Response of the mosquito immune system and symbiotic bacteria to pathogen infection.

Mosquitoes are the deadliest animal in the word, transmitting a variety of insect-borne infectious diseases, such as malaria, dengue fever, yellow fever, and Zika, causing more deaths than any other vector-borne pathogen. Moreover, in the absence of effective drugs and vaccines to prevent and treat insect-borne diseases, mosquito control is particularly important as the primary measure. In recent decades, due to the gradual increase in mosquito resistance, increasing attention has fallen on the mechanisms and effects associated with pathogen infection. This review provides an overview of mosquito innate immune mechanisms in terms of physical and physiological barriers, pattern recognition receptors, signalling pathways, and cellular and humoral immunity, as well as the antipathogenic effects of mosquito symbiotic bacteria. This review contributes to an in-depth understanding of the interaction process between mosquitoes and pathogens and provides a theoretical basis for biological defence strategies against mosquito-borne infectious diseases.

Screening and analysis of immune-related genes of Aedes aegypti infected with DENV2.

Dengue virus type Ⅱ (DENV2) is a primary serotype responsible for the dengue fever epidemic, and Aedes aegypti is the main DENV2 vector. Understanding the Aedes aegypti immune mechanism against DENV2 is the basis for research on immune blockade in mosquitoes. Some preliminary studies lack validation in the literature, so this study was performed to further study and validate the potential target genes to provide a further basis for screening key target genes. We screened 51 genes possibly related to Aedes aegypti infection and immunity from the literature for further verification. First, bioinformatic methods such as GO, KEGG and PPI analysis were used, and then RT-qPCR was used to detect the changes in mRNA expression in the midguts and salivary glands of Aedes aegypti infected with DENV2.Bioinformatic analysis showed that mostly genes of the glucose metabolism pathway and myoprotein were influenced. In salivary glands, the Gst (xa) and Toll (xb) expression levels were significantly correlated with DENV2 load (y, lg[DENV2 RNA copies]), y = -3436xa+0.2287xb+3.8194 (adjusted R2 = 0.5563, F = 9.148, PF = 0.0045). In midguts, DENV2 load was significantly correlated with the relative Fba(R2 = 0.4381, t = 2.497, p < 0.05, df = 8), UcCr(R2 = 0.4072, t = 2.344, p < 0.05, df = 8) and Gbps1(R2 = 0.4678, t = 2.652, p < 0.05, df = 8) expression levels, but multiple regression did not yield significant results. This study shows that genes related to glucose metabolism and muscle proteins contribute to the interaction between Aedes aegypti and dengue virus. It was confirmed that SAAG-4, histone H4, endoplasmin, catalase and other genes are involved in the regulation of DENV2 infection in Aedes aegypti. It was revealed that GST and Toll in salivary glands may have antagonistic effects on the regulation of DENV2 load. Fba, UcCr and Gbps1 in the midgut may increase DENV2 load. These study results further condensed the potential target gene range of the Aedes aegypti immune mechanism against DENV2 infection and provided basic information for research on the Aedes aegypti in vivo blockade strategy against DENV2.

Transcriptome Analysis of Responses to Dengue Virus 2 Infection in Aedes albopictus (Skuse) C6/36 Cells.

Dengue virus (DENV), a member of the Flavivirus genus of the Flaviviridae family, can cause dengue fever (DF) and more serious diseases and thus imposes a heavy burden worldwide. As the main vector of DENV, mosquitoes are a serious hazard. After infection, they induce a complex host-pathogen interaction mechanism. Our goal is to further study the interaction mechanism of viruses in homologous, sensitive, and repeatable C6/36 cell vectors. Transcriptome sequencing (RNA-Seq) technology was applied to the host transcript profiles of C6/36 cells infected with DENV2. Then, bioinformatics analysis was used to identify significant differentially expressed genes and the associated biological processes. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was performed to verify the sequencing data. A total of 1239 DEGs were found by transcriptional analysis of Aedes albopictus C6/36 cells that were infected and uninfected with dengue virus, among which 1133 were upregulated and 106 were downregulated. Further bioinformatics analysis showed that the upregulated DEGs were significantly enriched in signaling pathways such as the MAPK, Hippo, FoxO, Wnt, mTOR, and Notch; metabolic pathways and cellular physiological processes such as autophagy, endocytosis, and apoptosis. Downregulated DEGs were mainly enriched in DNA replication, pyrimidine metabolism, and repair pathways, including BER, NER, and MMR. The qRT-PCR results showed that the concordance between the RNA-Seq and RT-qPCR data was very high (92.3%). The results of this study provide more information about DENV2 infection of C6/36 cells at the transcriptome level, laying a foundation for further research on mosquito vector-virus interactions. These data provide candidate antiviral genes that can be used for further functional verification in the future.

Dispersal patterns and population genetic structure of Aedes albopictus (Diptera: Culicidae) in three different climatic regions of China.

BACKGROUND: Aedes albopictus is an indigenous primary vector for dengue and Zika viruses in China. Compared with its insecticide resistance, biology and vector competence, little is known about its genetic variation, which corresponds to environmental variations. Thus, the present study examines how Ae. albopictus varies among different climatic regions in China and deciphers its potential dispersal patterns. METHODS: The genetic variation and population structure of 17 Ae. albopictus populations collected from three climatic regions of China were investigated with 11 microsatellite loci and the mitochondrial coxI gene. RESULTS: Of 44 isolated microsatellite markers, 11 pairs were chosen for genotyping analysis and had an average PIC value of 0.713, representing high polymorphism. The number of alleles was high in each population, with the ne value increasing from the temperate region (3.876) to the tropical region (4.144). Twenty-five coxI haplotypes were detected, and the highest diversity was observed in the tropical region. The mean Ho value (ca. 0.557) of all the regions was significantly lower than the mean He value (ca. 0.684), with nearly all populations significantly departing from HWE and displaying significant population expansion (p value < 0.05). Two genetically isolated groups and three haplotype clades were evaluated via STRUCTURE and haplotype phylogenetic analyses, and the tropical populations were significantly isolated from those in the other regions. Most genetic variation in Ae. albopictus was detected within populations and individuals at 31.40 and 63.04%, respectively, via the AMOVA test, and a relatively significant positive correlation was observed among only the temperate populations via IBD analysis (R2 = 0.6614, p = 0.048). Recent dispersions were observed among different Ae. albopictus populations, and four major migration trends with high gene flow (Nm > 0.4) were reconstructed between the tropical region and the other two regions. Environmental factors, especially temperature and rainfall, may be the leading causes of genetic diversity in different climatic regions. CONCLUSIONS: Continuous dispersion contributes to the genetic communication of Ae. albopictus populations across different climatic regions, and environmental factors, especially temperature and rainfall, may be the leading causes of genetic variation.

Vertical Transmission of Zika Virus by Jiegao and Mengding Aedes aegypti (Diptera: Culicidae) Strains in Yunnan Province in China.

Zika virus (ZIKV) is a mosquito-borne pathogen classified in the genus Flavivirus of the family Flaviviridae. Vertical transmission is considered to be the primary way to maintain some arboviruses under adverse natural conditions, which play a critical epidemiological role in arbovirus spread and maintenance. Aedes aegypti is the primary vector for ZIKV. In this study, we demonstrated vertical transmission in two Ae. aegypti strains from Jiegao (JG) and Mengding (MD) in the border area of Yunnan province. The minimum infection rate of F1 adult progeny from JG Ae. aegypti strain was significantly higher than that of MD Ae. aegypti strain in the second gonotrophic cycle (1:14.29 and 1:200, respectively, p < 0.05). The cytopathic effect was observed in C6/36 cells after infection of ZIKV isolated from the progeny. The results suggest that Ae. aegypti mosquitoes from JG and MD play potential roles in ZIKV spread and maintenance. Therefore, more adult and eggs control methods should be implemented to control mosquitoes if a Zika epidemic occurs.

Transcriptome analysis of Aedes aegypti Aag2 cells in response to dengue virus-2 infection.

BACKGROUND: Dengue virus (DENV) is a flavivirus transmitted by mosquitoes that is prevalent in tropical and subtropical countries and has four serotypes (DENV1-4). Aedes aegypti, as the main transmission vector of DENV, exhibits strong infectivity and transmission. With the aim of obtaining a better understanding of the Ae. aegypti-DENV interaction, the transcriptome changes in DENV-2-infected Aag2 cells were studied to describe the immune responses of mosquitoes using the Ae. aegypti Aag2 cell line as a model. METHODS: RNAseq technology was used to sequence the transcripts of the Ae. aegypti Aag2 cell line before and after infection with DENV-2. A bioinformatics analysis was then performed to assess the biological functions of the differentially expressed genes, and the sequencing data were verified by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). RESULTS: The transcriptome analysis generated 8866 unigenes that were found in both groups, 225 unigenes that were only found in the infection group, and 683 unigenes that only existed in the control group. A total of 1199 differentially expressed genes, including 1014 upregulated and 185 downregulated genes, were identified. The bioinformatics analysis showed that the differentially expressed genes were mainly involved in the longevity regulating pathway, circadian rhythm, DNA replication, and peroxisome, purine, pyrimidine, and drug metabolism. The qRT-PCR verification results showed the same trend, which confirmed that the expression of the differentially expressed genes had changed, and that the transcriptome sequencing data were reliable. CONCLUSIONS: This study investigated the changes in the transcriptome levels in the DENV-2-infected Ae. aegypti Aag2 cell line, which provides a faster and effective method for discovering genes related to Ae. aegypti pathogen susceptibility. The findings provide basic data and directions for further research on the complex mechanism underlying host-pathogen interactions.

[Effects of benthic macro-invertebrate on decomposition of Acer buergerianum leaf litter in streams].

By using composite mesh bag method, the effects of benthic macro-invertebrate in an undisturbed stream and an ecologically restored stream on the decomposition process of Acer buergerianum leaf litter from the Purple Mountain of Nanjing in winter were studied. After 112 days of decomposition, the remaining rate of A. buergerianum leaf litter based on ash-free dry mass was 31-62%, and the decomposition rate followed a declined exponential equation (P < 0.05). In the flowing water of the undisturbed and ecologically restored streams, the decomposition rate of leaf litter was 0.0064 d(-1) and 0.0030 d(-1); while in the still water of the streams, it was 0.0016 d(-1) and 0. 0018 d(-1), respectively. The abundance and biomass of benthic macro-invertebrate were significantly higher in the flowing water of undisturbed stream than in that of ecologically restored stream (P < 0.05), but had no significant differences in the still water of the two streams. Shredders (mainly Asellus sp.) had the highest abundance (70.4%) in the flowing water of undisturbed stream, while filterers (mainly Tanytarsus sp.) were dominant (37.8%) in the flowing water of ecologically restored stream. The decomposition rate of the leaf litter was significantly correlated with the richness and abundance of shredder species in flowing water (P < 0.01), but had less correlation with the biomass of the shredders, suggesting that the decomposition of A. buergerianum leaf litter in streams in winter was more dependent on the richness and abundance of shredders.

[Effects of sediments on submerged macrophytes growth].

With mesocom experiment, this paper studied the effects of different sediments (sandstone, clay and mud) in Taihu Lake of China on the growth of four submerged macrophytes Vallisneria natans, Potamogeton malaianus, Hydrilla verticillata, and Ceratophyllum demersum. The results showed that among the test sediments, mud was more available. When growing on sandstone, clay and mud, the mean biomass of V. natans, P. malaianus, H. verticillata and C. demersum was 72.37, 126.25 and 134.10 g, 40.0, 72.10 and 90.70 g, 0.27, 6.58 and 73.64 g, and 0.17, 3.26 and 84.42 g, respectively. V. natans and P. malaianus had a stronger adaptability to the clay with lower nutrients contents, while H. verticillata and C. demersum didn't. All the test species grown on sandstone had the lowest biomass and shoot height, and H. verticillat and C. demersum were not able to survive by the end of the experiment. V. natans had a lower root activity (TTC) than P. malaianus, being 0, 0.16 +/- 0.05 and 0.36 +/- 0.33 mg x g(-1) x h(-1), and 2.68 +/- 0.34, 2.30 +/- 0.77 and 5.24 +/- 0.67 mg x g(-1) x h(-1) when growing on sandstone, clay, and mud, respectively. The oxygen release from the root systems of test submerged macrophytes was in the order of V. natans > P. malaianus > H. verticillata. The measurements of chlorophyll content, cell membrane permeability, and MDA also had the similar trends mentioned above.