphologically modified digestive system, which is adapted to the defense behavior. This species presents a pouch-shaped diverticulum, that is called abdominal pouch, which allows the expansion of the celomatic cavity and the temporary storage of food. Although it is used to store food, macroscopically the abdominal pouch does not show gastric folds. However, this absence is compensated by a small intestine containing innumerable villi.

The effect of Peruvian small-scale fisheries on sunfishes (Molidae).

Reports were compiled of sunfish (family Molidae) by-catch in Peruvian small-scale fisheries and sunfish by-catch rates were estimated using data from shore-based and onboard monitoring programmes. A total of 114 sunfishes were reported in the longline and gillnet fisheries along the Peru coast from 2005 to 2017. Systematic monitoring effort of small-scale gillnets leads to an estimate of between 23 and 352 individuals captured annually by the fleet fishing from the northern port of Salaverry and central ports of Ancon and Chorrillos and suggests that the actual number captured by the Peruvian gillnet fleet is in the thousands of individuals. Thus, Peruvian small-scale fisheries have the potential to greatly affect populations of these still poorly studied species. Moreover, new occurrence locations are reported for the newly described Mola tecta, which was only observed south of 11° S. Because of physical similarities among Mola species it was difficult to identify sunfishes to the species level and thus further studies (e.g., genetics) will be required to provide more detailed information on individual species vulnerability to by-catch in Peruvian waters.

Demographics of Lagocephalus inermis in the Arabian Sea unveils complex conservation challenges.

The first assessment of the demographics of Lagocephalus inermis, a species associated with pufferfish bites and fishing down the food web in the Arabian Sea, south-west coast of India, was performed based on length structured population dynamics of 1601 individuals caught in commercial fisheries. Analysis revealed that the current level of exploitation is 90% of the predicted exploitation producing maximum relative yield per recruit (Emax ), suggesting high levels of exploitation with potential for significant negative consequences for trophic cascades.

Including planocerid flatworms in the diet effectively toxifies the pufferfish, Takifugu niphobles.

Beginning with the larval stages, marine pufferfish such as Takifugu niphobles contain tetrodotoxin (TTX), an extremely potent neurotoxin. Although highly concentrated TTX has been detected in adults and juveniles of these fish, the source of the toxin has remained unclear. Here we show that TTX in the flatworm Planocera multitentaculata contributes to the toxification of the pufferfish throughout the life cycle of the flatworm. A species-specific PCR method was developed for the flatworm, and the specific DNA fragment was detected in the digesta of wild pufferfish adults. Predation experiments showed that flatworm larvae were eaten by the pufferfish juveniles, and that the two-day postprandial TTX content in these pufferfish was 20-50 µg/g. Predation experiments additionally showed flatworm adults were also eaten by pufferfish young, and after two days of feeding, TTX accumulated in the skin, liver and intestine of the pufferfish.