Development and Immature Mortality of the Sawtoothed Grain Beetle (Coleoptera: Silvanidae), on Different Sorghum Fractions and Different Temperatures.

Developmental ability of Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae) was examined on six different sorghum milling fractions: Bran, Shorts, Cgrits, Fgrits, Red dogs, Flour, and also on a standard diet of Oat Flakes. For this purpose, a 1-day-old egg was placed in a vial containing 1 g of one of the sorghum fractions and exposed to three temperatures: 25, 30, or 32°C. All vials were checked daily for pupal and adult emergence and mortality of immatures. The developmental time was significantly affected by the type of sorghum fraction. After two weeks, for most of the temperatures examined, the longest developmental times in most cases for both pupation and adult emergence were observed in Flour and Oat flakes. Increasing the temperature from 25 to 30 accelerated the development, while adult emergence time did not differ between 30 and 32°C for all fractions except Flour. Egg mortality ranged from 11 to 78%, while larval and pupal mortality ranged from 0 to 22 and 0 to 45%, respectively for all sorghum fractions and different temperatures tested. Moreover, the mean overall immature mortality occurred at 30°C was 49.2, 39.7, and 65.1% at 25, 30, and 32°C for all the diets examined. The findings of the present work show that O. surinamensis can develop and survive in sorghum milling fractions and that the optimal temperatures for growth enhancement are 30 and 32°C. The temperatures within sorghum milling facilities could support O. surinamensis development on milling fractions if they are not addressed through phytosanitary measures.

Wolbachia causes cytoplasmic incompatibility but not male-killing in a grain pest beetle.

The endosymbiotic Wolbachia is one of the most common intracellular bacteria known in arthropods and nematodes. Its ability for reproductive manipulation can cause unequal inheritance to male and female offspring, allowing the manipulator to spread, but potentially also impact the evolutionary dynamics of infected hosts. Estimated to be present in up to 66% of insect species, little is known about the phenotypic impact of Wolbachia within the order Coleoptera. Here, we describe the reproductive manipulation by the Wolbachia strain wSur harboured by the sawtoothed grain beetle Oryzaephilus surinamensis (Coleoptera, Silvanidae), through a combination of genomics approaches and bioassays. The Wolbachia strain wSur belongs to supergroup B that contains well-described reproductive manipulators of insects and encodes a pair of cytoplasmic incompatibility factor (cif) genes, as well as multiple homologues of the WO-mediated killing (wmk) gene. A phylogenetic comparison with wmk homologues of wMel of Drosophila melanogaster identified 18 wmk copies in wSur, including one that is closely related to the wMel male-killing homologue. However, further analysis of this particular wmk gene revealed an eight-nucleotide deletion leading to a stop-codon and subsequent reading frame shift midsequence, probably rendering it nonfunctional. Concordantly, utilizing a Wolbachia-deprived O. surinamensis population and controlled mating pairs of wSur-infected and noninfected partners, we found no experimental evidence for male-killing. However, a significant ~50% reduction of hatching rates in hybrid crosses of uninfected females with infected males indicates that wSur is causing cytoplasmic incompatibility. Thus, Wolbachia also represents an important determinant of host fitness in Coleoptera.

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.

Rearing method of Oryzaephilus surinamensis (L.) (Coleoptera, Silvanidae) on various wheat grain granulometry

Oryzaephilus surinamensis is one of most common insect pest of grains and a variety of stored products, and has been found in high numbers in almost all storage facilities. However, laboratory mass rearing of this insect for bioassays is not a simple task, mainly because of its feeding behavior, small size, and high mobility. Thus, the aim of this work was to develop a simple and efficient laboratory rearing method for O. surinamensis, using wheat kernels milled into different granulometry to obtain large number and standardized population at different life stages for bioassays. The adults were collected from storage grain facilities in the southern region of Brazil and 100 specimens were placed inside glass jars with wheat kernels milled at different grades and kept at 25±0.5ºC and 65±5 percent relative humidity. The insects were allowed to copulate and lay eggs for 10 days and then removed. The number of eggs, larvae, and pupae was counted at five-day intervals; longevity of the second generation adults was evaluated. The kernels milled at grade 20 were the best medium for offspring production: 89 percent of eggs by the 5th day; 30.5 percent larvae by the 10th day; 43 percent pupae by the 30th day and 63.4 percent adults at the 46th day. The adults survived up to 450 days. Culturing O. surinamensis under the described conditions, transferring the parental adults by the 10th day after infestation and replacing the media when population builds up will produce enough insects of each stage for various laboratory bioassays.

Oryzaephilus surinamensis é uma das espécies de insetos-praga mais comuns em grãos e outros produtos armazenados, e tem ocorrido em grandes populações em praticamente todas as unidades de armazenamento de grãos. Contudo, a criação massal deste inseto, em laboratório, para bioensaios diversos, não tem sido uma tarefa simples, em função de seu comportamento alimentar, tamanho reduzido e alta mobilidade. Assim, o objetivo deste trabalho foi desenvolver um método simples e eficiente para a criação de O. surinamensis, usando grãos de trigo moídos a diversas granulometrias para obter grandes populações e estágios padronizados para bioensaios. Insetos adultos foram coletados em diversos armazéns na região sul do Brasil; 100 espécimes foram colocados em frascos de vidro, com grãos de trigo moído a diferentes graus, mantidos a 25±0.5ºC e 65±5 por cento de umidade relativa. Os insetos foram mantidos por 10 dias para cópula e postura dos ovos, sendo então removidos. Ovos, larvas e pupas foram contados a intervalos de cinco dias; a longevidade dos adultos da segunda geração foi avaliada. Os grãos moídos a grau 20 representaram o melhor meio para a produção de progênie: 89 por cento dos ovos foram produzidos no 5º dia; 30,5 por cento das larvas no 10º; 43 por cento das pupas no 30º e aproximadamente 63 por cento dos adultos no 46º. Os adultos sobreviveram até 450 dias. A criação de O. surinamensis, nas condições descritas e transferindo os adultos após 10 dias e repondo o meio quando a população aumenta muito, produzirá um número de insetos de cada estágio suficiente para diversos bioensaios de laboratório.