Most individual fish in wild and farmed populations can be infected with parasites. Fish intestines can harbour protozoans, myxozoans and helminths, which include several species of digeneans, cestodes, nematodes and acanthocephalans. Enteric parasites often induce inflammation of the intestine; the pathogen provokes changes in the host physiology, which will be genetically selected for if they benefit the parasite. The host response to intestinal parasites involves neural, endocrine and immune systems and interaction among these systems is coordinated by hormones, chemokines, cytokines and neurotransmitters including peptides. Intestinal fish parasites have effects on the components of the enteric nervous and endocrine systems; mechanical/chemical changes impair the activity of these systems, including gut motility and digestion. Investigations on the role of the neuroendocrine system in response to fish intestinal parasites are very few. This paper provides immunohistochemical and ultrastructural data on effects of parasites on the enteric nervous system and the enteric endocrine system in several fish–parasite systems. Emphasis is on the occurrence of 21 molecules including cholecystokinin-8, neuropeptide Y, enkephalins, galanin, vasoactive intestinal peptide and serotonin in infected tissues.

In teleosts, vitellogenin (Vtg) is a phospholipoglycoprotein synthesized by the liver, released into the blood circulation and incorporated into the oocytes via endocytosis mediated by the Vtg receptor (VTGR) to form the yolk granules. The VTGR is crucial for oocyte growth in egg-laying animals but is also present in non-oviparous vertebrates, such as human. The VTGR belongs to the low-density lipoprotein receptor superfamily (LDLR) and is also named very-low-density lipoprotein receptor (VLDLR). In this study, we identified and phylogenetically positioned the VTGR of a basal teleost, the European eel, Anguilla anguilla. We developed quantitative real-time PCR (qRT-PCR) and investigated the tissue distribution of vtgr transcripts. We compared by qRT-PCR the ovarian expression levels of vtgr in juvenile yellow eels and pre-pubertal silver eels. We also analyzed the regulation of ovarian vtgr expression throughout vitellogenesis in experimentally matured eels. The Vtg plasma level was measured by homologous ELISA experimental maturation. Our in silico search and phylogenetical analysis revealed a single vtgr in the European eel, orthologous to other vertebrate vtgr. The qRT-PCR studies revealed that vtgr is mainly expressed in the ovary and also detected in various other tissues such as brain, pituitary, gill, fat, heart, and testis, suggesting some extra-ovarian functions of VTGR. We showed that vtgr is expressed in ovaries of juvenile yellow eels with no higher expression in pre-pubertal silver eels nor in experimentally matured eels. This suggests that vtgr transcription already occurs during early pre-vitellogenesis of immature eels and is not further activated in vitellogenic oocytes. European eel Vtg plasma level increased throughout experimental maturation in agreement with previous studies. Taken together, these results suggest that vtgr transcript levels may not be a limiting step for the uptake of Vtg by the oocyte in the European eel.

Literature about fish kidney peroxisomes is scarce. To tackle this caveat, a stereological approach on renal peroxisome morphological parameters was performed for the first time in a fish, establishing correlations with maturation stages as it was previously done in brown trout liver. Three-year-old brown trout males and females were collected at the major seasons of their reproductive cycle. Trunk kidney was fixed and processed for catalase cytochemistry. Classical stereological methods were applied to electromicrographs to quantitate morphological parameters. Different seasonal variation patterns were observed between genders, and between renal proximal tubule segments I and II. In males, peroxisomes from proximal tubule segment II had a relatively higher volume and number in May, being individually bigger in February. Females presented similar trends, though with less marked variations. Overall, males and females did not show exactly the same seasonal patterns for most peroxisomal parameters, and no correlations were found between the latter and the gonado-somatic index (GSI). Hence, and despite the variations, the morphology of renal peroxisomes is not strictly correlated with gonad maturation kinetics, therefore suggesting that kidney peroxisome morphology is not seasonally modulated by sex steroids, like estradiol, as it seems to happen in liver peroxisomes.

The urohyal is incorporated within the hyoid and branchial arches and performs a significant role in the mouth opening-closing mechanism of fishes, and is considered a synapomorphy of teleostean fishes. Morphological variation of the urohyal, in terms of size and shape parameters, can allow species identification. Morphology of the urohyal in 49 species belonging to 43 genera and 29 families from the Persian Gulf and from the Oman Sea were compared using size and shape measurements. The results examine the suitability of using the urohyal morphology in differentiating fish species from this region; highlighting the taxonomic value of the urohyal, which until now, had been studied little in terms of use as a diagnostic feature in the classification of teleosts.

Studies that address fish welfare before slaughter have concluded that many of the traditional systems used to stun fish including CO2 narcosis are unacceptable as they cause avoidable stress before death. One system recommended as a better alternative is electrical stunning, however, the welfare aspects of this method are not yet fully understood. To assess welfare in aquaculture both behavioural and physiological measurements have been used, but few studies have examined the relationship between these variables. In an on-site study aversive behaviours and several physiological stress indicators, including plasma levels of cortisol and ions as well as blood physiological variables, were compared in Arctic char (Salvelinus alpinus) stunned with CO2 or electricity. Exposure to water saturated with CO2 triggered aversive struggling and escape responses for several minutes before immobilization, whereas in fish exposed to an electric current immobilization was close to instant. On average, it took 5 min for the fish to recover from electrical stunning, whereas fish stunned with CO2 did not recover. Despite this, the electrically stunned fish had more than double the plasma levels of cortisol compared with fish stunned with CO2. This result is surprising considering that the behavioural reactions were much more pronounced following CO2 exposure. These contradictory results are discussed with regard to animal welfare and stress physiological responses. The present results emphasise the importance of using an integrative and interdisciplinary approach and to include both behavioural and physiological stress indicators in order to make accurate welfare assessments of fish in aquaculture.

Most animals are concurrently infected with multiple parasites, and interactions among them may influence both disease dynamics and host fitness. However, the sublethal costs of parasite infections are difficult to measure and the effects of concomitant infections with multiple parasite species on individual physiology and fitness are poorly described for wild hosts. To understand the costs of co-infection, we investigated the relationships among 189 European eel (Anguilla anguilla) from Mar Menor, parasites (richness and intensity) and eel's ‘health status’ (fluctuant asymmetry, splenic somatic index and the scaled mass index) by partial least squares regression. We found a positive relationship with 44% of the health status variance explained by parasites. Contracaecum sp. (Nematoda: Anisakidae) was the strongest predictor variable (44·72%) followed by Bucephalus anguillae (Platyhelminthes: Bucephalidae), (29·26%), considered the two most relevant parasites in the analysis. Subsequently, 15·67 and 12·01% of the response variables block were explained by parasite richness and Deropristis inflata (Platyhelminthes: Deropristiidae), respectively. Thus, the presence of multiple parasitic exposures with little effect on condition, strongly suggests that eels from Mar Menor tolerate multiparasitism.

The biocrystallization process and the prescence of manganese in sagitta otoliths is described for the first time via electron paramagnetic spectroscopy. Spectra of Mn2+ are applied to verify inter-species differences in 12 Sciaenidae species and geographic variations among three stocks of banded croaker Paralonchurus brasiliensis and rake stardrum Stellifer rastrifer. Similar spectra shape and zero-field splitting D (22.4 mT) indicated that the biocrystallization process is the same for all fish species, at any stage of development and under different environmental conditions, corresponding to Mn2+ in the aragonite (CaCO3). Inter-species differences in Mn2+ concentration were detected regarding the otolith shape. Elongated otoliths have higher manganese concentrations compared to rounded otoliths. Geographic variations among stocks of P. brasiliensis and S. rastrifer could be detected. Fish stocks under riverine influence have less Mn2+ concentration than ones under marine influence. Manganese detection via electron paramagnetic spectroscopy is a non-destructive approach that can be useful in fish stocks studies.

Horizontal cells of the vertebrate retina have large receptive fields as a result of extensive gap junction coupling. Increased ambient illumination reduces horizontal cell receptive field size. Using the isolated goldfish retina, we have assessed the contribution of nitric oxide to the light-dependent reduction of horizontal cell receptive field size. Horizontal cell receptive field size was assessed by comparing the responses to centered spot and annulus stimuli and from the responses to translated slit stimuli. A period of steady illumination decreased the receptive field size of horizontal cells, as did treatment with the nitric oxide donor (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl)amino]diazen-1-ium-1,2-diolate (100 μM). Blocking the endogenous production of nitric oxide with the nitric oxide synthase inhibitor, NG-nitro-l-arginine methyl ester (1 mM), decreased the light-induced reduction of horizontal cell receptive field size. These findings suggest that nitric oxide is involved in light-induced reduction of horizontal cell receptive field size.

The large variability in kidney morphology among fish makes it difficult to build a “universal” model on its function and structure. Therefore, a morphological study of brown trout trunk kidney was performed, considering potential seasonal and sex effects. Three-year-old specimens of both sexes were collected at four stages of their reproductive cycle. Kidney was processed for light and electron microscopy. The relative volumes of renal components, such as renal corpuscles and different nephron tubules, were estimated by stereological methods. Qualitatively, the general nephron structure of brown trout was similar to that described for other glomerular teleost species. Quantitatively, however, differences in the relative volume of some renal components were detected between sexes and among seasons. Particularly, highest values of vacuolized tubules and new growing tubules were observed after spawning, being more relevant in females. Despite seasonal changes, more linear correlations were found between those parameters and the reno-somatic index than the gonado-somatic index. Thus, we verified that some brown trout renal components undergo sex dependent seasonal variations, suggesting a morphological adaptation of the components to accomplish physiological needs. These findings constitute a baseline for launching studies to know which factors govern the morphological variations and their functional consequences.

Extracellular recordings were made from the isolated goldfish retina during superfusion with various serotonergic agonists and antagonists to determine the effects of these drugs on the maintained activity and response properties of the ganglion cells. Superfusion of the retina with serotonin (25–500 μM) increased the maintained activity of OFF-center ganglion cells and decreased the maintained activity of ON-center ganglion cells. In addition, serotonin also attenuated the excitatory responses to annular stimuli, suggesting a decrease in the strength of surround input to the ganglion cells. The effects of serotonin on OFF-center ganglion cells were mimicked by the nonselective 5-HT1, agonist 5-MeOT and the 5-HT1 receptor agonist 8-OH-DPAT, while only 5-MeOT mimicked the action of serotonin on ON-center ganglion cells. The effects of exogenously applied serotonin on the ganglion cells could be blocked by the mixed 5-HT1/5-HT2 receptor antagonist methysergide but not by the 5-HT2 receptor antagonist mianserin or the dopamine receptor antagonist haloperidol.

These results support previous anatomical and biochemical evidence that serotonin functions in a neurotransmitter or neuromodulatory role in the teleost retina and suggest that serotonin may be involved in modulating the maintained activity and surround input to the ganglion cells. The results also indicate that two different types of receptors may mediate the actions of serotonin in the ON and OFF pathways, respectively.

The retinae of lower vertebrates undergo a number of structural changes during light adaptation, including the photomechanical contraction of cone myoids and the dispersion of melanin granules within the epithelial pigment. Since the application of dopamine to dark-adapted retinae is known to produce morphological changes that are characteristic of light adaptation, dopamine is accepted as a causal mechanism for such retinomotor movements. However, we report here that in the teleost fish, Aequidens pulcher, the intraocular injection of 6-hydroxydopamine (6-OHDA), a substance known to destroy dopaminergic retinal cells, has no effect on the triggering of light-adaptive retinomotor movements of the cones and epithelial pigment and only slightly depresses the final level of light adaptation reached. Furthermore, the retina continues to show circadian retinomotor changes even after 48 h in continual darkness that are similar in both control and 6-OHDA injected fish. Biochemical assay and microscopic examination showed that 6-OHDA had destroyed dopaminergic retinal cells. We conclude, therefore, that although a dopaminergic mechanism is probably involved in the control of light-induced retinomotor movements, it cannot be the only control mechanism, nor can it be the cause of circadian retinomotor migrations. Interestingly, 6-OHDA injected eyes never reached full retinomotor dark adaptation, suggesting that dopamine has a role to play in the retina's response to darkness.

We investigated the attributes of transduction and light-adaptation in rods, single cones, and twin cones isolated from the retina of striped bass (Morone saxatilis). Outer-segment membrane currents were measured with suction electrodes under voltage clamp provided by tight-seal electrodes applied to the cell’s inner segment. Brief flashes of light transiently reduced the outer-segment current with kinetics and sensitivity characteristic of each receptor type. In all cells, the responses to dim lights increased linearly with light intensity. The amplitude-intensity relation for rods and single cones were well described by an exponential saturation function, while for twin cones it was best described by a Michaelis-Menten function. At the wavelength of maximum absorbance, the average intensity necessary to half-saturate the peak photocurrent in dark-adapted rods was 28 photons/μm2 and in single cones it was 238 photons/μm2. Among twin cones, the common type (88% of all twins recorded) half-saturated at an average of 1454 photons/μm2, while the fast type reached half-saturation at an average of 9402 photons/μm2. The action spectrum of the photocurrent in the three receptor types was well fit by a nomogram that describes the absorption spectrum of a vitamin A2-based photopigment. The wavelength of maximum absorbance for rods was 528 nm, for single cones it was 542 nm and for twin cones it was 605 nm. Both members of the twin pair contained the same photopigment and they were electrically coupled. Under voltage clamp, the response to dim flashes of light in both single and twin cones was biphasic. The initial peak was followed by a smaller amplitude undershoot. Single cones reached peak in 86 ms and common twins in 50 ms. Background light desensitized the flash sensitivity in all photoreceptor types, but was most effective in rods and least effective in fast twins. In the steady state, the desensitizing effect of a background intensity, Ib, at the respective optimum wavelength for each cell was well described by the Weber-Fechner law (1/(1 + Ib/Ibo)), where Ibo was, on average (in units of photons/μm2/s), 1.45 for rods, 1.81 x 103 for single cones, 4.56 x 103 for common twins, and 6.79 x 104 for fast twins.

The yolk syncytial layer (YSL) has been regarded as one of the main obstacles for a successful cryopreservation of fish embryos. The purpose of this study was to identify and characterize the YSL in Prochilodus lineatus, a fish species found in southeastern Brazil and considered a very important fishery resource. Embryos were obtained through artificial breeding by hormonal induction. After fertilization, the eggs were incubated in vertical incubators with a controlled temperature (28 °C). Embryos were collected in several periods of development up to hatching and then fixed with 2% glutaraldehyde and 4% paraformaldehyde in 0.1 M sodium phosphate buffer (pH 7.3). Morphological analyses were carried out under either light, transmission or scanning electron microscopy. The formation of the YSL in P. lineatus embryos starts at the end of the cleavage stage (morula), mainly at the margin of the blastoderm, and develops along the embryo finally covering the entire yolk mass (late gastrula) and producing a distinct intermediate zone between the yolk and the endodermal cells. The YSL was characterized by the presence of microvilli on the contact region with the yolk endoderm. A cytoplasmic mass, full of mitochondria, vacuoles, ribosomes, endomembrane nets and euchromatic nuclei, indicated a high metabolic activity. This layer is shown as an interface between the yolk and the embryo cells that, besides sustaining and separating the yolk, acts as a structure that makes it available for the embryo. The structural analyses identified no possible barriers to cryoprotectant penetration.

The general consensus is that brain serotonin (5-HT) inhibits feed intake in teleost fishes and other vertebrates. Dietary manipulations with the 5-HT precursor tryptophan (TRP) have, however, yielded contradictory effects on feed intake, while studies of the endocrine response to stress indicate that the effects of TRP-enriched feed are context dependent. A characteristic behavioural response to stress is a reduction in feed intake, and in the present study we investigated whether pre-treatment with TRP-enriched feed affected stress-induced changes in feeding behaviour in brown trout (Salmo trutta). After acclimatisation in observation aquaria, isolated fish were fed control or TRP-supplemented feed for 7 d, whereupon they were transferred to a novel environment, in which all fish were fed control feed. Transfer to a new environment resulted in decreased feeding in both the TRP pre-treated and the control-treated group. However, this decrease was more pronounced in the control-treated group. Previous experiments have concluded that stimulation of brain 5-HT systems by TRP enhancement does not affect feed intake in salmonid fishes, but in these studies food intake was observed in unstressed animals only. The present study suggests that pre-treatment with dietary TRP attenuates stress-induced anorexia. Hence, it appears that the effect of dietary manipulations of TRP on feeding behaviour is dependent on the stress levels experienced by experimental animals. These behavioural data are discussed in the context of the involvement of 5-HT in appetite regulation.

This survey was performed to characterize the embryogenesis of Prochilodus lineatus. Seven stages of embryo development were identified – zygote, cleavage, blastula, gastrula, segmentation, larval and hatching – after a period of incubation of 22 h (24 °C) or 14 h (28 °C). The following cleavage pattern was identified: the first plane was vertical (2 blastomeres); the second was vertical and perpendicular to the first (4 blastomeres); the third was vertical and parallel to the first (4 × 2); the fourth cleavage was vertical and parallel to the second (4 × 4); the fifth was vertical and parallel to the first (4 × 8); and the sixth cleavage was horizontal (64 blastomeres). At the blastula stage (3.0–4.0 h (24 °C); 1.66–2.0 h (28 °C)) irregular spaces were detected and periblast structuring was initiated. At the gastrula stage (4.0–8.0 h (24 °C); 3.0–6.0 h (28 °C)) the epiboly, convergence and cell movements, as well as the formation of embryonic layers, had begun. The segmentation stage (10.0–15.0 h (24 °C); 7.0–10.0 h (28 °C)) was characterized by a rudimentary formation of organs and systems (somites, optic vesicle and intestinal delimitation). The embryo at the larval stage (16.0–21.0 h (24 °C); 11.0–13.0 h (28 °C)) showed a free tail, more than 25 somites, an optic vesicle and a ready-to-hatch larval shape. The blastomeres at cleavage stage had disorganized nuclei indicating high mitotic activity. At gastrula, the blastomeres and the periblast had euchromatic nuclei and a large number of mitochondria and vesicles. The yolk was organized into globose sacs, which were dispersed into small pieces prior to absorption.

Visually evoked extracellular neural activity was recorded from the nucleus isthmi (NI) of goldfish and bluegill sunfish. When moving anywhere within the right eye's visual field, three-dimensional checkered balls or patterns on a computer screen evoked bursts of spikes in the left NI. Object motion parallel to the longitudinal body axis gave responses that habituated markedly upon repetition, but movement into recently unstimulated regions of the visual field gave vigorous responses. Thus, while NI's response is not visuotopic, its habituation is. An object approaching the animal's body generated a rising spike density, whereas object recession generated only a transient burst. During the approach of a checkered stimulus ball, average NI spike density rose linearly as the ball-to-eye distance decreased and at a rate proportional to the ball's speed (2.5–30 cm/s). Increasing ball size (2.2–9.2 cm) did not affect the rate of activity rise at a given speed, but did increase overall activity levels. NI also responded reliably to expanding textures of fixed overall size. The results suggest that NI signals changes in motion of objects relative to the fish, and estimates the proximity of approaching objects.

Following the break-up of Gondwana, the drift of Antarctica to its present position and the establishment of the Antarctic Convergence, fish evolution was characterized by adaptation to progressive cooling of the environment. The decrease of erythrocyte number and haemoglobin concentration in the blood of Antarctic teleosts raises several questions concerning the physiology of respiration and the enzymatic role of erythrocytes. Our study of the molecular basis of cold adaptation includes the relationship between molecular structure and biological function of haemoglobins. Species of the suborder Notothenioidei, largely confined within the Convergence, have only one major haemoglobin, which displays the Root effect in oxygen binding; on the other hand, Zoarcidae (a family found at all latitudes) have four or five haemoglobins, only one of which displays the Root effect. In addition, our data indicate that the physiological relevance of erythrocyte-like cells, present in very small number in the blood of haemoglobinless Channichthyidae, may be linked to higher content of enzymes, such as glucose-6-phosphate dehydrogenase, in comparison with erythrocytes of red-blooded fishes.

This work studies the afferent connectivity to different functionally identified tectal zones in goldfish. The sources of afferents contributed to different degrees to the functionally defined zones. The dorsocentral area of the telencephalon was connected mainly with the ipsilateral anteromedial tectal zone. At diencephalic levels, neurons were found in three different regions: preoptic, thalamic, and pretectal. Preoptic structures (suprachiasmatic and preoptic nuclei) projected mainly to the anteromedial tectal zone, whereas thalamic (ventral and dorsal) and pretectal (central, superficial, and posterior commissure) nuclei projected to all divisions of the tectum. In the mesencephalon, the mesencephalic reticular formation, torus longitudinalis, torus semicircularis, and nucleus isthmi were, in the anteroposterior axis, topographically connected with the tectum. In addition, neurons in the contralateral tectum projected to the injected zones in a symmetrical point-to-point correspondence. At rhombencephalic levels, the superior reticular formation was connected to all studied tectal zones, whereas medial and inferior reticular formations were connected with medial and posterior tectal zones. The present results support a different quantitative afferent connectivity to each tectal zone, possibly based on the sensorimotor transformations that the optic tectum carries out to generate orienting responses.

This work is aimed at demonstrating the influence of insulin in the triggering of hepatic cholestasis in young gilt-head sea bream larvae reared under experimental conditions. The results are based on an immunological assay of insulin and an ultrastructural study of the yolk syncytial layer and the liver. Compared to fed larvae, fasting gilt-head sea bream larvae displayed a substantial increase in insulin level correlated with dysfunction of the yolk syncytial layer and hepatic histopathology. This correlation was verified in larvae reared in sea water containing insulin. Ultrastructural observations suggested that insulin affects lipoprotein secretion by the yolk syncytial layer. The failure of plasmatic lipoproteins would then cause dysfunction of the biliary lipid secretion mechanisms and, hence, trigger hepatic cholestasis. These results in gilt-head sea bream show for the first time the influence of insulin in the triggering of hepatic cholestasis during the early development of a vertebrate.

The ability to accurately quantify ovarian condition in cultured species of fish is of paramount importance in the understanding of their reproductive biology and the development of appropriate culture conditions. Here, we discuss stereological analysis, a mathematically proven technique that determines 3-dimensional topology and geometry from 2-dimensional thin sections, as a safer and more precise replacement for existing methodology.