Endosymbiosis allows Sitophilus oryzae to persist in dry conditions.

Insects frequently associate with intracellular microbial symbionts (endosymbionts) that enhance their ability to cope with challenging environmental conditions. Endosymbioses with cuticle-enhancing microbes have been reported in several beetle families. However, the ecological relevance of these associations has seldom been demonstrated, particularly in the context of dry environments where high cuticle quality can reduce water loss. Thus, we investigated how cuticle-enhancing symbionts of the rice-weevil, Sitophilus oryzae contribute to desiccation resistance. We exposed symbiotic and symbiont-free (aposymbiotic) beetles to long-term stressful (47% RH) or relaxed (60% RH) humidity conditions and measured population growth. We found that symbiont presence benefits host fitness especially under dry conditions, enabling symbiotic beetles to increase their population size by over 33-fold within 3 months, while aposymbiotic beetles fail to increase in numbers beyond the starting population in the same conditions. To understand the mechanisms underlying this drastic effect, we compared beetle size and body water content and found that endosymbionts confer bigger body size and higher body water content. While chemical analyses revealed no significant differences in composition and quantity of cuticular hydrocarbons after long-term exposure to desiccation stress, symbiotic beetles lost water at a proportionally slower rate than did their aposymbiotic counterparts. We posit that the desiccation resistance and higher fitness observed in symbiotic beetles under dry conditions is due to their symbiont-enhanced thicker cuticle, which provides protection against cuticular transpiration. Thus, we demonstrate that the cuticle enhancing symbiosis of Sitophilus oryzae confers a fitness benefit under drought stress, an ecologically relevant condition for grain pest beetles. This benefit likely extends to many other systems where symbiont-mediated cuticle synthesis has been identified, including taxa spanning beetles and ants that occupy different ecological niches.

Nutritional symbionts enhance structural defence against predation and fungal infection in a grain pest beetle.

Many insects benefit from bacterial symbionts that provide essential nutrients and thereby extend the hosts' adaptive potential and their ability to cope with challenging environments. However, the implications of nutritional symbioses for the hosts' defence against natural enemies remain largely unstudied. Here, we investigated whether the cuticle-enhancing nutritional symbiosis of the saw-toothed grain beetle Oryzaephilus surinamensis confers protection against predation and fungal infection. We exposed age-defined symbiotic and symbiont-depleted (aposymbiotic) beetles to two antagonists that must actively penetrate the cuticle for a successful attack: wolf spiders (Lycosidae) and the fungal entomopathogen Beauveria bassiana. While young beetles suffered from high predation and fungal infection rates regardless of symbiont presence, symbiotic beetles were able to escape this period of vulnerability and reach high survival probabilities significantly faster than aposymbiotic beetles. To understand the mechanistic basis of these differences, we conducted a time-series analysis of cuticle development in symbiotic and aposymbiotic beetles by measuring cuticular melanisation and thickness. The results reveal that the symbionts accelerate their host's cuticle formation and thereby enable it to quickly reach a cuticle quality threshold that confers structural protection against predation and fungal infection. Considering the widespread occurrence of cuticle enhancement via symbiont-mediated tyrosine supplementation in beetles and other insects, our findings demonstrate how nutritional symbioses can have important ecological implications reaching beyond the immediate nutrient-provisioning benefits.

Metabolic Cost of a Nutritional Symbiont Manifests in Delayed Reproduction in a Grain Pest Beetle.

Animals engage in a plethora of mutualistic interactions with microorganisms that can confer various benefits to their host but can also incur context-dependent costs. The sawtoothed grain beetle Oryzaephilus surinamensis harbors nutritional, intracellular Bacteroidetes bacteria that supplement precursors for the cuticle synthesis and thereby enhance desiccation resistance of its host. Experimental elimination of the symbiont impairs cuticle formation and reduces fitness under desiccation stress but does not disrupt the host's life cycle. For this study, we first demonstrated that symbiont populations showed the strongest growth at the end of metamorphosis and then declined continuously in males, but not in females. The symbiont loss neither impacted the development time until adulthood nor adult mortality or lifespan. Furthermore, lifetime reproduction was not influenced by the symbiont presence. However, symbiotic females started to reproduce almost two weeks later than aposymbiotic ones. Thus, symbiont presence incurs a metabolic and context-dependent fitness cost to females, probably due to a nutrient allocation trade-off between symbiont growth and sexual maturation. The O. surinamensis symbiosis thereby represents an experimentally amenable system to study eco-evolutionary dynamics under variable selection pressures.

Biochar alleviates the toxicity of imidacloprid and silver nanoparticles (AgNPs) to Enchytraeus albidus (Oligochaeta).

The present study investigated the use of biochar for the alleviation of the toxic effects of a nanosilver colloidal dispersion and a chloronicotinyl insecticide. The survival and reproduction of the potworm Enchytraeus albidus were assessed after exposure to imidacloprid and silver nanoparticles (AgNPs). E. albidus was exposed to 0, 25, 50, 100, 200, and 400 mg imidacloprid/kg and 0, 5, 25, 125, and 625 mg Ag/kg for 21 days in 10% biochar amended and non-biochar amended OECD artificial soil. In both exposure substrates, the effects of imidacloprid on survival were significant in the two highest treatments (p < 0.01). No biochar effect was observed as survival was statistically similar in both soils after exposure to imidacloprid. In the case of AgNPs, significant mortality was only observed in the highest AgNP treatments in both the amended and non-amended soils (p < 0.05). Nevertheless, statistically greater survival occurred in the biochar-amended treatment (p < 0.05). Reproduction results showed a more pronounced biochar effect with an EC50 = 22.27 mg imidacloprid/kg in the non-amended soil and a higher EC50 = 46.23 mg imidacloprid/kg in the biochar-amended soil. This indicated a 2-fold decrease in imidacloprid toxicity due to biochar amendment. A similar observation was made in the case of AgNPs where a reproduction EC50 = 166.70 mg Ag/kg soil in the non-amended soil increased to an EC50 > 625 mg Ag/kg soil (the highest AgNP treatment) in the amended soil. This indicated at least a 3.7-fold decrease in AgNPs toxicity due to biochar amendment. Although more studies may be needed to optimize the easing effects of biochar on the toxicity of these chemicals, the present results show that biochar could be useful for the alleviation of the toxic effects of imidacloprid and silver nanoparticles in the soil.