晋西北酸粥源短乳杆菌(Levilactobacillus brevis)M-10安全性评估

从晋西北酸粥发酵液中分离到一株短乳杆菌M-10,发现其具有耐酸、耐胆盐、抗氧化、抗菌、降胆固醇等多种益生特性。为了更好地应用该菌株,该研究评价了其体外和体内的安全性,包括溶血活性检测、明胶液化能力测定、硝基还原酶活性测定、吲哚和生物胺生成能力测定、SD大鼠急性毒性和亚急性毒性实验。经检测发现,短乳杆菌M-10缺乏溶血活性,不能液化明胶,缺乏硝基还原酶活性,不会生成吲哚,不能产生生物胺,包括酪胺、组胺、尸胺、腐胺等;该菌对SD大鼠的一般体征和血常规指标均无明显不良影响。以上研究结果表明,短乳杆菌M-10具有很好的安全性。该研究可为后续短乳杆菌M-10作为发酵剂或益生菌的开发奠定基础。

The protective effects of Levilactobacillus brevis FZU0713 on lipid metabolism and intestinal microbiota in hyperlipidemic rats

Levilactobacillus brevis FZU0713, a potential probiotic previously isolated from the traditional brewing process of Hongqu rice wine, may have the beneficial effects on improving lipid metabolism. This study aimed to evaluate the in vivo protective effects and possible mechanism of L. brevis FZU0713 on the disturbance of lipid metabolism in hyperlipidemic rats fed a high-fat diet(HFD). Results showed that oral administration of L. brevis FZU0713 could significantly inhibit obesity, ameliorate the lipid metabolism disorder, including serum/liver biochemical parameters and hepatic oxidative stress in HFD-fed rats. Histopathological result also indicated that dietary intervention of L. brevis FZU0713 could reduce the accumulation of lipid droplets in liver induced by 8 weeks HFD feeding. Furthermore, L. brevis FZU0713 intervention significantly increased the fecal levels of short-chain fatty acids(SCFAs, including acetate, propionate, butyrate, isobutyrate, valerate and isovalerate)in HFD-fed rats, which may be closely related to the changes of intestinal microbial composition and metabolic function. Intestinal microbiota profiling by 16S rRNA gene sequencing revealed that L. brevis FZU0713 intervention significantly altered the relative abundance of Coprococcus, Butyricicoccus, Intestinimonas, Lachnospiraceae FCS020 group, Ruminococcaceae_NK4A214 group, Ruminococcaceae_UCG-005 and UCG-014 at genus levels. Based on Spearman's rank correlation coefficient, serum and liver lipid metabolism related biochemical parameters were positively correlated with genera Ruminococcus, Pediococcus and Lachnospiraceae, but negatively correlated with genera Pseudoflavonifractor, Butyricicoccus and Intestinimonas. Furthermore, liver metabolomics analysis demonstrated that L. brevis FZU0713 had a significant regulatory effect on the composition of liver metabolites in hyperlipidemic rats, especially the levels of some important biomarkers involved in the metabolic pathways of arachidonic acid metabolism, primary bile acid biosynthesis, amino sugar and nucleotide sugar metabolism, taurine and hypotaurine metabolism, biosynthesis of unsaturated fatty acid, fructose and mannose metabolism, tyrosine metabolism, etc. Additionally, oral administration of L. brevis FZU0713 significantly regulated the mR NA levels of liver genes(including Acat2, Acox1, Hmgcr, Cd36, Srebp-1c and Cyp7a1)involved in lipid metabolism and bile acid homeostasis. In conclusion, our findings provide the evidence that L. brevis FZU0713 has the potential to improve disturbance of lipid metabolism by regulating intestinal microflora and liver metabonomic profile. Therefore, L. brevis FZU0713 may be used as a potential probiotic strain to produce functional food to prevent hyperlipidemia.

菌株LPF-1(Brevibacillus brevis LPF-1)降解猪粪过程中降解物质的GS-MS分析

利用气-质联用(GS-MS)分析猪粪污降解菌LPF-1(Brevibacillus brevis LPF-1)降解猪粪过程中挥发性物质的变化。以匹配度大于80(最大为100),含量大于5%为标准,在3个处理中共检测到各类物质23种,其中菌株LPF-1发酵液检测到的物质种类最多,有16种;猪粪+水溶液处理检测到12种,其中苯类物质3种(1,2-二甲苯、对二甲苯、间二甲苯),酚类物质4种(2,4-二叔丁基苯酚、苯酚、2,5-二叔丁基酚、4-甲基苯酚);猪粪+LPF-1发酵液处理检测到物质10种,其中苯类物质1种(对二甲苯),酚类物质3种(2,4-二叔丁基苯酚、苯酚、4-甲基苯酚),菌株LPF-1能有效减少猪粪降解过程中苯类和酚类等有害物质种类的产生,具有进一步开发成粪污降解微生物菌剂潜力。

Brevibacillus brevis B011次级代谢物中抗青枯菌活性物质的纯化鉴定及生物合成基因簇分析

青枯病是世界上最为严重的植物细菌性病害之一,生物防治是防控其危害的热点研究领域,而分泌抑菌活性物质被认为是生防菌抑制病原菌的主要作用机制。本研究在前期筛选到对青枯菌有良好拮抗活性的短短芽孢杆菌B011(Brevibacillus brevis B011)基础上,采用色谱法分离纯化到抗青枯菌的主要活性物质,通过串联质谱和核磁波谱分析方法鉴定了其结构,并根据B011菌株全基因组序列,预测了活性化合物生物合成相关基因及其合成途径。结果表明,B.brevis B011次生代谢物中抑制青枯菌的主效活性物质为伊短菌素A(edeine A),次效活性物质为N-乙酰基色胺[N-(2-(1H-indol-3-yl)ethyl)acetamide]。这两种化合物对青枯菌的抑制活性为首次报道。基因簇预测结果表明,B011菌株基因组中存在完整的edeine合成基因簇,共包括17个基因,即edeA~edeQ,且基因簇中的基因结构完整,基因序列高度保守。B011基因组中有多个N-乙酰基色胺的生物合成相关基因,包括17个脱羧酶编码基因和54个乙酰转移酶编码基因,可能分别参与色氨酸脱羧反应和色胺的乙酰基转移反应,但具体由哪些基因参与还需进一步的研究。研究结果可为该生防菌的应用提供理论指导,也可为后续解析其生物合成途径及通过合成生物学方法进行定向改造奠定基础。

亚硝酸盐影响Lactobacillus brevis 4903发酵的研究

通过研究可知,亚硝酸盐对Lactobacillusbrevis4903发酵有抑制作用,环境中亚硝酸盐一旦分解掉,这种抑制作用就会被解除。分析其原因:①亚硝酸盐抑制了乳酸菌生长,从而抑制了乳酸发酵;②在发酵初期可能因亚硝酸盐还原酶的作用,使亚硝酸盐酶解生成NH3,NH3中和了乳酸菌生成的酸(H+),从而使环境pH值的下降和酸的积累变得缓慢。

短乳杆菌(Lactobacillus brevis)去除亚硝酸盐的研究

短乳杆菌有较强去除亚硝酸盐能力 ,亚硝酸盐含量在 2 5 0mg L以内 ,接种短乳杆菌48h亚硝酸盐全部去除。短乳杆菌处理亚硝酸盐主要处于亚硝酸还原酶还原亚硝酸盐阶段。短乳杆菌去除亚硝酸盐的最适pH值为 5 0～ 6 0 ,最适温度为 3 0℃ ;在其它条件不变的情况下 ,发酵初期 (1 0～ 2 6h)亚硝酸盐去除量随接种量的增加而增加 ,最适接种量为5 %。亚硝酸盐含量在 2 0 0mg L以内 。

短短芽孢杆菌(Brevibacillus brevis)XDH菌株发酵培养基配方的研究

从泰山土壤中分离获得一株对多种动物病原细菌具有强烈拮抗作用的细菌—短短芽孢杆菌(Brevibacillus bre-vis)XDH。采用摇瓶培养法对发酵培养基配方进行了研究。通过单因素试验和均匀设计试验确定了XDH菌株发酵培养基最佳组成为:葡萄糖1.5%、黄豆饼粉3.0%、蛋白胨0.2%、NaC l 0.1%、(NH4)2SO40.5%、CaCO30.6%、MgSO40.6%。发酵液效价达到1456.8μg/mL,比基础培养基效价提高近10倍。

短短芽孢杆菌JK-2(Brevibacillus brevis)胞外物质抗香蕉枯萎病菌的稳定性

研究温度、光照、紫外辐射、蛋白酶K、pH对生防菌短短芽孢杆菌（Brevibacillus brevis）JK-2胞外物质抗香蕉枯萎病菌稳定性的影响,为香蕉枯萎病生防菌剂的研制提供理论依据。在30℃,170r/min的条件下,恒温振荡培养36h采样制备生防菌短短芽孢杆菌（Brevibacillus brevis）JK-2胞外物质,以香蕉枯萎病菌为指示菌,对胞外物质的抑菌特性进行研究。结果表明：该胞外物质具有一定的热稳定性,60~80℃温育1h保持88.5%以上活性,在100℃放置1h后无抑菌活性;在酸性pH3.0~5.0和碱性pH9.0~11.0条件下,抑菌活性保持在77.2%以上,对光照、蛋白酶K、紫外照射均不敏感。其抑菌作用的最适温度为30℃,最适pH为7.0。

Optimization of Solid-State Fermentation with Lactobacillus brevis and Aspergillus oryzae for Trypsin Inhibitor Degradation in Soybean Meal

The aim of the present study was to optimize trypsin inhibitor degradation in soybean meal by solid-state fermentation （SSF） with Lactobacillus brevis and Aspergillus oryzae, and to determine the effect of SSF on phytic acid, crude protein, crude fat, and amino acid profile. Response surface methodology （RSM） with Box-Behnken design was used to optimize SSF. The optimal conditions derived from RSM for L. brevis fermentation were： pH=5. 1; inoculum size=10%; duration=72 h; substrate to water ratio=1.5. The minimum content of trypsin inhibitors was 6.4 mg g^-1 dry matter. The optimal conditions derived from RSM for A. oryzae fermentation were： substrate to water ratio= 0.8 1; inoculum size=4%; duration=120 h. The minimum content of trypsin inhibitors was 1.6 mg g^-1 dry matter. Both L. brevis and A. oryzae decreased trypsin inhibitors dramatically （57.1 and 89.2% respectively）. L. brevis fermentation did not affect phytic acid （0.4%） and crude fat （5.2%） considerably, whereas A. oryzae fermentation degraded phytic acid （34.8%） and crude fat （22.0%） contents to a certain extent. Crude protein content was increased after both fermentation （6.4 and 12.9% for L. brevis and A. oryzae respectively）. Urease activity was reduced greatly （83.3 and 58.3% for L. brevis and A. oryzae respectively）. In conclusion, SSF with A. oryzae and L. brevis reduced trypsin inhibitor content and modified major macronutrients in soybean meal.

龙眼保鲜功能微生物FJAT-0809-GLX(Brevibacillus brevis)功能成分的分析

从保鲜功能微生物短短芽胞杆菌FJAT-0809-GLX(Brevibacillus brevis FJAT-0809-GLX)乙醇提取物功能成分中共鉴定出24种化合物,其中12种为匹配度在90%以上的高匹配度成分,相对含量较高的2种成分,分别为羟苯乙酯(24.28%)和邻苯二甲酸单(2-乙基己基)酯(19.78%),这2种物质成分相对含量总和占所有成分的44.06%,很有可能是其主要活性成分。保鲜功能微生物功能成分对大肠杆菌K88具有一定的抑菌效果,其中,羟苯乙酯的抑菌圈直径为17.81mm。同时对龙眼果皮过氧化物酶活性也具有一定的抑制效果。保鲜功能微生物功能成分按其功能可以划分为以下2种类型,一种为具有防腐杀菌(或抑菌)作用的物质,另一种为挥发性或者芳香类物质。

短短芽孢杆菌(Brevibacillus brevis)对鲤鱼养殖水质及其生长特性的影响

试验共设单因素4水平,分别对应水体中含0 mL、20 mL、60 mL、100 mL4种体积短短芽孢杆菌的试验组。通过测定养殖水体COD、NH_4^+-N、DO、pH等指标,鲤鱼生长性能及消化酶活性等指标,研究短短芽孢杆菌(Brevibacillus brevis)对鲤鱼养殖水体水质及鲤鱼生长特性的影响。结果显示,短短芽孢杆菌能有效降低养殖水体中COD和NH_4^+-N浓度(P<0.05),维持DO在较高水平(P<0.05),保持pH稳定,改善养殖水质。试验组中鲤鱼的肥满度、增重率及特定生长率与对照组相比均显著提高(P<0.05),鲤鱼肠道蛋白酶、淀粉酶和肝胰脏淀粉酶活性与对照组相比显著升高(P<0.05)。

Dawazisaurus Brevis, A New Eosauropterygian From the Middle Triassic of Yunnan, China

Dawazisaurus brevis（gen. et sp. nov.） is a newly discovered Triassic marine reptile, represented by a complete skeleton from Member II of the Guanling Formation of Luoping, Yunnan Province, China. This paper aims to（1） present a thorough description of the species,（2） make a detailed comparison to demonstrate if the species can be referred to any known sauropterygian taxa, and（3） conduct phylogenetic analyses to establish the internal relationships of the species with other sauropterygians. In addition, the discovery of Dawazisaurus provides a chance not only to test the phylogenetic patterns of the Sauropterygia obtained by previous studies but also to evaluate the previous hypotheses on the origin of the sauropterygian groups at different levels. D. brevis is an eosauropterygian, characterized by a unique combination of derived features such as a pair of large nasals joining in the formation of the internarial septum, a short trunk with 16 dorsal vertebrae; the zygapophyses of the trunk vertebrae very small or weakly developed; the posterior margin of the skull roof deeply V-shaped, and an ossified distal carpal 5. Phylogenetic analysis indicates that D. brevis appears to be more closely related to the Nothosauroidea than the Pistosauroidea within the Eosauropterygia.

Kinetic modeling of gamma-aminobutyric acid production by Lactobacillus brevis based on pH-dependent model and rolling correction

Gamma-aminobutyric acid(GABA)is a natural non-protein functio nal amino acid,which has potential for fermentation industrial production by Lactobacillus brevis.This work investigated the batch fermentation process and developed a kinetic model based on substrate restrictive model established by experimental data from L25(5~6)orthogonal experiments.In this study,the OD600 value of fermentation broth was fixed to constant after reaching its maximum because the microorganism death showed no effect on the enzyme activity of glutamate decarboxylase(GAD).As pH is one of the key parameters in fermentation process,a pH-dependent kinetic model based on radial basis function was developed to enhance the practicality of the model.Furthermore,as to decrease the deviations between the simulated curves and the experimental data,the rolling correction strategy with OD600 values that was measured in real-time was introduced into this work to modify the model.Finally,the accu racy of the rolling corrected and pH-dependent model was validated by good fitness between the simulated curves and data of the initial batch fermentation(pH 5.2).As a result,this pH-dependent kinetic model revealed that the optimal pH for biomass growth is 5.6-5.7 and for GABA production is about 5,respectively.Therefore,the developed model is practical and convenient for the instruction of GABA fermentation production,and it has instructive significance for the industrial scale.

Heat-Killed <i>Lactobacillus brevis</i>SBC8803 Induces Serotonin Release from Intestinal Cells

Previously, we reported that changes induced in autonomic neurotransmission in rats by Lactobacillus brevis SBC8803 may be mediated by serotonin 3 (5-HT3) receptors. In this study, we evaluated the effects of heat-killed L. brevis SBC8803 on serotonin (5-HT) releasing from intestinal cells. In the in vitro study, L. brevis SBC8803 stimulated 5-HT release from cultured rat endocrine RIN-14B cells (SBC8803 vs. sterile water;P in vivo study, 2 mg of heat-killed L. brevis SBC8803 was administered using a stomach sonde (feeding needle) to C57BL/6J mice. Analysis of plasma by ELISA showed gradually increase in 5-HT concentrations (0 min vs. 60 min;P ex vivo cultured intestinal loops composed of duodenum and part of the jejunum, from C3H/HeN and C57BL/6J male mice indicated that L. brevis SBC8803 effectively induced 5-HT release (SBC8803 vs. sterile water;P L. brevis SBC8803 may stimulate 5-HT release from mouse intestinal cells such as enterochromaffin cells.

Verification of Lactobacillus brevis tolerance to simulated gastric juice and the potential effects of postbiotic gamma-aminobutyric acid in streptozotocin-induced diabetic mice

The therapeutic effect of gamma-aminobutyric acid(GABA)on diabetes was spread as one of the alarming epidemics worldwide.The study aims to investigate the function of Lactobacillus brevis KLDS_(1.0727) and KLDS_(1.0373) strains as glutamic acid decarboxylase 65(GAD65)carriers capable of generating GABA by comparing in vitro free and freeze-dried models and GABA intervention in vivo.PCR amplification of gad and in vitro i.e.,(growth rate,viability at different pH,bile tolerance,and survivability in simulated gastric juice)were performed.In vivo experiments were conducted in 7 groups of C57BL/6J mice.Each group was injected with streptozotocin(Cont_(STZ),INSSTZ,LAC1_(STZ),LAC_(1MFDSTZ),LAC_(2STZ),LAC_(2MFDSTZ))daily except for the control(Cont).One group was injected with insulin(INSSTZ).The body weight and hyperglycemia in the blood were assessed weekly,post-euthanasia blood plasma parameters,insulin,and histological examination were evaluated.Results indicated L.brevis strains demonstrated a great tolerance to bile and simulated gastric juice in vitro(P<0.05).Cont_(STZ) had the highest average glucose level(6.84±6.46)mmol/L while INS_(STZ) expressed dramatically decreed in glucose level and displayed a significant decline in the average of weekly blood glucose(−5.74±3.08)mmol/L.The lowest body weight(ContSTZ)was(19.30±0.25)g.Based on the blood plasma analysis,L.brevis strains improved good cholesterol properties,liver and kidney functions,where most of these parameters fall within the average the reference range and prevent the development of symptoms of type 1 diabetes in vivo.As recommended,L.brevis should be commonly distributed as a postbiotic GABA in pharmaceutical and nutritional applications.

Bacillus brevis DX01 Labeling by Green Fluorescent Protein Gene and Its Colonization in Rice Seedling Roots

A constitutive expression vector pHY300-F1gfp was constructed to test the function of a promoter F1 subcloned from a rice epiphyte Bacillus brevis strain DX01. The DX01 cells harboring plasmid pHY300-F1gfp were detected to produce bright green fluorescence. Subsequently, the gfp-tagged B. brevis strain was released into the soil and its survival was investigated by PCR and the detection of green fluorescence. The spatial location of in situ gfp-tagged bacterial cells on the root surface of rice seedlings was visualized. All these results indicated that green fluorescent protein is an ideal molecular marker for detection of the activities of promoter F1, and it is also a reliable probe to monitor specific B. brevis bacteria in the environment.

Extensor pollicis brevis tendon can hyperextend thumb interphalangeal joint in absence of extensor pollicis longus:Case report and review of the literature

We are reporting a case of extensor pollicis longus tendon rupture which did not require tendon transfer owing to the ability of the intact extensor pollicis brevis(EPB) to fully hyperextend the thumb interphalangeal joint. The thumb metacarpophalangeal joint was also able to be fully actively extended by the EPB. Previous anatomical studies have demonstrated that the insertional anatomy of the EPB tendon is highly variable and sometimes inserts onto the extensor hood and distal phalanx, which is likely the mechanism by which our patient was able to fully extend the thumb interphalangeal joint. Despite the potential for the EPB to extend the IP joint of the thumb, virtually all previously reported cases of extensor pollicis longus(EPL) tendon rupture had deficits of thumb IP extension requiring tendon transfer. This case highlights the potential ability of the EPB tendon to completely substitute for the function of the EPL tendon in providing thumb IP joint extension.

Effect of <i>Lactobacillus brevis</i>SBC8803 on Gamma-Glutamyl Transferase in Japanese Habitual Drinkers: A Double-Blind, Placebo-Controlled Study

A randomized, double-blind, placebo-controlled clinical trial in Japanese habitual drinkers was conducted to evaluate the efficacy of Lactobacillus brevis SBC8803 to alleviate adverse effect of alcohol. Subjects who drank habitually and had moderately higher levels of gamma-glutamyl transferase (GGT) (50 - 100 IU/L) were enrolled. The levels of transaminases in these subjects were almost within normal levels (aspartate transaminase (AST) <30 IU/L and alanine transaminase (ALT) <40 IU/L). Either the capsules containing placebo (n = 23) or 130 mg (4.0 × 1010 colony-forming units) of live L. brevis SBC8803 (n = 22) per day were administered for the continuous eight weeks (56 days). During the period, the subjects both in test group and placebo groups have kept each drinking behavior as usual. Regarding lipid metabolism, triacylglycerol (TG) levels in the male test group significantly decreased at week 4 as compared with week 0. Biomarkers of hepatocytes-damage;AST and ALT levels showed no significant differences between the pla- cebo and test groups at both weeks 4 and 8. Oxidative stress marker;GGT at weeks 4 was significantly lower in the test group than that in the placebo group (p = 0.017), but not at weeks 8. However, taking a reduced rate of GGT at weeks 8 comparing with that at week 0, that in the test group showed larger value comparing with that in the placebo group. These data about TG and GGT suggest that, although efficacy of L. brevis SBC8803 is limited in this study, intake of the probiotic may alleviate alcoholic influence in lipid metabolism and oxidative stress.

Influence of Glyphaea brevis twig extract on nucleus, tight junctions and expression of inhibin-β, stem cell factor, and androgen binding protein in TM4 Sertoli cells

Objective: To examine the influence of Glyphaea (G.) brevis twig extract on the mitochondrial dehydrogenase activity, integrity of the tight junctions between adjacent cells, mitochondria, apoptosis, nucleus and expression of inhibin-β, stem cell factor, and androgen binding protein in TM4 Sertoli cells. Methods: TM4 cell line was used in this study as it exhibited properties similar to the Sertoli cells. TM4 Sertoli cells were exposed to G. brevis twig extract (0.1, 1.0, 10.0, 100.0, or 1000.0μg/mL) for 24, 48 and 72 h. Parameters studied included cell viability [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay], mitochondrial membrane potential (tetra methyl rhodamine ethyl ester dye), transepithelial electrical resistance, apoptosis (Annexin V Alexa Fluor?488/propidium iodide assay) and mRNA expression (quantitative reverse transcription polymerase chain reaction). Results: G. brevis twig extract had no cytotoxic impact on cell viability, thus, considerably increasing the activity of mitochondrial dehydrogenase enzyme after 24 and 72 h exposure. Transepithelial electrical resistance values revealed substantial (P<0.05) rise in treated groups, especially after 72 h of treatment. Moreover, there was a significant decrease in mitochondrial depolarization of TM4 Sertoli cells exposed to G. brevis twig extract when compared to controls. In addition, G. brevis twig extract significantly reduced necrosis and apoptosis of TM4 Sertoli cells when compared to control. Nevertheless, fluorescence microscopy revealed that the nuclei were egg-shaped and marked uniformly with consistent cell shape at the middle of the TM4 Sertoli cells. Significant stimulatory effects were observed on mRNA levels of inhibin-β, androgen binding protein and stem cell factor. Conclusions: G. brevis twig extract may increase the secretory roles of TM4 Sertoli cells, cells proliferation, as well as cell-cell tight junction integrity. Thus, G. brevis twig may enhance spermatogenesis.

Flavonoid Glycosides from the Leaves of Glyphaea brevis

A new flavonoid, acacetin-7-O-β-D-glucopyranosyl-(1 → 2)-β-D-glucuronopyranoside (1), together with four known flavonoids, apigenin-7-O-β-D-glucopyranosyl-(1 → 2)-β-D-glucuronopyranoside (2), acacetin (3), acacetin-7-O-β-D-glucuronide (4) and genkwanin-5-O-primveroside (5) were isolated from the leaves of Glyphaea brevis. Their structures were elucidated by spectroscopic techniques. The isolated compounds (1 - 5) were tested for their antioxidant activity using the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay. The tested compounds showed slight negative antioxidant activities against DPPH radicals.