A histological study of the organogenesis of the digestive system in bay snook Petenia splendida Günther, 1862 from hatching to the juvenile stage

In bay snook (Petenia splendida) larvae the histological development of the digestive system and swim bladder, and their relative timing of differentiation were studied from hatching to 45 days post‐hatch (dph) at 29°C. Newly hatched larvae showed a simple digestive tract, which appeared as a straight undifferentiated tube lined by a single layer of columnar epithelial cells (future enterocytes). The anatomical and histological differentiation of the digestive tract and accessory glands was a very intense, asynchronous process, proceeding from the distal to the anterior part. The intestine was the first region to differentiate (9 days post‐hatch – dph, 6.5 mm SL), and the oesophagus the last (21 dph, 8.4 mm SL). At the onset of feeding, the digestive system was organized into different functional and histologically differentiated sections, such as the buccopharynx, oesophagus, glandular stomach, and anterior and posterior intestine. This organization resembled that of the juveniles, with the exception of pharyngeal teeth and buccopharyngeal as well as oesophageal goblet cells, which proliferated later during the mixed feeding period. Histological observations revealed that bay snook larvae retained endogenous yolk reserves until 24 dph (8.9 ± 0.4 mm SL), which might be helpful for weaning this species onto a compound diet. The important lipidic accumulation observed in the intestinal mucosa, liver, and pancreas in fish fed a compound trout diet indicated that although fish were able to digest and absorb lipids, the diet formulation did not fit the nutritional requirements of early juveniles of this species. The ontogeny of the digestive system followed the same general pattern as in most cichlid species described to date. However, we detected species‐specific differences in the timing of differentiation that were related to their reproductive guild. According to the histological results, some recommendations regarding the intensive culture of this species are also provided.     

Sex steroid changes during temperature‐induced gonadal differentiation in Paralichthys olivaceus (Temminck & Schegel, 1846)

Sex steroid changes during temperature‐induced gonadal differentiation were evaluated in the olive flounder, Paralichthys olivaceus. Larvae were reared at 21 ± 0.5°C, 24 ± 0.5°C and 28 ± 0.5°C from day 40 post‐hatching (dph) to 90 dph. The proportion of males was 61.1, 76.7, 87.8 and 47.8% in 21°C, 24°C, 28°C and in control groups, respectively. Gonadal differentiation was circa 65 dph, when fishes were a mean 39 mm total length (TL). The gonads developed faster when fishes were reared in higher temperatures. Radioimmunoassay (RIA) analyses indicated that the level of estradiol‐17β (E2) changed during the period of gonadal differentiation and peaked at an onset of ovarian differentiation in all groups. Compared with fish in control groups, the levels of E2 were lower in thermal‐treated groups, especially in the highest temperature groups. The present results indicate that E2 plays a major role in the process of ovarian differentiation, and suggest that temperature‐induced masculinization in P. olivaceus is mainly due to a decrease in the E2 level during the period of ovarian differentiation.     

Comparative larval ontogeny of two fish species (Characiformes and Siluriformes) endemic to the São Francisco River in Brazil

The aim of the present study was to perform comparative histological analyses of the ontogenetic development of two fish species endemic to the São Francisco River in Brazil: Prochilodus argenteus and Lophiosilurus alexandri. Histological analyses were performed every 24 h from the moment of hatching until 14 days post-hatching (dph) for the observation of larval development and until 39 dph for the observation of gonadal development. Whole larvae were fixed in Bouin's solution and the histological slides were stained with haematoxylin–eosin. Lophiosilurus alexandri larvae had a larger body size compared with P. argenteus larvae since hatching. Lophiosilurus alexandri larvae had mouth opening and pigmentation of the eyes upon hatching, whereas these events were observed at 1 dph in P. argenteus larvae. The visualisation and the inflation of the swim bladder occurred at 1 and 3 dph, respectively, in the P. argenteus, whereas these events occurred at 2 and 8 dph, respectively, in L. alexandri. Yolk granules were absorbed at 4 dph in P. argenteus and the 10 dph in L. alexandri. At 7 dph, the digestive tube was more differentiated in L. alexandri than P. argenteus and at 14 dph, the digestive system of both species had features of their eating habits: broad stomach and short intestine in L. alexandri, typical of carnivorous habits; stomach with a mechanical function and long intestine in P. argenteus, typical of detritivorous habits. The epithelial lining tissue, formed by a single layer of cells in the newly hatched larvae (0 dph), differentiated throughout the study, exhibiting scales in P. argenteus and numerous club cells in the middle epithelial region of L. alexandri at 39 dph. Undifferentiated gonads with somatic cells and primordial germ cells were observed at 39 dph, with caudal-cranial migration since 1 dph in both species. The anatomic changes during the ontogeny of P. argenteus and L. alexandri larvae are directly associated with the evolutionary history of each species, which explains their feeding habits, behaviour and distribution in the environment: Prochilodus argenteus is detritivorous and actively swims in the water column, whereas L. alexandri is carnivorous and inhabits bottom regions. At 39 dph neither species exhibited sexual differentiation.     

THE REPRODUCTIVE CYCLE AND SEXUALITY OF THE GREEN MUSSEL PERNA VIRIDIS (L.) (BIVALVIA: MYTILACEA) IN VICTORIA HARBOUR, HONG KONG

The reproductive cycle and sexuality of the green mussel Pernaviridis (L.) in Victoria Harbour, Hong Kong, were investigatedfrom July 1982 to May 1984. Histological studies showed thatthe cyde could be divided into 4 stages occurring in a highlyseasonal pattern. The very low (<0.1%) occurrence of functionalhermaphrodites indicates that P. viridis is gonochoristic. Pastreports of protandric rhythmic hermaphroditism may have resultedfrom reliance on fluctuations in sex ratio and sex identificationby gonad colour. These criteria are, however, considered invalidfor P. viridis because differences exist between the developmentalpatterns of the male and female gonads and, further, colourof the female gonad at the early proliferation stage resemblesthe male. Temperature was found to correlate positively withgonad development and with the lower threshold at 24°C.Temperature was not, however, limiting at some sites in HongKong. A joint Principal Component Analysis-Stepwise multipleregression procedure suggested that hypertrophication mightinhibit gonadal development in Victoria Harbour (Received 2 November 1987; accepted 30 December 1987)     

Age,growth and hatch dates of ingressing larvae and surviving juveniles of Atlantic menhaden Brevoortia tyrannus

Ages, growth and hatch dates of ingressing Brevoortia tyrannus larvae were determined in a 3 year sampling survey at the mouth of the Chesapeake Bay, U.S.A. To determine if otolith‐aged cohorts had variable relative survival, hatch dates of summer‐caught young‐of‐the‐year (YOY) juveniles collected throughout the Chesapeake Bay were compared with hatch dates of ingressing larvae. Modal total length of ingressing larvae was similar among years: 28 mm in 2005–2006 and 2007–2008, and 30 mm in 2006–2007. Ages of ingressing larvae ranged from 9 to 96 days post hatch (dph); mean ages were similar among years, but significantly older in 2006–2007 (50 dph) than in 2005–2006 (44 dph) and 2007–2008 (46 dph). Larval growth rates differed among years. Earliest growth, when larvae were offshore (0–20 dph), was faster in 2006–2007 (0·62 mm day^?1), than in 2005–2006 and 2007–2008 (0·55 mm day^?1 in these years). Subsequently, from 30 to 80 dph, growth was slowest in 2006–2007. Hatch dates of ingressing larvae occurred from September to March and 90% (2007–2008) to 98% (2006–2007) had hatched prior to 31 December. In contrast, most surviving YOY juvenile B. tyrannus had hatched in January to February, suggesting selective mortality of early‐hatched individuals, apparently during the overwinter, larval to juvenile transition period.     

Gonadal morphogenesis and sex differentiation in intraovarian embryos of the viviparous fish Zoarces viviparus (Teleostei, Perciformes, Zoarcidae): a histological and ultrastructural study

It is essential to know the timing and process of normal gonadal differentiation and development in the specific species being investigated in order to evaluate the effect of exposure to endocrine-disrupting chemicals on these processes. In the present study gonadal sex differentiation and development were investigated in embryos of a viviparous species of marine fish, the eelpout, Zoarces viviparus, during their intraovarian development (early September to January) using light and electron microscopy. In both sexes of the embryos at the time of hatching (September 20) the initially undifferentiated paired bilobed gonad contains primordial germ cells. In the female embryos, ovarian differentiation, initiated 14 days posthatch (dph), is characterized by the initial formation of the endoovarian cavity of the single ovary as well as by the presence of some early meiotic oocytes in a chromatin-nucleolus stage. By 30 dph, the endoovarian cavity has formed. By 44 dph and onward, the ovary and the oocytes grow in size and at 134 dph, just prior to birth, the majority of the oocytes are at the perinucleolar stage of primary growth and definitive follicles have formed. In the presumptive bilobed testis of the male embryos, the germ cells (spermatogonia), in contrast to the germ cells of the ovary, remain quiescent and do not enter meiosis during intraovarian development. However, other structural (somatic) changes, such as the initial formation of the sperm duct (30 dph), the presence of blood vessels in the stromal areas of the testis (30 dph), and the appearance of developing testicular lobules (102 dph), indicate testicular differentiation. Ultrastructually, the features of the primordial germ cells, oogonia, and spermatogonia are similar, including nuage, mitochondria, endoplasmic reticulum, and Golgi complexes.     

Growth in captivity and aspects of reproductive biology of the Antarctic fish parasite Aega antarctica (Crustacea,Isopoda)

Summary Specimens of the fish parasite Aega antarctica caught in the Weddell Sea and off the South Shetlands and South Orkneys were kept in aquaria for more than 2 years. These isopods were fed with plaice (Pleuronectes platessa) from the North Sea. Growth in captivity was very slow. For the first growth phase before reaching sexual maturity of females the value K = 0.12 was calculated for the Bertalanffy growth constant. Mature males are markedly smaller than breeding females (average lengths: 16.0 and 22.5 mm). An age of more than 5 years was calculated for an average-sized male, females spawn at an age of more than 10 years. No evidence for protandric hermaphroditism could be found.     

Cloning of gonadal aromatase gene <Emphasis Type="Italic">Cyp19a</Emphasis> in an endemic fish (<Emphasis Type="Italic">Schizothorax kozlovi</Emphasis>) of the Upper Yangtze River,and temperature effects on its expression

In order to elucidate the role of Cyp19a in sex differentiation of Schizothorax kozlovi, the full length cDNA of Cyp19a was cloned from the mature ovary of S. kozlovi by using rapid amplification of cDNA ends method, and then its relative mRNA expression levels among tissues and temperature groups were determined by using quantitative real-time PCR. The complete Cyp19a cDNA of 1795 bp of S. kozlovi was obtained, which encoded 517 amino acids and belonged to gonadal aromatase. Its deduced amino acid sequence had the above 70 % identity compared with gonadal aromatase genes of teleost fishes, but only 62–67 % when compared with brain aromatase genes of fishes. It was expressed only in heart and gonad, but no expression in other tissues, presenting relatively high tissue specificity. It also exhibited sex-specific expression pattern in gonads, but no sex differences in heart. Comparing with the Cyp19a expression levels at 12 days post hatching (dph), significant temperature effects were revealed in low temperature group (10 °C) at 18 dph, and in high temperature group (26 °C) at 40 dph. It suggested that gonadal aromatase Cyp19a gene may play important roles on the feminization or masculinization of S. kozlovi affected by temperature during the early developmental stage.     

Differentiation of ambisexual gonads and immunohistochemical localization of P450 cholesterol side-chain cleavage enzyme during gonadal sex differentiation in the protandrous anemonefish, Amphiprion clarkii

To clarify the relationship between steroid hormones and sex differentiation of the protandrous anemonefish Amphiprion clarkii, we histologically examined its gonadal differentiation. From hatching to 30 days post hatching (dph), all of the gonads surveyed were sexually undifferentiated. The gonads of all fish first differentiated into ovaries at 60 dph, and the oocytes gradually developed and increased in number as the ovaries grew up until 183 dph. Some cysts of differentiated spermatogenic germ cells appeared in the ovaries at 214 dph, and ambisexual gonads with both ovarian and testicular tissues formed by 273 dph. Using immunohistochemistry, we then investigated the expression of cytochrome P450 cholesterol side-chain cleavage enzyme (P450scc), during gonadal sex differentiation. P450scc-immunopositive reactions first appeared in sexually undifferentiated gonads at 30 dph. Beginning at 60 dph, the number of strongly positive cells increased throughout the differentiation of the ovaries and continued to increase during the testicular differentiation until 210 dph. Immunopositive cells were observed more frequently in ovarian tissue than in testicular tissue in the ambisexual gonads at 270 dph. These results suggest that endogenous steroid hormones are important for the sex differentiation, including the primary sex differentiation and subsequent testicular differentiation, of the anemonefish.     

GENETIC DETERMINANTS OF PROTANDRIC SEX IN THE PACIFIC OYSTER,CRASSOSTREA GIGAS THUNBERG

A unique feature of sex in Crassostrea oysters is the coexistence of protandric sex change, dioecy, and hermaphroditism. To determine whether such a system is genetically controlled, we analyzed sex ratios in 86 pair-mated families of the Pacific oyster, Crassostrea gigas Thunberg. The overall female ratios of one-, two-, and three-year-old oysters were 37%, 55%, and 75%, respectively, suggesting that a significant proportion of oysters matured first as males and changed to females in later years. Detailed analysis of sex ratios in factorial and nested crosses revealed significant paternal effects, which corresponded to two types of sires. No major maternal effects on sex were observed. Major genetic control of sex was further indicated by the distribution of family sex ratios in two to four apparently discreet groups. These and other data from the literature are compatible with a single-locus model of primary sex determination with a dominant male allele (M) and a protandric female allele (F), so that MF are true males and FF are protandric females that are capable of sex change. The rate of sex change of FF individuals may be influenced by secondary genes and/or environmental factors. Strong maternal and weak paternal effects on sexual maturation or time of spawning were also suggested.     

Captive hybridization of two geographically isolated pygmy angelfish species,Centropyge fisheri and Centropyge resplendens

This study documents the rearing of two pygmy angelfish species, Centropyge fisheri and Centropyge resplendens, and the early life history and reproduction of their hybrid offspring. A C. fisheri female, collected from Hawaii, and a C. resplendens male, captive‐bred from parental stock collected from Ascension Island, were maintained at the hatchery facility for 7 months. Continuous spawning was achieved at a photoperiod cycle of 14L:10D and a water temperature of 26·5° C, range ±1° C. Over the 110 day period, the C. fisheri female spawned 102 times, 57% of which resulted in embryos (fertilized eggs). The mean ±s.d. fecundity per spawn was 730 ± 459 eggs (range 52–1967). Fertility (% eggs that developed into embryos) of all eggs that were preserved was 22·4 ± 25·6%. A total of 235 hybrid juveniles were raised through metamorphosis with an average larval survival of 16·4%. Eight F₁ hybrid juveniles isolated for further study began to display signs of reproductive behaviour c. 300 days post‐hatch (dph). Spawn resulting in non‐fertile eggs were first obtained 319 dph, and fertilized eggs developing into embryos were obtained after 411 dph from at least two female individuals. While no attempt was made at rearing the F₂ larvae, embryo and larval development were normal up to 8 dph. Reproduction and development observed for all hybrid generations in this study were normal, similar to other Centropyge species and indicates a very close phylogenetic relationship between what are currently considered distinct species, e.g. C. fisheri and C. resplendens.     

Activation of brain steroidogenesis and neurogenesis during the gonadal differentiation in protandrous black porgy,Acanthopagrus schlegelii

The early brain development, at the time of gonadal differentiation was investigated using a protandrous teleost, black porgy. This natural model of monosex juvenile fish avoids the potential complexity of sexual dimorphism. Brain neurogenesis was evaluated by histological analyses of the diencephalon, at the time of testicular differentiation (in fish between 90 and 150 days after hatching). Increases in the number of both Nissl‐stained total brain cells, and Pcna‐immunostained proliferative brain cells were observed in specific area of the diencephalon, such as ventromedialis thalami and posterior preoptic area, revealing brain cell proliferation. qPCR analyses showed significantly higher expression of the radial glial cell marker blbp and neuron marker bdnf. Strong immunohistochemical staining of Blbp and extended cellular projections were observed. A peak expression of aromatase (cyp19a1b), as well as an increase in estradiol (E₂) content were also detected in the early brain. These data demonstrate that during gonadal differentiation, the early brain exhibits increased E₂ synthesis, cell proliferation, and neurogenesis. To investigate the role of E₂ in early brain, undifferentiated fish were treated with E₂ or aromatase inhibitor (AI). E₂ treatment upregulated brain cyp19a1b and blbp expression, and enhanced brain cell proliferation. Conversely, AI reduced brain cell proliferation. Castration experiment did not influence the brain gene expression patterns and the brain cell number. Our data clearly support E₂ biosynthesis in the early brain, and that brain E₂ induces neurogenesis. These peak activity patterns in the early brain occur at the time of gonad differentiation but are independent of the gonads. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 121–136, 2016     

Histology of the reproductive tissue of the clam Ruditapes decussatus from the gulf of Gabès (Tunisia)

Summary

The gametogenic cycle of the clam Ruditapes decussatus was investigated during one year. Samples were collected each month in two different sites located at the gulf of Gabès area. Gametogenic activity was characterized on the basis of histological analysis of the gonads. A classification and a scoring system were then proposed. According to our study, Ruditapes decussatus gametogenesis can be divided in four major steps: (1) a sexual rest (December- February), (2) starting of gametogenesis (February-March), (3) development (March-May), (4) ripe (from April onwards). This study revealed that the gonad of the clam Ruditapes decussatus was surrounded by muscular tissue which could represent the principal storage tissue. Two kinds of hermaphroditism appeared since maturity of gonads: simultaneous one showing gonadal tubules of both males and females in regionally distinct and separate zones and a “superposed” one showing a gonad consisting essentially of mature male gametes and few residual oocytes.     

The social feather duster worm Bispira brunnea (Polychaeta: Sabellidae): aggregations,morphology and reproduction

The fan worm Bispira brunnea is one of the most attractive sabellid polychaetes from Caribbean coral reef areas and it is exploited for ornamental purposes. An understanding of the structure of its aggregations, morphology and reproductive biology will provide information required to facilitate artificial propagation of this species. Ten aggregations were collected in October 2013, February and March 2014 in the Majahual reef lagoon, Mexican Caribbean. Whole aggregations were examined under light microscopy and scanning electron microscopy and the histology of oogenesis was determined. Aggregations were composed of 24–56 individuals and included juveniles and adults. The adults (5–20?mm) did not display any noticeable form of sexual dimorphism. In B. brunnea both sexual (hermaphroditism or gonochorism) and asexual reproduction occurred at the same time within the population: 92.71% reproduced sexually and 52% by architomy. The buds produced by architomy were inside the parental tube, at three regenerative stages. The sex ratio was 36.75% males, 33.11% females, 22.84% hermaphrodites and 7.28% unknown (gametes not seen). Gametes were distributed in the last thoracic segments and throughout the abdomen. Oogenesis was extra-ovarian, development followed four discrete stages and the oocytes were small and asynchronous (60.97 μm in diameter). Sperm morphology was adapted to external fertilization in the water column. Sequential (protandrous) hermaphroditism is suggested to occur in B. brunnea. The pyramidellid ectosymbiont mollusc Odostomia (Eulimastoma) caniculatum is recorded here for the first time as being associated with a sabellid worm.     

Same sexual system but variable sociobiology: evolution of protandric simultaneous hermaphroditism in Lysmata shrimps

The sexual system of the decapod (caridean) shrimp Lysmata isprotandric simultaneous hermaphroditism (PSH). Individuals firstmature as males (male phase = MP) and then when older (larger)change to the external phenotype of female carideans (femalephase = FP). However, unlike purely protandric carideans, LysmataFPs retain reduced male gonadal tissues and ducts, and are ableto mate nonreciprocally as males as well as to reproduce asfemales. Thus, FPs of Lysmata species are functional simultaneoushermaphrodites although most reproductive effort is devotedto embryo production and incubation. The question explored hereis, given the propensity of carideans to protandry, the apparentlow cost, and high reproductive advantage of PSH, why has notPSH evolved more frequently? The mating systems and sexual selectionof caridean shrimps, the original sex of protandric individuals,the cost of maleness, and sex allocation theory are discussedin relation to protandry and PSH. None of these factors adequatelyexplains the evolution of PSH of Lysmata species. Lysmata hasat least 2 species groups with very different sociobiologies;these groups do not appear to share current selective pressuresthat would explain PSH in both. A historical contingency hypothesis,testable in part with a phylogenetic analysis, may explain theevolution of PSH in Lysmata.     

Growth, sex differentiation and gonad and plasma levels of sex steroids in male‐ and female‐dominant populations of Dicentrarchus labrax obtained through repeated size grading

Starting from 66 days post hatching (dph), European sea bass Dicentrarchus labrax were graded successively to create a fast growing (L‐extreme) and a slow growing (S‐extreme) population. The L‐extreme population grew significantly larger (ANOVA, n = 89–101, P < 0·01) attaining twice the wet mass of the S‐extreme population at 300 dph (130·9 ± 1·8 v. 66·7 ± 0·9 g, mean ± s .e .). When the two populations were sexed, the L‐extreme consisted of 96·5% and the S‐extreme of 30·2% females, while the ungraded control had 59·2% females. Sex differentiation began first in females at a total length (L_T) of 97 ± 4 mm and wet mass of 9·4 ± 1·2 g (150 dph), and was completed when fish reached 166 ± 6 mm and 53·4 ± 6·4 g (250 dph) in both sexes. Precocious maturation in males was positively correlated to growth. Gonad oestradiol (E₂) was significantly higher in the female‐dominant population at the onset of ovarian differentiation (ANOVA, n = 10, P < 0·05) and in the plasma after the appearance of the first primary oocytes (P < 0·01). Gonad testosterone (T) increased in both populations after sex differentiation (ANOVA, n = 10, P < 0·05), while plasma levels were significantly higher in the male‐dominant population (P < 0·001). Both gonad and plasma 11‐keto testosterone (11‐KT) were significantly higher in the male‐dominant population (ANOVA, n = 10, P < 0·01) reaching maximal values at spermiation. The results suggest that E₂ is closely related with ovarian differentiation and the onset of oogenesis, while T and 11‐KT is more related to spermatogenesis and precocious maturation.     

Reproductive output of the ornamental shrimp Lysmata vittata (Stimpson, 1860) (Decapoda: Caridea) in wild populations and under different maturation diets

ABSTRACT

Reproductive output parameters (fecundity and egg volume) of Lysmata vittata were examined in a population in northeastern Brazil. Effect of maturation diets on the reproductive output of these shrimp under laboratory conditions was evaluated. Reproductive output was estimated for 25 shrimp collected in the wild. Another 45 pairs of shrimp were used for diet experiments, 15 pairs per treatment (T1: industrialized food, T2: fresh food, T3: mixed diets). For wild population, mean fecundity and egg volume were differed between developmental stages of the eggs. Fecundity was significantly lower in the specimens subjected to T1 diet (267 ± 141 eggs) compared with shrimp from the wild population (393 ± 183 eggs). Egg volume was significantly lower in shrimp subjected to three diets tested compared with those from wild population. Egg loss during embryonic development in L. vittata may be caused by several factors (e.g. aborted development and maternal cannibalism). Fresh food proved to be important for improving reproductive output in L. vittata reared in culture. We emphasize the significance of improving nutritional value and palatability of diets to improve cultivation efficiency. Lysmata vittata can be used as a model organism for future studies aiming to improve the cultivation techniques for shrimp of the genus.

Abbreviations PSH: protandric simultaneous hermaphroditism; ARS: artificial refuge structures; CL: carapace length; EV: egg volume; L: largest diameter of the egg; S: smallest diameter of the egg; T1: industrialized pellets; T2: fresh ingredients; T3: mixed diets; ANCOVA: analysis of covariancr; SD: standard deviation.