Transgenerational effects decrease larval resilience to ocean acidification and warming but juvenile European sea bass could benefit from higher temperatures in the NE Atlantic

The aim of this study was to investigate the effect of ocean acidification (OA) and warming (OW) as well as the transgenerational effect of OA on larval and juvenile growth and metabolism of a large economically important fish species with a long generation time. Therefore we incubated European sea bass from Brittany (France) for two generations (>5 years in total) under current and predicted OA conditions (PCO2: 650 and 1700 μatm). In the F1 generation both OA condition were crossed with OW (temperature: 15-18 °C and 20-23 °C). We found that OA alone did not affect larval or juvenile growth and OW increased developmental time and growth rates, but OAW decreased larval size at metamorphosis. Larval routine metabolic rate (RMR) and juvenile standard metabolic rate (SMR) were significantly lower in cold compared to warm conditioned fish and also lower in F0 compared to F1 fish. We did not find any effect of OA on RMR or SMR. Juvenile PO2crit was not affected by OA, OW or OAW in both generations. We discuss the potential underlying mechanisms resulting in beneficial effects of OW on F1 larval growth and RMR and in resilience of F0 and F1 larvae and juveniles to OA, but on the other hand resulting in vulnerability of F1, but not F0 larvae to OAW. With regard to the ecological perspective, we conclude that recruitment of larvae and early juveniles to nursery areas might decrease under OAW conditions but individuals reaching juvenile phase might benefit from increased performance at higher temperatures.

Effect of Temperature on the Daily Increment Deposition in the Otoliths of European Sardine Sardina pilchardus (Walbaum, 1792) Larvae

Otolith microstructure analysis is a valuable tool to evaluate the relationship between larval age and growth and how it relates to environmental variability. Otolith growth and daily increment deposition were analyzed in sardine (Sardina pilchardus) larvae reared in the laboratory under different temperatures (13, 15, and 17 °C), with a diet rich in microalgae, rotifers, and copepods Acartia grani. The number and width of growth increments, first-check and otolith diameter were determined in the otoliths and then related to larval age and total length. At hatching, the sagittal otoliths consisted of a lenticular core with a diameter of 10.56 μm (±1.07 μm SD). Somatic growth increased with the increasing temperature and the growth rate of larvae reared at 13 and 15 °C was significantly lower than for larvae reared at 17 °C. At 17 °C, otoliths exhibited a higher diameter with wider increments than at 13 °C. There was a high variability of increment counts-at-age for larvae reared within the same temperature treatment. The increase of growth increments with larval size was higher for larvae reared at 17 °C until 35 days post-hatching than those growing at 15 °C. Scanning electronic microscopy confirmed that increments are deposited daily, with an average width smaller than 1 µm and as low as 0.33 μm, therefore impossible to distinguish using light microscopy. At colder temperatures, larval otoliths had thinner and less marked increments and lower growth rates, which can lead to incorrect age determinations. The effect of temperature on the otolith microstructure can help in identifying strong temperature gradients experienced by wild sardine larvae.

Neuropeptide Bursicon Influences Reproductive Physiology in Tribolium Castaneum

Bursicon is a neuropeptide belonging to the cystine knot family and is composed of burs and partner of burs (pburs) subunits. It can form heterodimers or homodimers to execute different biological functions. Bursicon heterodimers regulate cuticle sclerotization and wing maturation, whereas bursicon homodimers mediate innate immunity and midgut stem cell proliferation. A recent study has shown that bursicon potentially induces the expression of vitellogenin (Vg) in the black tiger shrimp Penaeus monodon; however, the underlying mechanism remains unknown. In this study, we investigated the role of bursicon in the reproductive physiology of the red flour beetle, Tribolium castaneum. The knockdown of burs, pburs, or its receptor T. castaneum rickets (Tcrk) in 2-day pupae significantly downregulated the expression levels of Vg1, Vg2, and Vg receptor (VgR) genes in females 3- and 5-day post-adult emergence, leading to abnormal oocytes with limited Vg content. The silencing of burs repressed the number of eggs laid and completely inhibited egg hatch, whereas the silencing of pburs dramatically decreased the number of eggs laid, hatch rate, and offspring larval size, and this RNA interference (RNAi) effects persisted to the next generation. Furthermore, the knockdown of burs or pburs downregulated the expression of the insulin/insulin-like signaling/target of rapamycin (TOR) signaling genes encoding insulin receptor (InR), protein kinase B (Akt), TOR, and ribosomal protein S6 kinase (S6K). Most importantly, the injection of recombinant pburs (r-pburs) protein was able to upregulate the expression of Vg, VgR, InR, Akt, TOR, S6K, JH synthesis (JHAMT), Methoprene-tolerant (Met), and Taiman (Tai) in normal females and rescue the expression of Vg and VgR in pburs RNAi females but failed to rescue Vg and VgR in Tcrk knockdown females. We infer that bursicon homodimers influence Vg expression via the receptor Tcrk, possibly by mediating the expression of the juvenile hormone (JH) and IIS/TOR pathway genes, thereby regulating reproduction in T. castaneum.

Both consumptive and non-consumptive effects of predators impact mosquito populations and have implications for disease transmission

Predator-prey interactions influence prey traits through both consumptive and non-consumptive effects, and variation in these traits can shape vector-borne disease dynamics. Meta-analysis methods were employed to generate predation effect sizes by different categories of predators and mosquito prey. This analysis showed that multiple families of aquatic predators are effective in consumptively reducing mosquito survival, and that the survival of Aedes, Anopheles, and Culex mosquitoes is negatively impacted by consumptive effects of predators. Mosquito larval size was found to play a more important role in explaining the heterogeneity of consumptive effects from predators than mosquito genus. Mosquito survival and body size were reduced by non-consumptive effects of predators, but development time was not significantly impacted. In addition, Culex vectors demonstrated predator avoidance behavior during oviposition. The results of this meta-analysis suggest that predators limit disease transmission by reducing both vector survival and vector size, and that associations between drought and human West Nile virus cases could be driven by the vector behavior of predator avoidance during oviposition. These findings are likely to be useful to infectious disease modelers who rely on vector traits as predictors of transmission.

Contrasting Manual and Automated Assessment of Thermal Stress Responses and Larval Body Size in Black Soldier Flies and Houseflies

Within ecophysiological and genetic studies on insects, morphological and physiological traits are commonly assessed and phenotypes are typically obtained from manual measurements on numerous individuals. Manual observations are, however, time consuming, can introduce observer bias and are prone to human error. Here, we contrast results obtained from manual assessment of larval size and thermal tolerance traits in black soldier flies (Hermetia illucens) and houseflies (Musca domestica) that have been acclimated under three different temperature regimes with those obtained automatically using an image analysis software (Noldus EthoVision XT). We found that (i) larval size estimates of both species, obtained by manual weighing or by using the software, were highly correlated, (ii) measures of heat and cold tolerance using manual and automated approaches provided qualitatively similar results, and (iii) by using the software we obtained quantifiable information on stress responses and acclimation effects of potentially higher ecological relevance than the endpoint traits that are typically assessed when manual assessments are used. Based on these findings, we argue that automated assessment of insect stress responses and largescale phenotyping of morphological traits such as size will provide new opportunities within many disciplines where accurate and largescale phenotyping of insects is required.

Effect of dietary astaxanthin on reproductive performance, egg quality and larvae of clowfish Amphiprion ocellaris (Cuvier, 1830)

This study was designed to evaluate the effects of astaxanthin in broodfish diets on reproductive performance, egg quality and larvae quality parameters of clownfish (Amphirion ocellaris). Five treatments were tested with 5 levels of astaxanthin (Carophyll Pink 10% CWS) of 0, 50, 100, 150 and 200 mg/kg added to the feed. Each treatment was repeated in triplicate and the supplemental feeding trials were arranged for 13 months. The results showed that there were significant differences in hatching rate of egg, malformed rate and survival rate of larvae in 3 days post-hatch (p<0.05) among the feeding trials of astaxanthin supplements. The highest hatching rate of egg and survival rate and the lowest malformed rate of larvae were observed in the treatment that was supplemented with astaxanthin 150 mg/kg feed, respectively 92.14 %; 93.57 % and 0.55 %. However, the astaxanthin supplementary diets did not affect the re-maturation and spawning period, spawning frequency, fecundity, egg diameter and larval size of nemo fish among the treatments. The results also suggested that astaxanthin requirement for clownfish broodstock to improve reproductive performance was 150 mg/kg feed.

Embryonic and larval development is conditioned by water temperature and maternal origin of eggs in the sea urchin Arbacia dufresnii (Echinodermata: Echinoidea)

Introduction: Embryonic and larval development in sea urchins is highly dependent on maternal nutritional status and on the environmental conditions of the seawater. Objective: To compare the development of Arbacia dufresnii in two different water temperatures and in progeny with varying maternal origins. Methods: We induced A. dufresnii females and males from Nuevo Gulf to spawn, collected the eggs of each female individually (progeny), separated them into two seawater temperatures (12 and 17 °C), and fertilized them. We recorded the percentage of fertilized eggs and embryos per developmental stage according to time, temperature and progeny. We measured larval growth by total length (TL) and midline body length (M) according to time post fecundation (DPF), temperature, and progeny. Results: Temperature did not affect fertilization, but embryo development was faster and more synchronized in the high temperature treatment. The generalized linear models indicate that embryo development depends on a quadruple interaction between the embryonic stage, time (h), seawater temperature and progeny. Larval growth was faster, producing larger larvae at the highest temperature. Larval growth depends on a triple interaction between time (DPF), seawater temperature and progeny. Conclusions: We found a temperature and progeny impact during embryonic and larval development and, in both cases, these factors generate a synergistic effect on developmental timing and larval size. This probably provides a survival advantage as a more rapid speed of development implies a decrease in the time spent in the water column, where the sea urchins are vulnerable to biotic and abiotic stressors.

Nano-CT characterization reveals coordinated growth of a rudimentary organ necessary for soldier development in the antPheidole hyatti

The growth of imaginal discs in holometabolous insects is coordinated with larval growth to ensure the symmetrical and proportional development of the adult appendages. In ants, the differential growth of these discs generates distinct castes – the winged male and queen castes and the wingless worker caste. In the hyperdiverse ant genusPheidole, the worker caste is composed of two morphologically distinct subcastes: small minor workers and larger, big-headed soldiers. Although these worker subcastes are completely wingless, soldier larvae develop rudimentary forewing discs that are necessary for generating the disproportionate head-to-body scaling of the big-headed soldier subcaste. However, it remains unclear whether rudimentary forewing discs in soldier larvae are coordinated with other imaginal discs, and whether disc growth and coordination patterns vary between the minor worker and soldier subcastes. Here we show, using quantitative nano-CT three-dimensional analyses, that growth of the soldier rudimentary forewing discs is coordinated with the increase in volume of the leg and eye-antennal (head) discs as well as with larval size. We found that the growth rate of the rudimentary forewing discs differs from the leg discs but is similar to the growth of the head (eye-antennal) discs relative to larval size, suggesting that growth of each type of imaginal disc may be differentially regulated. In addition to their larger size, the soldier eye-antennal discs increase in width as they undergo morphogenesis to generate the characteristic shape of the large soldier head, suggesting that the rudimentary forewing discs may regulate their patterning in addition to their growth. Finally, we observe little growth of the leg and eye-antennal discs during the bipotential stage, while in minor worker development these discs grow at similar rates to one another in coordination with larval size to generate the smaller minor worker subcaste. Our results suggest that rudimentary organs with regulatory functions may participate in new patterns of inter-organ coordination and regulation to produce novel phenotypes and complex worker caste systems. We provide characterization of larval development and imaginal disc growth and morphogenesis with the aim of highlighting this as an emerging system for the study of rudimentary organs during development and evolution.

Growth, winter preparations and timing of emergence in temperate zone Odonata: control by a succession of larval response patterns

As warm-adapted insects of tropical origin, Odonata cope with cold periods by seasonal regulation and diapause. A model for larval-overwintering species is proposed with three response patterns related to the timing of emergence, which can be predicted from seasonal cues during the last few stadia. For emergence during the present season, there is an often time-constrained preemergence development, accelerated by long days and higher temperatures. In regulatory development, emergence is postponed to the next season, and a complex of diapause-like delays controlled by photoperiod and temperature prevents premature emergence. Instead, development converges on a winter diapause in sizes suitable for emergence during the following year. Long days are particularly delaying, and thermal responses are variable, sometimes inverted. In early development, with rapid growth, emergence is usually not predicted to season, but short-day winter diapauses may occur, and precocious preparations for a penultimate winter may be predictive. Thermal responses are steep, extremely so if a short-day diapause is suppressed by higher temperatures. Other physiological and also behavioural properties may differ between response patterns. Changes in photoperiod and temperature control the timing of seasonal events, and the transition from regulatory to pre-emergence development follows the increase in temperature and photoperiod after winter, which is an important time-setter. Interactions of larval size, photoperiod, temperature and previous changes affect development rate, and long-term constant conditions often end in regulatory diapauses. Proximate mechanisms of cohort splitting and the implications of the model for design and interpretation of experiments are discussed.