Muscle cellularity, growth performance and growth-related gene expression of juvenile climbing perch Anabas testudineus in response to different eggs incubation temperature.

Although indigenous climbing perch (Anabas testudineusis) is a highly valuable species, slow growth pattern during the culture period impeding its commercial success in aquaculture. In many fish species, it has been demonstrated that incubation temperature of eggs influenced the muscle development and growth rates, which persisted throughout the subsequent larval and juvenile phases. Therefore, this study aimed to investigate whether different incubation temperature of eggs prior to hatching can stimulate the muscle development, growth, and growth-related gene expression of the slow-growing indigenous species of climbing perch. The fertilized eggs of A. testudineus from an artificial breeding program were incubated under control temperature of 24 °C (IT24), 26 °C (IT26), 28 °C (IT28), and 30 °C (IT30) in 10L glass aquaria with four replicated units for each temperature treatment. After hatching, the larvae from each incubated temperature were separately reared at ambient temperature for 10 days in aquarium, 20 days in hapas, and the next 42 days in cages, totaling 72 days post-hatching (dph). The hatching rates were found significantly (P < 0.05) higher in IT28 compared to the other incubation temperature treatments. After 72 dph, the growth performances (%length gained, %weight gained and SGR) were found to be significantly highest (P < 0.05) in the IT28, followed by the treatments IT30, IT26, and IT24, respectively. Survival rate (73 ± 1.257%) was also found to be highest in the same treatment. The rate of new muscle fiber formation was identified to be significantly highest (P < 0.05) in IT28 followed by the IT26, IT30 and IT24, respectively. The relative mRNA expression level of GHRH, IGF1, IGF2 and PRL was also significantly highest in the IT28 (P < 0.05) compared to other treatments. Results from the present study clearly suggested that 28 °C is the optimum eggs incubation temperature of the native strain of A. testudineus for its highest growth performances in captive breeding condition.

Transmission electron microscopy of the gastrointestinal tract of Nile perch Lates niloticus / Uso de microscopía electrónica de transmisión para visualizar el tracto gastrointestinal de la perca del Nilo Lates niloticus

The ultrastructure of the gastrointestinal tract of Nile perch was described using Transmission Electron Microscopy standard procedures. Investigations revealed the presence of mucous cells, blood vessels and oil droplets plus several nerve cells and muscle bundles in the oral cavity. Further observations revealed columnar epithelial cells in the oesophagus, with a ragged surface, high electron dense cytoplasm, intercellular spaces, mitochondria and mucus granules. The lamina propria of the oesophagus was composed of loose connective tissue, blood vessels, nerve cells and several leucocytes, while the muscularis externa was composed of striated muscle bundles with the a Z line, A band and I band. The stomach was characterized by mucosal glandular cells with electron dense granules, a prominent nucleus, mitochondria, and secretory canaliculi. The liver had an isotropic parenchyma composed of several hepatocytes with a single spherical euchromatic nucleus. The exocrine pancreatic cells in the liver occurred around large blood vessels, had large centrally positioned nuclei with electron dense nucleolus, electron dense granules, Rough Endoplasmic Reticulum, and mitochondria which dominated the cytoplasm of pancreatic cells.

La ultraestructura del tracto gastrointestinal de la perca del Nilo fue descrita utilizando un procedimiento estándar de Microscopía Electrónica de Transmisión (MET). Las investigaciones revelaron la presencia de células mucosas, vasos sanguíneos y pequeñas gotas de aceite, además de varias células nerviosas y haces musculares en la cavidad oral. Nuevas observaciones revelaron células epiteliales columnares en el esófago, con una superficie irregular, citoplasma elevado y electrodenso, espacios intercelulares, mitocondrias y gránulos mucosos. La lámina propia del esófago se compone de tejido conectivo laxo, vasos sanguíneos, células nerviosas y numerosos leucocitos, mientras que la muscular externa se compone de haces musculares estriados, con presencia de la línea Z, banda A y banda I. El estómago se caracteriza por presentar células glandulares mucosas con gránulos electrodensos , un núcleo prominente, mitocondrias y canalículos secretores. El hígado presentó un parénquima isotrópico compuesto de varios hepatocitos con un solo núcleo eucromático y esférico. Las células pancreáticas exocrinas se encontraron alrededor de grandes vasos sanguíneos, presentando grandes núcleos posicionados centralmente, con nucleólos densos, gránulos electrodensos en el retículo endoplasmático rugoso, y mitocondrias, que dominaron el citoplasma de las células pancreáticas.

Scanning electron microscopy of the gastrointestinal tract of nile perch (lates niloticus, linneaus, 1758) / Microscopía electrónica de barrido del tracto gastrointestinal de la perca del nilo (lates niloticus, linnaeus, 1758)

The morphology of the gastrointestinal tract of adult Nile perch was described using standard SEM procedures. Investigations revealed the presence of cardiform teeth in the oral cavity, goblet cells and finger print-like microridges on the hard palate and oesophagus lumenal surface. Elaborate patterns and bacterial cells were observed on the stomach lumenal surface and intense foldings in the intestinal region. These observations provide a better understanding of the morphology of the gut in Nile perch and how it is suited for its digestive function.

El objetivo fue describir la morfología del tracto gastrointestinal de la perca del Nilo adulta mediante microscopía electrónica de barrido estándar. La investigación reveló la presencia de dientes cardiformes en la cavidad oral, células caliciformes y microcrestas como huellas digitales en el paladar duro y la superficie luminal del esófago. Se observaron patrones elaborados, así como bacterias en la superficie luminal del estómago, y plegamientos marcados en la región intestinal. Estas observaciones proporcionan una mejor comprensión de la morfología del intestino de la perca del Nilo y como se adapta para su función digestiva.

Catch me if you can--predation affects divergence in a polyphenic species.

Predation is a major driving force in evolution. Predation has been shown to select for size, morphology, and camouflage. Many animals use camouflage to reduce predation risk. In some cases, individuals can adjust their pigmentation, enabling them a higher survival in a heterogeneous environment. Here, we show that the difference in pigmentation between juvenile perch individuals (Perca fluvuiatilis) occupying different environments (open water and vegetated habitats of lakes) is likely a consequence of predator selection. Lightly pigmented individuals have a higher chance of survival in open water whereas darker pigmented individuals survive better in vegetation. As a response to predators, individuals forced into the vegetation by predators developed darker skin whereas the skin of individuals forced into open water became lighter. In a common garden experiment, in the absence of predation, we found that pigmentation in juvenile perch is only due to plasticity and not to genetic variation. However, contrary to predictions, individuals raised in open water developed darker skin compared to individuals raised in vegetation. This may be a response to UV-stress. Overall, our results suggest that predation can be a strong selective agent on pigmentation differences among conspecifics occupying different habitats.

Morphometric differentiation of two Nigerian fish species (Oreochromis niloticus and Lates niloticus) using principal components and discriminant analysis / Diferenciación morfométrica de dos especies de peces nigerianos (Oreochromis niloticus y Lates niloticus) utilizando análisis de componentes principales y análsis discriminante

Morphological differences between two close fish populations were investigated in Doma Dam, north central Nigeria using principal component and discriminant analyses. The purpose was to identify the two species correctly and assign them into distinct genetic groups. To achieve this, seven morphometric measurements (body weight, standard length, total length, head length, body depth, dorsal fin length and caudal fin length) were made on each of the studied 252 fish specimens (153 Oreochromis niloticus and 99 Lates niloticus). The analysis of variance showed that significantly (P<0.05) higher values were recorded for Lates niloticus in all the body traits examined, except dorsal fin length. From the principal component analysis, the differences between the species resulted mainly from dorsal fin length, body weight, caudal fin length and head length. The stepwise discriminant analysis revealed that standard length, dorsal fin length and caudal fin length were the three most discriminating variables useful to distinguish clearly the two species at mature age. These three variables with strong discriminating power correctly classified 98.0 percent of individuals from the sample of known-fish populations. The classification accuracy was cross-validated using split-sample method, and indicated a 97.6 percent success rate (98.0 percent of Oreochromis niloticus and 97.0 percent of Lates niloticus were correctly assigned). The results can provide a sound scientific basis for the effective management and sustainable exploitation of the genetic resources of the two species under subtropical conditions.

Las diferencias morfológicas entre dos poblaciones de peces similares se han investigado en Doma Dam, centro-norte de Nigeria utilizando análisis de componentes principales y análisis discriminante. El propósito fue identificar correctamente las dos especies y asignarlas a los distintos grupos genéticos. Para lograr esto, siete medidas morfométricas (peso corporal, longitud estándar, longitud total, longitud de la cabeza, profundidad del cuerpo, longitud de la aleta dorsal y la longitud de la aleta caudal) se realizaron en cada una de las muestras de los 252 peces estudiados (153 Oreochromis niloticus y 99 Lates niloticus) . El análisis de varianza mostró que de forma significativa (p<0,05) los valores más altos se registraron en Lates niloticus, en todos los rasgos del cuerpo examinado con excepción de la longitud de la aleta dorsal. A partir del análisis de componentes principales, las diferencias entre las especies se observó principalmente en la longitud de la aleta dorsal, el peso corporal, la longitud de la aleta caudal y longitud de la cabeza. El análisis discriminante reveló que la longitud estándar, longitud aleta dorsal y caudal son las tres variables más discriminatorias de utilidad para distinguir claramente las dos especies en la edad madura. Estas tres variables con fuerte poder de discriminación clasificaron correctamente el 98,0 por ciento de los individuos de la muestra de las poblaciones conocidas de peces. La exactitud de la clasificación fue sometida a una validación cruzada utilizando el método de la muestra dividida, la que indicó una tasa de éxito del 97,6 por ciento (98,0 por ciento de Oreochromis niloticus y 97,0 por ciento de Lates niloticus asignados correctamente). Los resultados pueden proporcionar una base científica sólida para el manejo eficaz y la explotación sostenible de los recursos genéticos de las dos especies en condiciones subtropicales.

Molecular, morphological, and biogeographic resolution of cryptic taxa in the Greenside Darter Etheostoma blennioides complex.

The systematic identity and genetic divergence of cryptic taxa and morphological subspecies in the Greenside Darter Etheostoma blennioides complex are analyzed from mitochondrial and nuclear DNA sequence data, along with morphological characters. We sequenced the mtDNA cytochrome b gene and control region and is the nuclear S7 intron 1 for 345 Greenside Darters from 19 locations across their distribution including areas of sympatry and allopatry, in comparison to putative sister species and relatives. Results provide the first genetic evidence that E. gutselli is a separate species and is the sister species of E. blennius, which together with E. rupestre comprise the sister group to the Greenside Darter complex; separating approximately 4.0 mya. MtDNA results show that the complex comprises 6 clades and supports only the morphological subspecies Etheostoma blennioides blennioides, distinguished by theta(ST)=0.94, approximately 1.7 my, scale counts, and ventral squamation. The former E. b. pholidotum and E. b. newmanii are polyphyletic and are invalid taxa, together comprising 5 differentiated clades that diverged approximately 0.90-1.7 mya. Nuclear DNA results recover some of the mtDNA clades, which are distinguished morphologically by subtle meristic count differences. Populations of E. b. blennioides genetically diverge, with diversity increasing to the southwest; attributed to restricted gene flow and genetic isolation with geographic distance. Samples of the former E. b. pholidotum from the Great Lakes/Wabash River clade are less divergent, with diversity increasing to the southwest, reflecting allopatric fragmentation and isolation by distance.

Hormonal, morphological, and physiological responses of yellow perch (Perca flavescens) to chronic environmental metal exposures.

The effects of a chronic environmental exposure to metals on the hormonal, physiological, and reproductive status were assessed in yellow perch (Perca flavescens) sampled in six lakes situated along a contamination gradient of Cd, Zn, Cu, Pb, and Ni in the mining region of Rouyn-Noranda, Québec. Fish were captured in the summer and fall, and sampled before or after a confinement of one hour. Metal concentrations in the kidneys and the interrenal tissues (homologous to mammalian adrenals) were measured to compare tissue-specific metal accumulation. An exposure-related decrease of condition factor, gonadosomatic index (GSI), branchial Na+/K(+)-ATPase activity, plasma thyroxine (T4), triiodothyronine (T3), and 17 beta-estradiol and an impaired capacity to enhance cortisol levels after confinement were observed. Fish from the metal-contaminated lakes possessed gonads at less mature stages and exhibited structural alterations of their gills, interrenal cells, and thyroid follicle epithelium. A comparison of the morphological, biochemical, and physiological endpoints measured in the present study revealed that plasma concentrations of hormones and parameters of gill function were the most affected by metal contamination. The results of this study indicate that lifelong exposures to sublethal concentrations of metals alter the physiological functions of fish and delay reproduction.

Immunocytochemical localization of different cell types in the adenohypophysis of the cichlid fish Cichlasoma dimerus (Heckel, 1840).

The adenohypophysis of the cichlid fish Cichlasoma dimerus was studied using the avidin-biotin-peroxidase method with antisera raised against piscine pituitary hormones and heterologous antisera against mammalian pituitary hormones. Antiserum raised against rabbit ACTH recognized a group of cells bordering the neurohypophysis (NH) in the rostral pars distalis (RPD). Anti-chum salmon prolactin (PRL) identified a compact group of cells in the periphery of the RPD. Gonadotropin II (GTH II), thyrotropin (TSH) and growth hormone (GH)-ir cells were localized in the proximal pars distalis. Ir-GTH II cells were also observed in the dorsal area of the pars intermedia (PI). Ir-GTH I cells could not be identified using anti-chum salmon GTH I, this may be due either to a failure of the antisera to recognize the gonadotropin or to a low expression of the hormone in adults of this species. PAS positive cells from the PI bound specifically with three different antisera raised against somatolactin (SL) of four different fish species. These cells surrounded deep branches of the NH in the PI.

GnRH-systems in the forebrain of cichlid fish.

The neuronal systems that contain gonadotropin-releasing hormone (GnRH) were quantitatively examined in the forebrain of two cichlid species (Oreochromis niloticus Lin. and Tilapia maria Boulenger) using tracing methods and immunostaining. In both species the nucleus olfacto-retinalis and a population of neurons in the basal preoptic region contained GnRH. The nucleus olfacto-retinalis was divided into several neuronal subpopulations which differed with respect to cytology, projection pattern, and peptide content. GnRH cell number increased with body size, the rates being different for the different subpopulations. A sexual dimorphism was found in the basal preoptic region of Tilapia; males had significantly fewer GnRH-containing cells than females. Selective staining of the basal preoptic region demonstrated that this cell group is the main source of GnRH-innervation of the pituitary gland and indicate that the molecular forms of GnRH expressed in the two cell populations studied may be different.

Ectopic ossicles associated with metacercariae of Apophallus brevis (Trematoda) in yellow perch, Perca flavescens (Teleostei): development and identification of bone and chondroid bone.

This paper describes the development and tissues in mineralized ossicles in the musculature of Perca flavescens infected with metacercariae of the trematode Apophallus brevis. Analysis involved light microscopy, transmission and scanning electron microscopy, X-ray scanning electron microprobe analysis, and tetracycline labelling. Two to 14 days post-infection, fibroblast-like host cells stream towards the parasite cyst forming a fusiform cellular capsule. By 14 days post-infection the capsule differentiates into an inner hypertrophied layer, an extensive middle layer of fibroblast-like cells, and a thin outer layer of flattened fibroblast-like cells forming a fibrous sheath at the capsule/muscle interface. From 21-35 days post-infection, a bony tissue is deposited periosteally in an equatorial ring around the cyst. With time, additional tissue is secreted over the ring increasing its thickness and advancing the matrix front towards the poles of the ossicle. Plump osteoblast-like cells cover the developing ossicle and may become trapped within the matrix in lacunae encapsulated by collagen. By 63 days post-infection, medium-sized ossicles are morphologically similar to large cysts from perch captured in the wild; ovoid with two polarized canals, but lacking acellular or lamellar bone-like tissue. Mineralized ossicles contain calcium, phosphorus and oxygen. Large ossicles retrieved from perch given multiple doses of tetracycline revealed discrete fluorescent bands, indicative of incremental growth. Fully developed ossicles are composed of two skeletal tissues, an inner region of chondroid bone and an outer region of acellular, lamellar bone.

Gonadotropin-releasing hormone-containing neurons change size with reproductive state in female Haplochromis burtoni.

In the preoptico-hypothalamic area (POA) of teleost fish, neurons containing gonadotropin-releasing hormone (GnRH) regulate reproduction through direct projections to pituitary gonadotropes. Here we show that these GnRH-containing cells change size depending on the reproductive and maturational state in female Haplochromis burtoni. We selected animals prior to, during, and after the reproductive portion of their life history, in both brooding and spawning states. Immunocytochemical staining of GnRH-containing neurons in the POA revealed that these cells are up to twice as large in females that have never spawned or are in the act of spawning than they are in females that are carrying broods. Older, postreproductive females have the largest cell sizes. Previous work on male H. burtoni has shown that soma sizes of the homologous neurons change according to social status, with dominant fish having larger cells than subordinates. Since reproductively active females have no apparent social hierarchy and are all exposed to approximately the same external stimuli, the primary factor(s) controlling GnRH-immunoreactive (irGnRH) neuron size appears to be internal reproductive state. Thus, while irGnRH neurons are pleiomorphic in both males and females, cell size change is differently regulated in each.

Ipsilateral retinofugal projections in a percomorph bony fish: their experimental induction, specificity and maintenance.

Adult bony fish possess only a small ipsilateral retinofugal projection, if any. Experimental manipulation, such as unilateral enucleation, can lead to an enhancement of this projection. We examined the patterns of, as well as the conditions for the development and maintenance of an enhanced ipsilateral retinofugal projection (EIRP) after nerve crush, after enucleation, and after various combinations of both types of surgery in juvenile and adult Haplochromis burtoni (Cichlidae). Retinal projections were labeled either unilaterally with horseradish perixodase, or with the lipophilic fluorescent dye DiI in aldehyde-fixed animals, or bilaterally with differently colored fluorescent dextran amines. Unilateral nerve crush always leads to the regeneration of retinofugal fibers to the contralateral tectum but spares some contralateral diencephalic nuclei. In addition, unilateral or bilateral nerve crush in many cases, and unilateral enucleation in some cases, leads to the development of an EIRP to the ipsilateral diencephalon and tectum. This EIRP persists (4 months and longer postoperatively) in only 10% of the unilaterally enucleated animals, in none of the animals subjected to unilateral nerve crush and in 79% of the animals subjected to bilateral nerve crush. All unilaterally enucleated animals in which the remaining, contralateral optic nerve was crushed develop and maintain an EIRP. These data suggest that nerve crush alone is sufficient to cause regenerating fibers to project, at least transiently, to the ipsilateral side of the brain. When the normal contralateral projection is either absent or in the process of regeneration, an EIRP can be maintained. In the latter case, alternate bands or patches of ipsi- and contralateral fibers in the tectum may result. Ipsilateral fibers follow unusual pathways by recrossing at the rostral diencephalon. Likewise, regenerating contralateral retinal fibers grow differently in this area; here, where the optic-nerve projection is reorganized into the optic tract, many regenerating fibers are deflected to the ipsilateral side of the brain. Despite atypical routes taken by some fibers, the EIRP nevertheless ends only in specific retinorecipient areas. An EIRP develops independently of the age of the animal, independently of the time lapse between enucleation and nerve lesion, and independently of persisting debris. However, in animals receiving an optic nerve lesion a long time after unilateral enucleation, the size of the EIRP and its tectal extent are reduced compared to that in animals enucleated around the same time as receiving the crush of the contralateral optic nerve.

Neuromast morphology and lateral line trunk canal ontogeny in two species of cichlids: an SEM study.

A study of neuromast ontogeny and lateral line canal formation in Oreochromis aureus and Cichlasoma nigrofasciatum reveals the existence of two classes of neuromasts: those that arise just before hatching (presumptive canal neuromasts, dorsal superficial neuromasts, gap neuromasts, and caudal fin neuromasts) and pairs of neuromasts that arise on each lateral line scale lateral to each canal segment at the same time as canal formation. In the anterior trunk canal segment, each presumptive canal neuromast is accompanied by a dorsoventrally oriented superficial neuromast forming an orthogonal neuromast pair. It is suggested that each of these dorsoventrally oriented superficial neuromasts is homologous to the transverse superficial neuromast row described by Münz (Zoomorphology 93:73-86, '79) in other cichlids. It is further suggested that the longitudinal lines described by Münz (Zoomorphology 93:73-86, '79) are derived from the pair of superficial neuromasts that arise during canal formation. Distinct changes in neuromast topography are documented. Neuromast formation, scale formation, and lateral line canal formation are three distinct and sequential processes. The distribution of neuromasts is correlated with myomere configuration; there is always one presumptive canal neuromast on each myomere. A single scale forms beneath each presumptive canal neuromast. Canal segment formation is initiated with the enclosure of each presumptive canal neuromast by an epithelial bridge which later ossifies. The distinction of these three processes raises questions as to the causal relationships among them.

Chondroid bone on the upper pharyngeal jaws and neurocranial base in the adult fish Astatotilapia elegans.

Serial cross sections of several adult specimens of the cichlid Astatotilapia elegans were used to investigate the fate and structure of the chondroid bone on the articulation between upper pharyngeal jaws and neurocranial base. The tissue persists in the adult on the three elements on which it previously developed, i.e., infrapharyngobranchial III-IV, parasphenoid, and basioccipital bones. It consists of haphazardly arranged, large vesicular cells without a canalicular system, embedded in a matrix histologically indistinguishable from bone matrix. Except for a narrow zone at the distal side, it is mineralized throughout. As in younger stages, the fibrous covering of the chondroid bone forms the articular tissue proper on each of the three elements. Acellular bone, found at the basal margin of the chondroid bone, it is argued, does not result from endochondral replacement of the latter but rather from dermal ossification projecting from the marrow cavity. Although lacunae may be filled in this way with bone, true obliteration of cells does not occur, so that there is no metaplasia from chondroid bone to bone. The part played by the chondroid bone in the outgrowth of the joint apophyses is discussed.