Mesophilic and thermophilic anaerobic digestion of swine manure with sulfamethoxazole and norfloxacin:Dynamics of microbial communities and evolution of resistance genes

The role of norfloxacin(NOR)and sulfamethoxazole(SMX)in mesophilic and thermophilic anaerobic digestion(AD)of pig manure,with respect to methane production and variations in the microbial community and resistance genes,including antibiotic resistance genes(ARGs),class I integrase(intIl),and heavy metal resistance genes(MRGs),was investigated.The results indicated that NOR exerted little influence on the microbial community,whereas SMX negatively affected the acetoclastic methanogens.The abundance of two sulfonamide resistance genes(sul1 and sul2),three quinolone resistance genes(qnrS,parC,and aac(6′)-Ib-cr),and intI1 decreased by 2–3 orders of magnitude at the end of thermophilic AD.In contrast,mesophilic AD was generally ineffective in reducing the abundance of resistance genes.According to the results of redundancy analysis,the abundance of ARGs was affected primarily by microbial community dynamics(68.5%),rather than the selective pressure due to antibiotic addition(13.3%).Horizontal gene transfer(HGT)through intI1 contributed to 26.4%of the ARG variation.The archaeal community also influenced the changes in the resistance genes,and ARG reduction was significantly correlated with enhanced methane production.Thermophilic AD presented a higher methane production potential and greater reduction in resistance gene abundance.

Enhancing sensitivity of microbial fuel cell sensors for low concentration biodegradable organic matter detection:Regulation of substrate concentration,anode area and external resistance

The relatively low sensitivity is an important reason for restricting the microbial fuel cell(MFC)sensors'application in low concentration biodegradable organic matter(BOM)detection.The startup parameters,including substrate concentration,anode area and external resistance,were regulated to enhance the sensitivity of MFC sensors.The results demonstrated that both the substrate concentration and anode area were positively correlated with the sensitivity of MFC sensors,and an external resistance of 210Ωwas found to be optimal in terms of sensitivity of MFC sensors.Optimized MFC sensors had lower detection limit(1 mg/L)and higher sensitivity(Slope value of the linear regression curve was 1.02),which effectively overcome the limitation of low concentration BOM detection.The essential reason is that optimized MFC sensors had higher coulombic efficiency,which was beneficial to improve the sensitivity of MFC sensors.The main impact of the substrate concentration and anode area was to regulate the proportion between electrogens and nonelectrogens,biomass and living cells of the anode biofilm.The external resistance mainly affected the morphology structure and the proportion of living cells of the anode.This study demonstrated an effective way to improve the sensitivity of MFC sensors for low concentration BOM detection.

Antimicrobial Resistance Profile of Bacteria Isolated from Boreholes and Hand Dug Wells Water in Ngaoundere Municipality of Adamawa Region in Cameroon

Uncontrolled uses of antibiotics have led to rapid evolution of antibiotic-resistance bacteria and antibiotic resistance gene transfer, especially in a pool of aquatic system where resistance, intermediate and susceptible bacteria to some antibiotics strive together. Consequently, there is a transfer of resistance genes. In this study, bacteria of the Enterobacteriaceae family and some gram positive bacteria isolated from some boreholes and hand dug wells water of public use were tested on 19 antibiotics of different classes. This was achieved through a disk diffusion technique to determine the antimicrobial resistance profile of the said bacteria, microbial resistance index of the drugs used (and their ability to produce Beta-lactamase). These isolates were shown to demonstrate a very high resistance to the drugs used in the area. The resistance was highest in Escherichia coli 1 (73.68%) and lowest in Streptococcus pneumoniae (47.82%). These isolates also indicated very high levels of multi-drug resistance. The minimum resistance index was 0.47, indicating that bacteria isolates were of fecal origin. It is evident from the present study that multiple antibiotic-resistant bacteria can thrive in water as an environmental reservoir, and can therefore provide a route to multidrug-resistant pathogens to enter human and animal population.

Enzymatic pretreatment mitigates the dissemination of antibiotic resistance genes via regulating microbial populations and gene expressions during food waste fermentation

Food waste(FW)has been recognized as essential reservoir for resource recovery via anaerobic fermentation,which could also bring the potential risk of antibiotic resistance genes(ARGs)dissemination.Although the structural deficiency of FW could be stimulated by enzymatic pretreatment to enhance fermentation efficiency,the influences of enzymatic pretreatment on ARGs fate and microbial metabolic pathways involved in ARGs dissemination have rarely been reported.This work proved that enzymatic pretreatment could effectively decrease the total abundance of ARGs(reduced by 13.8%-24.5%)during long-term FW fermentation.It was found that enzymatic pretreatment significantly reduced the ARGs belonging to the efflux pump,which might be ascribed to its ability to increase membrane permeability.Furthermore,enzymatic pretreatment was in favor of reducing microbial diversity and various potential ARGs host(e.g.,Methanosarcina,Clostridium,Prevotella,Parabacteroides).Also,this pretreatment remarkably up-regulated the genetic expressions involved in ABC transporter(e.g.,eryF and mntA)and down-regulated the genetic expressions that participated in DNA replication,two-component systems(e.g.,uphA and cckA),and quorum sensing(e.g.,rpfF and lsrG),thereby decreasing ARGs transmission.This study would expand the insight of the influences of pretreatment method on ARGs fate during FW fermentation,and offer practical guidance on the sustainable management of FW.

A global perspective on microbial risk factors in effluents of wastewater treatment plants

Effective monitoring and management of microbial risk factors in wastewater treatment plants(WWTPs)effluents require a comprehensive investigation of these risks.A global survey on microbial risk factors in WWTP effluents could reveal important insights into their risk features.This study aims to explore the abundance and types of antibiotic resistance genes(ARGs),virulence factor genes(VFGs),the vector of ARG/VFG,and dominant pathogens in global WWTP effluents.We collected 113 metagenomes of WWTP effluents from the Sequence Read Archive of the National Center for Biotechnology Information and characterized the microbial risk factors.Our results showed that multidrug resistance was the dominant ARG type,while offensive virulence factors were the most abundant type of VFGs.The most dominant types of ARGs in the vector of plasmid and phage were both aminoglycoside resistance,which is concerning as aminoglycosides are often a last resort for treating multi-resistant infections.Acinetobacter baumannii was the most dominant pathogen,rather than Escherichia coli,and a weak negative correlation between Escherichia coli and two other dominant pathogens(Acinetobacter baumannii and Bacteroides uniformis)suggests that using Escherichia coli as a biological indicator for all pathogens in WWTP effluents may not be appropriate.The Getah virus was the most dominant virus found in global WWTP effluents.Our study presents a comprehensive global-scale investigation of microbial risk factors in WWTP effluents,providing valuable insights into the potential risks associated with WWTP effluents and contributing to the monitoring and control of these risks.

Helicobacter pylori acquisition of metronidazole resistance by natural transformation in vitro

AIM To study whether Helicobacter pylori is naturally transformable. METHODS Transformation was performed in BHI broth supplemented with horse serum and yeast extract. Genomic DNA extracted from a metronidazole resistant H. pylori strain was added to H. pylori broth culture. The mixture was incubated at microaerophilic atmosphere. The DNA treated cells were plated on blood agar containing 8mg/L metronidazole to select for transformants. Sterile distilled water was used as a negative DNA control. The DNA profiles of transformants were compared with that of their parent strains by randomly amplified polymorphic DNA (RAPD) fingerprinting. RESULTS Transformation of H. pylori with DNA from a metronidazole resistant strain as a marker was demonstrated. Out of the 12 strains of H. pylori tested, 9 (75%) strains were found to be transformable. The transformation frequencies ranged from 3 4×10 -6 to 2 4×10 -4 . By RAPD, DNA fingerprints of the transformants and their parent strains showed no change in DNA profiles though transformants were all resistant to metronidazole as compared with their metronidazole sensitive parent strains. CONCLUSION Helicobacter pylori is naturally transformable which might be one of the ways that H. pylori develops resistance to metronidazole.

Microbial diversity of soil bacteria in agricultural field contaminated with heavy metals

In this study we evaluated the bacterial diversity in a soil sample from a site next to a chemical industrial factory previously contaminated with heavy metals. Analysis of 16S rDNA sequences amplified from DNA directly extracted from the soil revealed 17 different bacterial types （genera and/or species）. They included Polyangium spp., Sphingomonas spp., Variovorax spp., Hafina spp., Clostridia, Acidobacteria, the enterics and some uncultured strains. Microbes able to tolerate high concentrations of cadmium （500μmol/L and above） were also isolated from the soil. These isolates included strains of Acinetobacter （strain CD06）, Enterobacter sp. （strains CD01, CD03, CD04 and CD08） （similar strains also identified in culture-independent approach） and a strain of Stenotrophomonas sp. The results indicated that the species identified from direct analysis of 16S rDNA of the soil can be quite different from those strains obtained from enrichment cultures and the microbial activities for heavy metal resistance might be more appropriately addressed by the actual isolates.

Seasonal variations of microbial community and antibiotic resistome in a suburb drinking water distribution system in a northern Chinese city

Antibiotic resistance genes(ARGs)are an emerging issue for drinkingwater safety.However,the seasonal variation of ARGs in drinking water distribution systems(DWDS)is still unclear.This work revealed the tempo-spatial changes of microbial community,ARGs,mobile genetic elements(MGEs)co-occurring with ARGs,ARG hosts in DWDS bulk water by means of metagenome assembly.The microbial community and antibiotic resistome varied with sampling season and site.Temperature,ammonia,chlorite and total plate count(TPC)drove the variations of microbial community structure.Moreover,environmental parameters(total organic carbon(TOC),chlorite,TPC and hardness)shifted antibiotic resistome.ARGs and MGEs co-occurring with ARGs showed higher relative abundance in summer and autumn,which might be attributed to detached pipe biofilm.In particular,ARG-bacitracin and plasmid were the predominant ARG and MGE,respectively.ARG hosts changed with season and site and were more diverse in summer and autumn.In winter and spring,Limnohabitans and Mycobacterium were the major ARG hosts as well as the dominant genera in microbial community.In addition,in summer and autumn,high relative abundance of Achromobacter and Stenotrophomonas were the hosts harboring many kinds of ARGs and MGEs at site in a residential zone(0.4 km from the water treatment plant).Compared with MGEs,microbial community had a greater contribution to the variation of antibiotic resistome.This work gives new insights into the dynamics of ARGs in full-scale DWDS and the underlying factors.

Detection of Helicobacter pylori resistance to clarithromycin and fluoroquinolones in Brazil:A national survey

AIM To evaluate bacterial resistance to clarithromycin and fluoroquinolones in Brazil using molecular methods.METHODS The primary antibiotic resistance rates of Helicobacter pylori(H. pylori) were determined from November 2012 to March 2015 in the Southern,South-Eastern,Northern,North-Eastern,and Central-Western regions of Brazil. Four hundred ninety H. pylori patients [66% female,mean age 43 years(range: 18-79)] who had never been previously treated for this infection were enrolled. All patients underwent gastroscopy with antrum and corpus biopsies and molecular testing using Geno Type Helico DR(Hain Life Science,Germany). This test was performed to detect the presence of H. pylori and to identify point mutations in the genes responsible for clarithromycin and fluoroquinolone resistance. The molecular procedure was divided into three steps: DNA extraction from the biopsies,multiplex amplification,and reverse hybridization. RESULTS Clarithromycin resistance was found in 83(16.9%) patients,and fluoroquinolone resistance was found in 66(13.5%) patients. There was no statistical difference in resistance to either clarithromycin or fluoroquinolones(P = 0.55 and P = 0.06,respectively) among the different regions of Brazil. Dual resistance to clarithromycin and fluoroquinolones was found in 4.3%(21/490) of patients. The A2147 G mutation was present in 90.4%(75/83),A2146 G in 16.9%(14/83) and A2146 C in 3.6%(3/83) of clarithromycin-resistant patients. In 10.8%(9/83) of clarithromycin-resistant samples,more than 01 mutation in the 23 S r RNA gene was noticed. In fluoroquinolone-resistant samples,37.9%(25/66) showed mutations not specified by the Geno Type Helico DR test. D91 N mutation was observed in 34.8%(23/66),D91 G in 18.1%(12/66),N87 K in 16.6%(11/66) and D91 Y in 13.6%(9/66) of cases. Among fluoroquinolone-resistant samples,37.9%(25/66) showed mutations not specified by the Geno Type Helico DR test. CONCLUSION The H. pylori clarithromycin resistance rate in Brazil is at the borderline(15%-20%) for applying the standard triple therapy. The fluoroquinolone resistance rate(13.5%) is equally concerning.

High level arsenic resistance in bacteria present in biooxidation tanks used to treat gold-bearing arsenopyrite concentrates:A review

在连续流动的、搅动的坦克过程使用到金忍受的毒砂集中的对待的微生物引起的协会变得在很多年适应了砷的高集中。主导的微生物， Acidithiobacillus caldus 和 Leptospirillum ferriphilum，被发现包含砷抵抗基因的二个集合。艺术基因的一个集合在所有是在场的不管他们是否是，种孤立高度砷抵抗。艺术基因的第二个集合在象 Tn21 一样 transposons 上是在场的并且在测试的所有紧张被发现那被使适应了砷的高集中。在在场的砷抵抗 transposons 在。caldus 和 L。ferriphilum 为他们密切相关，却足够地不同独立地被获得了而非从一个细菌被传递了给其它。transposons 在 Escherchia coli 是置换地活跃的并且被显示比测试这是可能的定位 chromosomally 的艺术基因授与砷抵抗的高水平。相同的 Transposons 包含砷抵抗基因或仔细与从 L 的 transposon 有关。ferriphilum，原来在南非发现，也在两 L 被发现。ferrooxidans 和 L。ferriphilum 从南美洲和欧洲孤立。一个 arsB 基因大美人在。caldus 被两个都表明了 chromosomal 艺术基因的能力授与砷抵抗的底层在的相应再结合生产在。caldus 和为猛烈异种的创造的一个基因系统的发展。

One cell at a time:droplet-based microbial cultivation,screening and sequencing

Microbes thrive and,in turn,influence the earth's environment,but most are poorly understood because of our limited capacity to reveal their natural diversity and function.Developing novel tools and effective strategies are critical to ease this dilemma and will help to understand their roles in ecology and human health.Recently,droplet microfluidics is emerging as a promising technology for microbial studies with value in microbial cultivating,screening,and sequencing.This review aims to provide an overview of droplet micro flu idics techniques for microbial research.First,some critical points or steps in the microfluidic system are introduced,such as droplet stabilization,manipulation,and detection.We then highlight the recent progress of droplet-based methods for microbiological applications,from high-throughput single-cell cultivation,screening to the targeted or whole-genome sequencing of single cells.

Detection of presumed genes encoding beta-lactamases by sequence based screening of metagenomes derived from Antarctic microbial mats

Analysis of environmental samples for bacterial antibiotic resistance genes may have different objectives and analysis strategies. In some cases, the purpose was to study diversity and evolution of genes that could be grouped within a mechanism of antibiotic resistance. Different protocols have been designed for detection and confirmation that a functional gene was found. In this study, we present a sequence-based screening of candidate genes encoding beta-lactamases in 14 metagenomes of Antarctic microbial mats. The samples were obtained from different sites, representing diverse biogeographic regions of maritime and continental Antarctica. A protocol was designed based on generation of Hidden Markov Models from the four beta-lactamase classes by Ambler classification, using sequences from the Comprehensive Antibiotic Resistance Database (CARD). The models were used as queries for metagenome analysis and recovered contigs were subsequently annotated using RAST. According to our analysis, 14 metagenomes analyzed contain A, B and C beta-lactamase genes. Class D genes, however, were identified in 11 metagenomes. The most abundant was class C (46.8%), followed by classes B (35.5%), A (14.2%) and D (3.5%). A considerable number of sequences formed clusters which included, in some cases, contigs from different metagenomes. These assemblies are clearly separated from reference clusters, previously identified using CARD beta-lactamase sequences. While bacterial antibiotic resistance is a major challenge of public health worldwide, our results suggest that environmental diversity of beta-lactamase genes is higher than that currently reported, although this should be complemented with gene function analysis.

In vitro antimicrobial and synergistic effect of essential oil from the red macroalgae Centroceras clavulatum (C. Agardh) Montagne with conventional antibiotics

Objective:To study the chemical profile,antimicrobial properties,and synergistic effect with known antibiotics of essential oil extracted from the marine red macroalgae Centroceras clavulatum(C.Agardh)Montagne,collected in Morocco.Methods:The chemical composition of the oil was analyzed by gas chromatography-mass spectrometry.The oil was evaluated for antibacterial(Pseudomonas aeruginosa,Escherichia coli,Bacillus subtilis,Micrococcus luteus,Staphylococcus aureus,and Klebsiella pneumoniae),and antifungal activity(Candida albicans,Candida glabrata,Candida krusei,and Candida parapsilosis),by the disc diffusion method.The minimum inhibitory and minimum microbicidal concentrations of the oil were determined,as well as the synergistic effects of its application combined with the antibiotics ciprofloxacin and fluconazole,by the checkerboard method.Results:Thirty molecules were identified in the essential oil,comprising 96.27%of the total oil composition.Monoterpenes such as carvacrol(36.06%)were the most abundant compounds,followed by caryophyllene(14.67%),endo-borneol(9.04%),pyroterebic acid(3.23%)and caryophyllene oxide(3.13%).The oil exhibited a moderate antimicrobial activity with inhibition zone diameters ranging from 9.0 to 15.0 mm.The minimum inhibitory concentration values varied between 0.9 and 14.7 mg/mL,and Bacillus subtilis and Escherichia coli were the more sensitive bacteria with 0.9 and 1.9 mg/mL,respectively.The minimum microbicidal concentration values ranged from 0.4 to 14.7 mg/mL.A significant synergic action was observed when the oil was applied in combination with ciprofloxacin and fluconazole,with fractional inhibitory concentration index values ranging from 0.31 to 0.50.Synergy was found in 80%of the combinations and a 2 to 16-fold reduction of antibiotics MIC was observed.Conclusions:Our findings suggest that the essential oil of Centroceras clavulatum should be further appraised for its potential use in the management of multi-drug resistant microorganisms,with the purpose to restore the activity of standard antimicrobial drugs.

微塑料和多种抗生素胁迫下土壤环境因子的响应特性

为了深入探究微塑料(microplastics,MPs)、抗生素胁迫下土壤环境因子产生的响应特性,以聚乙烯(polyethylene,PE)、四环素(tetracycline,TC)、环丙沙星(ciprofloxacin,CIP)为研究对象,通过单独、联合施用到土壤中4周后,开展了土壤的理化性质、酶活性、抗生素残留、抗生素抗性菌(antibiotics resistant bacteria,ARB)抗性、抗生素抗性基因(antibiotic resistance genes,ARGs)、微生物群落的多样性等方面研究.结果显示,施用MPs与抗生素的容重比对照组分别增大了12.3%、16.9%、 21.8%.有机质含量由39.96g·kg^(-1)变化为53.21g·kg^(-1),与对照组相比分别增大了9.16%、12.39%、14.09%、18.47%、32.03%、33.16%、36.04%.阳离子交换量由对照组的44.36 cmol·kg^(-1)显著变化为62.45 cmol·kg^(-1),与对照组相比分别增大了24.06%、30.09%、33.97%、36.49%、47.14%、50.93%、56.1%.实验各组pH值在7.58—8.12之间变化.实验各组的过氧化氢酶含量分别为1.653、 1.559、1.421、1.486、1.376、1.545、1.524、1.453 IU·g^(-1);脲酶含量分别为89.56、78.32、64.65、66.79、57.27、72.31、 71.26、 61.56IU·g^(-1);蔗糖酶含量分别为158.69、 149.61、 134.56、 131.87、 123.65、 137.26、136.83、 126.34 IU·g^(-1). MPs-TC-CIP组的过氧化氢酶活性、脲酶活性、蔗糖酶活性分别下降12.1%、32.3%、26.7%. MPs-TC、MPs-CIP组与TC、CIP实验组相比,抗生素残留量有所降低,分别为187.1%、189.3%;MPs-TC-CIP实验组的抗生素残留则低于单一施用CIP、TC组,分别为182.6%、178.7%.筛选的TC抗性菌(TC resistant bacteria,TCRB)和CIP抗性菌(CIP resistant bacteria,CIPRB)的抗性增加2倍以上. MPs-TC-CIP组与对照组相比,tet W、tet O的相对丰度比值分别为1.82、1.78;qnr A、qnr S的相对丰度比值分别为1.68、1.71.各组中相对丰度较高菌群依次为变形菌(Proteobacteria),放线菌(Actinobacteria)、酸杆菌(Acidobacteria)、芽单胞菌(Gemmatimonadetes)、厚壁菌(Fimicutes).施用抗生素、MPs后Proteobacteria、Actinobacteria相对丰度增加、myxococcus相对丰度减少.研究结果表明,施用MPs能促进抗生素的富集;在共同胁迫下,对土壤理化特性和生物特性影响更为显著,进一步探明了MPs对抗生素残留、ARGs传播、微生物群落演化特征等潜在影响,以期为消减污染物的危害、构建综合智能修复体系提供依据.

广州市“十二五”与“十三五”期间利福平耐药肺结核患者发现与治疗情况分析

目的:分析广州市“十二五”与“十三五”结核病防治规划期间利福平耐药肺结核(RR-PTB)患者发现与治疗情况,为进一步制定本地区RR-PTB防治规划提供科学依据。方法:通过“中国疾病预防控制信息系统”子系统“结核病信息管理系统”,按照登记时间导出2011年1月1日至2020年12月31日,即“十二五”(2011—2015年)和“十三五”(2016—2020年)规划期间广州市登记的肺结核患者耐药病案数据(包括性别、年龄、民族、职业、户籍、耐药类型、治疗分类等相关信息),筛选出利福平耐药患者病案,分析患者登记、人群特征、耐药筛查和治疗转归情况。结果:2011—2020年,RR-PTB患者年均登记率为0.71/10万(1152/16286.08万),从2011年的0.31/10万(42/1346.32万)上升至2015年的0.38/10万(60/1594.95万)和2020年的0.97/10万(182/1874.03万),呈逐年上升趋势(χ_(趋势)^(2)=256.395,P<0.001)。其中,“十二五”期间年均登记率为0.34/10万(250/7358.06万),不同年份登记率的差异无统计学意义(χ_(趋势)^(2)=4.674,P=0.322);“十三五”期间年均登记率为1.01/10万(902/8928.02万),不同年份登记率的差异有统计学意义(χ_(趋势)^(2)=38.439,P<0.001)。1152例患者中,以男性(851例,73.87%)、25~34岁青壮年(257例,22.31%)和家政家务及待业(364例,31.60%)为主;流动人口、初治、RR-PTB(除异烟肼耐药)、广泛耐药肺结核比例分别从“十二五”的8.80%(22/250)、11.20%(28/250)、0.00%(0/250)和0.00%(0/250)上升到“十三五”的54.43%(491/902)、37.14%(335/902)、19.84%(179/902)和0.78%(7/902),差异均有统计学意义(χ^(2)=91.370、298.740、97.915、34.096,P值均<0.001)。广州市耐药肺结核高危人群筛查率由2017年的60.91%(148/243)上升至2020年的98.95%(568/574),新发/初治病原学阳性肺结核耐药应筛查率由2018年的83.93%(1410/1680)提高到2020年的94.99%(3222/3392),差异均有统计学意义(χ_(趋势)^(2)=425.043、269.670,P值均<0.001)。纳入治疗、完成治疗和治疗成功的患者比例分别从“十二五”的81.20%(203/250)、2.46%(5/203)和45.81%(93/203)提高到“十三五”的91.02%(821/902)、33.62%(276/821)和67.48%(554/821),治疗失败患者比例从17.73%(36/203)降低至2.68%(22/821),差异均有统计学意义(χ^(2)=19.112、86.809、46.636、58.572,P值均<0.001)。结论:在“十二五”与“十三五”规划期间,广州市RR-PTB的防治工作取得了显著的成效。下一步工作中需继续坚持政府主导、多部门合作和全社会共同参与的原则,切实落实结核病防治规划要求,加强结核病防治服务体系建设。

Evaluation of rational use of antimicrobial agents in a Brazilian intensive care unit

The present study sought to assess the rational use of antimicrobial agents in a Brazilian intensive care unit (ICU) and its association with antimicrobial resistance in elderly patients admitted to the unit. Results: Choice of empiric and sensitivity-guided therapy was inadequate in > 80% and 59% of cases respectively. Inadequate antimicrobial therapy, whether empiric or sensitivity-guided, was positively correlated with bacterial resistance (r = 0.316;p = 0.001). Sensitivity testing revealed a 46.5% resistance rate to eight out of the ten most commonly used antibiotics. Multiple drug-resistant organisms were found in 40.8% of patients. A significant increase was observed in the number of multidrug-resistant samples between 2006/2007 and 2008 (r = 0.41, p = 0.006), (r = 0.598, p = 0.001), (r = 0.688, p = 0.00). Conclusion: We found a high rate of antibiotic misuse in the study sample. Inadequate therapy was correlated with resistance to antimicrobial agents.

抗枯萎病香蕉品种宿根连作根际土壤微生物群落结构特征变化分析

【目的】分析抗枯萎病香蕉品种宿根连作根际土壤微生物群落结构特征变化,为深入研究抗枯萎病香蕉品种宿根连作抗性增强的作用机制及利用生防微生物进行香蕉枯萎病绿色防控提供理论依据。【方法】以抗枯萎病香蕉品种宝岛蕉为试验材料,在枯萎病发病严重土壤上宿根连续种植3造:组培苗第一代(第1造)、宿根第一代(第2造)和宿根第二代(第3造),调查各造枯萎病发病率,四分法采集各造土壤样本并测定土壤化学性质;采集第1造健康和发病植株、第2和第3造健康植株的根际土壤样本,分别编号为S1、V、S2和S3,重复3次,利用Illumina高通量测序平台对12个土壤样本的细菌16Sr RNA和真菌ITS区进行测序分析;采用样本复杂度(Alpha多样性指数)、主坐标(PCoA)和UPGMA聚类分析土壤样本细菌和真菌群落的丰富度和多样性、门和属水平上的群落构成及相对丰度差异;通过Spearman分析优势微生物群落与镰刀菌属的相关性。【结果】抗枯萎病香蕉品种宝岛蕉连续种植3造根际土壤化学性质未发生明显改变,种植第3造的香蕉枯萎病发病率显著低于第1和第2造(P<0.05)。随着种植年限的增加,根际土壤细菌群落的多样性整体呈逐年下降趋势,而真菌群落的多样性整体呈逐年上升趋势。连作改变了香蕉根际土壤微生物的群落构成,S2和S3细菌和真菌的群落结构相近,明显与S1和V群落结构分离。S2和S3的壶菌门(Chytridiomycota)的相对丰度较S1分别低88.02%和89.51%,Fungi_phy_Incertae_sedis的相对丰度分别高41.56%和82.81%,S3镰刀菌属(Fusarium)的相对丰度较S1、S2分别低45.60%和50.47%,与留芽种植宿根蕉抗性表现增强相关。发病植株根际土壤中变形菌门(Proteobacteria)和担子菌门(Basidiomycota)的相对丰度较高,而酸杆菌门(Acidobacteriota)的相对丰度较低。球囊菌门(Glomeromycota)、毛霉门(Mucoromycota)、隐真菌门(Rozellomycota)和壶菌门(Chytridiomycota)的相对丰度均与镰刀菌呈负相关。【结论】宿根第3造蕉的发病率最低,其根际土壤与第1造发病植株根际土壤的细菌和真菌群落差异明显,推测根际土壤微生物群落丰富度、多样性和群落构成改变是抗枯萎病香蕉品种宿根连作抗性增强的原因。

施肥对我国黑土农田土壤微生物群落多样性影响的研究及展望

东北黑土是珍贵的土壤资源,由于长期的重用轻养,农田黑土退化问题已是一个不争的事实。施肥是保持/提高黑土农田生产力、保障作物产量的一项重要的农艺措施。本文综述了长期施用有机肥、无机肥、有机无机肥配施,以及无机肥氮(N)、磷(P)、钾(K)及其不同的组合施肥对黑土农田土壤细菌、真菌、固氮菌、反硝化细菌等微生物的数量、群落alpha多样性和beta多样性等方面影响的最新研究进展。发现研究结果因试验地点、季节气候、试验处理、目标微生物种类、甚至研究者的采样年限而不同。多数研究发现,长期施用化肥,特别是氮肥显著地降低了细菌丰度和alpha多样性指数,但对真菌丰度和alpha多样性指数影响不显著。不同施肥明显改变了黑土微生物beta多样性,但这种改变是有限的,相同施肥处理没有驱动不同试验地点土壤微生物群落结构同质化演替,表明东北黑土微生物分布具有很强的地域性。对长期施用不同的无机肥研究发现,N素较P素和K素在驱动黑土农田土壤微生物群落变化上作用更为明显。在论述长期施用有机肥改变土壤微生物群落结构的同时,也阐述了有机肥施用带来潜在的抗生素抗性基因污染的生态风险及对抗性基因的阻控措施。文章最后总结了目前施肥影响黑土农田微生物研究中存在的不足之处,并提出5点未来应加强的研究建议。