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One-third of all mosquitoes belong to the Aedini, a tribe comprising common vectors of viral zoonoses such as Aedes aegypti and Aedes albopictus. To improve our understanding of their evolution, we present an updated multigene estimate of Aedini phylogeny and divergence, focusing on the disentanglement between nuclear and mitochondrial phylogenetic signals. We first show that there are some phylogenetic discrepancies between nuclear and mitochondrial markers which may be caused by wrong taxa assignment in samples collections or by some stochastic effect due to small gene samples. We indeed show that the concatenated dataset is model and framework dependent, indicating a general paucity of signal. Our Bayesian calibrated divergence estimates point toward a mosquito radiation in the mid-Jurassic and an Aedes radiation from the mid-Cretaceous on. We observe, however a strong chronological incongruence between mitochondrial and nuclear data, the latter providing divergence times within the Aedini significantly younger than the former. We show that this incongruence is consistent over different datasets and taxon sampling and that may be explained by either peculiar evolutionary event such as different levels of saturation in certain lineages or a past history of hybridization throughout the genus. Overall, our updated picture of Aedini phylogeny, reveal a strong nuclear-mitochondrial incongruence which may be of help in setting the research agenda for future phylogenomic studies of Aedini mosquitoes.

Background The psyllid genus Cacopsylla includes several species that act as vectors for phytoplasma-associated diseases affecting plantations across Europe. Among them, Cacopsylla melanoneura and Cacopsylla picta are the primary vectors of ‘ Candidatus Phytoplasma mali’, the phloem-restricted bacterium responsible for Apple Proliferation disease in Europe. To explore whether vector competence in these species reflects shared ancestry or independent evolution, we assembled mitochondrial and draft nuclear genomes of Italian populations of C. melanoneura and C. picta and reconstructed time-calibrated phylogenies using 13 mitochondrial protein-coding genes from 12 Cacopsylla species. Results Phylogenetic analyses revealed two major Cacopsylla clades (Clade I and II) whose divergence times range from the Early Miocene (18.4 MYA; 95% HPD: 10.8–27.5) to the Middle Miocene (12.7 MYA; 95% HPD: 9.7–16.0). Both C. melanoneura and C. picta are within Clade I, which is predominantly composed of univoltine species that overwinter on conifers. Within this clade, Cacopsylla melanoneura is more closely related to the plum psyllid Cacopsylla pruni than to the apple-associated Cacopsylla picta and Cacopsylla mali , the latter belonging to Clade II. Draft nuclear genomes revealed significant differences in size (438 Mb in C. melanoneura vs. 631 Mb in C. picta ), largely attributed to repetitive elements. Comparative analyses of repetitive elements across Cacopsylla species revealed a recent expansion of transposable elements, particularly LINE elements, which were slightly more abundant in Clade I and contributed to the larger genome size observed in C. picta . Conclusions Collectively, our findings provide the first genomic resources for C. melanoneura , C. picta , and several other phytoplasma-vectoring Cacopsylla species. We established a robust mitogenomic phylogeny with divergence estimated for this genus showing the presence of two clades with the representatives predominantly associated with different overwintering strategies. Our results further indicate that vectorial capacity in Cacopsylla reflects an independent evolutionary trajectory rather than a shared ancestral origin. This evolutionary framework advances our understanding of the biology and origin of vector competence in this agriculturally important group.

The leafhopper genus Arboridia includes several species that feed on Vitis vinifera and cause leaf chlorosis. We report the first alien Arboridia infestation in Italy in 2021 in an Apulian vineyard. To confirm the taxonomic status of the species responsible for crop damage, and reconstruct its demographic history, we barcoded individuals from Apulia together with Arboridia spp. from Crete (Greece), A. adanae from Central Turkey and other specimens of the presumed sister species, A. dalmatina from Dalmatia (Croatia). Molecular phylogenies and barcoding gap analysis identified clades not associated with sampling locations. This result is incongruent with classical specimen assignment and is further supported by morphological analyses, which did not reveal significant differences among the populations. Therefore, we propose A. dalmatina as a junior synonym of A. adanae , which would become the only grapevine-related Arboridia species in the eastern Mediterranean. To further characterise A. adanae evolution, we performed a molecular clock analysis that suggested a radiation during the Pleistocene glaciations. Finally, to assess whether the Apulian individuals carried microorganisms of agricultural relevance, we sequenced their bacterial microbiota using 16S rRNA amplicon sequencing identifying three phytopathogens not generally associated with Arboridia activities as well as Wolbachia in one Apulian haplogroup. We discuss the agricultural implications of this infestation.

We report our decennial experience with 1161 newly-placed long-term central venous catheters inserted in 919 hematology-oncology patients for a total of 413,901 CVC-days of observation. Most of the CVCs were partially-implanted, open-ended, Broviac-Hickman type of CVC (95 %). One thousand and twenty-four complications were recorded equal to 2.47 per 1000 CVC-days. The frequency of complications per CVC, the rate of episodes per 1000 CVC-days, and removal rate were malfunction/occlusion 42 %, 1.18/1000, and 2.3 %; mechanical (dislodgement/rupture/kinking) 18.3 %, 0.51/1000, and 77.4 %; bacteremia 14.8 %, 0.42/1000, and 18.6 %; exit-site/tunnel infection 11.5 %, 0.32/1000, and 9.7 %; thrombosis 0.86 %, 0.02/1000, and 30 %; pneumothorax 0.52 %, 0.01/1000, and 0. In multivariate analysis, the risk factors were for mechanical complications, a younger age <6.1 years at CVC insertion (HR 1.8, p  = 0.0006); for bacteremia, a double lumen CVC (HR 3.1, p  < 0.0001) and the surgical modality of CVC insertion (HR 1.5, p  = 0.03); for exit-site/tunnel infection, a double lumen CVC (HR 2.1, p  = 0.0003) and a diagnosis of leukemia or lymphoma (HR 1.8, p  = 0.01); for malfunction/occlusion, an age <6.1 years (HR 1.6, p  = 0.0003), the diagnosis of leukemia or lymphoma (HR 1.9, p  < 0.0001) and double lumen CVC (HR 1.33, p  = 0.023). The cumulative incidence of premature CVC removal was 29.2 % and the risk factors associated with this event were the surgical modality of CVC insertion (HR 1.4, p  = 0.0153) and an age at CVC positioning less than 6.1 years (HR 1.6, p  = 0.0025). We conclude that a best-practice set of rules resulted in reduced CVC complications.

Aedes japonicus and Aedes koreicus are two invasive mosquitoes native to East Asia that are quickly establishing in temperate regions of Europe. Both species are vectors of arboviruses, but we currently lack a clear understanding of their evolution. Here, we present new short-read, shallow genome sequencing of A. japonicus and A. koreicus individuals from northern Italy, which we used for downstream phylogenetic and barcode analyses. We explored associated microbial DNA and found high occurrences of Delftia bacteria in both samples, but neither Asaia nor Wolbachia. We then assembled complete mitogenomes and used these data to infer divergence times estimating the split of A. japonicus from A. koreicus in the Oligocene, which was more recent than that previously reported using mitochondrial markers. We recover a younger age for most other nodes within Aedini and other Culicidae. COI barcoding and phylogenetic analyses indicate that A. japonicus yaeyamensis, A. japonicus amamiensis, and the two A. koreicus sampled from Europe should be considered as separate species within a monophyletic species complex. Our studies further clarify the evolution of A. japonicus and A. koreicus, and indicate the need to obtain whole-genome data from putative species in order to disentangle their complex patterns of evolution.

The picornavirus named ‘Ljungan virus’ (LV, species Parechovirus B) has been detected in a dozen small mammal species from across Europe, but detailed information on its genetic diversity and host specificity is lacking. Here, we analyze the evolutionary relationships of LV variants circulating in free-living mammal populations by comparing the phylogenetics of the VP1 region (encoding the capsid protein and associated with LV serotype) and the 3Dpol region (encoding the RNA polymerase) from 24 LV RNA-positive animals and a fragment of the 5′ untranslated region (UTR) sequence (used for defining strains) in sympatric small mammals. We define three new VP1 genotypes: two in bank voles (Myodes glareolus) (genotype 8 from Finland, Sweden, France, and Italy, and genotype 9 from France and Italy) and one in field voles (Microtus arvalis) (genotype 7 from Finland). There are several other indications that LV variants are host-specific, at least in parts of their range. Our results suggest that LV evolution is rapid, ongoing and affected by genetic drift, purifying selection, spillover and host evolutionary history. Although recent studies suggest that LV does not have zoonotic potential, its widespread geographical and host distribution in natural populations of well-characterized small mammals could make it useful as a model for studying RNA virus evolution and transmission.

Several true fruit flies (Tephritidae) cause major damage to agriculture worldwide. Among them, species of the genus Bactrocera are extensively studied to understand the traits associated with their invasiveness and ecology. Comparative approaches based on a reliable phylogenetic framework are particularly effective, but several nodes of the Bactrocera phylogeny are still controversial, especially concerning the reciprocal affinities of the two major pests B. dorsalis and B. tryoni. Here, we analyzed a newly assembled genomic-scaled dataset using different models of evolution to infer a phylogenomic backbone of ten representative Bactrocera species and two outgroups. We further provide the first genome-scaled inference of their divergence by calibrating the clock using fossil records and the spontaneous mutation rate. The results reveal a closer relationship of B. dorsalis with B. latifrons than to B. tryoni, contrary to what was previously supported by mitochondrial-based phylogenies. By employing coalescent-aware and heterogeneous evolutionary models, we show that this incongruence likely derives from a hitherto undetected systematic error, exacerbated by incomplete lineage sorting and possibly hybridization. This agrees with our clock analysis, which supports a rapid and recent radiation of the clade to which B. dorsalis, B. latifrons and B. tryoni belong. These results provide a new picture of Bactrocera phylogeny that can serve as the basis for future comparative analyses.

Objective As far as the ventriculoatrial shunt placement in children is concerned, the percutaneous approach to the internal jugular vein under ultrasonographic control has been hitherto strongly recommended. Unfortunately, children still represent a challenge, having them peculiar characteristics for which the internal jugular vein cannulation shows some disadvantages. Methods In this manuscript, we describe a percutaneous placement of ventriculoatrial shunt via right brachiocephalic vein under intraoperative ultrasonographic control. Conclusions Brachiocephalic vein cannulation in surgery provides notable advantages in paediatric population and it may be considered as the first choice in younger children.

A prospective pediatric survey on the incidence of central venous catheter (CVC) complications was performed aimed at identifying risk factors of premature CVC removal. The study comprised 129 Broviac-Hickman CVCs inserted during a 13-month period in 112 children. The total number of CVC days was 19,328 (median: 122 days, range: 1-385). The overall rate of complications was 6.2/1000 CVC days, i.e., 4.5/1000 and 1.7/1000 CVC days for mechanical and infectious complications, respectively. Interestingly, only two CVC-related cases of septicemia and no thrombotic events were documented. At the end of the study period, 38 of 129 CVC (29.5%) had been removed: 20 due to CVC-related complications (dislocation18, rupture 2), 10 due to the patient's death, and 8 due to completion of therapy. Age at CVC insertion <4.9 years was a significant predictor of premature CVC removal ( p=0.01). Mechanical complications, especially in younger children, are the main cause of premature loss of CVC. These data underline the importance of more effectively securing the CVC to subcutaneous tissue in pediatric patients to reduce accidental dislocations.

Comparative approaches are widely used to investigate the traits that underlie particular biological and ecological traits. They are effective, however, only if we know the correct relationships among the species under consideration. Here, we aimed at reconstructing a robust phylogeny for selected true fruit flies of the genus Bactrocera, which are well known as important agricultural pests worldwide. Existing phylogenetic inferences are still ambiguous, especially concerning the relationship between the two major pests B. dorsalis and B. tryoni. In this study, we employed a genome-scaled dataset and different models of molecular evolution to reconstruct the phylogenetic relationships of ten Bactrocera species and two outgroups and further date their divergence times. The resulting phylogeny fully supports B. dorsalis as more closely related to B. latifrons than to B. tryoni, opposite to what was supported by previous works. This incongruence likely derives from a fast divergence of these lineages, as revealed by our clock analysis, which can lead to conflicting results when using few genetic markers. Our results thus highlight the utility of using large datasets and of exploring different evolutionary models to study the evolutionary history of species of economic importance. Several true fruit flies (Tephritidae) cause major damage to agriculture worldwide. Among them, species of the genus Bactrocera are extensively studied to understand the traits associated with their invasiveness and ecology. Comparative approaches based on a reliable phylogenetic framework are particularly effective, but several nodes of the Bactrocera phylogeny are still controversial, especially concerning the reciprocal affinities of the two major pests B. dorsalis and B. tryoni. Here, we analyzed a newly assembled genomic-scaled dataset using different models of evolution to infer a phylogenomic backbone of ten representative Bactrocera species and two outgroups. We further provide the first genome-scaled inference of their divergence by calibrating the clock using fossil records and the spontaneous mutation rate. The results reveal a closer relationship of B. dorsalis with B. latifrons than to B. tryoni, contrary to what was previously supported by mitochondrial-based phylogenies. By employing coalescent-aware and heterogeneous evolutionary models, we show that this incongruence likely derives from a hitherto undetected systematic error, exacerbated by incomplete lineage sorting and possibly hybridization. This agrees with our clock analysis, which supports a rapid and recent radiation of the clade to which B. dorsalis, B. latifrons and B. tryoni belong. These results provide a new picture of Bactrocera phylogeny that can serve as the basis for future comparative analyses.