Creating transgenic insects is a key technology in insect genetics and molecular biology. A widely used instrument in insect transgenesis is the piggyBac transposase, resulting in essentially random genomic integrations. In contrast, site-specific recombinases allow the targeted integration of the transgene construct into a specific genomic target site. Both strategies, however, often face limitations due to low transgenesis efficiencies. We aimed to enhance transgenesis efficiencies by utilizing capped mRNA as a source of transposase or recombinase instead of a helper plasmid. A systematic comparison of transgenesis efficiencies in Aedes mosquitoes, as models for hard-to-transform insects, showed that suppling piggyBac transposase as mRNA increased the average transformation efficiency in Aedes aegypti from less than 5% with the plasmid source to about 50% with mRNA. Similar high activity was observed in Ae. albopictus with pBac mRNA. No efficiency differences between plasmid and mRNA were observed in recombination experiments. Furthermore, a hyperactive version of piggyBac transposase delivered as a plasmid did not improve the transformation efficiency in Ae. aegypti or the agricultural pest Drosophila suzukii. We believe that the use of mRNA has strong potential for enhancing piggyBac transformation efficiencies in other mosquitoes and important agricultural pests, such as tephritids.

Acetic acid bacteria (family Acetobacteraceae) are found in the gut of most insects. Two clades are currently recognized: Commensalibacter-Entomobacter and Bombella-Oecophyllibacter. The latter group is only found in hymenopteran insects and the described species have been isolated from bees and ants. In this study, two new strains DDB2-T1T (=KACC 21507T=LMG 31759T) and DM15PD (=CCM 9165=DSM 112731=KACC 22353=LMG 32454) were isolated from wasps collected in the Republic of Korea and Germany, respectively. Molecular and phenotypic analysis revealed that the strains are closely related, with 16S rRNA gene sequences showing 100 % identity and genomic average nucleotide identity (ANI) values ≥99 %. The closest related species based on type strain 16S rRNA gene sequences are Swingsia samuiensis, Acetobacter peroxydans, Bombella favorum and Bombella intestini (94.8-94.7% identity), whereas the closest related species based on type strain genome analysis are Saccharibacter floricola and Bombella intestini (ANI values of 68.8 and 68.2 %, respectively). The reconstruction of a phylogenomic tree based on 107 core proteins revealed that the branch leading to DDB2-T1T and DM15PD is localized between Oecophyllibacter and Saccharibacter-Bombella. Further genomic distance metrics such as ANI, percentage of conserved proteins and alignment fraction values were consistent with these strains belonging to a new genus. The key phenotypic characteristics were one MALDI-TOF-MS peak (m/z=4601.9±2.0) and the ability to produce acid from d-arabinose. Based on this polyphasic approach, including phylogenetics, phylogenomics, genome distance calculations, ecology and phenotypic characteristics, we propose to name the novel strains Aristophania vespae gen. nov., sp. nov., with the type strain DDB2-T1T (=KACC 21507T=LMG 31759T).

Metarhizium anisopliae and Beauveria bassiana are commonly used entomopathogenic fungi, but their non-target effects over generalist predatory insects, which can contribute to pest control, are not well known. We studied the capacity of the social wasp Polistes myersi to detect the pathogens in either a powdered form or in Galleria mellonella larvae infected with either of the two pathogens offered as prey. The effects of these treatments were compared considering wasp behaviors such as prey preference, frequency, duration and transitions of both hunting and grooming behaviors. Additionally, the effects of each entomopathogenic fungus on the wasp's mortality were measured. Wasps seem not to detect the pathogens in powdered form but preferred healthy over infected larvae. Seventeen behavioral units for hunting and 34 for grooming were recognized. There were no differences in grooming frequency but there were significant differences on grooming duration, hunting behaviors and the patterns of transitions. Exposure of wasp colonies to either B. bassiana or M. anisopliae had no detectable impact on the mortality of adults, but mortality of larvae increased. For the first time, this study documented behavioral changes that indicated the capacity of social wasps to detect pathogens before physical contact and the display of hygienic strategies once contact occurs. The study also suggested a potential non-target effect of these entomopathogenic fungi on a generalist predator.

The adult male of Xenos colombiensis n. sp. is described from a specimen extracted from the pupal case of its host, Polistes meyersi. While several species of Strepsiptera are known to parasitze species of Polistes in South America, this is the first species known to parasitize P. meyersi. An identification key is provided for adult males of Xenos species parasitizing Polistes species in the New World.

Information about parasitoids of neotropical vespids is scarce. Parasitoids collected from 43 colonies of Polistes myersi Bequaert, 1934 and one ofPolistes erythrocephalus Latreille, 1813 are reported from an Andean region of Colombia. Colony parasitism rates in P. myersi ranged from 35 % to 57 %, being higher in colonies with more cells; however, the number of parasitized colonies did not differ when considering the mean number of adult wasps (8.2 vs. 8.1 respectively). Parasitoidism ranged from one up to four species per colony. P. myersi parasitoids were: Seminotalaeviceps (Cresson, 1879) (Trigonalidae); Signiphora polistomyiella Richards, 1935 (Signiphoridae); Elasmuspolistis Burks, 1971 (Eulophidae, Elasminae); and a new species of Xenos(Strepsiptera, Xenidae). The latter three are first records for Colombia. P. myersi and P. erythrocephalus are the first host reports for the trigonalid S.laeviceps. We also report an unknown Tachinid fly species of the tribe Blondeliini attacking P. myersi.