Effect of Six Bacterial Strains on the Activity of Second Instar Larvae and Egg Hatching of Meloidogyne incognita

[Objectives]The paper was to screen effective biocontrol strains against Meloidogyne incognita.[Methods]The effect of six bacterial strains sourced from the research group s strain library on the activity of second instar larvae of M.incognita,as well as on egg hatching,was evaluated.[Results]The treatment of fermentation supernatant derived from the X-2 strain exhibited a pronounced lethal effect on M.incognita,with a corrected mortality rate reaching 97%within 72 h.Additionally,this treatment significantly inhibited egg hatching,achieving an inhibition rate of 94.69%at a 20-fold dilution.The strain was identified as Bacillus velezensis,belonging to the genus Bacillus,and was designated as RKN1111.[Conclusions]This study presents alternative strains and a theoretical framework for the biological control of M.incognita.

Characterization of Congolese Strains of <i>Xanthomonas axonopodis</i>pv. <i>manihotis</i>Associated with Cassava Bacterial Blight

Cassava bacterial blight (CBB) caused by Xanthomonas axonopodis pv. manihotis has been reported in several African countries since 1970. Knowledge of the virulence and diversity of Xanthomonas axonopodis pv. manihotis strains is important for an integrated control of CBB. The main objective of the present study was to characterize strains of Xanthomonas axonopodis collected from various regions in the DR-Congo. There was variability among strains for shape (form), contour (margin) and elevation. Bacterial cell size for the strains analyzed varied from 24.1 μm × 11.3 μm to 11.4 μm × 4.2 μm. All the Xanthomonas axonopodis pv. manihotis strains but one was motile. Two distinctive groups were identified based on radial growth of their colonies. The first group grows faster (7.8-10.5 mm/d) compared to the second group (4.8-6.9 mm/d). Five strains (Gandajika, Inera/Stat, Kansasa, Mulumba and Musakatshi) were classified as virulent with a damage rating ≤ 1 and four were aggressive (Luputa, M'vuazi, Boketa and Kiyaka) with a damage rating > 1. Significant differences were also observed among strains for disease onset, incidence and plant mortality. The highest incidence (33%) of bacterial blight 21 days after infestation (DAI) resulted from the Boketa strain inoculation and the lowest (0 % disease incidence) from INERA/STAT and Musakatshi strains. There was no clear association between geographic origin of the strains and their aggressiveness.

Optimization of Hydrocarbons Biodegradation by Bacterial Strains Isolated from Wastewaters in Ouagadougou, Burkina Faso: Case Study of SAE 40/50 Used Oils and Diesel

Environmental pollution with petroleum and petrochemical products such as diesel and used oils has been recognized as one of the most serious current problem in the world, especially in developing countries. These petrochemical products devastate the soil, surface and underground waters and alter the microbial population at the polluted sites. Thus, the present work aims to optimize the biodegradation of diesel and two used oils (SAE 40 and SAE 50) by bacterial strains namely Acinetobacter S2 and Pseudomonas S7 using either nutrient factors (yeast extract, peptone or trace elements) or surfactants (tween 80 or Sodium Dodecyl Sulfate: SDS). The strains are incubated alone or together with the used oils or diesel supplemented or not with nutrient factors or surfactants for 14, 28, 42 and 56 days, respectively. For all the incubation period, the hydrocarbons degradation rates are determined by gravimetric assay. The results obtained show that nutrient factors increase significantly SAE 50 used oil biodegradation (p = 0.009). Similarly, tween 80 increases SAE 50 and SAE 40 used oils biodegradation but not diesel one. The results also show a significant difference between biodegradation rates at 14, 28, 42 and 56 days for all the hydro-carbons tested (p Acinetobacter S2 and Pseudomonas S7 increases the degradation over the one of the strains alone.

Evaluatien of Near-lsogenic Rice Lines with 8 Genes for Bacterial Blight Resistance to Strains in China

A set of near-isogenic rice lines withmonogenic resistance to bacterial blight weredeveloped by IRRI.The Cultivar IR24 wasused as the recurred parent.They wereevaluated with 6 races of Xanthomonascampestris pv.oryzae(Xco)in the Philip-pines at the maximum tillering and the bootingstages by ZHANG and MEW at IRRI in 1989.

Partial Characteristics of Hydrogen Production by Fermentative Hydrogen-producing Bacterial Strain B49

To investigate the characteristics of hydrogen production by a novel fermentative hydrogen-producing bacterial strain B49 (AF481148 in EMBL), batch experiments are conducted under different conditions. Hydrogen production has a correlation with cell growth and the consumption of glucose and soluble protein. The optimum pH for cell growth is 4.5±0.15. At acidic pH 4.0±0.15, the bacteria has the maximum accumulated hydrogen volume of 2382 ml/L culture and the maximum hydrogen evolution rate of 339.9 ml/L culture·h with 1% glucose. The optimum temperature for cell growth and hydrogen production is 35℃. In addition, fermentative hydrogen-producing bacterial strain B49 can generate hydrogen from the decomposition of other organic substrates such as wheat, soybean, corn, and potato. Moreover, it can also produce hydrogen from molasses wastewater and brewage wastewater, and hydrogen yields are 137.9 ml H 2/g COD and 49.9 ml H 2/g COD, respectively.

Screening, Identification and Biocontrol Effect of Antagonistic Streptomyces Strain Tra69 against Tobacco Bacterial Wilt

[Objective] The paper was to screen the biocontrol agents with good control effects against tobacco bacterial wilt. [Method] The strains with antagonistic activity against tobacco bacterial wilt were isolated from 36 copies of rhizosphere soil sample. The strain with best inhibition effect was identified, and carried out pot test and growth-promoting experiment. [Result] The strain with best inhibtion effect was Tra69, which was identified to be Streptomyces flavogriseus. The fermentation liquid of Tra69 had good control effect against Ralstonia solanacarum, and also had good growth-promoting effect on tobacco. [Conclusion] Tra69 was the biocontrol agent with excellent control effect against R. solanacarum, which could be further developed and used in biological control against tobacco diseases.

Magnesium improves hydrogen production by a novel fermentative hydrogen-producing bacterial strain B49

Batch experiments were conducted to investigate the effects of magnesium on glucose metabolism, including growth and hydrogen-producing capacity of fermentative hydrogen-producing bacterial strain B49. These abilities were enhanced with an increase in magnesium concentration. At the end of fermentation from （10 g/L） glucose, for 10 mg/L MgCl2·6H2O the cell growth in terms of optical density （OD） at 600nm was 0.46, the ratio of ethanol amount （mg/L） to acetate amount （mg/L） was 1.1, and the accumulated hydrogen volume was 934.9 mL H2/L culture; for 200 mg/L of MgCl2·6H2O OD600 nm was increased to 1.34. The accumulated hydrogen volume was increased to 2 360.5 mL H2/L culture, the ratio of ethanol amount （mg/L） to acetate amount （mg/L） was increased to 1.3 and polysaccharide was decreased to 2.5 mg/L. Moreover, the magnesium solution addition to the medium at different fermentation times affected hydrogen-producing ability. However, the later the addition time was postponed, the less the effect was on hydrogen evolution. Further experiments confirmed the enhancement was dependent on magnesium ions and not on the other inorganic ions such as SO42- or Cl-, which constituted the magnesium salts.

Occurrence of <i>N</i>-Acyl Homoserine Lactones in Extracts of Bacterial Strain of <i>Pseudomonas aeruginosa</i>and in Sputum Sample Evaluated by Gas Chromatography–Mass Spectrometry

This study presents a fast, accurate and sensitive technique using gas chromatography-mass spectrometry (GC-MS) for the identification and quantification of N-acyl homoserine lactones (AHLs) in the extracts of bacterial strain of Pseudomonas aeruginosa and sputum sample of a cystic fibrosis patient. This method involves direct separation and determination of AHLs by using GC-MS as simultaneous separation and characterization of AHLs were possible without any prior derivatiza-tion. Electron ionization resulted in a common fragmentation pattern with the most common fragment ion at m/z 143 and other minor peaks at 73, 57 and 43. The limit of detection for N-butanoyl, N-hexanoyl, N-octanoyl, N-decanoyl, N-dodecanoyl and N-tetradecanoyl homoserine lactones was 2.14, 3.59, 2.71, 2.10, 2.45 and 2.34 μg/L, respectively. The presence of AHLs in the culture of P. aeruginosa strain and spu-tum of a cystic fibrosis patient was achieved in selected ion monitoring (SIM) mode by using the prominent fragment at m/z 143.

Preliminary Characterization of a Cellulase Producing Bacterial Strain Isolated from a Romanian Hypersaline Lake

Cellulases are a group of enzymes that are used in many biotechnological processes. Since most of the enzymes synthesised by mesophilic microorganisms are unstable in industrial environments, it is necessary to direct research towards extremophile cellulolytic microorganisms because the enzymes synthesised by them are stable and active even in harsh physicochemical conditions. In the present investigation, our aim was to isolate and identify some microbial cellulolytic strains from a hypersaline lake located in Romania and to determine their optimal growth conditions. Of a total of 25 microbial strains isolated, only one extreme halotolerant bacterial strain was able to produce an endoglucanase. Based on molecular identification, we identified this cellulolytic strain as a species of Bacillus genus, most closely related to Bacillus zhangzhouensis. Optimal growth conditions were found to be at 15&deg;C, pH 7.5 and 2 M NaCl. Endoglucanase activity of this bacterial strain is influenced by both salinity and temperature. The most significant endoglucanase activity was detected in the presence of 3 M NaCl, after 72 h of incubation at 15&deg;C. In this situation, the amount of glucose released from a volume of 0.5 mL of 2% (w/v) carboxymethyl cellulose substrate is equivalent to 2.05 mg. In conclusion, this study represents the first preliminary characterization of a B. zhangzhouensis strain that has the ability to degrade cellulose and that demonstrates tolerance to high salt concentrations.

Antibacterial activity of Lawsonia inermis Linn(Henna) against Pseudomonas aeruginosa

Objective:To investigate the antibacterial activity of henna(Lawsonia inermis Linn) obtained from different regions of Oman against a wide array of micro-organisms.Methods:fresh henna samples were obtained from different regions of Oman as leaves and seeds,100 g fresh and dry leaves and SO g of fresh and dry seeds were separately soaked in 500 mL of ethanol for three days,respectively,with frequent agitation.The mixture was filtered,and the crude extract was collected.The crude extract was then heated,at 48 ℃ in a water bath to evaporate its liquid content.The dry crude henna extract was then tested for its antibacterial activity using well-diffusion antibiotic susceptibility technique.Henna extracts were investigated for their antibacterial activity at different concentrations against a wide array of different micro-organisms including a laboratory standard bacterial strain of Pseudomonas aeruginosa(NCTC 10662)(A aeruginosa) and eleven fresh clinical isolates of P.aeruginosa obtained from patients attending the Sultan Qaboos University Hospital(SQUH).2-Hydroxy-p-Nathoqinone-Tech(2-HPNT, MW=174.16,C_(10)H_40_3) was included as control(at 50%concentration) along with the henna samples tested.Results:Henna samples demonstrated antibacterial activity against all isolates but the highest susceptibility was against P.aeruginosa with henna samples obtained from Al-sharqyia region.Conclusions:Omani henna from Al-sharqyia region demonstrates high in vitro anti-P. aeruginosa activity compared with many henna samples from different regions of Oman.

Morphology controlled synthesis of ZnO nanoparticles for in-vitro evaluation of antibacterial activity

Novel hierarchical flower-and nanorod-shaped ZnO nanoparticles with uniform morphological features were successfully synthesized through controlled precipitation method in aqueous media without using any surfactant or template.To elucidate the growth mechanism of the synthesized nanoparticles,the effects of pH,reaction time and temperature were studied systematically.Selected ZnO samples were then subjected to SEM,FT-IR and XRD analysis.XRD patterns confirmed well crystalline nature of the as-synthesized powders.Furthermore,synthesized nanoparticles(hierarchical flowers as ZnO-1 and nanorods as ZnO-2),as well as commercial ZnO(ZnO-Com),were then investigated for in-vitro evaluation of antibacterial activity against various bacterial strains of clinical importance.Results showed that ZnO-2 exhibited higher antibacterial activity to all tested strains than ZnO-1,while ZnO-Com showed no antibacterial response in the applied experimental conditions.In addition,ZnO concentration-dependent antibacterial study unfolded that size of inhibition zones increased significantly from^30 to 33 mm against Streptococcus mutans and from^28 to 30 mm against Escherichia coli with increasing ZnO-2 concentration from 0.25 to 0.75μg/μL.The present study,therefore,suggests that the application of synthesized ZnO nanoparticles as the antibacterial agent may be effective for inhibiting certain pathogenic bacteria in biomedical sides.

Assessment of Expression Cassettes and Culture Media for Different Escherichia coli Strains to Produce Astaxanthin

Astaxanthin is a value-added ketocarotenoid with great potential in nutraceutical and pharmaceutical industries.Genetic engineering of heterologous hosts for astaxanthin production has attracted great attention.In this study,we assessed some key factors,including codon usage of the expressed genes,types of promoters,bacterial strains,and culture media,for engineered Escherichia coli to produce astaxanthin.The effect of codon usage was shown to be related to the types of promoters.E.coli DH5a was superior to other strains for astaxanthin production.Different culture media greatly affected the contents and yields of astaxanthin in engineered E.coli.When the expression cassette containing GadE promoter and its driving genes,HpCHY and CrBKT,was inserted into the plasmid pACCAR16DcrtX and expressed in E.coli DH5a,the engineered strain was able to produce 4.30±0.28 mg/g dry cell weight(DCW)or 24.16±2.03 mg/L of astaxanthin,which was a sevenfold or 40-fold increase over the initial production of 0.62±0.03 mg/g DCW or 0.61±0.05 mg/L.

Strain-specific effects of Akkermansia muciniphila on the regulation of intestinal barrier

Akkermansia muciniphila, one of the most promising next-generation probiotics, was reported to exhibit beneficial modulatory effects on the gut barrier. However, the strain-specific and underlying regulatory mechanisms of this species on gut barrier function were not well studied. Therefore, this study evaluated the protective effect of A. muciniphila strains on the intestinal barrier and investigated the mode of action and material basis of this modulatory effect. We first confirmed the strain-specific effects of A. muciniphila on intestinal barrier regulation and found that this phenomenon may be explained by the different abilities of strains to affect tight junction protein expression in enterocytes. Comparative genomic analysis proved that the ability of A. muciniphila to regulate the intestinal barrier was exerted in part by the functional genes(such as COG0438, COG0463, and COG2244)related to the synthesis of cellular surface proteins. The role of these surface proteins in intestinal barrier regulation was further verified by strain-comparative experiments in animal and cell models and surface protein removal trials. This study confirmed the different effects of A. muciniphila strains on gut barrier modulation and provided molecular and genetic targets for the screening of A. muciniphila strains with superior protection against gut barrier dysfunction.

新型产甲烷菌系提高极限含水油藏采收率技术

我国东部老油田已整体进入特高含水开发阶段,呈含水上升快、采油速度低、水驱效益低等开发特征,现有提高采收率技术已无法实现原油的经济采出,亟需建立接替技术。为此,以胜利油田某聚驱后油藏为试验区,开展了油藏菌群结构分析、新型产甲烷菌系的激活产气、油藏适应性及驱油性能研究,探索新型产甲烷菌系在这类油藏的应用潜力。研究结果显示,试验区油藏具有丰富的石油烃降解菌,有利于生物气化技术的实施。模拟试验区油藏条件下,新型产甲烷菌系与油藏内源微生物有较好的相容性,90 d每克原油的产气量达到3.12 mmol,是单独激活油藏微生物产气量的4.5倍,且产生的气体中甲烷气占比达到78%。菌群结构分析显示,新型产甲烷菌系占比达到35.9%,是产气速率大幅提升的关键。适应性研究结果显示,在油藏温度低于65℃、原油黏度小于1 356 mPa·s条件下,新型产甲烷菌系均展示了良好的产气性能。利用实验室设计的物理模型,评价了该菌系产气提高驱油性能,结果显示,注入该菌系后产气作用有效动用了模型顶部的剩余油,极限含水条件下驱油效率提高5.4个百分点;在此基础上提出了生物气化技术提高极限含水油藏采收率的机理。

Screening of Antagonistic Bacteria against Phytophthora infestans and Its Inhibition Effect

[ Objective ] The paper was to screen bacterial strain with significant antagonistic effect against Phytophthora infestans, so as to provide basis for further development and utilization of antagonistic bacteria to inhibit P. infestans and control potato late bright. [ Method] Plate dual culture and filter paper method were used to determine the inhibition effect of strains in vivo, fermentation broth and bacterial liquid of 61 strains against P. infestans and the resistance-induction effect of SR13-2 strain. [ Result] The inhibition rate of 24 strains among 61 tested strains against mycelial growth of P. infestans was greater than 60%, and the inhibi- tion effect of HT-6 strain was the strongest with the inhibition rate of 89.92%. However, fermentation broth of all tested strains had no significant inhibition effect against P. infestans, while the inhibition effect of bacterial liquid of most strains was significantly higher than strain in vivo; the inhibition effect of $34-1 strain was the strongest with inhibition rate of 91.50%. The bacterial liquid of SR13-2 strain was found to have significant resistance-induction effect with protective rate of 60%. [ Conclusion] The inhibition effect of strains in vivo and fermentation broth of antagonistic strains S34-1 and SR13-2 had no relationship with each other, while bacterial liquid had great application potential in controlling potato late bright.

菌酶协同发酵蛋白质饲料及在畜禽生产上应用的研究进展

菌酶协同发酵是一种利用微生物和外源酶共同作用于饲料的改良技术,可以降低饲料中的抗营养因子,促进氨基酸平衡和提高蛋白质利用率,可有效改善蛋白质品质。蛋白质饲料是畜牧业发展的重要基础,但目前我国的蛋白质饲料存在品质低、供应不足等问题。文章介绍了菌酶协同发酵技术应用在蛋白质饲料中的常用酶类与菌种,以及在畜禽生产中的应用进展,并指出菌酶协同发酵蛋白质饲料存在的不足和发展方向,以期为菌酶协同发酵蛋白质饲料的实际生产应用提供参考。

产单核细胞李氏杆菌Lm4b_02325/26双基因缺失株构建及部分生物学特性试验

旨在构建食源性产单核细胞李氏杆菌毒力岛4(LIPI-4)中的抗转录终止子(Lm4b_02325)和未知蛋白(Lm4b_02326)基因的双缺失株,研究Lm4b_02325/26基因对产单核细胞李氏杆菌(Listeria monocytogenes,LM)的部分生物学特性的影响。利用同源重组技术构建LM928ΔLm4b_02325/26双缺失株,测定LM928和LM928ΔLm4b_02325/26双缺失株在脑心浸出液肉汤(brain heart infusion broth, BHI)、不同EtOH浓度、不同NaCl浓度、不同pH、不同温度下的D_(600 nm),绘制生长曲线;RT-qPCR检测双基因缺失前后体外BHI培养环境下部分毒力因子转录水平;平板计数测定其对鼠巨噬细胞RAW264.7的黏附、侵袭与胞内增殖情况;感染C57BL/6小鼠后检测肝、脾、脑组织中的细菌数量。结果表明,成功构建具有遗传稳定性的双缺失株LM928ΔLm4b_02325/26。在含0.5%~10%NaCl或3%~4.5%EtOH的培养基中,双缺失株与野毒株生长没有明显差异,在pH=5条件下,双缺失株的生长率显著高于野毒株,pH=9条件下双缺失株的生长率显著低于野毒株。在不同温度(4℃、37℃、42℃)BHI培养基中,双缺失株与野毒株生长没有明显差异。Lm4b_02325/26基因双缺失株在BHI培养基中毒力因子hly、inlC和PlcA极显著上调(P<0.01),mpl、inlB、actA、inlP、PlcB、prfA、SigB、iap和inlA极显著下调(P<0.01);双缺失株对RAW264.7细胞的黏附率(14.86%)和侵袭率(2.23%)明显低于野毒株的黏附率(22.93%)和侵袭率(4.28%)(P<0.01);3、6、12 h时胞内增殖数量极显著低于野毒株(P<0.01);双缺失株与野毒株感染小鼠后,载菌量在肝、脾中没有显著差异,在脑组织中双缺失株载菌量为10~4,高于野毒株的10~(3.07),差异极显著(P<0.01)。综上认为,LIPI-4的Lm4b_02325和Lm4b_02326基因参与LM毒力因子的表达和脑组织的定植能力,与弱酸强碱条件下适应力及对RAW264.7细胞的黏附、侵袭和胞内增殖有关。本研究为LIPI-4的功能研究奠定了基础。

北京平谷区桃细菌性穿孔病病原菌鉴定及其拮抗菌筛选

为筛选对平谷区桃穿孔病有抑制作用的菌株,对桃穿孔病病原菌进行DNA测序鉴定后筛选拮抗菌。应用组织分离法从平谷区发病桃果实分离桃穿孔病原菌并进行致病性测定,根据菌株形态特征和16S rDNA序列相似率分析,初步确定病原菌为树生黄单胞菌桃李致病变种Xanthomomonas arboricola pv. pruni。采用平板对峙法筛选出病原菌的拮抗菌5株,其中菌株xj39抑菌效果最为显著,抑菌圈直径达到54.7 mm,菌株xj12次之,抑菌圈直径50.8 mm;经16S rDNA序列和rpoB基因序列比对及相似率分析,确定拮抗菌株xj39为大孢子链霉菌Streptomyces fimbriatus,菌株xj12为生二素链霉菌Streptomyces ambofaciens。田间防验结果表明,拮抗菌xj39发酵液对桃细菌性穿孔病防治效果为33%,与化学药剂春雷霉素处理无显著差异。

废弃烟叶中产细菌纤维素菌株的筛选鉴定及混菌发酵工艺优化

以废弃烟叶为来源,分离筛选产细菌纤维素菌株,对筛选菌株进行形态学观察、生理生化试验及分子生物学鉴定。以废弃烟叶浸提液为碳源和氮源,采用筛选菌株与酿酒酵母(Saccharomyces cerevisiae)以混菌(1∶1)发酵,考察废弃烟叶浸提液添加量、混菌接种量、pH、发酵温度及发酵时间对细菌纤维素产量的影响,并利用单因素、Plackett-Burman和Box-Benhnken试验优化混菌发酵工艺条件。结果表明,筛选得到一株产细菌纤维素菌株YC1,其被鉴定为氧化葡萄糖酸杆菌(Gluconobacter oxydans)。最佳混菌发酵工艺条件为:废弃烟叶浸提液添加量10%,菌株YC1与酿酒酵母(1∶1)混菌接种量1.0%,初始pH值5.5,发酵温度30℃,发酵时间6 d。在该优化条件下,细菌纤维素产量最大,为2.71 g/L,比菌株YC1单独发酵细菌纤维素产量(1.18 g/L)提高了1.29倍。

大曲中酵母菌组成及优势酵母的功能特性

酵母菌是酿酒大曲的主要功能菌之一,研究不同种类大曲的酵母菌群及优势酵母的发酵特性,有助于制曲工艺控制及酿酒功能菌的筛选与应用。以成熟高温大曲、中高温大曲和低温大曲为试材,对大曲中的酵母菌进行分离纯化和分子鉴定,初步探索优势酵母菌的耐温、耐酸、耐酒精及产酶特性。从三类大曲中各获得1株优势酵母菌,经鉴定分别为酿酒酵母(编号:GY1)和异常威克汉姆酵母种(编号:ZY1,DY1)。3株优势酵母菌均对酒精有一定的耐受能力,均具备产生果胶酶、淀粉酶、纤维素酶、蛋白酶及酯化酶的能力,可为酿酒产香功能酵母菌的筛选及生产应用提供菌株资源。