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Introduction: Symbiotic bacteria play key roles in a variety of important life processes of insects such as development, reproduction and environmental adaptation, and the elucidation of symbiont population structure and dynamics is crucial for revealing the underlying regulatory mechanisms. The marmalade hoverfly (Episyrphus balteatus) is not only a remarkable aphid predator, but also a worldwide pollinator second to honeybees. However, its symbiont composition and dynamics remain unclear. Methods: Herein, we investigate the symbiotic bacterial dynamics in marmalade hoverfly throughout whole life cycle, across two sexes, and in its prey Megoura crassicauda by 16S rRNA sequencing. Results: In general, the dominant phyla were Proteobacteria and Firmicutes, and the dominant genera were Serratia and Wolbachia. Serratia mainly existed in the larval stage of hoverfly with the highest relative abundance of 86.24% in the 1st instar larvae. Wolbachia was found in adults and eggs with the highest relative abundance of 62.80% in eggs. Significant difference in species diversity was observed between the adults feeding on pollen and larvae feeding on M. crassicauda, in which the dominant symbiotic bacteria were Asaia and Serratia, respectively. However, between two sexes, the symbionts exhibited high similarity in species composition. In addition, our results suggested that E. balteatus obtainded Serratia mainly through horizontal transmission by feeding on prey aphids, whereas it acquired Wolbachia mainly through intergeneration vertical transmission. Taken together, our study revealed the effects of development stages, diet types and genders of E. balteatus on symbionts, and explored transmission modes of dominant bacteria Serratia and Wolbachia. Discussion: Our findings lay a foundation for further studying the roles of symbiotic bacteria in E. balteatus life cycle, which will benefit for revealing the co-adaptation mechanisms of insects and symbiotic bacteria. [ABSTRACT FROM AUTHOR]

Introduction: Excessive application of nitrogen fertilizer in cotton field causes soil and water pollution as well as significant increase of aphid population. Reasonable fertilization is an important approach to improve agricultural production efficiency and reduce agriculture-derived pollutions. This study was aimed to explore the effects of nitrogen fertilizer on the Bt cotton physiological characteristics and the growth and development of A. gossypii, a sap-sucking cotton pest. Methods: Five different levels of Ca(NO3)2 (0.0 g/kg, 0.3 g/kg, 0.9 g/kg, 2.7 g/kg and 8.1 g/kg) were applied into vermiculite as nitrogen fertilizer in order to explore the effects of nitrogen fertilizer on the growth and development of Bt cotton and aphids. Results: The results showed that the medium level of nitrogen fertilizer (0.9 g/kg) effectively facilitated the growth of Bt cotton plant and suppressed the population expansion of aphids, whereas high and extremely high nitrogen application (2.7 and 8.1 g/kg) significantly increased the population size of aphids. Both high and low nitrogen application benefited aphid growth in multiple aspects such as prolonging nymph period and adult lifespan, enhancing fecundity, and improving adult survival rate by elevating soluble sugar content in host Bt cotton plants. Cotton leaf Bt toxin content in medium nitrogen group (0.9 g/kg) was significantly higher than that in high (2.7 and 8.1 g/ kg) and low (0.3 g/kg) nitrogen groups, but Bt toxin content in aphids was very low in all the nitrogen treatment groups, suggesting that medium level (0.9 g/kg) might be the optimal nitrogen fertilizer treatment level for promoting cotton seedling growth and inhibiting aphids. Discussion: Overall, this study provides insight into trophic interaction among nitrogen fertilizer levels, Bt cotton, and cotton aphid, and reveals the multiple effects of nitrogen fertilizer levels on growth and development of cotton and aphids. Our findings will contribute to the optimization of the integrated management of Bt cotton and cotton aphids under nitrogen fertilization. [ABSTRACT FROM AUTHOR]

Cotton bollworm (Helicoverpa armigera) poses a global problem, causing substantial economic and ecological losses. Endosymbionts in insects play crucial roles in multiple insect biological processes. However, the interactions between H. armigera and its symbionts have not been well characterized to date. We investigated the symbionts of H. armigera in the whole life cycle from different geographical locations. In the whole life cycle of H. armigera, Proteobacteria, Firmicutes, Bacteroidetes, and Actinobacteria were the dominant bacteria at the phylum level, while Enterococcus, Enterobacter, Glutamicibacter, and Bacillus were the four dominant bacteria at the genus level. Furthermore, high similarity in symbiotic bacterial community was observed in different stages of H. armigera, which were dominated by Enterococcus and Enterobacter. In fields, the dominant bacteria were Proteobacteria and Bacteroidetes, whereas, in the laboratory, the dominant bacteria were Proteobacteria. At the genus level, the dominant bacteria in cotton bollworm eggs of wild populations were Enterobacter, Morganella, Lactococcus, Asaia, Apibacter, and Enterococcus, and the subdominant bacteria were Bartonella, Pseudomonas, and Orbus. Moreover, the symbionts varied with geographical locations, and the closer the geographical distance, the more similar the microbial composition. Taken together, our study identifies and compares the symbiont variation along with geographical gradients and host development dynamic and reveals the high flexibility of microbiome communities in H. armigera, which probably benefits for the successful survival in a complicated changing environment. [ABSTRACT FROM AUTHOR]

The relationships between hosts and symbionts are complex and dynamic. Not only can symbionts significantly impact host phenotypes, but host interactions with symbionts may influence parasitoids. However, few studies to date have investigated the microbiomes of parasitized host offspring and considered the ecological implications of longitudinal microbiome dynamics. In this study, the parasitoid wasp Lysiphlebia japonica and its aphid host, Aphis gossypii, were used to examine the impact of parasitism on host offspring. Parental parasitism reduced the weight and reproductive capacity of the offspring; furthermore, the offspring showed reduced L. japonica parasitism rates. Moreover, the microbial composition of the parasitized and unparasitized aphid offspring were characterized with high-throughput 16S rRNA gene sequencing. This revealed that L. japonica parasitism strongly altered the microbial community in the offspring of parasitized aphids, particularly, the offspring of aphids parasitized for day 4. Overall, these results indicated that parasitoids can influence host--microbiome interactions by altering the microbiome of the host offspring. Our findings support the importance of further investigations into host--parasitoid--microbiome interactions. [ABSTRACT FROM AUTHOR]

Pardosa astrigera wolf spiders are important natural enemies of insect pests in cotton fields. Genetically modified (GM) crops expressing Bacillus thuringiensis (Bt) Cry proteins - toxic to target insect pests - are increasingly cultivated for insect pest management. To evaluate the potential negative effects of Bt-GM crops on beneficial non-target organisms, we assessed the effect of the Cry1Fa protein on P. astrigera, including on its symbiotic microbial communities. No adverse effects were detected on the survival rate and body weight of P. astrigera during growth and development. However, P. astrigera adult microbiome exhibited a reduced diversity as well as a shift in the microbial community structure. The dominant bacterial phyla in P. astrigera shifted from Actinobacteria to Firmicutes after feeding on prey enriched in Cry1Fa protein. Furthermore, Bacillus became the most abundant genus in P. astrigera females, while the dominant genus (Corynebacterium_1) in P. astrigera males did not change. Both males and females exhibited lower abundances of Wolbachia after feeding on Cry1Fa-contaminated prey. Overall, consuming Cry1Fa-contaminated prey changed the microbial community in P. astrigera, with higher impact in female microbiome. These findings provide new insights for environmental safety assessments of GM crops and to understand the importance of the potential effects of GM crops on microbiota of non-target organisms of importance for biocontrol ecosystem services. [ABSTRACT FROM AUTHOR]

Introduction: Reproductive polymorphism and symbiotic bacteria are commonly observed in aphids, but their interaction remains largely unclear. In polymorphic aphid species (Aphis gossypii), offspring of parthenogenetic females (PFs) develops into sexuparae which produces gynoparae and males successively. Gynoparae further produces sexual females (SFs), and these sexual females mate with males to produce offspring. Methods: In this study, we investigated the dynamic changes of symbiotic bacteria during the above-mentioned five reproductive morph switch in A. gossypii via 16S rRNA sequencing technology. Results: The results showed that species richness and community diversity of symbiotic bacteria in males were the highest. Proteobacteria was absolutely dominant bacterial phylum (with relative abundance of more than 90%) in the five reproductive morphs of A. gossypii, and Buchnera was absolutely dominant genus (with relative abundance of >90%), followed by Rhodococcus, Pseudomonas, and Pantoea. Male-killing symbiont Arsenophonus presented the highest relative abundance in gynoparae, a specific morph whose offsprings were exclusively sexual females. Both principal component analysis (PCA) and clustering analysis showed trans-generation similarity in microbial community structure between sexuparae and sexual females, between PFs and gynoparae. PICRUSt 2 analysis showed that symbiotic bacteria in the five reproductive morphs were mainly enriched in metabolic pathways. Discussion: Reproductive morph switch induced by environmental changes might be associated with bacterial community variation and sexual polymorphism of aphids. This study provides a new perspective for further deciphering the interactions between microbes and reproductive polymorphism in host aphids. [ABSTRACT FROM AUTHOR]

Introduction: Bacteria play critical roles in the reproduction, metabolism, physiology, and detoxification of their insect hosts. The ladybird beetle (Harmonia axyridis) harbors a myriad of endosymbiotic microbes. However, to date, little is known about how the microbial composition of H. axyridis varies throughout its life cycle. Methods: In this study, 16S rRNA amplicon sequencing and quantitative PCR were employed to investigate the diversity and dynamics of bacterial symbionts across the egg, larval, pupae, and adults stages of H. axyridis. Results: Higher bacterial community richness and diversity were observed in eggs, followed by those in adults and pupae. The community richness index differed significantly between second-instar larvae and other developmental stages. Proteobacteria, Firmicutes, and Actinobacteria were the dominant phyla. Staphylococcus, Enterobacter, Glutamicibacter, and Acinetobacter were the dominant bacteria genera; however, their relative abundances fluctuated across host developmental stages. Interestingly, the larval stage harbored high proportions of Firmicutes, whereas the adult microbial community largely consisted of Proteobacteria. Discussion: This study is the first to determine the symbiotic bacterial composition across key life stages of H. axyridis. These outcomes can foster the development of environmental risk assessments and novel biological control strategies. [ABSTRACT FROM AUTHOR]

Jian-Ti-Kang-Yi decoction (JTKY) is widely used in the treatment of COVID-19. However, the protective mechanisms of JTKY against pneumonia remain unknown. In this study, polyinosinic-polycytidylic acid (poly(I:C)), a mimic of viral dsRNA, was used to induce pneumonia in mice; the therapeutic effects of JTKY on poly(I:C)-induced pneumonia model mice were evaluated. In addition, the anti-inflammatory and anti-oxidative potentials of JTKY were also investigated. Lastly, the metabolic regulatory effects of JTKY in poly(I:C)-induced pneumonia model mice were studied using untargeted metabolomics. Our results showed that JTKY treatment decreased the wetto-dry ratio in the lung tissue, total protein concentration, and total cell count of the bronchoalveolar lavage fluid (BALF). Hematoxylin and Eosin (HE) and Masson staining indicated that the JTKY treatment alleviated the pathological changes and decreased the fibrotic contents in the lungs. JTKY treatment also decreased the expression of pro-inflammatory cytokines [interleukin (IL)-1ß, IL-6, and tumor necrosis factor-alpha (TNF-a)] and increased the levels of immunomodulatory cytokines (IL-4 and IL-10) in the BALF and serum. Flow cytometry analysis showed that the JTKY treatment lowered the ratio of CD86+/CD206+ macrophages in the BALF, decreased inducible nitric oxide synthase (iNOS) level, and increased arginase 1 (Arg-1) level in lung. JTKY also lowered CD11b+Ly6G+ neutrophils in BALF and decreased myeloperoxidase (MPO) activity in lung. Moreover, it also elevated superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities and decreased methane dicarboxylic aldehyde (MDA) level in lung. Untargeted metabolomic analysis showed that the JTKY treatment could affect 19 metabolites in lung, such as L-adrenaline, L-asparagine, ornithine, and alphaketoglutaric acid. These metabolites are associated with the synthesis and degradation of ketone bodies, butanoate, alanine, aspartate, and glutamate metabolism, and tricarboxylic acid (TCA) cycle processes. In conclusion, our study demonstrated that treatment with JTKY ameliorated poly(I:C)-induced pneumonia. The mechanism of action of JTKY may be associated with the inhibition of the inflammatory response, the reduction of oxidative stress, and the regulation of the synthesis and degradation of ketone bodies, TCA cycle, and metabolism of alanine, aspartate, glutamate, and butanoate processes in lung. [ABSTRACT FROM AUTHOR]

The complete mitochondrial genome of Aphis gossypii Glover cucumber biotype was sequenced using traditional PCR amplification coupled with Sanger sequencing. The genome is 15,870 bp long, with 83.7% AT content (MW048625). The genome encodes 37 typical mitochondrial genes, including 13 protein-coding genes, 2 ribosomal RNA genes, 22 transfer RNAs, a repeat region of 784 bp, and a control region of 627 bp. The base composition of the genome is A (45.4%), T (38.3%), C (10.5%), and G (5.8%). An analysis of two biotypes A. gossypii mitogenomes identified 77 single nucleotide polymorphisms and 1 insertion and deletion. [ABSTRACT FROM AUTHOR]

Increasing evidence shows that mechanical stresses are critical in regulating cell functions, fate, and diseases. However, no methods exist that can quantify isotropic compressive stresses. Here we describe fluorescent nanoparticle-labeled, monodisperse elastic microspheres made of Arg-Gly-Asp-conjugated alginate hydrogels (elastic round microgels, ERMGs). We generate 3D displacements and calculate strains and tractions exerted on an ERMG. Average compressive tractions on an ERMG are 570 Pa within cell layers and 360 Pa in tumor-repopulating cell (TRC) colonies grown in 400-Pa matrices. 3D compressive tractions on a 1.4-kPa ERMG are applied by surrounding cells via endogenous actomyosin forces but not via mature focal adhesions. Compressive stresses are substantially heterogeneous on ERMGs within a uniform cell colony and do not increase with TRC colony sizes. Early-stage zebrafish embryos generate spatial and temporal differences in local normal and shear stresses. This ERMG method could be useful for quantifying stresses in vitro and in vivo. [ABSTRACT FROM AUTHOR]

Understanding the population genetic structure and demographic dynamics of important agricultural pest insects is very important in studying insecticide resistance and control strategies. The plant bug Adelphocoris fasciaticollis (Reuter) (Hemiptera: Miridae) is one of the main insect pests on cotton in Northern China. We studied large-scale genetic structure and genetic diversity patterns and explored the demographic history. We also employed a phylogeographical approach on A. fasciaticollis samples using sequence data from mitochondrial DNA (mtDNA) and nuclear DNA (ncDNA). MtDNA showed a significant correlation between genetic and geographical distances, but ncDNA did not support this correlation. Both datasets revealed that A. fasciaticollis populations had experienced recent expansion and lacked a geographical structure. One reason for lack of a genetic structure is a high level of gene flow among the populations. We suggest that the strong flight capability of A. fasciaticollis has been aided by the East Asian monsoon and cold fronts to accelerate the rate of range expansion. In addition, decreased insecticide usage and disturbances from human activities may all have contributed to population expansion. The low effective population number was based on two datasets. It is suggested that the host crop use, local climate conditions, and the recent population expansion might be the main influencing factors. [ABSTRACT FROM AUTHOR]

Transgenic cotton (Gossypium hirsutum L.) varieties, adapted to China, have been bred that express two genes for resistance to insects, the CrylAc gene from Bacillus thuringiensis (Berliner) (Bt), and a trypsin inhibitor gene from cowpea (CpTI). Effectiveness of the double gene modification in conferring resistance to cotton bollworm, Helicouerpa arinigera (Hübner) (Lepidoptera: Noctuidae), was studied in laboratory and field experiments. In each experiment, performance of Bt+CpTI cotton was compared with Bt cotton and to a conventional nontransgenic variety. Larval survival was lower on both types of transgenic variety, compared with the conventional cotton. Survival of first-, second-, and third-stage larvae was lower on Bt+CpTI cotton than on Bt cotton. Plant structures differed in level of resistance, and these differences were similar on Bt and Bt+CpTI cotton. Likewise, seasonal trends in level of resistance in different plant structures were similar in Bt and Bt+CpTI cotton. Both types of transgenic cotton interfered with development of sixth-stage larvae to adults, and no offspring was produced by H. armigera that fed on Bt or Bt+CpTI cotton from the sixth stage onward. First-, second-, and third-stage larvae spent significantly less time feeding on transgenic cotton than on conventional cotton, and the reduction in feeding time was significantly greater on Bt+CpTI cotton than on Bt cotton. Food conversion efficiency was lower on transgenic varieties than on conventional cotton, but there was no significant difference between Bt and Bt+CpTI cotton. In 3-yr field experimentation, boliworm densities were greatly suppressed on transgenic as compared with conventional cotton, but no significant differences between Bt and Bt+CpTI cotton were found. Overall, the results from laboratory work indicate that introduction of the CpTI gene in Bt cotton raises some components of resistance in cotton against H. armigera, but enhanced control of H. armigera under field conditions, due to expression of the CpTI gene, was not demonstrated. [ABSTRACT FROM AUTHOR]

Transgenic insect-resistant cotton expressing the Cry1Ac and/or CpTI protein has been released into the environment for commercial cultivation for over a decade in China. In this consecutive 3-year field study (2007-2009), we characterized the microbial populations in two different transgenic cotton fields and one non-transgenic cotton field to assess the environmental impact of long-term cultivation of transgenic insect-resistant plants in credible field conditions. We obtained samples at four different growth stages of cotton (seedling, budding, boll forming and boll opening). The results showed significant seasonal variation in the number of bacteria, fungi, azotobacter, denitrifying bacteria and ammonia-oxidizing bacteria and in diversity indices of microorganisms, but no significant difference in the number of each microbial population or diversity indices attributable to long-term cultivation of transgenic cotton. [ABSTRACT FROM AUTHOR]