Detection of Arsenophonus in Glycaspis brimblecombei (Hemiptera: Aphalaridae) populations in Brazil.

Eucalyptus is the most intensively managed tree genus in the world. Different factors, including damage by insect pests, affect its growth and productivity. Among these pests is Glycaspis brimblecombei Moore (Hemiptera: Aphalaridae), an exotic insect of Australian origin. The evolutionary success of this insect depends on symbiotic associations with microorganisms. The influence of these microorganisms on insect pests and their natural enemies is important for integrated management tactics. Within this context, this work aimed to detect Arsenophonus in populations of G. brimblecombei in Brazil. Eucalyptus branches infested with G. brimblecombei nymphs were collected in commercial eucalyptus plantations in six Brazilian states. Specimens of this pest were sampled soon after emergence and frozen for molecular analysis. The genomic DNA of G. brimblecombei adults from each population was extracted and used to detect the endosymbiont Arsenophonus by polymerase chain reaction (PCR) employing specific primers that target its 23 S rRNA gene. This endosymbiont was identified in all of the studied G. brimblecombei populations. This is the first report on the association between Arsenophonus and G. brimblecombei in Brazil.

Entomopathogenic Fungi, Isolated From Soils and Bemisia tabaci (Hemiptera: Aleyrodidae) Adults, to Manage the Eucalyptus Red Gum Lerp Psyllid Glycaspis brimblecombei (Hemiptera: Aphalaridae).

The parasitoid Psyllaephagus bliteus Riek (Hymenoptera: Encyrtidae), entomopathogenic fungi, and chemical insecticides are the main strategies to manage the eucalypts pest Glycaspis brimblecombei Moore. The objective of this study was to isolate and to identify entomopathogenic fungi, collected from Bemisia tabaci Gennadius adults in soybean and tomato crops, and from soil samples in eucalypts, soybean, and native forest areas to evaluate their potential to manage G. brimblecombei. Twelve Beauveria and Cordyceps isolates were selected and compared with the commercial products Boveril Beauveria bassiana Bals. (Hypocreales: Cordycipitaceae), Metarril Metarhizium anisopliae Metschn. (Hypocreales: Clavicipitaceae), and Octane Cordyceps fumosorosea Wize (Hypocreales: Cordycipitaceae) and their respective strains. The fungal isolates were diluted in 0.1% aqueous Tween 80 at a concentration of 1.0 × 108 conidia/ml and sprayed on the G. brimblecombei nymphs with or without lerps. Pest mortality was higher and the TL50 and TL90 lower with the isolates LCBPF 11 C. javanica Frieder. & Bally (Hypocreales: Cordycipitaceae), LCBPF 12 (C. fumosorosea), and LCBPF 67 (C. fumosorosea) from B. tabaci adults. Fungi of the genera Beauveria and Cordyceps developed and caused high mortality of G. brimblecombei nymphs with lerps. The B. bassiana, C. cateniannulata Liang (Hypocreales: Cordycipitaceae), C. fumosorosea, C. javanica, and M. anisopliae isolates showed potential to manage G. brimblecombei. The lerp of this insect enhances entomopathogenic fungus development as a source of inoculum accelerating G. brimblecombei nymph mortality. Entomopathogenic fungi isolated from insects and soils are effective against G. brimblecombei and the presence of the lerp of this insect increases the effectiveness of its control.

First record of a new microsporidium pathogenic to Gonipterus platensis in Brazil.

Microsporidia are naturally occurring fungal-related parasites that can infect nearly all animal hosts, but their biocontrol potential of insect pests is routinely overlooked in agriculture and forestry. This research brings the first report describing the natural occurrence of a microsporidium causing disease in field-collected populations of the invasive eucalyptus snout beetle, Gonipterus platensis (Coleoptera: Curculionidae), a major destructive pest of eucalyptus plantations in Brazil. Adult beetles were collected during field surveys in commercial eucalyptus plantations in southern Brazil to be examined and dissected with typical symptoms to verify presence of microsporidian spores in haemolymph. From 14 plantations in different sites, the natural infection occurrence in these populations ranged from 0 to 65%, while a lab colony exhibited an infection incidence of 70%. Spore density in haemolymph of symptomatic insects averaged 2.1 (± 0.4) × 107 spores/beetle. Symptoms in infected adults were identified by an abnormal abdomen with malformation of the second pair of wings, impairing their flight activity. Electron transmission microscopy of the pathogen showed morphological features similar to species belonging to the genus Nosema or Vairimorpha. Phylogenetic analysis of the full-length small subunit ribosomal RNA gene suggests this pathogen's placement in the genus Vairimorpha, but with a sequence identity of ~ 94% with the nearest neighbours. The low level of sequence identity suggests this pathogen may represent a novel taxon in the genus and further requires whole genome sequencing for definitive taxonomic resolution. These findings provide insights on the natural occurrence of this novel pathogen of this invasive pest in Eucalyptus plantations in Brazil. Further studies are needed to determine potential of this microsporidium in the design of conservative or augmentative biological control programs for this invasive pest.

Mayaro Virus: The Potential Role of Microbiota and Wolbachia.

The Mayaro virus (MAYV) is an arbovirus that circulates mainly in tropical forests or rural areas in Latin America and is transmitted mainly by Haemagogus mosquitoes. The objective of this study was to evaluate the vector competence, microbiome, and the presence of Wolbachia in three Aedes albopictus populations infected with MAYV. The vector competence was assessed based on viral infection and transmission by RT-qPCR. In addition, the microbiome was evaluated by amplification of the 16S rRNA V4 region and PCR to detect the presence of Wolbachia (strain wAlbA/wAlbB). Our results show that all three populations were susceptible to MAYV infection. The potential transmission of the MAYV was consistent in all populations of naïve mosquitoes injected (more than 50%). The microbiome analysis revealed 118 OTUs (operational taxonomic unit) from the three populations, 8 phyla, 15 classes, 26 orders, 35 families, 65 genera, and 53 species. All populations had Pseudomonas and Wolbachia as predominant genera. There was no difference between the variables for MAYV and Wolbachia (wAlbA or wAlbB) in the abdomen. However, in the head + thorax samples at 14 dpi, there was a difference between the two populations, indicating a possible correlation between the presence of Wolbachia (wAlbB) and infection. Overall, we show evidence that Ae. albopictus displays significant infection and transmission competence for the MAYV in the laboratory, and its bacterial microbiota play an important role in the host, mainly the strains of Wolbachia. The influence of the intestinal microbiota of Ae. albopictus is poorly known, and a better understanding of these interactions would open new perspectives for disease control through the manipulation of microbial communities. The exact contribution of this mosquito species to the transmission of the MAYV in the field remains to be confirmed.

Identification and genome sequencing of RNA viruses in the eucalyptus snout beetle Gonipterus spp. (Coleoptera: Curculionidae).

The genomes of two putative new RNA viruses (macula-like virus and bunya-like virus) were identified in total RNA extracted from dead eucalyptus snout beetles (Gonipterus spp.) from a laboratory colony. However, only bunya-like virus was detected in field-collected insects. The macula-like virus has a monopartite single-stranded RNA genome that contains three open reading frames (ORFs) encoding an RNA-dependent RNA polymerase (RdRp), a capsid protein (CP), protein with unknown function. The bunya-like virus genome was predicted to consist of two RNA segments: a large segment (L) encoding a single protein (RdRp) and a small segment (S) encoding a putative nucleocapsid protein.

Steinernema diaprepesi (Rhabditida: Steinernematidae) parasitizing Gonipterus platensis (Coleoptera: Curculionidae).

Entomopathogenic nematodes (EPNs) can control pests due to mutualistic association with bacteria that reproduce and kill the host from septicemia, making the environment favourable for nematode development and reproduction. The objective of this study was to identify an EPN isolate collected in eucalyptus cultivation and to determine its pathogenicity with regard to Gonipterus platensis Marelli (Coleoptera: Curculionidae). Four steel-mesh traps with two seventh-instar Galleria mellonella larvae were buried 5 cm deep in the soil in a commercial Eucalyptus plantation. After 7 days, the traps were packed in plastic bags and transported to laboratory to isolate the EPNs using White traps. The obtained nematodes were multiplied in G. mellonella larvae and identified by sequencing their D2/D3 expansion of the 28S rDNA region by polymerase chain reaction (PCR) and specific primers for ITS regions. Steinernema diaprepesi was identified and inoculated into G. platensis pupae at doses of 500, 1000 and 5000 infective juveniles (IJs) to determine its pathogenicity to this pest. At 8 days after inoculation, the mortality rate of the G. platensis pupae was 80% with the lowest concentration and 100% with the others. The emergence of nematodes and the rapid degradation of G. platensis pupae were observed in those inoculated with IJs. The pathogenicity to the G. platensis pupae indicates potential for using this nematode in the integrated management of this insect.

Effects of negatively and positively charged Ti metal surfaces on ceramic coating adhesion and cell response.

This manuscript reports an evaluation of the effects of simple chemical-heat treatments on the deposition of different ceramic coatings, i.e., TiO2, CaTiO3 and CaP, on commercially pure titanium (cp-Ti) and Ti6Al4V and the influence of the coatings on cells interaction with the surfaces. The ceramic materials were prepared by the sol-gel method and the coating adhesion was analyzed by pull-off bending tests. The wettability of positively or negatively charged surfaces was characterized by contact angle measurements, which also enabled the calculation of the surface free energy through the polar-apolar liquids approach. Both acid and alkaline treatments activated the cp-Ti, whereas Ti6Al4V was only activated by the alkaline treatment. Such treatment led to increased hydrophilicity with inhibition of the fibroblastic response on Ti6Al4V. On the other hand, osteoblastic cells adhered to and proliferated on the positively and negatively charged surfaces. The maximum adhesion strength (~ 3400 N) was obtained with a negative Ti6Al4V-CaTiO3-CaP multilayer surface.