Dynamic changes of gut bacterial communities present in larvae of Anoplophora glabripennies collected at different developmental stages.

The Asian long-horned beetle, Anoplophora glabripennies (Motschulsky), is a destructive wood-boring pest that is capable of killing healthy trees. Gut bacteria in the larvae of the wood-boring pest is essential for the fitness of hosts. However, little is known about the structure of the intestinal microbiome of A. glabripennies during larval development. Here, we used Illumina MiSeq high-throughput sequencing technology to analyze the larval intestinal bacterial communities of A. glabripennies at the stages of newly hatched larvae, 1st instar larvae and 4th instar larvae. Significant differences were found in larval gut microbial community structure at different larvae developmental stages. Different dominant genus was detected during larval development. Acinetobacter were dominant in the newly hatched larvae, Enterobacter and Raoultella in the 1st instar larvae, and Enterococcus and Gibbsiella in the 4th instar larvae. The microbial richness in the newly hatched larvae was higher than those in the 1st and 4th instar larvae. Many important functions of the intestinal microbiome were predicted, for example, fermentation and chemoheterotrophy functions that may play an important role in insect growth and development was detected in the bacteria at all tested stages. However, some specific functions are found to be associated with different development stages. Our study provides a theoretical basis for investigating the function of the intestinal symbiosis bacteria of A. glabripennies.

Similar gut bacteria composition in Apriona germari on two preferred host plants.

Apriona germari is one of the most serious wood-boring pests that cause damage to economic and landscaping trees and has adapted to a wide range of plants as diet. Gut bacteria play an important role in biology and ecology of herbivores, especially in growth and adaptation. To investigate how plant hosts shape A. germari gut microbiota, A. germari larvae were collected from Populus tomentosa and Malus pumilal, and gut microbiomes were sequenced based on 16S rDNA high-throughput sequencing technology. A total of 853,424 high-quality reads were obtained and clustered into 196 operational taxonomic units under a 97% similarity cutoff, which were annotated into 8 phyla, 10 classes, 21 orders, 34 families, 59 genera, and 39 species. Gibbsiella was the most dominant genus of intestinal bacteria, followed by Enterobacter and Acinetobacter. No significant difference was observed in larvae gut bacterial richness and diversity of A. germari collected from two hosts, though alpha diversity showed that the richness of gut bacteria in A. germari larvae collected on P. tomentosa was slightly higher than that in A. germari on M. pumilal, and beta diversity showed little difference between two host plants. The functional abundance analysis of the detected bacteria revealed fermentation, chemoheterotrophy, symbionts, and nitrate relative functions that highly possibly support wood-boring beetles to feed on woody tissues. Our study provided a theoretical basis for investigating the function of intestinal symbiosis bacteria of A. germari.

Influence of dietary aconitine and nicotine on the gut microbiota of two lepidopteran herbivores.

The gut microbiota plays an important role in pheromone production, pesticide degradation, vitamin synthesis, and pathogen prevention in the host animal. Therefore, similar to gut morphology and digestive enzyme activity, the gut microbiota may also get altered under plant defensive compound-induced stress. To test this hypothesis, Dendrolimus superans larvae were fed either aconitine- or nicotine-treated fresh leaves of Larix gmelinii, and Lymantria dispar larvae were fed either aconitine- or nicotine-treated fresh leaves of Salix matsudana. Subsequently, the larvae were sampled 72hr after diet administration and DNA extracted from larval enteric canals were employed for gut microbial 16S ribosomal RNA gene sequencing (338 F and 806 R primers). The sequence analysis revealed that dietary nicotine and aconitine influenced the dominant bacteria in the larval gut and determined their abundance. Moreover, the effect of either aconitine or nicotine on D. superans and L. dispar larvae had a greater dependence on insect species than on secondary plant metabolites. These findings further our understanding of the interaction between herbivores and host plants and the coevolution of plants and insects.

Diel pattern in the structure and function of the gut microbial community in Lymantria dispar asiatica (Lepidoptera: Lymantriidae) larvae.

In the present study, diel pattern in gut microbial communities in insects were evaluated. Lymantria dispar asiatica fourth instar larvae (72 ± 2 hr after molting) at noon (LdD) and midnight (LdN) were used for a comparative analysis of the gut microbial community. Ten bacterial operational taxonomic units (OTUs) were shared between LdD and LdN samples. One bacterial OTU was specific to LdD. The dominant gut microbes were OTU72 in LdD and OTU75 in LdN. A linear discriminant analysis effect size cladogram suggested that ten bacterial OTUs maintain significant differences in relative abundances between LdD and LdN. These results agreed with the discrete ellipses between LdD and LdN in principal coordinates analysis plots. Additionally, using phylogenetic investigation of communities by reconstruction of unobserved states, the gut microbial community was assigned to 23 functional terms, among which 22 exhibited significant differences between LdD and LdN. To conclude, the present study documented a diel pattern in the gut microbial community of L. dispar asiatica larvae.

Variation in the pH of experimental diets affects the performance of Lymantria dispar asiatica larvae and its gut microbiota.

To study dietary pH effects on Lymantria dispar asiatica larvae and provide a theoretical basis for its control in different forests, phosphate buffers (PBs) of pH 6, 7, and 8 were used to prepare experimental diets. The diet prepared with pH 6 PB was named as DPB6, with pH 8 PB as DPB8, and with pH 7 PB as DPB7 (control). The dietary pH was 5.00 in DPB6, 6.05 in control, and 6.50 in DPB8. After feeding on the diets with different pH values for 84 hr, fourth-instar caterpillars were randomly collected. Growth and various physiological traits were determined and 16S recombinant DNA sequencing was performed using the intestinal microflora of surviving larvae. Results showed that the mortality was 30% in DPB6, and 10% in DPB8, while no mortality was observed in control. The partial least squares discriminant analyses suggested that diets prepared with PB of different pH resulted in different food intake, amount of produced feces, weight gain, digestive enzyme activities, and antioxidant enzyme activities in larvae. Interestingly, both the highest weight gain and the lowest total antioxidant capacities were seen in control larvae. Results also showed that the larval gut microbiota community structure was significantly affected by dietary pH. Moreover, linear discriminant analysis effect size suggested that the family Acetobacteraceae in control, genus Prevotella in DPB8, and genus Lactococcus, family Flavobacteriaceae, family Mitochondria, and family Burkholderiaceae in DPB6 contributed to the diversity of the larval gut microbial community.

Avermectin stress varied structure and function of gut microbial community in Lymantria dispar asiatica (Lepidoptera: Lymantriidae) larvae.

Lymantria dispar asiatica is a globally distributed herbivorous pest. Avermectin is a highly effective, broad-spectrum insecticide. In this study, fourth instar L. dispar asiatica larvae were exposed to a LC30 dose of avermectin. The structure and function of larval gut microbial community was analyzed to examine how gut microbiota in L. dispar asiatica larvae responded to avermectin stress. Results showed that the structure and function of gut microbial community in L. dispar asiatica larvae were varied by avermectin stress. To be precise, more than half quantity of the observed Optical Taxonomic Units (OTUs) showed significantly different abundances under avermectin stress. Linear discriminant analysis effect size (LEfSe) suggested nine bacterial genera and 12 fungal genera contributed to the different gut microbial community structure in L. dispar asiatica larvae. Gut microbial function classification (PICRUSt and FUNGuild) suggested that three bacterial function categories and a fungal function guild were significantly increased, and two fungal function guilds were significantly decreased by avermectin stress. This study furthers our understanding of the physiology of L. dispar asiatica larvae under avermectin stress, and is an essential step towards future development of potential pesticide targets.

Ambient temperature-mediated enzymic activities and intestinal microflora in Lymantria dispar larvae.

To understand how ambient temperature affect the gypsy moth larvae, and provide a theoretical basis for pest control in different environments. Fourth instar gypsy moth larvae were incubating for 3 hr at 15℃, 20℃, 25℃, 30℃, 35℃, and 40℃, respectively. Afterward, digestive and antioxidant enzyme activities, total antioxidant capacity, and intestinal microflora community were analyzed to reveal how the caterpillars respond to ambient temperature stress. Results showed that both digestive and antioxidant enzymes were regulated by the ambient temperature. The optimum incubation temperatures of protease, amylase, trehalase, and lipase in gypsy moth larvae were 30℃, 25℃, and 20℃, respectively. When the incubation temperature was deviated optimum temperatures, digestive enzyme activities would be downregulated depending on the extent of temperature stress. In addition, glutathione S-transferase, peroxidase, catalase, and polyphenol oxidase would be activated under a sufferable temperature stress, but superoxide dismutase and carboxylesterase (CarE) would be inhibited. In addition, results showed that the top two abundant phyla were Proteobacteria and Firmicutes. The phylum Firmicutes abundance was decreased and phylum Proteobacteria abundance was increased by ambient temperature stress. Moreover, it suggested that gypsy moth caterpillars at different ambient temperature mainly differed from each other by Escherichia-Shigella and Bifidobacterium in control, Acinetobacter in T15, and Lactobacillus in T40, respectively.

Cu/ZnSOD always responded stronger and rapider than MnSOD in Lymantria dispar larvae under the avermectin stress.

Metalloenzyme SODs play important roles in insects dealing with environmental stress. Here, we cloned the Cu/ZnSOD (LdCZS) and MnSOD (LdMS) mRNA of Lymantria dispar by rapid amplification of cDNA ends (RACE). Afterwards their expression patterns were detected by quantitative real-time polymerase chain reaction (qPCR) after bioinformatic analysis. We found that both LdCZS and LdMS were widely detected in all gypsy moth larvae and all five tissues that we analyzed, and both of them were up-regulated after larvae were fed with avermectin of sublethal concentration and LC10. The LdCZS expression value are always higher than LdMS after treating with avermectin of sublethal concentrations. In addition, temporal expression profile in avermectin treated larvae showed that LdCZS expressed highest at 2nd hour, and LdMS expressed highest at 6th hour. The cuticulas transcribed LdCZS and LdMS significantly higher than heads, fat bodies, Malpighian tubes, and midguts after spraying avermectin of sublethal concentration. These results suggested that both Cu/ZnSOD and MnSOD are important antioxidant enzymes in L. dispar defensing against pesticide stress, and LdCZS always responded rapider and stronger than LdMS.

Dynamic Changes in Liver Stiffness Measured by Transient Elastography Predict Clinical Outcomes Among Patients With Chronic Hepatitis B.

OBJECTIVES: We aimed to evaluate the validity of transient elastography in monitoring the antiviral outcomes in patients with chronic hepatitis B. METHODS: This study included 108 patients treated with nucleos(t)ide analogues and 67 patients treated with interferon (IFN). Liver biopsies were evaluated by the METAVIR score. Transient elastography was performed initially at baseline, 48 weeks, and 96 weeks. Liver tissue was obtained before and after 96 weeks of treatment. The area under the receiver operating characteristic curve was used to examine the diagnostic value of transient elastography in predicting and monitoring outcomes of antiviral treatment. RESULTS: The liver stiffness value correlated well with the baseline alanine aminotransferase level (r = 0.33; P < .001) and was significantly different among various stages of liver fibrosis (P < .001). In the nucleos(t)ide analogue group, the mean pretreatment and posttreatment liver stiffness values ± SD were 8.7 ± 3.1 and 5.9 ± 1.6 kPa, respectively (P < .001), and they were 9.2 ± 3.7 and 7.2 ± 1.9 kPa (P < .001) in the IFN group. Although the liver stiffness values at baseline between the groups were similar (P = .45), they were 5.9 ± 1.6 kPa in the nucleos(t)ide analogue group and 7.2 ± 1.9 kPa in the IFN group after 48 weeks of treatment (P < .001). With the decreased magnitude liver stiffness for predicting the improvement in liver fibrosis, the area under the receiver operating characteristic curve was 0.68 (P = .029). When the decreased magnitude of liver stiffness was 4.1 kPa or higher, the sensitivity and specificity for predicting a histologic response were 88.2% and 50.0%. CONCLUSIONS: Our findings suggest that transient elastography is an effective measurement tool for diagnosing and monitoring the histologic response in patients with chronic hepatitis B during antiviral treatment and can help avoid multiple liver biopsies.

Potential effects of telbivudine and entecavir on renal function: a systematic review and meta-analysis.

BACKGROUND: To assess the potential effects of telbivudine (LdT) and entecavir (ETV) on renal function in patients with chronic hepatitis B (CHB), we performed a meta-analysis of the relevant data available on these agents to evaluate their effects on the estimated glomerular filtration rate (eGFR) during treatment. METHODS: The PubMed, EMBASE, Scopus, CNKI (China National Knowledge Infrastructure), Cochrane Library, and WanFang databases were searched for relevant articles appearing in the literature up to July 1, 2015. A total of 6 studies (1960 CHB patients) with 1-year eGFR outcomes were retrieved and analyzed. RESULTS: Generally, the results of the 6 studies analyzed showed that eGFR was improved after LdT treatment, but was decreased after ETV treatment. Using a fixed-effects approach, the change in eGFR was found to be significantly different between LdT and ETV treatment (Z = 3.64; P = 0.0003). Whereas the eGFR was slightly decreased with ETV compared with baseline (-1.45 mL/min/1.73 m(2)), the eGFR was improved with LdT (2.99 mL/min/1.73 m(2)) after 1 year of treatment. An overall test of effect in the meta-analysis showed that the eGFR in LdT-treated patients was significantly improved after 1-year of treatment (Z = 3.71; P = 0.0002). CONCLUSION: This meta-analysis has confirmed that LdT has a renal protective effect whereas ETV does not. However, whether the benefit on renal function outweighs the occurrence of resistance in specific clinical situations is not yet clear.

Emerald Ash Borer Infestation-Induced Elevated Negative Correlations and Core Genera Shift in the Endophyte Community of Fraxinus bungeana.

Endophytes, prevalent in plants, mediate plant-insect interactions. Nevertheless, our understanding of the key members of endophyte communities involved in inhibiting or assisting EAB infestation remains limited. Employing ITS and 16S rRNA high-throughput sequencing, along with network analysis techniques, we conducted a comprehensive investigation into the reaction of endophytic fungi and bacteria within F. bungeana phloem by comparing EAB-infested and uninfected samples. Our findings reveal that EAB infestation significantly impacts the endophytic communities, altering both their diversity and overall structure. Interestingly, both endophytic fungi and bacteria exhibited distinct patterns in response to the infestation. For instance, in the EAB-infested phloem, the fungi abundance remained unchanged, but diversity decreased significantly. Conversely, bacterial abundance increased, without significant diversity changes. The fungi community structure altered significantly, which was not observed in bacteria. The bacterial composition in the infested phloem underwent significant changes, characterized by a substantial decrease in beneficial species abundance, whereas the fungal composition remained largely unaffected. In network analysis, the endophytes in infested phloem exhibited a modular topology, demonstrating greater complexity due to an augmented number of network nodes, elevated negative correlations, and a core genera shift compared to those observed in healthy phloem. Our findings increase understanding of plant-insect-microorganism relationships, crucial for pest control, considering endophytic roles in plant defense.

Distinct gut bacterial composition in Anoplophora glabripennis reared on two host plants.

Anoplophora glabripennis (Coleoptera: Cerambycidae: Lamiinae) is an invasive wood borer pest that has caused considerable damage to forests. Gut bacteria are of great importance in the biology and ecology of herbivores, especially in growth and adaptation; however, change in the gut bacterial community of this pest feeding on different hosts is largely unknown. In this study, we investigated the gut bacterial communities of A. glabripennis larvae fed on different preferred hosts, Salix matsudana and Ulmus pumila, using 16S rDNA high-throughput sequencing technology. A total of 15 phyla, 25 classes, 65 orders, 114 families, 188 genera, and 170 species were annotated in the gut of A. glabripennis larvae fed on S. matsudana or U. pumila using a 97% similarity cutoff level. The dominant phyla were Firmicutes and Proteobacteria and the core dominant genera were Enterococcus, Gibbsiella, Citrobacter, Enterobacter, and Klebsiella. There was significantly higher alpha diversity in the U. pumila group than in the S. matsudana group, and principal co-ordinate analysis showed significant differences in gut bacterial communities between the two groups. The genera with significant abundance differences between the two groups were Gibbsiella, Enterobacter, Leuconostoc, Rhodobacter, TM7a, norank, Rhodobacter, and Aurantisolimonas, indicating that the abundance of larval gut bacteria was affected by feeding on different hosts. Further network diagrams showed that the complexity of the network structure and the modularity were higher in the U. pumila group than in the S. matsudana group, suggesting more diverse gut bacteria in the U. pumila group. The dominant role of most gut microbiota was related to fermentation and chemoheterotrophy, and specific OTUs positively correlated with different functions were reported. Our study provides an essential resource for the gut bacteria functional study of A. glabripennis associated with host diet.

Soil fungal and bacterial communities in southern boreal forests of the Greater Khingan Mountains and their relationship with soil properties.

Little is known about the relationship between soil microbial communities and soil properties in southern boreal forests. To further our knowledge about that relationship, we compared the soil samples in southern boreal forests of the Greater Khingan Mountains-the southernmost boreal forest biome in the world. The forests can be divided into boardleaf forests dominated by birch (Betula platyphylla) or aspen (Populus davidiana) and coniferous forests dominated by larch (Larix gmelinii) or pine (Pinus sylvestris var. mongolica). Results suggested different soil microbial communities and soil properties between these southern boreal forests. Soil protease activity strongly associated with soil fungal communities in broadleaf and coniferous forests (p < 0.05), but not with soil bacterial communities (p > 0.05). Soil ammonium nitrogen and total phosphorus contents strongly associated with soil fungal and bacterial communities in broadleaf forests (p < 0.05), but not in coniferous forests (p > 0.05). Soil potassium content demonstrated strong correlations with both soil fungal and bacterial communities in broadleaf and coniferous forests (p < 0.05). These results provide evidence for different soil communities and soil properties in southern boreal forest, and further elucidate the explicit correlation between soil microbial communities and soil properties in southern boreal forests.

Effectiveness of tenofovir or telbivudine in preventing HBV vertical transmission for pregnancy.

To evaluate the efficacy and safety of telbivudine (LdT) and tenofovir (TDF) for preventing hepatitis B virus (HBV) vertical transmission for HBV-positive pregnant women.Pregnant women (n = 145) from January 2013 to June 2017 were enrolled when they met inclusion criteria, which included HBV DNA ≥1.0 × 10 copies/mL and increased alanine aminotransferase (ALT) levels. Groups A (n = 58) and B (n = 51) were treated with LdT and TDF, respectively. Group C (n = 36) received no antiviral treatment. All infants were vaccinated with hepatitis B immunoglobulin and HBV vaccine. Vertical transmission of HBV was indicated by the presence of hepatitis B surface antigen (HBsAg) in infants 6 months and 12 months after birth.There is no difference of clinical characteristics of patients among the 3 groups. Serum HBV DNA levels of the 3 groups were similar at baseline (Group A vs. Group B vs. Group C, 7.88 ±â€Š0.65 vs. 7.91 ±â€Š0.75 vs. 7.69 ±â€Š0.53 P = .25). In addition, the after anti-HBV treatment in Groups A and B were significantly decreased. Also, the serum HBV DNA levels in both Groups A and B were lower than that of Group C (P < .01, both). The HBV infection rate in Group A treated with LdT was not different from Group B treated with TDF. The dynamic changes of serum ALT level were similar. ALT levels were similar among the 3 Groups (P = .171), while there is statistically significant difference between A and C, and between B and C before delivery (P < .01). For the infants, there were no significant differences among body weight, height, head circumference, or Apgar score. However, the HBsAg positivity rates of infants in Groups A, B, C at postpartum 24 weeks and 48 weeks was 0%, 0%, and 11.1%, respectively (P < .001).Administration of LdT or TDF to HBV-infected mothers are effective and safe to block mother-to-infant HBV transmission.

Comparison of pegylated interferon monotherapy and de novo pegylated interferon plus tenofovir combination therapy in patients with chronic hepatitis B.

Background and aim: We aimed to evaluate the effectiveness of pegylated interferon (Peg-IFN) monotherapy (IFN group) and combination therapy with tenofovir (TDF) and Peg-IFN (IFN+TDF group) in chronic hepatitis B (CHB) patients. Patients and methods: Data of 143 CHB patients were analyzed in this study. All patients enrolled received liver biopsy. Virologic responses were defined as hepatitis B virus (HBV) DNA <100 IU/mL, biochemical responses were defined as normalization of alanine aminotransferse (ALT) levels, and HBeAg serological response was defined as HBeAg loss or HBeAg seroconversion to HBeAb. HBsAg serological response was defined as HBsAg loss or HBsAg seroconversion to HBsAb. Results: We observed that a total of 16.7% (11/66) and 33.8% (26/77) patients in IFN and IFN+TDF group achieved complete viral suppression after 48 weeks treatment (P=0.02). Although HBeAg levels in CHB patients in the IFN+TDF group decreased more rapidly during the 48-week treatment, we did not observe significant differences in HBeAg serological loss or seroconversion rates between the two groups at 24 and 48 weeks. HBsAg loss was observed in 13.0% (10/77) of CHB patients in the IFN+TDF group at 48 weeks, compared with only 3% (2/66) patients in the IFN group (P=0.032). No significant difference was observed in HBsAg seroconversion rate between the two groups during 48-week treatment. The biochemical response rate was also significantly higher in the IFN+TDF group than that in the IFN group at week 48 (P=0.015). Multivariate logistic analysis showed that IFN+TDF treatment (OR=4.41, P=0.003), severe baseline hepatic inflammation (OR=4.16, P<0.001), and lower baseline serum HBV DNA levels (OR=0.98, P=0.03) were strong predictors for the virological response. Younger age (OR=0.89, P=0.01), higher baseline ALT level (OR=1.01, P=0.038), and lower baseline HBeAg level (OR=0.99, P=0.008) were independent predictors for HBeAg sero-response after 48 weeks treatment. While only severe liver fibrosis (OR=1.69, P=0.028) and lower baseline HBsAg level (OR=0.22, P=0.005) were independent factors associated with HBsAg sero-response after 48 weeks treatment. Conclusion: Peg-IFN combined with TDF may increase the virological response rate, biochemical response rate, and HBsAg loss rate in patients with CHB infection. The combination treatment is more suitable for those patients who are likely to respond to the treatment.

Search for Nutritional Fitness Traits in a Biological Pest Control Agent Harmonia axyridis Using Comparative Transcriptomics.

Harmonia axyridis is an important natural predator used in the biological control of insect pests. Vitellogenin (Vg) supplementation to artificial diet can improve fecundity of H. axyridis, however, the effects of Vg on physiology of H. axyridis at the molecular level is unclear. This study investigated the effects of Vg on the physiology (digestive enzyme activities) and transcriptome patterns by feeding H. axyridis adults with treatment (artificial diet with Vg supplement) and control (artificial diet supplemented with bovine serum albumin (BSA). The transcriptome sequencing yielded 43.94 Gb of clean data, and 3,946 differentially expressed genes (DEGs) - including 93 upregulated and 3,853 downregulated genes between the treatment and control. Six DEGs related to development and digestive enzyme were chosen for quantitative real-time PCR (qRT-PCR) to validate the accuracy of the RNA-seq results and confirmed that the transcriptome analysis yielded reliable results. The Vg supplement has increased activities of digestive enzymes and related genes expression in H. axyridis. The transcript level of digestive enzyme genes (apolipoprotein D and phosphoenolpyruvate carboxykinase) were much higher in adults fed on diet supplemented with Vg compared with that of the control.