Genetic dissection of assortative mating behavior.

The evolution of new species is made easier when traits under divergent ecological selection are also mating cues. Such ecological mating cues are now considered more common than previously thought, but we still know little about the genetic changes underlying their evolution or more generally about the genetic basis for assortative mating behaviors. Both tight physical linkage and the existence of large-effect preference loci will strengthen genetic associations between behavioral and ecological barriers, promoting the evolution of assortative mating. The warning patterns of Heliconius melpomene and H. cydno are under disruptive selection due to increased predation of nonmimetic hybrids and are used during mate recognition. We carried out a genome-wide quantitative trait locus (QTL) analysis of preference behaviors between these species and showed that divergent male preference has a simple genetic basis. We identify three QTLs that together explain a large proportion (approximately 60%) of the difference in preference behavior observed between the parental species. One of these QTLs is just 1.2 (0-4.8) centiMorgans (cM) from the major color pattern gene optix, and, individually, all three have a large effect on the preference phenotype. Genomic divergence between H. cydno and H. melpomene is high but broadly heterogenous, and admixture is reduced at the preference-optix color pattern locus but not the other preference QTLs. The simple genetic architecture we reveal will facilitate the evolution and maintenance of new species despite ongoing gene flow by coupling behavioral and ecological aspects of reproductive isolation.

Assessing the potential of unmanned aerial vehicle spraying of aqueous ozone as an outdoor disinfectant for SARS-CoV-2.

The COVID-19 pandemic has revealed gaps in our understanding of safe, effective and efficient means of disinfecting high use public spaces. Whilst this creates an opportunity for development and application of innovative approaches such as unmanned aerial vehicle (UAV) based disinfection, unregulated outdoor disinfection using chlorine has led to environmental and public health risks. This study has quantified the efficiency, safety and efficacy of UAV-based spraying of aqueous ozone. Optimised UAV flight characteristics of 4.7 km/h at 1.7 m elevation spraying 2.4 L/min were able to provide >97% and >92% coverage of a 1 m and 2 m wide swath respectively. During spraying operations using 1 mg/L aqueous ozone, atmospheric concentrations of ozone remained within background levels (<0.04 ppm). Highly efficient inactivation of two different isolates of SARS-CoV-2 virus was achieved at aqueous ozone concentrations of 0.75 mg/L after an incubation period of only 5 min, with 0.375 mg/L achieving 82-91.5% inactivation in this time. Exposure of diamondback moth larvae and parasitic wasps to 1 mg/L aqueous ozone did not significantly affect their survivorship. These results indicate for the first time that aqueous ozone may provide the required balance between human and environmental safety and viral inactivation efficacy for targeted application in high risk outdoor settings.

Biodiversity of Coreoidea and Pentatomidae (Heteroptera) from Atlantic forest protected areas. Insights into their conservation.

Although the majority of threatened species are likely to be tropical insects, knowledge of the diversity, ecological role and impact of insect biodiversity loss on ecosystem processes is very limited. Specimens belonging to four families of Heteroptera: Pentatomidae, Coreidae, Alydidae and Rhopalidae, were collected from a protected area in the Paraná Forest, the largest ecoregion of the Atlantic Forest, in Argentina. The assemblages were characterized and the biodiversity estimated, and they were compared with the assemblages found in five other protected areas in the Brazilian Atlantic Forest. In our study area, Pentatomidae had the greatest richness and diversity; Coreidae was the second most diverse family, with highest sampling deficit, highest percentage of singletons, and lowest inventory completeness; and Rhopalidae was the best sampled family with asymptotic rarefaction curves. We explored the application of the Species Conservation Importance index, following four criteria, to evaluate the relative importance of the pentatomid species studied and its usefulness for assigning conservation values to areas. We found similar Site Conservation Values among the six areas and noted that the use of criteria was limited by the lack of information, being crucial to increase the knowledge of most of the species.

Strong extrinsic reproductive isolation between parapatric populations of an Australian groundsel.

Speciation with gene flow, or the evolution of reproductive isolation between interbreeding populations, remains a controversial problem in evolution. This is because gene flow erodes the adaptive differences that selection creates between populations. Here, we use a combination of common garden experiments in the field and in the glasshouse to investigate what ecological and genetic mechanisms prevent gene flow and maintain morphological and genetic differentiation between coastal parapatric populations of the Australian groundsel Senecio lautus. We discovered that in each habitat extrinsic reproductive barriers prevented gene flow, whereas intrinsic barriers in F1 hybrids were weak. In the field, herbivores played a major role in preventing gene flow, but glasshouse experiments demonstrated that soil type also created variable selective pressures both locally and on a greater geographic scale. Our experimental results demonstrate that interfertile plant populations adapting to contrasting environments may diverge as a consequence of concurrent natural selection acting against migrants and hybrids through multiple mechanisms. These results provide novel insights into the consequences of local adaptation in the origin of strong barriers to gene flow in plants, and suggest that herbivory may play an important role in the early stages of plant speciation.

Doxorubicin-sensitive and -resistant colorectal cancer spheroid models: assessing tumor microenvironment features for therapeutic modulation.

Introduction: The research on tumor microenvironment (TME) has recently been gaining attention due to its important role in tumor growth, progression, and response to therapy. Because of this, the development of three-dimensional cancer models that mimic the interactions in the TME and the tumor structure and complexity is of great relevance to cancer research and drug development. Methods: This study aimed to characterize colorectal cancer spheroids overtime and assess how the susceptibility or resistance to doxorubicin (Dox) or the inclusion of fibroblasts in heterotypic spheroids influence and modulate their secretory activity, namely the release of extracellular vesicles (EVs), and the response to Dox-mediated chemotherapy. Different characteristics were assessed over time, namely spheroid growth, viability, presence of hypoxia, expression of hypoxia and inflammation-associated genes and proteins. Due to the importance of EVs in biomarker discovery with impact on early diagnostics, prognostics and response to treatment, proteomic profiling of the EVs released by the different 3D spheroid models was also assessed. Response to treatment was also monitored by assessing Dox internalization and its effects on the different 3D spheroid structures and on the cell viability. Results and Discussion: The results show that distinct features are affected by both Dox resistance and the presence of fibroblasts. Fibroblasts can stabilize spheroid models, through the modulation of their growth, viability, hypoxia and inflammation levels, as well as the expressions of its associated transcripts/proteins, and promotes alterations in the protein profile exhibit by EVs. Summarily, fibroblasts can increase cell-cell and cell-extracellular matrix interactions, making the heterotypic spheroids a great model to study TME and understand TME role in chemotherapies resistance. Dox resistance induction is shown to influence the internalization of Dox, especially in homotypic spheroids, and it is also shown to influence cell viability and consequently the chemoresistance of those spheroids when exposed to Dox. Taken together these results highlight the importance of finding and characterizing different 3D models resembling more closely the in vivo interactions of tumors with their microenvironment as well as modulating drug resistance.

Continent-wide evidence that landscape context can mediate the effects of local habitats on in-field abundance of pests and natural enemies.

Landscape-scale factors known to influence in-field abundance of pest herbivores and their natural enemies, but little is known about effects that operate through the shorter-range influences exerted by habitats immediately adjacent to crop fields.This study first compared the abundance of brassica insect pests and their natural enemy arthropods in 24 spatially independent brassica vegetable fields across southern Australia. An 'edge effect' index was used to compare the abundance of each taxon in the field center with abundance in areas of the crop adjacent to differing habitats. Then, three landscape properties: landscape composition, edge density, and connectivity of diverse crop and non-crop habitats were analyzed at five scales up to 5 km from these focal field centers to assess longer-range influences on arthropod abundances in field centers and on the edge effects.Edge effect of adjacent woody vegetation promoted ladybirds and reduced diamondback moth and whiteflies. Conversely, the presence of crops and pastures immediately adjacent to focal crop fields reduced whiteflies and aphids but with no effect on natural enemies.Effect of landscape composition and connectivity on arthropod abundance at field center found promotion of aphids (cabbage aphid and green peach aphid) by woodland in the landscape.Effect of landscape properties on the edge effects of adjacent habitats was contrasting; strengthened (landscape composition and edge density on edge effect of crops, pasture and woody vegetation in reducing diamondback moth and whiteflies) as well as weakened (edge density and landscape connectivity on edge effect of crops, pasture and woody vegetation in reducing diamondback moth on diamondback moth, whiteflies and aphids, and promoting ladybirds). Synthesis and applications: Findings of this geographically extensive study help define the level of pest risk associated with sites as well as suggest potential interventions such as establishment or restorations of woody vegetation adjacent to crop fields that could reduce risk.

Generalist Predators Shape Biotic Resistance along a Tropical Island Chain.

Islands offer exclusive prisms for an experimental investigation of biodiversity x ecosystem function interplay. Given that species in upper trophic layers, e.g., arthropod predators, experience a comparative disadvantage on small, isolated islands, such settings can help to clarify how predation features within biotic resistance equations. Here, we use observational and manipulative studies on a chain of nine Indonesian islands to quantify predator-mediated biotic resistance against the cassava mealybug Phenacoccus manihoti (Homoptera: Pseudococcidae) and the fall armyworm Spodoptera frugiperda (Lepidoptera: Noctuidae). Across island settings, a diverse set of generalist lacewing, spider and ladybeetle predators aggregates on P. manihoti infested plants, attaining max. (field-level) abundance levels of 1.0, 8.0 and 3.2 individuals per plant, respectively. Though biotic resistance-as imperfectly defined by a predator/prey ratio index-exhibits no inter-island differences, P. manihoti population regulation is primarily provided through an introduced monophagous parasitoid. Meanwhile, resident predators, such as soil-dwelling ants, inflict apparent mortality rates up to 100% for various S. frugiperda life stages, which translates into a 13- to 800-fold lower S. frugiperda survivorship on small versus large islands. While biotic resistance against S. frugiperda is ubiquitous along the island chain, its magnitude differs between island contexts, seasons and ecological realms, i.e., plant canopy vs. soil surface. Hence, under our experimental context, generalist predators determine biotic resistance and exert important levels of mortality even in biodiversity-poor settings. Given the rapid pace of biodiversity loss and alien species accumulation globally, their active conservation in farmland settings (e.g., through pesticide phasedown) is pivotal to ensuring the overall resilience of production ecosystems.

On the synergy between silver nanoparticles and doxycycline towards the inhibition of Staphylococcus aureus growth.

In a previous paper (RSC Adv., 2015, 5, 66886-66893), we showed that the combination of silver nanoparticles (NanoAg) with doxycycline (DO) culminated in an increased bactericidal activity towards E. coli. Herein we further investigated the metabolic changes that occurred on Staphylococcus aureus upon exposure to NanoAg with the help of attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) coupled with multivariate data analysis. It has been discovered that the combination of DO with NanoAg produced metabolic changes in S. aureus that were not simply the overlap of the treatments with DO and NanoAg separately. Our results suggest that DO and NanoAg act synergistically to impede protein synthesis by the bacteria.

Inhibition of the hemolytic activity caused by Staphylococcus aureus alpha-hemolysin through isatin-Schiff copper(II) complexes.

A great number of pathogens secrete pore-forming proteins during infection. Such molecules, from either bacterial or viral origin, are considered important virulence factors, which makes them attractive targets in the study of new therapeutic agents. Thus, the inhibitory activity of isatin-Schiff base copper(II) complexes was evaluated against membrane damage activity of Staphylococcus aureus α-hemolysin (α-HL). For this purpose, a standard hemolysis assay with rabbit erythrocytes and micromolar concentrations of the compounds was employed. Additionally, planar artificial lipid membranes with a single α-HL ion channel and molecular docking studies were used to elucidate the molecular mechanism of the complexes. Accordingly, the compounds were observed to possess a significant anti-hemolytic activity, capable of interacting with the constriction region of α-HL channel and blocking it in a potential dependent manner. Based on these results, it is expected that such isatin-Schiff base Copper(II) complexes may be employed as cotherapeutic agents for the treatment of staphylococcal infections.

Biodiversity and distribution of lethaeine seed bugs (Heteroptera, Rhyparochromidae, Lethaeini) from Argentina.

BACKGROUND: The tribe Lethaeini has received little attention in Argentina. In 2014, Dellapé recorded 9 genera and 15 species from this country. RESULTS: A comprehensive study of the Lethaeini (Heteroptera, Rhyparochromidae) from Argentina is presented. Herein three new species of Cryphula Stål, one new species of Cistalia Stål, and the male of Cryphula australis Berg aredescribed. The genus PetissiusDistantand the species Cistaliabinotata Slater& Baranowski, Cistalianeotropicalis Slater & Baranowski, and Petissius spinipes Stål are reported for the first time from Argentina; also, the known distribution is extended for many of the previously recorded species. A generic key, keys to species, and distributional maps to the Argentinean species are also given. Dorsal habitus photographs of all species and the male and female genitalia of the new species are provided to facilitateidentification. CONCLUSIONS: The Lethaeini fauna from Argentina is increased to 10 genera and 22 species. The distribution of the tribe in the country is mainly Neotropical into the Chacoan Subregion, with most of the species distributed in the Chacoan and Pampean provinces (Chacoan Domain) and Parana Forest Province (Parana Domain). Only Rhaptus quadricollis appears to be an Andean element, with most of the known records in the South American Transition Zone (Monte Province).

Biodiversity of Coreoidea and Pentatomidae (Heteroptera) from Atlantic forest protected areas. Insights into their conservation

ABSTRACT Although the majority of threatened species are likely to be tropical insects, knowledge of the diversity, ecological role and impact of insect biodiversity loss on ecosystem processes is very limited. Specimens belonging to four families of Heteroptera: Pentatomidae, Coreidae, Alydidae and Rhopalidae, were collected from a protected area in the Paraná Forest, the largest ecoregion of the Atlantic Forest, in Argentina. The assemblages were characterized and the biodiversity estimated, and they were compared with the assemblages found in five other protected areas in the Brazilian Atlantic Forest. In our study area, Pentatomidae had the greatest richness and diversity; Coreidae was the second most diverse family, with highest sampling deficit, highest percentage of singletons, and lowest inventory completeness; and Rhopalidae was the best sampled family with asymptotic rarefaction curves. We explored the application of the Species Conservation Importance index, following four criteria, to evaluate the relative importance of the pentatomid species studied and its usefulness for assigning conservation values to areas. We found similar Site Conservation Values among the six areas and noted that the use of criteria was limited by the lack of information, being crucial to increase the knowledge of most of the species.

Convergence and divergence during the adaptation to similar environments by an Australian groundsel.

Adaptation to replicate environments is often achieved through similar phenotypic solutions. Whether selection also produces convergent genomic changes in these situations remains largely unknown. The variable groundsel, Senecio lautus, is an excellent system to investigate the genetic underpinnings of convergent evolution, because morphologically similar forms of these plants have adapted to the same environments along the coast of Australia. We compared range-wide patterns of genomic divergence in natural populations of this plant and searched for regions putatively affected by natural selection. Our results indicate that environmental adaptation followed complex genetic trajectories, affecting multiple loci, implying both the parallel recruitment of the same alleles and the divergence of completely different genomic regions across geography. An analysis of the biological functions of candidate genes suggests that adaptation to coastal environments may have occurred through the recruitment of different genes participating in similar processes. The relatively low genetic convergence that characterizes the parallel evolution of S. lautus forms suggests that evolution is more constrained at higher levels of biological organization.

Assortative mating preferences among hybrids offers a route to hybrid speciation.

Homoploid speciation generates species without a change in chromosome number via introgressive hybridization and has been considered rare in animals. Heliconius butterflies exhibit bright aposematic color patterns that also act as cues in assortative mating. Heliconius heurippa has a color pattern that can be recreated by introgression of the H. melpomene red band into an H. cydno genetic background. Wild H. heurippa males show assortative mating based on color pattern and we here investigate the origin of this preference by studying first-generation backcross hybrids between H. melpomene and H. cydno that resemble H. heurippa. These hybrids show assortative mating preferences, showing a strong preference for their own color pattern over that of either parental species. This is consistent with a genetic basis to wing pattern preference and implies, first, that assortative mating preferences would facilitate the initial establishment of a homozygous hybrid color pattern by increasing the likelihood that early generation hybrids mate among themselves. Second, once established such a lineage would inherit assortative mating preferences that would lead to partial reproductive isolation from parental lineages.

O oleo essencial de Lippia gracilis Schauer, Verbenaceae, em ratos diabeticos / The essential oil from Lippia gracilis Schauer, Verbenaceae, in diabetic rats

O óleo essencial das folhas de Lippia gracilis Schauer, Verbenaceae, foi examinado por CG e CG-MS. Quinze constituintes foram identificados, onde o carvacrol, p-cimeno e o γ-terpineno foram as substâncias majoritárias. No estudo in vitro, a solução a 5 por cento do óleo de L. gracilis Schauer apresentou atividade antibacteriana para Staphylococcus aureus isolado de úlcera infectada de paciente diabético. O estudo avaliou a atividade antibacteriana da solução a 5 por cento do óleo de L. gracilis Schauer em modelo experimental em diabetes utilizando ratos albinos Wistar machos com membro pélvico infectado com cepa de Staphylococcus aureus. No experimento foram utilizados 28 ratos Wistar distribuídos em quatro grupos (G1-branco, G2-controle negativo, G3-controle positivo, G4-teste) de sete ratos. Quando comparado o grupo G4 com G3, observou-se que a solução a 5 por cento promoveu uma redução nas CFU/mL após 24h da administração do inóculo (S.aureus sem L. gracilis Schauer 10(8) ± 313 versus S.aureus com L. gracilis Schauer 13,28 ± 4,03). Os resultados foram expressos através do cálculo da média±EPM) e análise de variança (ANOVA). A diferença entre a concentração inibitória mínima no estudo in vitro foi determinada pelo teste Turkey (p<0.05). O teste Newman-Keuls com nível de significância (p<0.05) foi utilizado para o cálculo dos resultados obtidos no experimento in vivo. A solução a 5 por cento do óleo essencial de Lippia gracilis Schauer apresentou boa atividade antibacteriana tanto no estudo in vivo como no in vitro.

The essential oil from Lippia gracilis Schauer (Verbenaceae) leaves was examined by GC and GC-MS. Fifteen constituents were identified. Carvacrol, p-cymene and γ-terpinene were found to be the major components. In the in vitro study, 5 percent solution of the Lippia gracilis Schauer oil presented antibacterial activity against Staphylococcus aureus isolated from diabetic patients with infected ulcers. The study evaluated the antibacterial activity of the 5 percent solution of the Lippia gracilis Schauer oil on the experimental model of diabetic adult male albino Wistar rats with leaft pelvic limb infected by Staphylococcus aureus strain. In this experiment, 28 diabetic Wistar rats were used, randomly distributed in four different groups of seven rats, (G1-white; G2-negative control; G3-positive control and G4-test). When comparing group G4 with G3, it was observed that the 5 percent solution presented a reduced CFU/mL level showing the antibacterial effect of the oil 24 hours after the administration of the inoculum (S .aureus without Lippia gracilis Schauer 108 ±313 versus S.aureus with Lippia gracilis Schauer 13.28±4.03). The results were expressed as mean±S.E.M. One-way analysis of the variance (ANOVA) was used. The differences between the minimum inhibitory concentration in vitro test were determined by the Tukey test (p<0.05). The Newman-Keuls test with level of significance (p<0.05) was used to measure the results in vivo. The findings have shown that 5 percent solution of the Lippia gracilis Schauer oil presented antibacterial activity in vitro and in vivo.

Hiperfosfatasemia transitória benigna na infância / Benign transient hyperphosphatasemia in childhood

A hiperfosfatasemia transitória benigna na infância (HTBI) é uma condição pouco conhecida, que se traduz por um estado benigno e de bom prognóstico. A HTBI resulta do aumento súbito e transitório na concentração sérica da fosfatase alcalina (FA) em crianças sadias. A sua etiopatogenia é ainda incerta, porém a diminuição do clearance da enzima na circulação tem sido considerada o mecanismo mais provável. Para divulgar os conhecimentos atuais acerca do diagnóstico da HTBI, foram selecionados, no Medline, os artigos de periódicos médicos internacionais mais relevantes desde a descrição inicial desta condição, em 1954. Os critérios para diagnóstico de HTBI são: acometer crianças com idade inferior a cinco anos, ausência de sintomas ou presença de sintomas associados a doenças não relacionadas, sem evidência clínica ou laboratorial de doença óssea ou hepática. A elevação da fosfatase alcalina varia entre 3 e 50 vezes o limite superior do valor para a idade. A análise das isoenzimas mostra elevação das formas óssea e hepática. Os níveis séricos da fosfatase alcalina retornam ao normal em quatro meses. A elevação sérica da fosfatase alcalina foi relatada em patologias como hiperfosfatasemia familiar persistente assintomßtica e sintomática e hiperfosfatasemia persistente não-familiar. É indispensável que a HTBI seja considerada no diagnóstico diferencial de um aumento considerável dos níveis séricos de fosfatase alcalina em crianças, sem que seja detectada uma causa óbvia, o que evitará procedimentos desnecessários e dispendiosos durante a investigação diagnóstica.