Conditions For Larval Development Research Articles

Fecundity, in vitro early larval development and karyotype of the zoonotic nematode Anisakis pegreffii

The in vitro life cycle of zoonotic helminths is an essential tool for -omic translational studies focused on disease control and treatment. Anisakiosis is an emerging zoonosis contracted by the ingestion of raw or undercooked fish infected with the third stage larvae (L3) of two sibling species Anisakis simplex sensu stricto (s.s.) and Anisakis pegreffii, the latter being the predominant species in the Mediterranean basin. Recently, in vitro culture of A. pegreffii has been developed to enable fast and large-scale production of fertile adults. However, the conditions for larval development from hatching to infective L3 were not fulfilled to complete the cycle. Herein, we used a Drosophila medium supplemented with chicken serum and adjusted different osmolarities to maintain the culture of L3 hatched from eggs for up to 17 weeks. The highest survival rate was observed in the medium with the highest osmolarities, which also allowed the highest larval exsheathment rate. Key morphological features of embryogenesis and postembryogenesis studied by transmission electron microscopy revealed that the excretory gland cell is differentiated already up to 48 h post-hatching. Extracellular vesicles and cell-free mitochondria are discharged between the two cuticle sheets of the second stage larvae (L2). Contemporarly cultivated, two populations of adult A. simplex s.s. and A. pegreffii reached an average production of 29,914.05 (± 27,629.36) and 24,370.96 (± 12,564.86) eggs/day/female, respectively. The chromosome spreads of A. pegreffii obtained from mature gonads suggests a diploid karyotype formula of 2n = 18. The development of a reliable protocol for the in vitro culture of a polyxenous nematode such as Anisakis spp. will serve to screen for much needed novel drug targets, but also to study the intricated and unknown ecological and physiological traits of these trophically transmitted marine nematodes.

Change of morphometric and allometric patterns on wings of banded demoiselle (Calopteryx splendens) males in case of ecologically different watercourse types

In the nature, larvae living in watercourses are exposed to a complex system of environmental influences. It is known that different watercourse types (creeks, brooks, streams, little rivers and medial rivers) provide different conditions for larval development (water depth, flow rate, temperature, oxygen content, substrate type, nutrient supply, etc.). These conditions can vary significantly between watercourse types, but be very similar within types. In this work, we examined the body sizes and wing morphometric characteristics of males of Calopteryx splendens reared from different watercourse types (brook, stream, creek, little river, medial river). Although there were no significant differences in body size among watercourse types, we found significant differences in the wing features. We found the most differences between the individuals reared from streams and creeks and between the individuals reared from stream and medial river. Our results show that the individuals reared from different watercourse types were clearly separated on the two wings. The results also suggest that there are significant differences in the number and pattern of allometric features on the wings of individuals reared from different watercourse types.

Regional variations in early life stages response to a temperature gradient in the northern shrimp Pandalus borealis and vulnerability of the populations to ocean warming

In order to define the relative vulnerability of northern shrimp (Pandalus borealis) populations to the ongoing global warming, we compared the thermal performance curves for survival and growth in the first three pelagic larval stages from three populations of the Northwest Atlantic. Egg carrying females were obtained from different regions characterized by distinct sea surface temperature (SST) conditions for larval development in spring. Two independent experiments were conducted in two different years. In spring 2012, larvae from females captured in the Lower St Lawrence Estuary (LE) and in the Northeast Gulf of St Lawrence (GSL) were compared. In spring 2014, larvae from females captured in the LE and on the Labrador–Newfoundland Shelf (Northwest Atlantic, NWA) were used. The LE larvae were used both years and served as the reference population for comparisons. In 2012 and 2014, groups of 25 newly hatched northern shrimp larvae from each source population were incubated at six temperatures (0, 3, 6, 9, 12, and 15°C) to monitor and compare survival and growth at moult. Northern shrimp larvae from the LE (warmer May–June SST) had a higher optimal temperature range for survival compared to larvae from the GSL and the NWA (colder May–June SST) populations. However, in 2012 growth performance at moult was reduced at higher temperatures for the LE population compared to the GSL population. The differences in thermal performance curves observed may suggest the presence of a certain level of local adaptation in response to the different regional SST regimes in spring–early summer. Northern shrimp larvae in the Northeast Gulf of St Lawrence and Northwest Atlantic shelf could benefit from warmer early-spring temperatures; however, larvae from the Lower Estuary may be closer to their upper tolerance limits and thus more likely at risk of negative impact of future warming of surface water masses.

Optimization of Methodology for Rearing Spodoptera albula on Artificial Diet.

Advances in techniques for rearing insects on artificial diets are fundamental to solving issues of basic and applied entomology. In this study, we evaluated the development of Spodoptera albula (Walker) (Lepidoptera: Noctuidae) on three artificial diets used for other species of Lepidoptera, at three larval densities, and two densities of adult couples housed in oviposition cages of two sizes, with the aim of optimizing methodology for rearing S. albula in the laboratory. Biological parameters were recorded from S. albula, and a fitness index was calculated based on the larval survival and duration and weight of pupae. The total and daily oviposition was recorded using 5 or 10 adult couples of S. albula housed in two cage sizes. Concentrations of total nitrogen and protein in the tested diets were determined. Development of S. albula was completed in all artificial diets; however, the diet used for rearing Anticarsia gemmatalis (Hübner) larvae was the most suitable for S. albula, yielding intermediate development time and higher survival relative to the other diets. Individualization of larvae favored S. albula development by producing overall greater weights of larvae and pupae, higher survival rates, and longer adult longevity. Cage size and number of couples per cage did not influence S. albula fecundity in the experiment conditions. Spodoptera albula can be satisfactorily reared on the artificial diet used for A. gemmatalis, using one larva per tube, and either density of adults at any cage size. Additional amendments are needed in the rearing methodology to achieve optimal conditions for larval development to adulthood.

Energetic adaptations to larval export within the brackish-water palaemonine shrimp, Palaemonetes varians

Decapod crustaceans have repeatedly colonised brackish, freshwater, and terrestrial environments. Many decapods, which inhabit brackish and freshwater habitats, export larvae into estuarine and coastal areas where conditions for larval development may be better. In this study, we assess the starvation resistance, biochemical composition and respiration rate during larval development, and the effects of temperature on these factors within the brackish living palaemonine shrimp, Palaemonetes varians. Our results demonstrate that P. varians is highly resistant to starvation and may be considered facultative lecithotrophic in its first and second larval instars and planktotrophic from its third instar. This high starvation resistance is associated with a relatively large size, high carbon content (~45%) and C:N ratio (~4.2), and visible yolk reserves at hatching. These energy reserves are interpreted as an adaptation to the exportation of larvae from peripheral adult environments into mid and lower estuarine waters. Respiration rates varied with the moult cycle and were similar between fed and unfed larvae, suggesting that starved larvae do not suppress their metabolism as an energy saving strategy. Despite higher respiration rates at higher temperatures, energy loss throughout development (estimated from respiration rates) increased with decreasing temperature, whilst larval growth and development rates decreased with increasing temperature. High energy reserves at hatching, as within Palaemonetes varians, is an important life history adaptation in the colonisation of brackish and freshwaters, initially enabling the exportation of larvae from adult environments and eventually enabling lecithotrophy and direct development.

Stable Transmission ofDirofilaria repensNematodes, Northern Germany

Stable Transmission of<i>Dirofilaria repens</i>Nematodes, Northern Germany

Building of Leaf Shelters by Stenoma scitiorella Walker (Lepidoptera: Elachistidae): Manipulation of Host Plant Quality?

Building of leaf shelters is a widespread behavior among microlepidopteran larvae. Apparently, these shelters provide refuge to caterpillars against bad weather and reduce the risks of predation and parasitism. Recent studies about nutritional ecology of insects have suggested that such behavior would actually be an strategy evolutionarily selected to maximize the quality of the host plant consumed by them. Stenoma scitiorella Walker caterpillars tie together young Xylopia aromatica (Lam.) Mart. (Annonaceae) leaves to build a shelter in which they retreat, feeding exclusively on inner leaves. The accumulation of damage and excrement in leaf shelters with several caterpillars may lead to a decline in larval development and survivorship. For providing the best conditions for larval development most leaf shelters (53,4%) are occupied by only one caterpillar. Although altering the water and soluble carbohydrates contents, the building of shelters did not affect either the tannins concentrations or the preference of caterpillars for external or internal leaves. Hence, probably this behavior does not denote an adaptative strategy of the insect to overcome the host plant chemical defenses. Thus, it is possible that the leaf shelters of S. scitiorella caterpillars are built for other purposes as, for example, to protect caterpillars from desiccation and predation.