Evaluation of Phosphine Resistance in Populations of Sitophilus oryzae, Oryzaephilus surinamensis and Rhyzopertha dominica in the Czech Republic.

Phosphine is globally the most widely adopted fumigant for the control of storage pests. Recently, an increase in the frequency of stored-product pest resistance has been observed with significant geographical and interspecific variations. In this context, there are available data for the occurrence of resistant populations from America, Asia, Africa, and Australia, but there are few data in the case of Europe. Therefore, the aim of this work was to evaluate phosphine efficacy in important beetle pests of stored products, i.e., Sitophilus oryzae (L.), Oryzaephilus surinamensis (L.), and Rhyzopertha dominica (F.) sampled from the Czech Republic, using a rapid diagnostic test that is based on the speed to knockdown after exposure. Apart from the standard laboratory populations, which were used as the controls, we tested 56 field populations of these three species, collected in Czech farm grain stores. The survey revealed that 57.1% of the tested field populations were classified as phosphine-susceptible, based on the knockdown method used. However, profound variations among species and populations were recorded. The species with the highest percentage of resistant populations was R. dominica (71.4% of the populations; resistance coefficient 0.5-4.1), followed by S. oryzae (57.1% of the populations; resistance coefficient 0.8-6.9), and O. surinamensis (9.5% of the populations; resistance coefficient 0.5-2.9). Regarding the intra-population variability in response to phosphine (slope of the knockdown time regression), the laboratory and slightly resistant populations of all species were homogenous, whereas the most resistant populations were strongly heterogeneous. Our data show that the occurrence of resistance in the Czech Republic is relatively widespread and covers a wide range of species, necessitating the need for the adoption of an action plan for resistance mitigation.

Carbon Dioxide-Enriched Atmosphere to Control Oryzaephilus surinamensis L. On Stored Saqie Date Fruits.

<b>Background and Objective:</b> Control of the stored date insects by an eco-friendly method is essential to maintain the fruit for prolonged periods, especially that dates are eaten fresh. The search for a safe method, as an alternative to commonly used chemical pesticides, is needed. The objective is to investigate CO₂ enriched atmosphere on the Mortality Percentage (MP) of the Saw-toothed grain beetle, <i>O. surinamensis</i>, life stages with special emphasis on reducing damage to stored dates. <b>Materials and Methods:</b> The effect of high levels of CO₂ as an alternative control method against the Saw-toothed grain beetle, <i>Oryzaephilus surinamensis</i> L., at different life stages was studied on infested 'Saqie' dates. Four CO₂ pressures (25, 50, 75 and 90 kPa, balance is nitrogen) were tested for 6, 12, 18, 24, 48, 72 and 96 hrs intervals. The response of different life stages of <i>O. surinamensis</i> to the different treatments varied according to CO₂ level, developmental stage and exposure period. <b>Results:</b> Mortality (%) was higher during the larval stage, followed by adults, pupae and eggs, in descending order. The larvae and adult stages were more sensitive to CO₂ treatment than the pupal and egg. Exposure time was more effective on eggs, larval and adult MP than the CO₂ atmosphere level. Mortality% at 96 hrs exposure time was almost 100% with CO₂ atmospheres of 50, 75 and 90 kPa. Mathematical equations were developed to model the relationship between mortality% and CO₂ treatments using multiple regression analyses for each life stage. <b>Conclusion:</b> The results confirmed that CO₂ could be applied to final food products during packaging to control the residual occurrence of insect pests after storage and before the packaging process to prevent further infestation in the final packages.

Characterization of the complete mitochondrial genome of Oryzaephilus surinamensis Linne (Insecta: Coleoptera: Silvanidae) from Xichuan.

The saw-toothed grain beetle, Oryzaephilus surinamensis Linné, is a well-known stored-product insect. Beetles were obtained from Xichuan County and the mitochondrial genome was characterized (GenBank accession number MN535903). The mitogenome consists of a circular DNA molecule of 15,941 bp, with only 27.36% GC content. It comprises 13 protein-coding, 22 tRNA, and 2 rDNA genes. The protein-coding genes have typical ATN (Met) initiation codons and are terminated by typical TAN stop codons.

A Novel, Extremely Elongated, and Endocellular Bacterial Symbiont Supports Cuticle Formation of a Grain Pest Beetle.

The saw-toothed grain beetle, Oryzaephilus surinamensis (Silvanidae), is a cosmopolitan stored-product pest. Early studies on O. surinamensis in the 1930s described the presence of peculiar bacteriomes harboring endosymbiotic bacteria in the abdomen. Since then, however, the microbiological nature of the symbiont has been elusive. Here we investigated the endosymbiotic system of O. surinamensis in detail. In the abdomen of adults, pupae, and larvae, four oval bacteriomes were consistently identified, whose cytoplasm was full of extremely elongated tubular bacterial cells several micrometers wide and several hundred micrometers long. Molecular phylogenetic analysis identified the symbiont as a member of the Bacteroidetes, in which the symbiont was the most closely related to the endosymbiont of a grain pest beetle, Rhyzopertha dominica (Bostrichidae). The symbiont was detected in developing embryos, corroborating vertical symbiont transmission through host generations. The symbiont gene showed AT-biased nucleotide composition and accelerated molecular evolution, plausibly reflecting degenerative evolution of the symbiont genome. When the symbiont infection was experimentally removed, the aposymbiotic insects grew and reproduced normally, but exhibited a slightly but significantly more reddish cuticle and lighter body mass. These results indicate that the symbiont of O. surinamensis is not essential for the host's growth and reproduction but contributes to the host's cuticle formation. Symbiont genome sequencing and detailed comparison of fitness parameters between symbiotic and aposymbiotic insects under various environmental conditions will provide further insights into the symbiont's biological roles for the stored-product pest.IMPORTANCE Some beetles notorious as stored-product pests possess well-developed symbiotic organs called bacteriomes for harboring specific symbiotic bacteria, although their biological roles have been poorly understood. Here we report a peculiar endosymbiotic system of a grain pest beetle, Oryzaephilus surinamensis, in which four oval bacteriomes in the abdomen are full of extremely elongated tubular bacterial cells. Experimental symbiont elimination did not hinder the host's growth and reproduction, but resulted in emergence of reddish beetles, uncovering the symbiont's involvement in host's cuticle formation. We speculate that the extremely elongated symbiont cell morphology might be due to the degenerative symbiont genome deficient in bacterial cell division and/or cell wall formation, which highlights an evolutionary consequence of intimate host-symbiont coevolution.