Swim Bladder Research Articles

Development of sound production in Danionella cerebrum.

Acoustic signalling, integral to intraspecific communication and reproductive behaviour, undergoes notable changes during an animal's ontogenetic development. The onset and progression of this maturation in fish remains poorly understood. Here, we investigated the ontogeny of acoustic communication in the miniature teleost Danionella cerebrum, one of the smallest known vertebrates and an emerging model organism. Its adult males produce audible clicks that appear in sequences with a repetition rate of ∼60 or ∼120 Hz, caused by consecutive unilateral or alternating bilateral compressions of the swim bladder. To investigate the maturation of this ability, we performed long-term sound recordings and morphological studies of the sound production apparatus in D. cerebrum throughout its ontogenetic development. We found that fish start producing clicks during the second month of their lives and continually increase their abundance and structured repetition over the course of the following 1 to 2months. The sound production machinery, including specialised bone and cartilage structures, starts to form in males after approximately 4weeks and prior to reaching sexual maturity. Although clicks increase in amplitude as animals mature, click repetition rates of 60 and 120 Hz are stable throughout development. This suggests fully mature pattern generation in juvenile males, yet a continued development of the drumming apparatus capable of creating louder sounds.

Post-smolt Atlantic salmon can regulate buoyancy in submerged sea-cages by gulping air bubbles

Salmon lice (Lepeophtheirus salmonis) are the most severe health challenge facing Atlantic salmon (Salmo salar) aquaculture. Infestations threaten farmed and wild salmonids, drive up production costs, and limit industry growth. Submersible sea-cages can reduce infestation pressure by holding salmon below the surface layers where lice larvae are most abundant. However, submergence leads to loss of buoyancy over time, as salmonids must periodically refill their swim bladders by gulping air. Pockets of air held at depth within ‘air domes’ enable salmon to gulp air, but air domes are large and unwieldy structures. A more intuitive solution may be to deliver a stream of air bubbles at depth, as bubbling is commonly used in sea-cages to promote vertical mixing, and salmon interact with these bubbles. Here, we conducted a pilot study to test whether newly sea-transferred salmon would use bubbles for buoyancy regulation. We conducted 3 trials in sequence, each involving 15–19 days of continuous submergence, and recorded buoyancy-related behaviors in 1 cage with full surface access, 1 cage submerged without air, and 2 cages submerged with air bubbling. Bubbling was provided continuously in the first two trials, before being reduced to 90 min day−1 in the third trial. Salmon submerged with bubbling maintained their swim bladder volume and swimming speed at similar levels to the cage with full surface access throughout all trials, indicating that they successfully refilled using bubbles. The same parameters deteriorated in salmon in the cage submerged without air. Acclimation to bubbles took ∼3 days whether at the surface or after submergence, indicating that several days of bubbling before submergence may be beneficial. Some fish accumulated fluid in the swim bladder during submergence (more severe in fish submerged without bubbling). Together, the results suggest that bubbling can improve outcomes for salmon undergoing prolonged submergence. Moreover, bubbling offers an alternative to air domes that is less technically demanding, provides a greater spatial distribution of air, and supports welfare through behavioral enrichment.

PHYSICAL STABILITY AND SAFETY TESTING OF COSMETIC FORMULATIONS CONTAINING COLLAGEN FROM CATFISH (PANGASIUS SP.) SWIM BLADDER

Objective: Collagen is a fibrous protein that confers strength and flexibility on bone and other body tissues, such as the skin and tendons, and is the main component of the extracellular matrix. It is used in cosmetics for its anti-aging activity. The primary source of commercial collagen is pig skin, but its use is imperfect due to halal concerns and the risk of biological contamination. Therefore, this research was conducted to identify alternate sources of collagen derived from fish. This study used the swim bladders of catfish that were not consumed and sought to separate the collagen from and use it in a safe anti-ageing serum for humans. Methods: Three formulations of 1%, 2%, and 3% collagen were developed in the laboratory using experimental methods. The three formulations were then evaluated for physical stability and in vivo irritation. Results: Physical quality testing showed that the 1%, 2%, and 3% formulations remained stable at room, low, and high temperatures. However, in the irritation test, the 2% and 3% formulations resulted in mild erythema after 24, 48, and 72 h under constant conditions. Conclusion: The 1% formula was confirmed stable and safe.

From hatching to juvenile: Larval development of Vieja fenestrata (Teleostei: Cichlidae).

This study delves into the early development of Vieja fenestrata (Cichlidae), with a specific focus on the description of external morphological and morphometric changes, and growth patterns from hatching to the loss of larval characters under controlled laboratory conditions at a temperature of 28°C. Asynchronous hatching was observed between 58 and 60 h postfertilization, with the posterior body emerging first. Over 14 days, significant morphological, physiological, and behavioral changes were observed, revealing a complex developmental trajectory. The initial developmental phases were characterized by rapid vascularization, fin differentiation, and heightened activity, and the subsequent days witnessed the flexion of the notochord, emergence of swim bladder functionality, and transition to exogenous feeding. Maturation progressed with the absorption of the yolk sac, regression of cement glands, and fin ray development, culminating in metamorphosis by 14 days post-hatching. Throughout this period, evolving pigmentation patterns and structural adaptations highlight the species' adaptive strategies. During the larval period of V. fenestrata, substantial changes in morphological proportions were observed. Before the inflection, tail length, trunk length, and body depth had negative allometric growth, and head length, eye diameter, and snout length had positive allometric growth. After the inflection, body depth and snout length showed positive allometric growth; head length and trunk length exhibited isometric growth, whereas tail length and eye diameter demonstrated negative allometric growth. These findings contribute insights into the intricate developmental dynamics of V. fenestrata. Moreover, further research may explore these developmental dynamics' ecological and evolutionary implications.

Species authentication and conservation challenges in Chinese fish maw market using Mini-DNA barcoding

Fish maws, derived from the swim bladders of fish, are valued for their high collagen content and nutritional benefits, leading to significant market demand, especially in Asia. This study investigates the species authenticity of fish maw products using mini-DNA barcoding technology to address issues of mislabeling and substitution. Out of 350 samples, 347 were successfully sequenced, revealing a high mislabeling rate of 51.01% among identified species. Species such as Diodon liturosus and Diodon hystrix were found, posing significant health risks due to the presence of tetrodotoxin, a potent neurotoxin. The study also highlights the conservation implications of this market, with endangered species like Totoaba macdonaldi being illicitly traded. These findings underscore the urgent need for accurate species identification, stringent labeling regulations, and enhanced fishery management to protect endangered species and ensure consumer safety. This research advocates for robust international cooperation and reporting to combat illegal fishing and trade, promoting the sustainable development of fish populations and marine ecosystem health.

Attenuation of arsenic-induced gut injury in mice by sulfated glycosaminoglycan from swim bladder via regulation of gut microbiota, metabolites, and intestinal barrier integrity

Attenuation of arsenic-induced gut injury in mice by sulfated glycosaminoglycan from swim bladder via regulation of gut microbiota, metabolites, and intestinal barrier integrity

Collagen extraction and physicochemical characterization of animal by-products

Collagen is the most abundant structural protein in animals, being the main component of the skin, and can be researched and utilized by the cosmetics, pharmaceutical, food, and packaging industries. The present study investigates the extraction and quantification of collagen from three different animal sources: beef mocotó, chicken feet, and tambaqui swim bladder. Using the method of extraction in soluble acid solution (acetic acid) and precipitation in sodium chloride saline solution, collagen yields and their physicochemical properties were determined. The results revealed that the tambaqui swim bladder, especially the outer layer, presented the highest yield (39.97%), followed by chicken feet (3.49%) and bovine mocotó (1.79%). This study highlights the viability of these sources for the production of high-quality collagen, with potential applications in the cosmetics, pharmaceutical, and biomaterials industries, promoting a sustainable and economical approach to the use of animal by-products.

Early life-history stages of the olive barb, Systomus sarana (Hamilton), and a key ontogenetic systematic character of Smiliogastrinae.

The current study aims to uncover the early life-history stages of Systomus sarana, a medium-sized smiliogastrin cyprinid important for aquaculture in South Asia. The fish were effectively bred in captivity by administering 0.25 mL of breeding hormone per kilogram of fish. The spawning occurred 8.54 ± 0.55 h after the injection, and the eggs were phyto-lithophilic with a pale yellow color and a diameter of 1.49 ± 0.04 mm. Hatching occurred 17 h after fertilization, and the yolk-sac larvae of 3.43 ± 0.08 mm total length (TL) were adhering to the plant parts and other substrata with the cement glands on the forehead. On the third day, with complete absorption of the yolk sac and the disappearance of the attachment organ, the pre-flexion larvae measured 5.3 ± 0.11 mm TL. On the eighth day, the flexion larvae measured 6 ± 0.4 mm TL with a well-inflated posterior swim bladder, and the post-flexion larvae, at 11 days post-hatching (dph), developed a two-chambered gas bladder. The juvenile stage, on day 21 post-hatching, was marked by the loss of the median finfolds and the appearance of black blotches on the caudal, subdorsal, and supra-anal regions. The commencement of squamation and the appearance of the rudiments of maxillary barbels distinguished the juvenile stage. The subadults measuring 4.6 ± 0.36 cm TL had finished squamation and completely lost the subdorsal and supra-anal blotches. We propose that the presence of subdorsal blotches is a distinctive ontogenetic and systematic feature of larval and juvenile forms of smiliogastrin barbs.

Using Zebrafish to Screen Developmental Toxicity of Per- and Polyfluoroalkyl Substances (PFAS).

Per- and polyfluoroalkyl substances (PFAS) are found in many consumer and industrial products. While some PFAS, notably perfluorooctanoic acid (PFOA) and perfluorooctanesulfonic acid (PFOS), are developmentally toxic in mammals, the vast majority of PFAS have not been evaluated for developmental toxicity potential. A concentration-response study of 182 unique PFAS chemicals using the zebrafish medium-throughput, developmental vertebrate toxicity assay was conducted to investigate chemical structural identifiers for toxicity. Embryos were exposed to each PFAS compound (≤100 μM) beginning on the day of fertilization. At 6 days post-fertilization (dpf), two independent observers graded developmental landmarks for each larva (e.g., mortality, hatching, swim bladder inflation, edema, abnormal spine/tail, or craniofacial structure). Thirty percent of the PFAS were developmentally toxic, but there was no enrichment of any OECD structural category. PFOS was developmentally toxic (benchmark concentration [BMC] = 7.48 μM); however, other chemicals were more potent: perfluorooctanesulfonamide (PFOSA), N-methylperfluorooctane sulfonamide (N-MeFOSA), ((perfluorooctyl)ethyl)phosphonic acid, perfluoro-3,6,9-trioxatridecanoic acid, and perfluorohexane sulfonamide. The developmental toxicity profile for these more potent PFAS is largely unexplored in mammals and other species. Based on these zebrafish developmental toxicity results, additional screening may be warranted to understand the toxicity profile of these chemicals in other species.

Scale-based landmark for internal organs assessments of common carp (Cyprinus carpio) during digital radiography and ultrasonography imaging.

Medical imaging techniques such as digital radiography and ultrasonography are non-invasive and provide precise results for examining internal organs and structures within fish. Their effectiveness can be further enhanced by using body parts like scales as markers for the organs beneath them. This study utilized the number of scales as landmarks in digital radiography and ultrasonography to non-invasively evaluate the muscles, bones, and images of internal and reproductive organs of common carp (Cyprinus carpio). Digital radiography was performed in the dorsoventral and lateral views of the fish, whereas ultrasonography was conducted in longitudinal and transverse views on sequence scale numbers with brightness and colour Doppler-modes. Digital radiography of the common carp revealed the whole-body morphology, including the bony parts from the head, pectoral fins, dorsal fins, pelvic fins, anal fins, and vertebrae to the tail that appeared radiopaque. Internal organs were also observed, with the swim bladder and heart appeared radiolucent, while the intestines, liver, testes, and ovaries appeared radiopaque. Ultrasonography in brightness mode displayed the digestive organs, reproductive organs, and muscle thickness. Additionally, colour Doppler mode demonstrated blood flow within the heart's ventricle.

Rheological properties of the swim bladder colloid from Atlantic cod (Gadus Morhua) and its digestive characteristics

The swim bladder colloid (SWG) was prepared by the swim bladder (SW) from Atlantic cod (Gadus Morhua), and its physicochemical, rheological and digestive properties were evaluated. It showed SW mainly consisted of collagens and chondroitin sulfate. The optimal condition of SWG was treated with acetic acid for 4.0 h, operated with high temperature and pressure time for 30 min and homogenized twice at 100 MPa pressure. The viscosity of SWG, containing the mixed system of proteins and polysaccharides, decreased with the increasing of shear rate, exhibiting shear thinning behavior, which conformed to pseudoplastic fluid. It demonstrated larger values for G″ than that for G′ at the frequency of 20 Hz at 25 °C, which reached 30.8 Pa and 5.15 Pa at 60% concentration, respectively. Moreover, when the concentrations increased from 20% to 80%, the gelation temperatures and the melting temperatures increased from 6.79 ± 0.01 °C and 12.61 ± 0.06 °C to 13.73 ± 0.06 °C and 18.31 ± 0.05 °C, respectively. All of G''/G′ were much less than 0.1, confirming the presence of solid-like elastic behavior at 4.0 °C. When the temperature increased from 40 °C to 85 °C, the viscosity at 20% concentration of SWG decreased from 34.50 mPa s to 8.07 mPa s at the shear rate of 100 s−1. Furthermore, it exhibited that SWG was digested and degraded after simulated digestion in vitro, and bioactive peptides were produced.

Exposure to thiourea during the early stages of development impedes the formation of the swim bladder in zebrafish larvae.

Thiourea, a widely used agrochemical, is known to inhibit the activity of thyroid peroxidase, a key enzyme in the biosynthetic pathway of thyroid hormones. Thyroid insufficiency compromises the basal metabolic rate in warm-blooded organisms and embryonic development in vertebrates. In this study, we looked for developmental defects by exposing the zebrafish embryos to an environmentally relevant dose of thiourea (3mg/mL). Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was performed to validate thiourea's presence in the treated zebrafish embryos. Structural anomalies like bent tail and pericardial edema were noticed in 96-h post-fertilization (hpf) larvae. On histological examination, underdeveloped swim bladder was noticed in 96 hpf larvae exposed to 3mg/mL thiourea. The treated larvae also failed to follow the characteristic swimming behavior in response to stimuli due to defective swim bladder. Swim bladder being homologous to the lung of tetrapod, the role of Bmp4, a major regulator of lung development, was studied along with the associated regulatory genes. Gene expression analysis revealed that thiourea administration led to the downregulation of bmp4, shh, pcna, anxa5, acta2, and the downstream effector snail3 but the upregulation of caspase3. The protein expression showed a similar trend, wherein Bmp4, Shh, and Pcna were downregulated, but Cleaved Caspase3 showed an increased expression in the treated group. Therefore, it is prudent to presume that exposure to thiourea significantly reduces the expression of Bmp4 and other key regulators; hence, the larvae fail to develop a swim bladder, a vital organ that regulates buoyancy.

Culture protocols of the Metallic Shiner for ornamental aquaculture

AbstractObjectiveWhile North American fishes are often overlooked by the ornamental fish industry, there is growing interest in fishes native to the southeastern USA, a hot spot for fish diversity. The Metallic Shiner Pteronotropis metallicus, a small‐bodied leuciscid native to Florida and Georgia, is sometimes available in the ornamental fish trade through wild collection and limited captive production. This species exhibits a distinctive color pattern straddling the lateral line: a dorsal pink to red line, fading to a black to blue stripe. This group of minnows is largely unknown to ornamental aquaculture and lacks defined culture protocols. Our objective was to develop culture protocols for the Metallic Shiner.MethodsFollowing collection of wild broodstock, we used routine culture methods to experimentally determine substrate preference, egg incubation, egg disinfection, larval feeding, and larval development.ResultMetallic Shiner spawning was more successful on yarn mops compared with other media, with average output of >50 embryos per tank per day. Hatching success was similar for static incubators (24%) and upwelling incubators (20%) and was also similar among egg disinfection treatments (iodine, hydrogen peroxide, and formalin). Larval feeding regimes altered survival, with Otohime A1 exhibiting higher survival (44%) over 14 days posthatch (dph) and better growth than other feeding regimes. During the developmental trial with water temperature at a relatively constant 25°C, length increased from 5 mm notochord length (4 dph) to 8.7 mm standard length (45 dph), with significant milestones occurring at 4 dph (swim bladder inflation), 16 dph (flexion), and 23 dph (hypural plate formation).ConclusionThese data provide basic culture information for the Metallic Shiner while also providing a place of reference to begin investigating culture protocols for related species, including species of conservation concern.

PEDOT-Integrated Fish Swim Bladders as Conductive Nerve Conduits.

Advanced artificial nerve conduits offer a promising alternative for nerve injury repair. Current research focuses on improving the therapeutic effectiveness of nerve conduits by optimizing scaffold materials and functional components. In this study, a novel poly(3,4-ethylenedioxythiophene) (PEDOT)-integrated fish swim bladder (FSB) is presented as a conductive nerve conduit with ordered topology and electrical stimulation to promote nerve regeneration. PEDOT nanomaterials and adhesive peptides (IKVAV) are successfully incorporated onto the decellularized FSB substrate through pre-coating with polydopamine. The obtained PEDOT/IKVAV-integrated FSB substrate exhibits outstanding mechanical properties, high electrical conductivity, stability, as well as excellent biocompatibility and bioadhesive properties. In vitro studies confirm that the PEDOT/IKVAV-integrated FSB can effectively facilitate the growth and directional extension of pheochromocytoma 12 cells and dorsal root ganglion neurites. In addition, in vivo experiments demonstrate that the proposed PEDOT/IKVAV-integrated FSB conduit can accelerate defective nerve repair and functional restoration. The findings indicate that the FSB-derived conductive nerve conduits with multiple regenerative inducing signals integration provide a conducive milieu for nerve regeneration, exhibiting great potential for repairing long-segment neural defects.

Potential prebiotic activity and improvement effect on intestinal injury of sulfated glycosaminoglycan from swim bladder

Recently, the exploration of bioactive substances for improving intestinal function has attracted wide attention. In this study, a sulfated glycosaminoglycan from the swim bladder (SBSG) was used to explore the prebiotic activities and its improvement effect on arsenic-induced intestinal injury. In vitro digestion experiments showed that SBSG was not degraded in the upper digestive tract and it could promote the growth of several lactic acid bacteria, which showed that it possessed potential prebiotic activities. In vivo study showed that SBSG could reduce cytokine (IL-2, IL-1β and TNF-α) to alleviate the level of inflammation, and decrease intestinal permeability by reducing endotoxin levels. SBSG greatly alleviated the macroscopic and microscopic pathological damage of the jejunum and colon induced by arsenic. Besides, SBSG could directly intervene in intestinal injury by reducing oxidative stress and acute inflammation. Our results indicated that SBSG could be developed as a new prebiotic resource against intestinal injury and regulate microecological balance.

Physoschistura longibulla, a new loach (Cypriniformes: Nemacheilidae) from Irrawaddy basin in central Myanmar

Physoschistura longibulla, a new species and the first species of the revised genus Physoschistura from Pyin Oo Lwin of Irrawaddy basin, can be distinguished from other congeners by the combination of following characters: posterior chamber of air bladder well developed, connecting anterior with a long and thin tube, long and oval; dorsal fin with 81/2 branched rays, its origin slightly anterior to pelvic-fin base vertically; lateral line complete and straight, with 93–102 pores; pectoral fin with 8–9 branched rays, reaching 1/2 to 2/3 of distance to pelvic-fin base; suborbital flap absent in male; axillary pelvic lobe present; caudal fin with 9 + 8 branched rays.

Development of a Zebrafish Embryo-Based Test System for Thyroid Hormone System Disruption: 3Rs in Ecotoxicological Research.

There is increasing concern regarding pollutants disrupting the vertebrate thyroid hormone (TH) system, which is crucial for development. Thus, identification of TH system-disrupting chemicals (THSDCs) is an important requirement in the Organisation for Economic Co-operation and Development (OECD) testing framework. The current OECD approach uses different model organisms for different endocrine modalities, leading to a high number of animal tests. Alternative models compatible with the 3Rs (replacement, reduction, refinement) principle are required. Zebrafish embryos, not protected by current European Union animal welfare legislation, represent a promising model. Studies show that zebrafish swim bladder inflation and eye development are affected by THSDCs, and the respective adverse outcome pathways (AOPs) have been established. The present study compared effects of four THSDCs with distinct molecular modes of action: Propylthiouracil (PTU), potassium perchlorate, iopanoic acid, and the TH triiodothyronine (T3) were tested with a protocol based on the OECD fish embryo toxicity test (FET). Effects were analyzed according to the AOP concept from molecular over morphological to behavioral levels: Analysis of thyroid- and eye-related gene expression revealed significant effects after PTU and T3 exposure. All substances caused changes in thyroid follicle morphology of a transgenic zebrafish line expressing fluorescence in thyrocytes. Impaired eye development and swimming activity were observed in all treatments, supporting the hypothesis that THSDCs cause adverse population-relevant changes. Findings thus confirm that the FET can be amended by TH system-related endpoints into an integrated protocol comprising molecular, morphological, and behavioral endpoints for environmental risk assessment of potential endocrine disruptors, which is compatible with the 3Rs principle. Environ Toxicol Chem 2024;00:1-18. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Toxicology of milrinone by zebrafish embryotoxicity test.

Milrinone, a phosphodiesterase III inhibitor with contractile and vasodilatory effects, is widely used in acute decompensated heart failure and medically refractory end-stage heart failure (HF). The adverse reactions of milrinone have been extensively explored clinically, but its possible toxicities and underlying molecular mechanisms in embryo development need further understanding as its clinical applications increase. Herein, we assessed the milrinone toxicity using the zebrafish embryotoxicity test (ZET), with a view of providing evidence and guidancefor gravidas medicine. We carried out ZET by exposing embryos to a milrinone culture with a series concentration gradients since 1.5 hours post fertilization (hpf) and observed and assessed mortality and hatching rates of drug-treated zebrafishes at 24, 48, 72, and 96 hpf. No significant lethal effect was found in milrinone-treated zebrafish, but hatching rate of eggs at 48 hpf was up-regulated with the increase of milrinone concentration. The impact of milrinone on embryogenesis was assessed through body length, eye area, yolk sac area, swim bladder inflation area, pericardial area and venous congestion area at 96hpf. 150 μg/mL or higher milrinone treatment showed significant effects in the indicators. Organ disorders including enlarged pericardium, liver atrophy and decreased blood vessels were observed in dysplasia individuals versus controls. TUNEL assay suggested the ability of milrinone to induce apoptosis in malformation embryos. Quantitative real-time PCR showed aberrant expressions of transcription factors associated with heartdevelopment and genes related to liver development and apoptosis regulation. Therefore, ZET is feasible for the milrinone toxicity test, and high-dose milrinone causes harm to the embryonic development of zebrafish, especially in embryonic carcinogenesis, vasculogenesis, and hepatogenesis.

Parental exposure to Cypermethrin causes intergenerational toxicity in zebrafish offspring

Cypermethrin (CYP), a synthetic pyrethroid pesticide, has been detected in agriculture and aquaculture. However, there is limited knowledge about the transgenerational impacts. This study aimed to investigate the developmental toxicity of CYP on F1 larvae offspring of adult zebrafish exposed to various CYP concentrations (5, 10, and 20 μg/L) for 28 days. The results indicated that CYP accumulated in parental zebrafish, and CYP was below the limit of quantification in offspring. Paternal exposure impacted the hatching rate and heart rate of the F1 generation. Furthermore, CYP significantly impacted the development of swim bladders in progeny and dysregulated the genes relevant to swim bladder development. The neutrophil migrated to the swim bladder. The mRNA levels of the inflammatory factors were also significantly elevated. According to network toxicology, PI3-AKT may be the signaling pathway for CYP-influenced bladder development. Subsequent molecular docking and Western blot analysis showed CYP affected the PI3-AKT signaling pathway. Notably, MK-2206, a specific Akt inhibitor, rescued the CYP-induced damage of swim bladder development in offspring. The present study highlights the potential risks of CYP on the development of offspring and lasting impact in aquatic environments.

The generation gap in endocrine disruption: Can the integrated fish endocrine disruptor test (iFEDT) bridge the gap by assessing intergenerational effects of thyroid hormone system disruption?

Thyroid hormones (THs) act early in ontogenesis, even prior to the differentiation of thyrocytes. Maternal transfer of THs is therefore known to play an essential role in early development. Current OECD test guidelines for the assessment of TH system disruption (THSD) do not address inter- or transgenerational effects. The integrated fish endocrine disruptor test (iFEDT), a test combining parental and developmental exposure of filial fish, may fill this gap. We tested the ability of the iFEDT to detect intergenerational effects in zebrafish (Danio rerio): Parental fish were exposed to propylthiouracil (PTU), an inhibitor of TH synthesis, or not exposed. The offspring was submitted to a crossed experimental design to obtain four exposure scenarios: (1) no exposure at all, (2) parental exposure only, (3) embryonic exposure only, and (4) combined parental and embryonic exposure. Swim bladder inflation, visual motor response (VMR) and gene expression of the progeny were analysed. Parental, but not embryonic PTU exposure reduced the size of the swim bladder of 5 d old embryos, indicating the existence of intergenerational effects. The VMR test produced opposite responses in 4.5 d old embryos exposed to PTU vs. embryos derived from exposed parents. Embryonic exposure, but not parental exposure increased gene expression of thyroperoxidase, the target of PTU, most likely due to a compensatory mechanism. The gene expression of pde-6h (phosphodiesterase) was reduced by embryonic, but not parental exposure, suggesting downregulation of phototransduction pathways. Hence, adverse effects on swim bladder inflation appear more sensitive to parental than embryonic exposure and the iFEDT represents an improvement in the testing strategy for THSD.