Is the Intergenic Region of Aedes aegypti Totivirus a Recombination Hotspot?

The genus totivirus in the family Totiviridae contains double-stranded RNA viruses. Their genome has two open reading frames (ORFs) that encode capsid protein (CP) and RNA-dependent RNA polymerase (RdRp). The toti-like viruses recently identified in Anopheles sp. and Aedes aegypti mosquitoes (AaTV) share the same genome organization as other totiviruses. The AaTVs that have been described in distinct geographical regions are monophyletic. In this study, we show that AaTV sequences can be grouped into at least three phylogenetic clades (named A, B, and C). Clades A and B are composed of AaTV sequences from mosquitoes collected in the Caribbean region (Guadeloupe), and clade C contains sequences from the USA. These clades may represent AaTV lineages that are locally adapted to their host populations. We also identified three recombinant AaTV strains circulating in mosquitoes in Guadeloupe. Although these strains have different chimeric patterns, the position of the recombination breakpoint was identical in all strains. Interestingly, this breakpoint is located in a hairpin-like structure in the intergenic region of the AaTV genome. This RNA structure may stall RNA polymerase processivity and consequently induce template switching. In vitro studies should be conducted to further investigate the biological significance of AaTV's intergenic region as a recombination hotspot.

Sequencial passages of the multiple nucleopolyhedrovirus of Anagrapha falcifera (Kirby) (Lepidoptera: Noctuidae) (AfMNPV) in Spodoptera cosmioides (Walker) (Lepidoptera, Noctuidae) / Passagens sequenciais do múltiplo nucleopoliedrovírus de Anagrapha falcifera (Kirby) (Lepidoptera: Noctuidae) (AfMNPV) em Spodoptera cosmioides (Walker) (Lepidoptera, Noctuidae)

The aim of this work was to evaluate the effects of Anagrapha falcifera multiple nucleopolyhedrovirus (AfMNPV) passages in the Spodoptera cosmioides caterpillar's biology at different times of infection and histological changes that the virus could cause in the caterpillar midgut, seeking correlate histopathologic effects to the effectiveness of this virus as a potential biological control of this pest. Larvae were infected with seven days of development, by using three different passages of AfMNPV on S. cosmioides (F1, F4 and F7, which is the first, fourth and seventh passages, respectively) and the control treatment. Compared biology assays with the same treatments for analyzing behavior and mortality of caterpillars were performed concomitantly. The midgut morphology was compared between infected and uninfected larvae. The digestive tubes were collected at 24, 72 and 144 hours of infection (20 tubes/treatment/time of infection). After collection, the digestive tubes were fixed in Karnovsky, processed, stained with Hematoxylin-Eosin, and examined under a light microscope. The biology results of F4 and F7treatments, showed a drastic reduction in locomotion and feeding from the fourth day after infection and higher cumulative mortality rate compared to the control and F1. All treatments caused morphological changes in the midgut of S. cosmioides, in the three times of infection, with the greatest changes occurring at the epithelium. The AfMNPV, in the three passages tested in S. cosmioides, caused behavioral and morphological changes in the midgut, indicating that it can be a promising agent for biological control of this pest.

O objetivo deste trabalho foi avaliar os efeitos de passagens do múltiplo nucleopoliedrovírus de Anagrapha falcifera (AfMNPV) na biologia de lagartas de Spodoptera cosmioides, em diferentes momentos de infecção, e as alterações histológicas que o vírus poderia causar no intestino médio da lagarta, buscando correlacionar os efeitos histopatológicos e a eficácia deste vírus, como um potencial controle biológico desta praga. As lagartas foram infectadas com sete dias de desenvolvimento, utilizando três passagens diferentes do AfMNPV em S. cosmioides (F1, F4 e F7, sendo a primeira, quarta e sétima passagens, respectivamente) e o tratamento controle. Foram realizados, concomitantemente, ensaios de biologia, comparados com os mesmos tratamentos, para analisar o comportamento e mortalidade das lagartas. A morfologia do intestino médio foi comparada entre as lagartas infectadas e as não infectadas. Os tubos digestivos foram coletados com 24, 72 e 144 horas de infecção (20 tubos/tratamento/ tempo de infecção). Após a coleta, os tubos foram fixados em Karnovsky, processados, corados com Hematoxilina-Eosina e analisados ao microscópio de luz. Os resultados da biologia dos tratamentos F4 e F7 mostraram uma redução drástica na locomoção e alimentação das lagartas, a partir do quarto dia pós-infecção e maior taxa de mortalidade cumulativa, em relação ao controle e F1. Morfologicamente, todos os tratamentos causaram alterações no intestino médio das lagartas de S. cosmioides, nos três tempos de infecção, sendo que as maiores alterações ocorreram no epitélio. O AfMNPV, nas três passagens testadas em S. cosmioides, provocou alterações comportamentais e morfológicas no intestino médio, indicando que pode ser um promissor agente de controle biológico desta praga

Entomopathogenic Viruses in the Neotropics: Current Status and Recently Discovered Species.

The market for biological control of insect pests in the world and in Brazil has grown in recent years due to the unwanted ecological and human health impacts of chemical insecticides. Therefore, research on biological control agents for pest management has also increased. For instance, insect viruses have been used to protect crops and forests around the world for decades. Among insect viruses, the baculoviruses are the most studied and used viral biocontrol agent. More than 700 species of insects have been found to be naturally infected by baculoviruses, with 90% isolated from lepidopteran insects. In this review, some basic aspects of baculovirus infection in vivo and in vitro infection, gene content, viral replication will be discussed. Furthermore, we provide examples of the use of insect viruses for biological pest control and recently characterized baculoviruses in Brazil.

Detection of RNA-Dependent RNA Polymerase of Hubei Reo-Like Virus 7 by Next-Generation Sequencing in Aedes aegypti and Culex quinquefasciatus Mosquitoes from Brazil.

Advancements in next-generation sequencing and bioinformatics have expanded our knowledge of the diversity of viruses (pathogens and non-pathogens) harbored by mosquitoes. Hubei reo-like virus 7 (HRLV 7) was recently detected by the virome analysis of fecal samples from migratory birds in Australia. We now report the detection of RNA-dependent RNA polymerase sequences of HRLV 7 in pools of Aedes aegypti and Culex quinquefasciatus mosquitoes species from the Brazilian Amazon forest. Phylogenetic inferences indicated that all HRLV 7 strains fall within the same independent clade. In addition, HRLV 7 shared a close ancestral lineage with the Dinovernavirus genus of the Reoviridae family. Our findings indicate that HRLV 7 is present in two species of mosquitoes.