Extrinsic nerves are not involved in branchial 5-HT dynamics or pulsatile urea excretion in Gulf toadfish, Opsanus beta.

Gulf toadfish (Opsanus beta) can switch from continuously excreting ammonia as their primary nitrogenous waste to excreting predominantly urea in distinct pulses. Previous studies have shown that the neurotransmitter serotonin (5-HT) is involved in controlling this process, but it is unknown if 5-HT availability is under central nervous control or if the 5-HT signal originates from a peripheral source. Following up on a previous study, cranial nerves IX (glossopharyngeal) and X (vagus) were sectioned to further characterize their role in controlling pulsatile urea excretion and 5-HT release within the gill. In contrast to an earlier study, nerve sectioning did not result in a change in urea pulse frequency. Total urea excretion, average pulse size, total nitrogen excretion, and percent ureotely were reduced the first day post-surgery in nerve-sectioned fish but recovered by 72h post-surgery. Nerve sectioning also had no effect on toadfish urea transporter (tUT), 5-HT transporter (SERT), or 5-HT2A receptor mRNA expression or 5-HT and 5-hydroxyindoleacetic acid (5-HIAA) abundance in the gill, all of which were found consistently across the three gill arches except 5-HIAA, which was undetectable in the first gill arch. Our findings indicate that the central nervous system does not directly control pulsatile urea excretion or local changes in gill 5-HT and 5-HIAA abundance.

Ontogenetic development of the inner ear saccule and utricle in the Lusitanian toadfish: Potential implications for auditory sensitivity.

Studies addressing structure-function relationships of the fish auditory system during development are sparse compared to other taxa. The Batrachoididae has become an important group to investigate mechanisms of auditory plasticity and evolution of auditory-vocal systems. A recent study reported ontogenetic improvements in the inner ear saccule sensitivity of the Lusitanian toadfish, Halobatrachus didactylus, but whether this results from changes in the sensory morphology remains unknown. We investigated how the macula and organization of auditory receptors in the saccule and utricle change during growth in this species. Inner ear sensory epithelia were removed from the end organs of previously PFA-fixed specimens, from non-vocal posthatch fry (<1.4 cm, standard length) to adults (>23 cm). Epithelia were phalloidin-stained and analysed for area, shape, number and orientation patterns of hair cells (HC), and number and size of saccular supporting cells (SC). Saccular macula area expanded 41x in total, and significantly more (relative to body length) among vocal juveniles (2.3-2.9 cm). Saccular HC number increased 25x but HC density decreased, suggesting that HC addition is slower relative to epithelial growth. While SC density decreased, SC apical area increased, contributing to the epithelial expansion. The utricule revealed increased HC density (striolar region) and less epithelial expansion (5x) with growth, contrasting with the saccule that may have a different developmental pattern due to its larger size and main auditory functions. Both macula shape and HC orientation patterns were already established in the posthatch fry and retained throughout growth in both end organs. We suggest that previously reported ontogenetic improvements in saccular sensitivity might be associated with changes in HC number (not density), size and/or molecular mechanisms controlling HC sensitivity. This is one of the first studies investigating the ontogenetic development of the saccule and utricle in a vocal fish and how it potentially relates to auditory enhancement for acoustic communication.

Optical micro-tomography "OPenT" allows the study of large toadfish Halobatrachus didactylus embryos and larvae.

Batrachoidids, which include midshipman and toadfish are less known among embryologists, but are common in other fields. They are characteristic for their acoustic communication, and develop hearing and sound production while young juveniles. They lay large benthic eggs (>5mm) with a thick chorion and adhesive disk and slow development, which are particularly challenging for studying embryology. Here we took advantage of a classical tissue clearing technique and the OPenT open-source platform for optical tomography imaging, to image a series of embryos and larvae from 3 to 30mm in length, which allowed detailed 3D anatomical reconstructions non-destructively. We documented some of the developmental stages (early and late in development) and the anatomy of the delicate stato-acoustic organs, swimming bladder and associated sonic muscles. Compared to other techniques accessible to developmental biology labs, OPenT provided advantages in terms of image quality, cost of operation and data throughput, allowing identification and quantitative morphometrics of organs in larvae, earlier and with higher accuracy than is possible with other imaging techniques.

Allometric larval growth of the bottom-dwelling catfish Lophiosilurus alexandri Steindachner, 1876 (Siluriformes: Pseudopimelodidae)

We investigated the early life growth pattern of pacamã, Lophiosilurus alexandri , a catfish endemic to the rio São Francisco basin, Brazil. Pacamã larvae were kept in a galvanized trough for 29 days after hatching. We collected approximately 10 larvae daily (total number = 285). We obtained from each specimen the total length (independent variable) and 11 other body measurements (dependent variables). We performed linear regressions on the log-transformed data to determine the growth pattern of the body measurements and the respective inflexion point on the growth curves. Except for trunk length, trunk width and yolk sac volume, the remaining body measurements showed a positive allometric growth coefficient up to the respective IP followed by a decrease in the rate of relative growth afterwards. Although trunk width presented a positive allometric growth, no significant inflexion point was detected. The dorsal-ventral head flattening seen in the adult pacamã was quickly expressed in the larvae as indicated by the large allometric growth of head width. Maxillary barbels showed the highest growth rate. The exceptionally high growth rates of eyes and maxillary barbels and the acquired ability to capture prey (as suggested by mouth length growth) strengthened the hypothesis of growth priorities in which the most essential organs develop first.

Investigamos o padrão de crescimento inicial do pacamã, Lophiosilurus alexandri , um bagre endêmico da bacia do rio São Francisco, Brasil. Larvas de pacamã foram mantidas numa canaleta horizontal de metal galvanizado durante 29 dias após a eclosão. Diariamente, coletamos circa 10 larvas (número total = 285) das quais obtivemos o comprimento total (variável independente) e 11 outras medidas corporais (variáveis dependentes). Fizemos regressões lineares nos dados log transformados para determinar o padrão de crescimento das medidas corporais e respectivo ponto de inflexão das curvas de crescimento. Exceto comprimento do tronco, largura do tronco e volume do saco vitelino, as demais medidas corporais apresentaram crescimento alométrico positivo até o respectivo ponto de inflexão, seguido de redução na taxa relativa de crescimento. Embora a largura do tronco apresentasse crescimento alométrico positivo, não foi detectado um ponto de inflexão significativo. O achatamento dorso-ventral da cabeça, presente nos adultos, mostrou-se evidente nas larvas com o acentuado crescimento alométrico de sua largura. Os barbilhões maxilares exibiram a maior taxa de crescimento. O extraordinário crescimento dos olhos e dos barbilhões maxilares e a habilidade adquirida para a captura de presas (como sugere o crescimento do comprimento bucal) reforçam a hipótese de prioridade de crescimento em que os órgãos essenciais são os primeiros a se desenvolverem.

Steroidogenesis by testis and accessory glands of the Lusitanian toadfish, Halobatrachus didactylus, during reproductive season.

In teleost fish sex steroids are essential for gonadal function and have marked effects in reproductive and agonistic behavior and in the expression of secondary sexual characteristics. The Lusitanian toadfish, Halobatrachus didactylus, has two male morphotypes: type I males are territorial nest-holders and have large accessory glands while type II males are smaller, have a relatively large testis and small accessory glands. In the present study, the steroidogenic activity of the testis and accessory testicular glands of the Lusitanian toadfish were examined in vitro as well as their presence in urine. The testis of type I males produced 11-ketotestosterone (11KT) and 11ß-hydroxy-4-androstene-3,17-dione (11ßA) from tritiated 17-hydroxyprogesterone, while those of type II males produced testosterone (T) and 11ß,17ß-dihydroxy-4-andosten-3-one (11ßT), but not 11KT. Additionally, the testis and accessory glands of both morphs produced mostly 5ß,3α-reduced and 17,20α-hydroxylated metabolites. Type I, but not of type II, males synthesised 5ß-reduced androgens in their accessory glands. The presence of 11ßA exclusively in the urine of type I males during reproductive season suggests an association with maintenance of secondary sexual characteristics and behavior in this morph. The urine of both types of males contained two 5α-androstane and 5ß-pregnane glucuronides. Among the latter steroids, those that are 17,21-dihydroxylated are potentially metabolites from cortisol and were found only in type I males during the spawning season. The diversity of metabolites produced by the testis and accessory glands and the presence of some in urine is suggestive of a potential role in chemical communication and reproductive behavior.

Vocal differentiation parallels development of auditory saccular sensitivity in a highly soniferous fish.

Vocal differentiation is widely documented in birds and mammals but has been poorly investigated in other vertebrates, including fish, which represent the oldest extant vertebrate group. Neural circuitry controlling vocal behaviour is thought to have evolved from conserved brain areas that originated in fish, making this taxon key to understanding the evolution and development of the vertebrate vocal-auditory systems. This study examines ontogenetic changes in the vocal repertoire and whether vocal differentiation parallels auditory development in the Lusitanian toadfish Halobatrachus didactylus (Batrachoididae). This species exhibits a complex acoustic repertoire and is vocally active during early development. Vocalisations were recorded during social interactions for four size groups (fry: <2 cm; small juveniles: 2-4 cm; large juveniles: 5-7 cm; adults >25 cm, standard length). Auditory sensitivity of juveniles and adults was determined based on evoked potentials recorded from the inner ear saccule in response to pure tones of 75-945 Hz. We show an ontogenetic increment in the vocal repertoire from simple broadband-pulsed 'grunts' that later differentiate into four distinct vocalisations, including low-frequency amplitude-modulated 'boatwhistles'. Whereas fry emitted mostly single grunts, large juveniles exhibited vocalisations similar to the adult vocal repertoire. Saccular sensitivity revealed a three-fold enhancement at most frequencies tested from small to large juveniles; however, large juveniles were similar in sensitivity to adults. We provide the first clear evidence of ontogenetic vocal differentiation in fish, as previously described for higher vertebrates. Our results suggest a parallel development between the vocal motor pathway and the peripheral auditory system for acoustic social communication in fish.

Darkedge midshipman Porichthys analis (Batrachoidiformes: Batrachoididae), a common shrimp-bycatch fish from the Gulf of California

Shrimp fishery bycatch of the Gulf of California constitutes a wide variety of highly unknown fish, crustacean, and mollusk species with very low or null economic value, in contrast to those of commercial interest. However, there are no studies yet on the role of these low economic valued species have in the community structure and function, together with their possible effect on commercial populations. With the aim of contributing to the knowledge of Darkedge Midshipman fish Porichthys analis, the most common waste species in this fishery, we estimated some population characteristics. For this purpose, we obtained shrimp-bycatch fish during the 2004-2005 fishing season and performed some research surveys. A total of 1 725 Darkedge Midshipman were captured from 350 bycatch samples. Individual size ranged from 43-352mm SL. The weight to standard length relationship was determined as W=0.000092SL3.0509. Von Bertalanffy’s growth coefficient indicated a moderate growth rate (K=0.5) with an estimated asymptotic length ofL∞=352mm standard length, and a longevity of 6yr. Natural mortality was estimated as 0.97 and total mortality as 4.67. The recruitment pattern, as estimated by ELEFAN II, was extended over the year, peaking during spring and summer seasons. Sexual proportion of male:female was 1.65:1. Mature organisms appeared from August to March, and length at maturity was 157mm SL, which is larger than the mean size at capture 135mm SL. We concluded that the species is indirectly protected by its own bathymetric distribution and the off-shrimping season. This is the first study that considers population characteristics from this common but still unknown species.

A diferencia de las especies de interés comercial, la historia de vida de los peces no comerciales es casi desconocida a pesar de su potencial importancia en la estructura y función de la comunidad. Para contribuir al conocimiento de estas especies se estimaron algunas características poblacionales del chupalodo Porcihchty sanalis. La fauna de acompañamiento de la pesquería del camarón en el Golfo de California está constituida por especies poco conocidas y de escaso valor comercial. Las especies de la familia Batrachoididae son frecuentemente utilizadas como animales experimentales, pero poco se sabe de P. analis. De esta manera a partir de la fauna de acompañamiento se recolectaron 1 725 organismos y se determinó la relación talla-peso W=0.000092SL3.0509. Mediante ELEFAN I, se estimó un crecimiento moderado (K=0.5/año) con una longitud asintótica mayor que la observada reportada anteriormente (L∞=352mm SL) y una longevidad de 6 años. La mortalidad natural se estimó en M=0.97 la total en Z=4.67. El patrón de reclutamiento fue mayor durante primavera-verano. La proporción sexual M:F fue 1.65:1 y la talla media de primera madurez fue de 157mm SL mayor que la talla media de captura 135mm SL. Este es el primer reporte que considera las características poblacionales de esta especie común, pero poco conocida.

Egg surface structure of the freshwater toadfish Thalassophryne amazonica (Teleostei: Batrachoididae) with information on its distribution and natural habitat

The egg surface structure of Thalassophryne amazonica, a freshwater toadfish from the Amazon basin is described. Eggs of this species show a remarkable, highly unusual system of parallel ridges and intermittent grooves that originate at the equator of the egg and lead to the micropylar pit, at which they end in a spiralling pattern. A similar egg surface structure has so far been described only from a group of Asian anabantoid percomorphs, obviously not closely related to Thalassophryne. This egg surface pattern may enhance fertilization success by guiding sperm to the micropyle. We review museum records for T. amazonica, present an updated map of its occurrence in the Amazon basin, and provide information on its habitat.

A estrutura da superfície do ovo de Thalassophryne amazonica um batracoidídeo de água doce da bacia amazônica é descrita. Os ovos dessa espécie apresentam um sistema notável e incomum de cristas paralelas e canais intermitentes, que se originam na porção equatorial do ovo e vão até a micrópila onde terminam em um padrão espiralado. Uma estrutura superficial de ovo semelhante foi até agora descrita apenas para um grupo de anabantóideos asiáticos, obviamente não relacionado a Thalassophryne. Esse padrão de superfície de ovo talvez aumente o sucesso da fertilização ao guiar o espermatozóide até à micrópila. Nós revisamos os registros de T. amazonica em museus, apresentamos um mapa atualizado de sua ocorrência na bacia Amazônica e fornecemos informações sobre seu habitat.

Darkedge midshipman Porichthys analis (Batrachoidiformes: Batrachoididae), a common shrimp-bycatch fish from the Gulf of California.

Shrimp fishery bycatch of the Gulf of California constitutes a wide variety of highly unknown fish, crustacean, and mollusk species with very low or null economic value, in contrast to those of commercial interest. However, there are no studies yet on the role of these low economic valued species have in the community structure and function, together with their possible effect on commercial populations. With the aim of contributing to the knowledge of Darkedge Midshipman fish Porichthys analis, the most common waste species in this fishery, we estimated some population characteristics. For this purpose, we obtained shrimp-bycatch fish during the 2004-2005 fishing season and performed some research surveys. A total of 1 725 Darkedge Midshipman were captured from 350 bycatch samples. Individual size ranged from 43-352mm SL. The weight to standard length relationship was determined as W=0.000092SL(3.0509). Von Bertalanffy's growth coefficient indicated a moderate growth rate (K=0.5) with an estimated asymptotic length of L(infinity) = 352mm standard length, and a longevity of 6yr. Natural mortality was estimated as 0.97 and total mortality as 4.67. The recruitment pattern, as estimated by ELEFAN II, was extended over the year, peaking during spring and summer seasons. Sexual proportion of male:female was 1.65:1. Mature organisms appeared from August to March, and length at maturity was 157mm SL, which is larger than the mean size at capture 135mm SL. We concluded that the species is indirectly protected by its own bathymetric distribution and the off-shrimping season. This is the first study that considers population characteristics from this common but still unknown species.

Ontogeny of auditory saccular sensitivity in the plainfin midshipman fish, Porichthys notatus.

The auditory system of the plainfin midshipman fish, Porichthys notatus, is an important sensory receiver system used to encode intraspecific social communication signals in adults, but the response properties and function of this receiver system in pre-adult stages are less known. In this study we examined the response properties of auditory-evoked potentials from the midshipman saccule, the main organ of hearing in this species, to determine whether the frequency response and auditory threshold of saccular hair cells to behaviorally relevant single tone stimuli change during ontogeny. Saccular potentials were recorded from three relative sizes of midshipman fish: small juveniles [1.9-3.1 cm standard length (SL), large juveniles (6.8-8.0 cm SL) and non-reproductive adults (9.0-22.6 cm SL)]. The auditory evoked potentials were recorded from the rostral, middle and caudal regions of the saccule while single tone stimuli (75-1,025 Hz) were presented via an underwater speaker. We show that the frequency response and auditory threshold of the midshipman saccule is established early in development and retained throughout ontogeny. We also show that saccular sensitivity to frequencies greater than 385 Hz increases with age/size and that the midshipman saccule of small and large juveniles, like that of non-reproductive adults, is best suited to detect low frequency sounds (<105 Hz) in their natural acoustic environment.

Age and growth of the gulf toadfish Opsanus beta based on otolith increment analysis.

In the present study, sagittal otoliths of confirmed male and female specimens of the gulf toadfish Opsanus beta that were collected monthly over the course of a year from Biscayne Bay, Florida, U.S.A. were analysed. The timing and frequency of O. beta spawning seasons are reported by examination of the gonado-somatic index. The estimated ages of males and females ranged from <1 year to 6 and 5 years, respectively. Strong sexual dimorphism in growth was apparent with von Bertalanffy parameter estimates for males of Linfinity=393.8 mm, K=0.30, t0=0.36 and females of Linfinity=201.1 mm, K=0.79, t0=0.47. Comparison with previously published growth trajectories of the more northerly distributed conspecific Opsanus tau showed that O. beta males had a higher growth rate. Female O. beta and O. tau growth trajectories appear similar, with an indication that the former becomes asymptotic at least a year before the latter. Results are discussed in the context of temperature regimes, reproductive energy allocation and waste urea excretion in the two species.

Evolutionary origins for social vocalization in a vertebrate hindbrain-spinal compartment.

The macroevolutionary events leading to neural innovations for social communication, such as vocalization, are essentially unexplored. Many fish vocalize during female courtship and territorial defense, as do amphibians, birds, and mammals. Here, we map the neural circuitry for vocalization in larval fish and show that the vocal network develops in a segment-like region across the most caudal hindbrain and rostral spinal cord. Taxonomic analysis demonstrates a highly conserved pattern between fish and all major lineages of vocal tetrapods. We propose that the vocal basis for acoustic communication among vertebrates evolved from an ancestrally shared developmental compartment already present in the early fishes.

Effects of temperature and diet on the growth rate of year 0 oyster toadfish, Opsanus tau.

The effects of temperature and diet on the growth of captive year 0 specimens of Opansus tau were examined for two consecutive year classes. The 2001 year class was raised at about 23, 26, or 29 degrees C and provided with live brine shrimp, frozen butterfish and squid, or commercial food pellets (45% protein, 19% fat, and 3% fiber) three times per week. Maximal growth was achieved with the pellet diet, and fish raised at 29 degrees C attained the highest mean wet weight (84.0 g +/- 14.6 g SE) and fastest instantaneous relative growth rate (IRGR, 1.79% body weight/d). The 2002 year class was raised on the same pellet diet at 31.5 degrees C and fed 3, 5, or 7 times per week. Although more frequent feedings led to significantly greater mean wet weight in the first half of the year, by month 12 there was no significant difference between the three feeding frequencies. These fish weighed approximately 68 g and had an average IRGR of 1.74% body weight/d. The pellet diet during both years was correlated with high survival (> 75%).

Ontogenetic changes in the response properties of individual, primary auditory afferents in the vocal plainfin midshipman fish Porichthys notatus Girard.

The auditory system of adult midshipman fish Porichthys notatus Girard is an important sensory receiver system used during intraspecific social communication to encode conspecific vocalizations, but the response properties and function of this system in the pre-adult stages are unknown. Midshipman fish, like other teleosts, use the saccule as the main acoustic end organ of the inner ear. In this study, we examined the discharge properties and the frequency response dynamics of auditory saccular afferent neurons in pre-adult midshipman (approximately 4-12 months of age) to determine whether encoding of auditory information, inclusive of conspecific vocalizations, changes across life history stages. Extracellular single unit recordings were made from saccular afferents while sound was presented via an underwater speaker. Comparisons with adult data show that the resting discharge rate and auditory threshold sensitivity increased with age/size, while temporal encoding of frequency did not show any significant shifts. The results indicate that the saccular afferents of juveniles, like those of non-reproductive adults, are best adapted to temporally encode the low frequency components (