Evolutionary change in the construction of the nursery environment when parents are prevented from caring for their young directly

Parental care can be partitioned into traits that involve direct engagement with offspring and traits that are expressed as an extended phenotype and influence the developmental environment, such as constructing a nursery. Here, we use experimental evolution to test whether parents can evolve modifications in nursery construction when they are experimentally prevented from supplying care directly to offspring. We exposed replicate experimental populations of burying beetles (Nicrophorus vespilloides) to different regimes of posthatching care by allowing larvae to develop in the presence (Full Care) or absence of parents (No Care). After only 13 generations of experimental evolution, we found an adaptive evolutionary increase in the pace at which parents in the No Care populations converted a dead body into a carrion nest for larvae. Cross-fostering experiments further revealed that No Care larvae performed better on a carrion nest prepared by No Care parents than did Full Care larvae. We conclude that parents construct the nursery environment in relation to their effectiveness at supplying care directly, after offspring are born. When direct care is prevented entirely, they evolve to make compensatory adjustments to the nursery in which their young will develop. The rapid evolutionary change observed in our experiments suggests there is considerable standing genetic variation for parental care traits in natural burying beetle populations—for reasons that remain unclear.

Biparental age effects in the burying beetle Nicrophorus vespilloides

Parental age at reproduction influences offspring size and survival by affecting prenatal and postnatal conditions in a wide variety of species, including humans. However, most investigations into this manifestation of ageing focus upon maternal age effects; the effects of paternal age and interactions between maternal and paternal age are often neglected. Furthermore, even when maternal age effects are studied, pre- and postnatal effects are confounded. Using a cross-fostered experimental design, we investigated the joint effects of paternal and pre- and postnatal maternal ages on numerous offspring outcomes in a laboratory population of a species of burying beetle, Nicrophorus vespilloides. When we correct our tests for significance for multiple comparisons, we found no clear evidence for any parental effect senescence acting on egg size, larval weight, or larval survival. Nor did we find a statistical effect of paternal or egg producer age on the outcomes of foster mothers as measured by weight change experienced during caregiving. These findings are consistent with recent negative results reported in a similar study of N. vespilloides maternal age effects while also expanding these to other offspring traits and to paternal age effects. We discuss how the peculiar life history of this species may promote selection to resist the evolution of parental age effects, and how this might have influenced our ability to detect senescence.

Effect of carcass contamination on necrophagous invertebrate performance

Background The breakdown of dead organic matter is driven by a diverse array of organisms and is an important process increasingly impacted by a range of contaminants. While many studies have documented how contaminants affect food webs that are fueled by decaying plant litters, much less is known about how contaminants affect organisms that rely on dead animal material. Here, we begin to explore the effects of food contamination—using silver nanoparticles (AgNPs) as a model contaminant—on the carrion beetle Nicrophorus vespilloides that buries carcasses of small vertebrates in soils as food source and larval nursing grounds. Results Our data show that a single ingestion of a non-lethal dose of 1 μg mL−1 AgNPs by adult female beetles does not affect overall gut microbial activity but results in shifts in the gut microbial community composition towards pathogens including Alcaligenes, Morganella, and Pseudomonas. While no effects were observed in offspring clutch size, some reductions were visible in clutch weight, number of larvae, and number of eclosing pupae in exposed N. vespilloides in comparison with controls. Repeated ingestion of AgNPs over several weeks led to a decrease in survival of adult beetles, suggesting that more environmentally realistic exposure scenarios can directly affect the success of carcass-feeding animals. Conclusions Sub-lethal carcass contamination with a model pollutant can affect the gut microbial composition in female beetles and reduce offspring fitness. This encourages consideration of currently overlooked propagation routes of contaminants through necrophagous food webs and inherent consequences for ecological and evolutionary processes.

Access to Resources Shapes Sex Differences Between Caring Parents

In species where both parents cooperate to care for their joint offspring, one sex often provides more care than the other. The magnitude of such sex differences often varies both between and within species and may depend on environmental conditions, such as access to resources, predation risk and interspecific competition. Here we investigated the impact of one such environmental variable – access to resources for breeding – on the magnitude of sex differences in parental care in the burying beetle Nicrophorus vespilloides. This species breeds on the carcasses of small vertebrates, which are the sole food source for parents and offspring during breeding. We manipulated access to resources by providing pairs with mouse carcasses from a broad mass range (3.65–26.15 g). We then monitored subsequent effects on the duration and amount of care provided by males and females, male and female food consumption and weight change during breeding, and larval traits related to offspring performance. We found that males increased their duration of care as carcass mass increased, while females remained with the brood until it had completed its development irrespective of carcass mass. There were thus more pronounced sex differences in parental care when parents had access to fewer resources for breeding. Overall, our findings show that sex differences between caring parents vary depending on access to resources during breeding. The finding that males extended their duration of care on larger carcasses suggests that access to more resources leads to a shift toward more cooperation between caring parents.

Effect of Short-Term Desiccation, Recovery Time, and CAPA–PVK Neuropeptide on the Immune System of the Burying Beetle Nicrophorus vespilloides

Environmental conditions, especially related to winter, are crucial for shaping activity of insect immune system. However, our previous research clearly indicates differences in the immune system functioning when the cold stress was induced in the laboratory conditions and when the beetles were collected from natural environment during winter. This is probably related to the multiplication of observed effects by simultaneous presence of different stress factors characteristic of winter, including desiccation. For these reasons, our next step was analysis of the effects of short-term desiccation and recovery time on the functioning of immune system of burying beetle Nicrophorus vespilloides. Also, the effect of Tenmo–PVK-2 (tenebrionid periviscerokinin), member of the CAPA–PVK neuropeptide family, was investigated to better understand observed changes. Short-term desiccation decreases the phagocytic activity of burying beetle haemocytes, which is correlated with a reduction in their adhesive ability. On the other hand, there was a significant increase in phenoloxidase (PO) activity and the level of proPO expression, which may suggest sealing the cuticula by melanin deposition and prevention of water loss. Additionally, the elevated level of defensin expression may be associated with the cross-talk between mechanisms, which participate in insect response to environmental stress, including pathogen infection. After 1 h of recovery time, the activity of tested cellular and humoral mechanisms was mostly back to the control level. However, inhibition of the activity of PO and down-regulation of proPO were noted. These results also indicate importance of melanin deposition during water loss. Moreover, it suggests that some changes in immune system functioning during stress conditions do not have an immune function. Interestingly, part of the effects characteristic of recovery time were also observed after the application of Tenmo–PVK-2, mainly related to haemocyte morphology. These results indicate that CAPA–PVK neuropeptides may also influence on activity of burying beetle immune system. It should be also highlighted that, because of the study of the effects of CAPA–PVK neuropeptides, homologs of vertebrate neuromedin U, the results may be interesting for search evolutionary similarities in the functioning of the neuroendocrine system of insects and vertebrates.

Genomic analysis of novel Yarrowia-like yeast symbionts associated with the carrion-feeding burying beetle Nicrophorus vespilloides

Background Mutualistic interactions with microbes can help insects adapt to extreme environments and unusual diets. An intriguing example is the burying beetle Nicrophorus vespilloides, which feeds and reproduces on small vertebrate carcasses. Its fungal microbiome is dominated by yeasts that potentially facilitate carcass utilization by producing digestive enzymes, eliminating cadaver-associated toxic volatiles (that would otherwise attract competitors), and releasing antimicrobials to sanitize the microenvironment. Some of these yeasts are closely related to the biotechnologically important species Yarrowia lipolytica. Results To investigate the roles of these Yarrowia-like yeast (YLY) strains in more detail, we selected five strains from two different phylogenetic clades for third-generation sequencing and genome analysis. The first clade, represented by strain B02, has a 20-Mb genome containing ~ 6400 predicted protein-coding genes. The second clade, represented by strain C11, has a 25-Mb genome containing ~ 6300 predicted protein-coding genes, and extensive intraspecific variability within the ITS–D1/D2 rDNA region commonly used for species assignments. Phenotypic microarray analysis revealed that both YLY strains were able to utilize a diverse range of carbon and nitrogen sources (including microbial metabolites associated with putrefaction), and can grow in environments with extreme pH and salt concentrations. Conclusions The genomic characterization of five yeast strains isolated from N. vespilloides resulted in the identification of strains potentially representing new YLY species. Given their abundance in the beetle hindgut, and dominant growth on beetle-prepared carcasses, the analysis of these strains has revealed the genetic basis of a potential symbiotic relationship between yeasts and burying beetles that facilitates carcass digestion and preservation.

Carry on caring: infected females maintain their parental care despite high mortality

Parental care is a key component of an organism’s reproductive strategy that is thought to trade-off with allocation toward immunity. Yet, it is unclear how caring parents respond to pathogens: do infected parents reduce care as a sickness behavior or simply from being ill or do they prioritize their offspring by maintaining high levels of care? To address this issue, we investigated the consequences of infection by the pathogen Serratia marcescens on mortality, time spent providing care, reproductive output, and expression of immune genes of female parents in the burying beetle Nicrophorus vespilloides. We compared untreated control females with infected females that were inoculated with live bacteria, immune-challenged females that were inoculated with heat-killed bacteria, and injured females that were injected with buffer. We found that infected and immune-challenged females changed their immune gene expression and that infected females suffered increased mortality. Nevertheless, infected and immune-challenged females maintained their normal level of care and reproductive output. There was thus no evidence that infection led to either a decrease or an increase in parental care or reproductive output. Our results show that parental care, which is generally highly flexible, can remain remarkably robust and consistent despite the elevated mortality caused by infection by pathogens. Overall, these findings suggest that infected females maintain a high level of parental care, a strategy that may ensure that offspring receive the necessary amount of care but that might be detrimental to the parents’ own survival or that may even facilitate disease transmission to offspring.

Evolved changes in DNA methylation in response to the sustained loss of parental care in the burying beetle

Levels of parental care critically influence the development environment with the capacity to impact the growth, survival, physiology, and behaviour of offspring. Plastic changes in DNA methylation have been hypothesised to modulate gene expression responses to parental environments. Moreover, these effects can be inherited and so may affect the process of adaptive evolution. In this study, using experimental evolution, we investigated how plastic changes in DNA methylation induced by the loss of parental care have evolved in a biparental insect (Nicrophorus vespilloides) using experimental evolution. We show that removal of care in a single generation is associated with changes in gene expression in stress-related pathways in 1st instar larvae. However, in larvae that have adapted to the loss of parental care after being deprived of care for 30 generations, gene expression is shifted from stress-related gene expression towards growth and brain development pathways. We found that changes in gene body methylation arose both as a direct response to the loss of parental care and stochastically as populations diverged. Overall, our results suggest that a complex interplay between transcription and DNA methylation shapes the molecular adaptation to environmental change.