Reporting form and content of research priorities identified in knee osteoarthritis clinical practice guidelines: a methodological literature analysis.

OBJECTIVES: Clinical practice guideline (CPG) developers conduct systematic summaries of research evidence, providing them great capacity and ability to identify research priorities. We systematically analysed the reporting form and content of research priorities in CPGs related to knee osteoarthritis (KOA) to provide a valuable reference for guideline developers and clinicians. DESIGN: A methodological literature analysis was done and the characteristics of the reporting form and the content of the research priorities identified in KOA CPGs were summarised. DATA SOURCES: Six databases (PubMed, Embase, China National Knowledge Infrastructure, VIP Database for Chinese Technical Periodicals, Wanfang and Chinese Biomedical Literature Database) were searched for CPGs published from 1 January 2017 to 4 December 2022. The official websites of 40 authoritative orthopaedic societies, rheumatology societies and guideline development organisations were additionally searched. ELIGIBILITY CRITERIA: We included all KOA CPGs published in English or Chinese from 1 January 2017 that included at least one recommendation for KOA. We excluded duplicate publications, older versions of CPGs as well as guidance documents for guideline development. DATA EXTRACTION AND SYNTHESIS: Reviewers worked in pairs and independently screened and extracted the data. Descriptive statistics were used, and absolute frequencies and proportions of related items were calculated. RESULTS: 187 research priorities reported in 41 KOA CPGs were identified. 24 CPGs reported research priorities, of which 17 (41.5%) presented overall research priorities for the entire guideline rather than for specific recommendations. 110 (58.8%) research priorities were put forward due to lack of evidence. Meanwhile, more than 70% of the research priorities reflected the P (population) and I (intervention) structural elements, with 135 (72.2%) and 146 (78.1%), respectively. More than half of the research priorities (118, 63.8%) revolved around evaluating the efficacy of interventions. Research priorities primarily focused on physical activity (32, 17.3%), physical therapy (30, 16.2%), surgical therapy (27, 14.6%) and pharmacological treatment (26, 14.1%). CONCLUSIONS: Research priorities reported in KOA CPGs mainly focused on evaluating non-pharmacological interventions. There exists considerable room for improvement for a comprehensive and standardised generation and reporting of research priorities in KOA CPGs.

Emphasis should be placed on identifying and reporting research priorities to increase research value: An empirical analysis.

OBJECTIVES: To compared the presentation of research priorities in the GRADE (Grading of Recommendations Assessment, Development, and Evaluation) clinical practice guidelines (CPGs) developed under the guidance of the GRADE working group or its two co-chair, and the Chinese CPGs. METHODS: This was a methodological empirical analysis. We searched PubMed, Embase, and four Chinese databases (Wanfang, VIP Database for Chinese Technical Periodicals, China National Knowledge Infrastructure and Chinese Biomedical Literature Database) and retrieved nine Chinese guideline databases or Society websites as well as GRADE Pro websites. We included all eligible GRADE CPGs and a random sample of double number of Chinese CPGs, published 2018 to 2022. The reviewers independently screened and extracted the data, and we summarized and analyzed the reporting on the research priorities in the CPGs. RESULTS: Of the 135 eligible CPGs (45 GRADE CPGs and 90 Chinese CPGs), 668, 138 research priorities were identified respectively. More than 70% of the research priorities in GRADE CPGs and Chinese CPGs had population and intervention (PI) structure. 99 (14.8%) of GRADE CPG research priorities had PIC structures, compared with only 4(2.9%) in Chinese. And 28.4% (190) GRADE CPG research priorities reflected comparisons between PICO elements, approximately double those in Chinese. The types of research priorities among GRADE CPGs and Chinese CPGs were mostly focused on the efficacy of interventions, and the type of comparative effectiveness in the GRADE research priorities was double those in Chinese. CONCLUSIONS: There was still considerable room for improvement in the developing and reporting of research priorities in Chinese CPGs. Key PICO elements were inadequately presented, with more attention on intervention efficacy and insufficient consideration given to values, preferences, health equity, and feasibility. Identifying and reporting of research priorities deserves greater effort in the future.

A Review on Traditional Uses, Phytochemistry, Pharmacology and Clinical Application of Tinospora sinensis (Lour.) Merr.

Tinospora sinensis (T. sinensis), whose Tibetan name is "Lezhe", as a traditional medicine, is widely distributed in China, India and Sri Lanka. It is used for the treatment of rheumatic arthralgia, sciatica, lumbar muscle strain and bruises. Research over the previous decades indicated that T. sinensis mainly contains terpenes, lignans, alkaloids, phenol glycosides and other chemical components. A wide range of pharmacologic activities such as anti-inflammatory, analgesic, immunosuppressive, anti-aging, anti-radiation, anti-leishmania and liver protection have been reported. However, the scholar's research on the pharmacodynamic material basis of T. sinensis is relatively weak. Data regarding many aspects such as links between the traditional uses and bioactivities, pharmacokinetics, and quality control standard of active compositions is still limited and need more attention. This review reports a total of 241 compounds, the ethnopharmacology and clinical application of T. sinensis, covering the literature which were searched by multiple databases including Web of Science, PubMed, Google Scholar, Science Direct, CNKI and other literature sources from 1996 to date, with a view to provide a systematic and insightful reference and lays a foundation and inspiration for the application and further in-depth research of T. sinensis resources.

[Analysis of metabolites in simulated gastric fluid of Saposhnikoviae Radix polysaccharides].

To fully understand whether Saposhnikoviae Radix polysaccharides(SP) can be metabolized in gastric fluid and the meta-bolic behavior, this study systematically analyzed the metabolites in simulated gastric fluid of SP by high-performance liquid chromatography-ion trap time-of-flight mass spectrometry(HPLC-IT-TOF-MS) technology in combination with zebrafish immune activity evaluation. Based on the obtained accurate relative molecular mass, chromatographic retention behavior, MS fragmentation patterns, refe-rence standards, and relevant literature reports, 19 metabolites were analyzed and identified. Among them, five monosaccharides and 14 oligosaccharides were generated as metabolites. Several reducing sugars, including mannose, glucose, rhamnose, and xylose, were accurately identified in the gastric fluid metabolites. Zebrafish pharmacological evaluation results indicated that SP maintained good immune activity after gastric fluid metabolism, with the most significant increase in immune cell density observed at W3(simulated gastric fluid metabolism for 2 hours). Among the gastric fluid metabolites, M1 and M3(Hex-Hex-Man) may be most closely related to pharmacological activity and could be further studied as potential active fragments.

Impact analysis of hydraulic residence time and dissolved oxygen on performance efficiency and microbial community in N, N-dimethylformamide wastewater treated by an AnSBR-ASBR.

Suitable operating parameters are one of the key factors to efficient and stable biological wastewater treatment of N, N-dimethylformamide (DMF) wastewater. In this study, an improved AnSBR-ASBR reactor (anaerobic sequencing batch reactor, AnSBR, and aerobic SBR, ASBR, run in series) was used to investigated the effects of operating conditions such as hydraulic residence time (HRT), AnSBR stirring speed and ASBR dissolved oxygen (DO) for DMF wastewater treatment. When HRT decreased from 24 h to 12 h, the average removal rates of COD by the AnSBR were 34.59% and 39.54%, respectively. Meanwhile, the removal rate of NH4+-N by ASBR decreased from 88.38% to 62.81%. The DMF removal rate reached the best at 18 h and the expression of dehydrogenase was the highest in the AnSBR. The abundance of Megasphaera, the dominant sugar-degrading bacteria in the AnSBR, continued to decline due to the decrease of HRT. The relative abundance of Methanobacterium gradually increased to 80.2% with the decrease of HRT and that hydrotrophic methanogenesis dominated the methanogenic process. The HRT decrease promoted butyrate and pyruvate metabolism in anaerobic sludge, but the proportion of glycolysis and methane metabolism decreased. The AnSBR-ASBR reactor had the best operation performance when HRT was 18 h, AnSBR speed was 220 r/min, and ASBR DO content was 3-4 mg/L. This study provided an effective reference for the reasonable selection of operating parameters in the treatment of DMF-containing wastewater by the AnSBR-ASBR.

The complete mitochondrial genome and gene rearrangements in a gall wasp species, Dryocosmus liui (Hymenoptera: Cynipoidea: Cynipidae).

Mitochondrial genomes (mitogenomes) have been widely used in comparative and evolutionary genomics, molecular evolution, phylogenetics, and population genetics, but very limited information is available for the family Cynipidae. In this report, we describe the mitogenome of Dryocosmus liui Pang, Su et Zhu, providing the first complete mitogenomic data for a cynipid gall wasp species. The mitogenome of D. liui is 16,819 bp in length, and contains the typical set of 37 genes. Two control regions were detected, with the second being a perfect inverted repeat of the major portion of the first. Gene rearrangements were found in transfer RNA (tRNA) genes, protein-coding genes (PCGs) and ribosomal RNA (rRNA) genes, compared with the putative ancestral mitogenome. Similar to two other Cynipidae species with mitogenome data available, D. liui has a novel tRNA gene cluster trnL1-trnI-trnL2-trnW-trnM-trnQ between nad1 and nad2. Phylogenetic analysis based on sequences of PCGs and rRNA genes with D. liui included obtained topologies identical to previous studies supporting the a relationship of (Cynipoidea , (Platygastroidea, Proctotrupoidea)) within the monophyletic Proctotrupomorpha and (Cynipidae, Figitidae), Ibaliidae) within the Cynipoidea.

Effects of microplastics on the properties of different types of sewage sludge and strategies to overcome the inhibition: A review.

Microplastics have been identified as an emerging pollutant. When microplastics enter wastewater treatment plants, the plant traps most of the microplastics in the sludge during sewage treatment. Therefore, the effects of microplastics on sludge removal performance, and on the physical and chemical properties and microbial communities in sludge, have attracted extensive attention. This review mainly describes the presence of microplastics in wastewater treatment plants, and the effects of microplastics on the decontamination efficiency and physicochemical properties of activated sludge, aerobic granular sludge, anaerobic granular sludge and anaerobic ammonium oxidation sludge. Further, the review summarizes the effects of microplastics on microbial activity and microbial community dynamics in various sludges in terms of type, concentration, and contact time. The mechanisms used to strengthen the reduction of microplastics, such as biochar and hydrochar, are also discussed. This review summarizes the mechanism by which microplastics influence the performance of different types of sludge, and proposes effective strategies to mitigate the inhibitive effect of microplastics on sludge and discusses removal technologies of microplastics in sewage. Biochar and hydrochar are one of the effective measures to overcome the inhibition of microplastics on sludge. Meanwhile, constructed wetland may be one of the important choice for the future removal of microplastics from sewage. The goal is to provide theoretical support and insights for ensuring the stable operation of wastewater treatment plants and reducing the impact of microplastics on the environment.

Performance and microbial metabolic mechanism of imidacloprid removal in a microbial electrolysis cell-integrated adsorption biological coupling system.

Coke used as a filler to treat imidacloprid (IMI) wastewater by both adsorption biological coupling and microbial electrolysis cells (MEC)-adsorption biological coupling technologies, the removal efficiencies on pollutions in wastewater containing IMI were investigated, and the key functional genes related to IMI degradation pathways were also revealed. Results showed that the removal rates of COD, ammonia nitrogen, TP, and IMI under the adsorption biological coupling treatment and MEC-adsorption biological coupling treatment were 94.61-95.54%, 93.37-95.79%, 73.69-83.80%, and 100%, respectively. MEC increased the relative abundance of Proteobacteria by 9.01% and transformed the dominant bacteria from Lysobacter and Reyranella to Brevundimonas and Aquincola. Moreover, MEC up-regulated the abundance of the coding genes PK (9.30%), narG (2.26%), pstS (3.63%), and phnD (1.32%), and converted the IMI degradation products to smaller molecular weight C6H8N2 and C6H6ClNO. This study provided an important reference information for efficient treatment of IMI wastewater using the MEC-adsorption biological coupling technology.

Effects of upward flow rate and modified biochar location on the performance and microecology of an anaerobic reactor treating kitchen waste.

Increasing the value of food waste through anaerobic digestion is an attractive strategy. Meanwhile, the anaerobic digestion of kitchen waste also faces some technical challenges. In this study, four EGSB reactors were equipped with Fe-Mg-chitosan bagasse biochar at different locations, and the reflux pump flow rate was increased to change the upward flow rate of the reactor. The effects of adding modified biochar at different locations under different upward flow rate on the efficacy and microecology of anaerobic reactors treating kitchen waste were investigated. Results showed that Chloroflexi was the dominant microorganism when the modified biochar was added to the lower, middle, and upper parts of the reactor and mixed in the reactor, accounting for 54%, 56%, 58%, and 47%, respectively, on day 45. With the increased upward flow rate, the abundance of Bacteroidetes and Chloroflexi increased, while Proteobacteria and Firmicutes decreased. It was worth noting that the best COD removal effect was achieved when the anaerobic reactor upward flow rate was v2 = 0.6 m/h and the modified biochar was added in the upper part of the reactor, during which the average COD removal rate reached 96%. In addition, mixing modified biochar throughout the reactor while increasing the upward flow rate provided the greatest stimulus for the secretion of tryptophan and aromatic proteins in the sludge extracellular polymeric substances. The results provided a certain technical reference for improving the efficiency of anaerobic digestion of kitchen waste and scientific support for the application of modified biochar to the anaerobic digestion process.

Four new diastereoisomeric neolignan glycosides from the root bark of Lycium chinense Miller. and their α-glucosidase inhibitory activity.

Four new diastereoisomeric neolignan glycosides (1-4) along with nine known lignan glycosides (5-13) were isolated from the root bark of Lycium chinense Mill. Their structures with absolute configurations were elucidated on the basis of NMR spectroscopy, ECD, Mo2(OAc)4-induced ECD spectra, enzymatic hydrolysis and acid hydrolysis. The isolated compounds were evaluated for their α-glucosidase inhibitory activity. Compounds 8 and 13 exhibited moderate inhibitory activities against α-glucosidase with IC50 values of 26.82 ± 2.71 and 43.14 ± 2.81 µg/mL.

Effects of perfluoroalkyl substances on the operational efficiency, microbial communities, and key metabolic pathways of constructed rapid infiltration system with coke as filler layer.

Influences of perfluoroalkyl substances on the performance and microbial metabolic pathways of constructed rapid infiltration systems are not fully understood. In this study, wastewater containing different concentrations of perfluorooctanoic acid (PFOA)/perfluorobutyric acid (PFBA) was treated in constructed rapid infiltration systems with coke as filler. The addition of 5 and 10 mg/L PFOA inhibited the removal of chemical oxygen demand (COD) (80.42%, 89.27%), ammonia nitrogen (31.32%, 41.14%), and total phosphorus (TP) (43.30%, 39.34%). Meanwhile, 10 mg/L PFBA inhibited TP removal of the systems. Based on X-ray photoelectron spectroscopy, the percentages of F- within the PFOA and PFBA groups were 12.91% and 48.46%, respectively. PFOA transformed Proteobacteria (71.79%) into the dominant phyla of the systems, whereas PFBA enriched Actinobacteria (72.51%). The PFBA up-regulated the coding gene of 6-phosphofructokinase by 14.44%, whereas PFOA down-regulated it by 4.76%. These findings provide insights into the toxicity of perfluoroalkyl substances on constructed rapid infiltration systems.

Fe(III) enhances Cr(VI) bioreduction in a MFC-granular sludge coupling system: Experimental evidence and metagenomics analysis.

The influence of Fe(III) on the bioreduction efficiency of Cr(VI) in a microbial fuel cell (MFC)-granular sludge coupling system using dissolved methane as an electron donor and carbon source was explored, and the mechanism of Fe(III) mediating enhancement in the bioreduction process of Cr(VI) in the coupling system was also investigated. Results showed that the presence of Fe(III) enhanced the ability of the coupling system to reduce Cr(VI). The average removal efficiencies of Cr(VI) in the anaerobic zone in response to 0, 5, and 20 mg/L of Fe(III) were 16.53±2.12%, 24.17±2.10%, and 46.33±4.41%, respectively. Fe(III) improved the reducing ability and output power of the system. In addition, Fe(III) enhanced the electron transport systems activity of the sludge, the polysaccharide and protein content in the anaerobic sludge. Meanwhile, X-ray photoelectron spectrometer (XPS) spectra demonstrated that Cr(VI) was reduced to Cr(III), while Fe2p participated in reducing Cr(VI) in the form of Fe(III) and Fe(II). Proteobacteria, Chloroflexi, and Bacteroidetes were the dominant phylum in the Fe(III)-enhanced MFC-granular sludge coupling system, accounting for 49.7%-81.83% of the microbial community. The relative abundance of Syntrophobacter and Geobacter increased after adding Fe(III), indicating that Fe(III) contributed to the microbial mediated anaerobic oxidation of methane (AOM) and bioreduction of Cr(VI). The genes mcr, hdr, and mtr were highly expressed in the coupling system after the Fe(III) concentration increased. Meanwhile, the relative abundances of coo and aacs genes were up-regulated by 0.014% and 0.075%, respectively. Overall, these findings deepen understanding of the mechanism of the Cr(VI) bioreduction in the MFC-granular sludge coupling system driven by methane under the influence of Fe(III).

Pangenomic analysis of Wolbachia provides insight into the evolution of host adaptation and cytoplasmic incompatibility factor genes.

Introduction: The genus Wolbachia provides a typical example of intracellular bacteria that infect the germline of arthropods and filarial nematodes worldwide. Their importance as biological regulators of invertebrates, so it is particularly important to study the evolution, divergence and host adaptation of these bacteria at the genome-wide level. Methods: Here, we used publicly available Wolbachia genomes to reconstruct their evolutionary history and explore their adaptation under host selection. Results: Our findings indicate that segmental and single-gene duplications, such as DNA methylase, bZIP transcription factor, heat shock protein 90, in single monophyletic Wolbachia lineages (including supergroups A and B) may be responsible for improving the ability to adapt to a broad host range in arthropod-infecting strains. In contrast to A strains, high genetic diversity and rapidly evolving gene families occur in B strains, which may promote the ability of supergroup B strains to adapt to new hosts and their large-scale spreading. In addition, we hypothesize that there might have been two independent horizontal transfer events of cif genes in two sublineages of supergroup A strains. Interestingly, during the independent evolution of supergroup A and B strains, the rapid evolution of cif genes in supergroup B strains resulted in the loss of their functional domain, reflected in a possible decrease in the proportion of induced cytoplasmic incompatibility (CI) strains. Discussion: This present study highlights for reconstructing of evolutionary history, addressing host adaptation-related evolution and exploring the origin and divergence of CI genes in each Wolbachia supergroup. Our results thus not only provide a basis for further exploring the evolutionary history of Wolbachia adaptation under host selection but also reveal a new research direction for studying the molecular regulation of Wolbachia- induced cytoplasmic incompatibility.

Role of coke media strategy in an adsorption-biological coupling technology for wastewater treatment performance, microbial community, and metabolic pathways features.

With the increase of wastewater discharge, the requirement of wastewater treatment technology is gradually increased. How to treat wastewater economically, while making the treatment process short, easy to manage and low running cost, is the focus of attention. Adsorption-biological coupling technology could make adsorption and biodegradation complement each other, which has coupled accumulation effect. In this study, with coke as the adsorbent, the efficiency of the adsorption-biological coupling reactor on the treatment of total phosphorus (TP), chemical oxygen demand (COD), and ammonia nitrogen (NH3-N) in domestic wastewater under different influent modes was investigated. Meanwhile, microbial community and metabolic pathways analysis of the reactor were carried out. Results showed that when the influent modes of the coupling reactor was once a day and the daily sewage treatment capacity was 2 L, the treatment efficiency of TP, COD, and NH3-N was the best. The removal rate of TP and NH3-N was 87.96% and 96.14%, respectively. The dominant phylum was Proteobacteria (39.84-44.49%), and the dominant genus was Sphingomonas (4.27-7.16%), and Gemmatimonas (1.27-3.58%). According to the metagenomic analysis, carbon metabolism process was evenly distributed in U (upper), M (middle), and L (lower) layers of the coupling reactor. Phosphate metabolism was mainly in the U layer at first, then in the M and L layers gradually. Carbon metabolism and phosphate metabolism provided sufficient energy for microbial degradation of pollutants. Nitrogen removal in the reactor mainly happened in the S and Z layers by nitrification (M00528) and denitrification (M00529), respectively.

Metagenomic and extracellular polymeric substances analysis reveals the mechanism of exogenous N-hexanoyl-L-homoserine lactone in alleviating the inhibition of perfluorooctanoic acid on anammox process.

To alleviate the negative effects of perfluorooctanoic acid (PFOA) on nitrogen removal via anaerobic ammonia oxidation (anammox), an exogenous signaling factor (N-hexanoyl-L-homoserine lactone, C6-HSL) was introduced into an anammox reactor. Results showed that 2 µmol/L C6-HSL promoted the nitrogen removal efficiency of the anammox reactor under PFOA stress, with the removal efficiencies of ammonia and nitrite increasing from 79.7 ± 4.8 % and 80.8 ± 3.8 %, to 94.4 ± 4.3 % and 97.1 ± 3.8 %. Exogenous C6-HSL enhanced the compactness of the extracellular proteins, and improved the sludge hydrophobicity. Meanwhile, C6-HSL resulted in a microbial shift, with the relative abundance of Planctomycetes increasing from 30.2 % to 49.5 %. Candidatus Kuenenia stuttgartiensis replaced Candidatus Brocadia sp. BL1 as the dominant species, while the available space for other nitrogen-removing bacteria was reduced. Exogenous C6-HSL promoted the expression of anammox-related genes, such as hzsB and hdh, while denitrifying genes were down-regulated. In addition, the relative abundance of HdtS, which synthesizes AHLs, increased by 0.02446%.

Use of iron-loaded biochar to alleviate anammox performance inhibition under PFOA stress conditions: Integrated analysis of sludge characteristics and metagenomics.

The negative effects of perfluorooctanoic acid (PFOA) on biological nitrogen removal performance in wastewater treatment plants, are receiving increasing attention due to the widespread reporting of this issue. In this study, pomelo peel iron-loaded biochar (Fe-PBC) was added to an anammox bioreactor to alleviate the negative effects of PFOA. Results showed that the addition of Fe-PBC increased the ammonia and nitrite removal efficiencies from 77.7 ± 9.6 % and 79.5 ± 5.6 % to 94.45 ± 5.1 % and 95.9 ± 5.0 %, respectively. In addition, Fe-PBC promoted the removal of PFOA from wastewater, increasing the PFOA removal efficiency from 5.2 % to 29.2 ± 4.3 % from 100 to 200 days. The introduction of iron-loaded biochar into the anammox bioreactor increased the CO ratio by 13.64 % by 150 days. In addition, a CO fitting peak was detected in the Fe-PBC, indicating that the Fe-PBC was loaded with microorganisms. Microbial community analysis showed a decrease in the relative abundances of Proteobacteria and Nitrospirae from 31 % and 3.4 % to 16.8 % and 0.9 %, respectively, while the relative abundance of Planctomycetes increased from 26.8 % to 44.1 %. Metagenomic analysis found that the functional genes hzsB and hdh increased from 98,666 ± 11,400 and 3190 ± 460 to 119,333 ± 15,534 and 138,650 ± 11,233 copy numbers/MLSS. The increase in anammox biomass may be attributed to the presence of iron, an essential element for the synthesis of key anammox enzyme. Furthermore, iron was also associated with the enhanced extracellular electron transfer in the anammox system induced by Fe-PBC.

[Two new monoterpenoid indole alkaloids from hook-bearing branches of Uncaria sessilifructus].

The present study investigated the chemical constituents from Uncaria sessilifructus and their neuroprotective activities. The compounds were separated and purified from the 90% ethanol extract of U. sessilifructus by various chromatographic methods, including silica gel, Sephadex LH-20, and semi-preparative HPLC. Seven compounds were obtained, and their structures were identified as uncanidine J(1), uncanidine K(2), 17-O-ethylhirsutine(3), tetrahydroalstonine(4), akuammigine(5), hirsutine(6), and hirsuteine(7) by physicochemical properties and various spectral techniques, including UV, IR, MS, and NMR. Compounds 1 and 2 are two new compounds. Compound 3 is a new natural product, and compound 4 was isolated from U. sessilifructus for the first time. In addition, the isolated compounds were evaluated for their neuroprotective effects on oxygen and glucose deprivation/reoxygenation(OGD/R) injury in primary cortical neurons in rats. The results showed that compounds 1-7 had different degrees of protective effects on OGD/R injury. The EC_(50) values of compounds 2-4 were(0.17±0.03),(1.70±0.38), and(1.79±0.23) µmol·L~(-1), respectively.

Effect of asparagine, corncob biochar and Fe(II) on anaerobic biological treatment under low temperature: Enhanced performance and microbial community dynamic.

To ensure the efficiency of anaerobic biological treatment technology at lower temperature will expand the application of anaerobic reactor in practical industrial wastewater treatment. Through a batch experiment, asparagine, corncob biochar and Fe2+ were selected as strengthening measures to analyze the effects on the anaerobic sludge characteristics, microbial community and functional genes in the low temperature (15 °C). Results showed that after 21 days, asparagine began to promote chemical oxygen demand (COD) removal by the anaerobic treatment, with highest COD removal rate (81.65%) observed when the asparagine concentration was 1 mmol/L. When adding 3 g biochar, 25 mg/L Fe2+, and the combination of biochar and Fe2+, the COD removal rates reached to 82%, 92% and 97%, respectively. In the presence of asparagine, both biochar and Fe2+ alone or in combination increased the activity of protease (16.35%-120.71%) and coenzyme F420 (5.63%-130.2%). The relative abundance of Proteobacteria and Methanobacterium increased in the presence of biochar and Fe2+. In addition, the KEGG results showed that the combined addition of biochar and Fe2+ enhanced bacterial replication and repair and promoted amino acid metabolism of archaea.

High Levels of Multiple Phage WO Infections and Its Evolutionary Dynamics Associated With Wolbachia-Infected Butterflies.

Wolbachia is a maternally inherited bacterium that is widely distributed among arthropods, in which it manipulates the reproduction of its hosts. Phage WO is the only bacteriophage known to infect Wolbachia, and may provide benefit to its host or arthropods. We screened for the presence of phage WO in Wolbachia-infected butterfly species for the first time, to investigate their diversity and evolutionary dynamics. All Wolbachia-infected butterfly species, including members of the families Hesperiidae, Lycaenidae, Nymphalidae, Papilionidae, and Pieridae, were found to harbor phage WO. Interestingly, 84% of 19 butterfly species, which were infected with a single Wolbachia strain harbored high levels of multiple phage types (ranging from 3 to 17 types), another three species harbored one or two phage types. For Wolbachia strains (ST-41, ST-19, ST-125 and ST-374) shared among various butterfly species, their host insects all harbored multiple phage types, while two Wolbachia strains (ST-297 and ST-wPcau) were found to infect one butterfly species, whose insect hosts harbored a single phage type, suggesting that horizontal transfer of Wolbachia between insects increased the likelihood of exposure to phages, resulting in increased phage genetic diversity. Twelve horizontal transmission events of phage WO were found, which shared common phage WO types among different Wolbachia strains associated with butterflies. Most horizontal transfer events involved different Wolbachia supergroups (A and B). Horizontal acquisition of phage WO might also occur between eukaryotes without Wolbachia transfer. Furthermore, 22 putative recombination events were identified in 13 of 16 butterfly species which harbored multiple phage types. These results showed that horizontal transfer of Wolbachia caused it to be exposed to the phage gene pool, and that horizontal transmission of phage WO, as well as intragenic recombination were important dynamics for phage WO genome evolution, which effectively promoted the high level of phage WO diversity associated with butterflies.

Effects of PFOA on the physicochemical properties of anaerobic granular sludge: Performance evaluation, microbial community and metagenomic analysis.

The impact of perfluorooctanoic acid (PFOA) on the anaerobic granular sludge was evaluated through a sequential batch experiment. Results showed that PFOA inhibited the chemical oxygen demand (COD) removal rate of the sludge and the dosage of 100 mg/L PFOA was more obvious. However, this negative effect would gradually weaken with the adaptation of microorganisms. For the 50 mg/L PFOA experimental group, the proteins content in the extracellular polymeric substances (EPS) of the anaerobic granular sludge increased from 1.53 mg/g to 3.65 mg/g. Meanwhile, PFOA inhibited the 2-(p-iodophenyl)-3-(p-nitrophenyl)-5-phenyltetrazolium chloride (INT) activity of the anaerobic granular sludge. Furthermore, 100 mg/L PFOA reduced the relative abundance of Proteobacteria by 5.99% and Longilinea by 1.11%. 100 mg/L PFOA mainly restricted COD removal by affecting the glycolysis process, with the abundances of glucokinase and pyruvate kinase reduced by 8% and 28.1%, respectively. Compared with the control group, the relative abundance of the methyl-coenzyme M reductase alpha subunit increased by 84%, respectively, under 100 mg/L PFOA.