Integrating morphology and physiology of the key endocrine organ during tadpole development: The interrenal gland.

Indirect development is widespread in anurans and is considered an ancestral condition. The metamorphosis of larvae into juveniles involves highly coordinated morphological, physiological, biochemical, and behavioral changes, promoted by the thyroid hormone and interrenal corticosteroids. Stress response to environmental changes is also mediated by corticosteroids, affecting the timing and rate of metamorphosis and leading to great developmental plasticity in tadpoles. Given the potential effect of interrenal gland ontogeny alterations on metamorphosis and the lack of studies addressing both the morphology and endocrinology of this gland in tadpoles, we present corticosterone (CORT) production and histological changes through the ontogeny of interrenal gland in the generalized pond-type tadpole of Rhinella arenarum (Anura, Bufonidae). This species shows the highest concentration of whole-body CORT by the early climax when drastic metamorphic changes begin. This is coincident with the morphological differentiation of steroidogenic cells and the formation of interrenal cords. By this stage, steroidogenic cells have a shrunken cytoplasm, with a significantly higher nucleus-to-cell diameter ratio. The lowest CORT concentration during premetamorphosis and late climax is associated with small undifferentiated cells with lipid inclusions surrounding large blood vessels between kidneys, and with cords of differentiated steroidogenic cells with a significantly lower nucleus-to-cell diameter ratio, respectively. Our study characterizes the morphological and physiological pattern of interrenal gland development, showing an association between certain histological and morphometric characteristics and CORT levels. Variations in this morpho-physiological pattern should be considered when studying the phenotypic plasticity or variable growth rates of tadpoles.

Brothers are better than nothing: first report of incestuous mating and inbreeding depression in a freshwater decapod crustacean.

This study aimed at evaluating the effect of one generation of full-sibling mating on traits related to the fitness of the gregarious freshwater shrimp Neocaridina davidi, both under optimal and stressful (i.e. starvation) laboratory conditions. Females were maintained either with their brothers (Inbreeding treatment) or non-brothers (Outbreeding treatment), and the first and second broods were used to evaluate egg production and juvenile quality, respectively. The latter was analyzed in a 60-day period following hatching under optimal rearing conditions, and in a 20-day period following hatching under food deprivation conditions. All surviving females from both treatments mated and spawned, indicating that mating with brothers was as likely as mating with non-brothers. With respect to offspring production, inbreeding had no effect on fecundity and fertilization success, but negatively affected the number of hatched juveniles. These results suggest that egg loss was higher in inbred clutches, possibly due to lower embryonic survival. On the other hand, the effect of inbreeding on growth was absent for embryos and for juveniles under optimal rearing conditions, while it was significant for juveniles starved for 10 consecutive days following hatching. Inbreeding depression for survival was only detected in juveniles from stages S8-S9 under optimal rearing conditions. Overall, the present results show that N. davidi fitness decreases after one generation of full-sibling mating. The use of a potentially weak food deprivation protocol and/or the evaluation of only one inbred generation could explain the absence or subtle inbreeding depression for some of the evaluated traits. No evident relationship between life-history traits and the existence and magnitude of inbreeding was found when comparing our results with those previously reported in high-fecundity marine crustaceans and low-fecundity terrestrial crustaceans.

Getting ready for mating: The importance of male touching as an accelerator of ovarian growth in a caridean shrimp.

The present study was aimed at evaluating the effect of male presence on ovarian maturation in juvenile females and the role of potential chemical, visual and tactile cues emitted by males in that physiological process. A highly gregarious caridean shrimp with sexual dimorphism, Neocaridina davidi, was used as experimental model. We tested the hypothesis that male presence accelerates ovarian maturation, mainly through chemical cues. Two experiments were performed. In Experiment 1, juvenile females were reared with adult males, adult females or alone, allowing full contact among shrimps. In Experiment 2, these treatments were evaluated allowing chemical and visual communication, only visual communication, or only chemical communication among shrimps. In both experiments juvenile females were observed once a week under a stereomicroscope to determine ovarian growth rate. Although male presence was not necessary for ovarian maturation, it clearly accelerated the rate of ovarian growth, particularly in the last maturation phase. This lead to relatively longer mature ovaries with higher lipid content. On the contrary, the presence of adult females delayed ovarian maturation in juvenile females, while females reared alone showed an intermediate ovarian growth. All these results suggest that adult males release certain cues that stimulate ovarian maturation, while adult females release cues that delay this physiological process. Neither visual cues nor chemical cues released at a distance from females were responsible, either alone or in combination, for the observed effects. Ovarian growth was only influenced when shrimps were allowed to interact freely, probably because of the "mounting" behavior of males towards females. Tactile cues and/or potential chemical cues released by males during this behavior may mediate male stimulatory effect on ovarian growth. Altogether, present results partially support our initial hypothesis and contribute to increase the limited amount of information available on the role of intraspecific multimodal communication in non-behavioral reproductive processes in invertebrate species.

Losing Reproduction: Effect of High Temperature on Female Biochemical Composition and Egg Quality in a Freshwater Crustacean with Direct Development, the Red Cherry Shrimp, Neocaridina davidi (Decapoda, Atyidae).

Maternal provisioning is particularly important in invertebrates with abbreviated development because large energy reserves must be provided for the developing embryo. In this context, the objective of the present study was to analyze in an aquatic invertebrate with direct development the effect of temperature on female biochemical composition and reserve allocation to maturing ovaries, which determine egg quality. A decapod crustacean, the freshwater shrimp Neocaridina davidi, was used as experimental model. Newly hatched juveniles were exposed to 28 °C or 33 °C. Females showed mature ovaries and spawned at 28 °C (control ovigerous females), but no ovigerous female was found at 33 °C. After a 200-day period, half of the females at 33 °C were transferred to 28 °C, where they rapidly showed mature ovaries and spawned (transferred ovigerous females). Ovigerous females and females that did not spawn at 28 °C (control non-ovigerous females) and at 33 °C (high-temperature non-ovigerous females) were sacrificed to determine their biochemical composition. The number, volume, weight, and biochemical composition of the eggs from transferred and control ovigerous females were also analyzed as indicators of their quality. Female biochemical composition was not influenced by temperature, because control and high-temperature non-ovigerous females had similar lipid, protein, and glycogen contents. However, ovarian maturation and spawning were inhibited at 33 °C, which indicates a negative effect of this temperature on nutrient transfer to the oocytes. This effect was rapidly reversed after females were moved to 28 °C; the eggs from control and transferred ovigerous females were of similar quality, except for a lower protein content in the latter. The present results provide valuable information on reserve allocation to reproduction under thermal stress.

Female Growth and Offspring Quality over Successive Spawnings in a Caridean Shrimp Neocaridina davidi (Decapoda, Atyidae) with Direct Development.

This study analyzed the quality of recently spawned eggs and of juveniles over five and six consecutive spawns, respectively, in a caridean shrimp Neocaridina davidi with direct development. The potential energetic antagonism between reproduction and somatic growth was also evaluated. The number of eggs per spawn per female was highest in the first spawn, while the number of recently hatched juveniles per spawn per female declined in the sixth spawn. Lower lipid concentration and energy content were detected in eggs of the fourth and fifth spawns, which may indicate for the first time a decrease in maternal provisioning as a result of multiple spawning in a decapod with direct development. This result had no effect on the size of eggs or of recently hatched juveniles, nor on the growth performance of juveniles during a 30-day period following hatching. Lipids were the most abundant biochemical component of eggs, followed by proteins and glycogen; the relative proportion of each component was probably related to embryonic development type. Egg volume was unsuitable as an indicator of nutrient content, as no correlation was found between these variables. The physiological costs of reproduction were evident from the lower energy content of females that reproduced versus females that remained virgin. The lower body weight of the reproductive females at the end of the experiment showed that allocation of resources to reproduction occurred at the expense of somatic growth. To our knowledge, this is the first empirical demonstration of a decapod with direct development.

Effect of temperature on biochemical composition, growth and reproduction of the ornamental red cherry shrimp Neocaridina heteropoda heteropoda (Decapoda, Caridea).

The effect of water temperature on biochemical composition, growth and reproduction of the ornamental shrimp, Neocaridina heteropoda heteropoda, was investigated to determine the optimum temperature for its culture. The effect of embryo incubation temperature on the subsequent performance of juveniles was also evaluated. Ovigerous females and recently hatched juveniles (JI) were maintained during egg incubation and for a 90-day period, respectively, at three temperatures (24, 28 and 32 °C). Incubation period increased with decreasing water temperature, but the number and size of JI were similar among treatments. At day 30 of the 90-day period, body weight and growth increment (GI) at 24 °C were lower than those at 28 and 32 °C. On subsequent days, GI at 24 °C exceeded that at 28 and 32 °C, leading to a similar body weight among treatments. These results suggest growth was delayed at 24 °C, but only for 30 days after hatching. The lipid concentration tended to be lowest, intermediate and highest at 28, 32 and 24 °C, respectively, possibly as a consequence of the metabolic processes involved in growth and ovarian maturation. Protein and glycogen concentrations were similar among treatments. Both the growth trajectory and biochemical composition of shrimps were affected by the temperature experienced during the 90-day growth period independently of the embryo incubation temperature. During the growth period, shrimps reached sexual maturity and mated, with the highest proportion of ovigerous females occurring at 28 °C. All the females that matured and mated at 32 °C lost their eggs, indicating a potentially stressful effect of high temperature on ovarian maturation. Based on high survival and good growth performance of shrimps at the three temperatures tested over the 90-day period it is concluded that N. heteropoda heteropoda is tolerant to a wide range of water temperatures, with 28 °C being the optimum temperature for its culture.

Effects of androgenic gland ablation on growth and reproductive parameters of Cherax quadricarinatus males (Parastacidae, Decapoda).

This work investigates the effects of androgenic gland (AG) ablation on the structure of the reproductive system, development of secondary sexual characters and somatic growth in Cherax quadricarinatus males. The AG ablation, which was performed at an early developmental stage (initial weight: 1.85±0.03 g), had no effect on the somatic growth parameters (specific growth rate and growth increment), but it prevented the re-formation of male gonopores and appendices masculinae. However, the red patch differentiation and chelae size were similar to those in control males. All the ablated animals developed a male reproductive system. Testis structure was macroscopically and histologically normal. The distal portion of the vas deferens (DVD) was enlarged in some animals, with histological alterations of the epithelium and the structure of the spermatophore. Results suggest that the higher growth in males than in females may be due to an indirect effect of the AG on energy investment in reproduction rather than to a direct effect of an androgen. This is the first report of a potential action of the AG on the secretory activity of the distal VD and the structural organization of the spermatophore. Although the AG may play a role in the development of male copulatory organs, its association with the red patch development deserves further research. The results obtained in the present study support and complement those from intersexes of the same species.