Bacterial community dynamics involved in Reactive Orange M2R dye degradation using a real time quantitative PCR and scale up studies using sequence batch reactor

Bacterial consortium volatile suspended soil (VSS) (Vatva soil sample) with a capability of azo dye Reactive Orange M2R (ROM2R) decolorization and degradation (shown in our earlier studies using Fourier transform infrared spectroscopy (FTIR) and phytotoxicity studies) was isolated from industrial wastewaters by enrichment culture technique. The present study was carried out to study bacterial population dynamics in consortium Vatva soil sample (VSS) during azo dye ROM2R degradation and to identify the consortium members that were actively involved in the degradation process. To achieve this goal, a real-time Polymerase chain reaction (PCR) assay targeting species-specific region of 16S rDNA of each consortial bacteria was developed to provide quantitative information about the bacterial abundance during azo dye degradation. The real-time PCR assay indicated that Pseudomonas aeruginosa (VSS-6) dominated consortium bacterial community during the active continuous bioremediation process. Attempt has been made to scale up from 100 ml volume to 10 L operation volume with intermittent additions (batch fed loadings) in a Sequence batch reactor (SBR). The development of VSS consortium biomass (MLVSS), changes in COD and biochemical oxygen demand, and the dye degradation were studied under conditions simulating the operations of biological effluent treatment in an attempt to develop a commercially applicable dye effluent treatment process unit.     

呼吸机相关性肺炎的痰培养细菌分布和细菌耐药性分析

目的:探讨呼吸机相关性肺炎痰培养的泛耐药鲍曼不动杆菌的分布情况,分析泛耐药鲍曼不动杆菌的耐药性与危险因素。方法:选择2016年1月到2016年12月在我院进行机械通气的患者3826例作为研究对象,其中发生泛耐药鲍曼不动杆菌呼吸机相关性肺炎98例为观察组,同期按照2:1的比例选择非泛耐药鲍曼不动杆菌呼吸机相关性肺炎患者49例作为对照组,两组都进行细菌耐药性分析,同时调查患者的临床资料,分析呼吸机相关性肺炎患者中泛耐药鲍曼不动杆菌感染发生的危险因素。结果:观察组对于头孢噻肟、哌拉西林、亚胺培南、庆大霉素、左氧氟沙星、四环素、头孢他啶都高度耐药;除哌拉西林、多粘菌素外,对照组对抗菌药物的耐药率均明显低于观察组(P0.05)。单因素分析结果显示合并糖尿病、机械通气时间、APACHEⅡ评分、GCS评分、深静脉置管等因素与呼吸机相关性肺炎患者中泛耐药鲍曼不动杆菌感染显著相关(P0.05);非条件Logistic回归分析结果显示糖尿病、机械通气时间、APACHEⅡ评分、GCS评分导致呼吸机相关性肺炎患者中泛耐药鲍曼不动杆菌感染的独立危险因素(P0.05)。结论:呼吸机相关性肺炎患者中泛耐药鲍曼不动杆菌感染比较常见,对绝大多数抗菌药物有耐药性,糖尿病、机械通气时间、APACHEⅡ评分、GCS评分为主要的危险因素。     

脑脊液检查在早产儿及足月儿细菌性脑膜炎的诊断价值

目的:评价脑脊液检查在早产儿及足月儿细菌性脑膜炎诊断中的价值。方法:选取2014年6月1日至2016年12月31日上海市儿童医院新生儿科收治的行腰椎穿刺检查的447例新生儿,回顾性分析新生儿的一般资料、脑脊液常规生化、培养等指标,根据胎龄将患儿分为早产儿167例与足月儿280例,再根据有无患发细菌性脑膜炎分为早产儿细菌性脑膜炎27例(早产儿观察组)、早产儿非细菌性脑膜炎140例(早产儿对照组)、足月儿细菌性脑膜炎38例(足月儿观察组)、足月儿非细菌性脑膜炎242例(足月儿对照组),采用受试者工作特征(ROC)曲线评估蛋白定量、白细胞计数、葡萄糖对早产儿及足月儿细菌性脑膜炎的诊断价值。结果:与同组对照组相比,足月儿观察组和早产儿观察组蛋白定量和白细胞计数均明显升高,而葡萄糖含量显著下降,且差异均具有统计学意义(P0.05);本研究65例细菌性脑膜炎患儿脑脊液培养分离出11株细菌(16.9%)。足月儿脑脊液白细胞计数、蛋白定量以及葡萄糖诊断细菌性脑膜炎的ROC曲线下面积分别为0.995、0.846、0.703。早产儿脑脊液白细胞计数、蛋白定量以及葡萄糖诊断细菌性脑膜炎ROC曲线下面积分别为0.970、0.711、0.705。结论:脑脊液白细胞计数、蛋白定量在足月儿和早产儿细菌性脑膜炎中具有较高的诊断价值。     

Up-regulated RxLR effector genes of Plasmopara viticola in synchronized host-free stages and infected leaves of hosts with different susceptibility

Fast recognition of host signals and early activation of infection mechanisms in Plasmopara viticola are decisive for successful infestation of Vitis vinifera. To better understand interactive processes at the first front line of combat between the pathogen and its host, a specific pre-infective stage was generated in a host-free system. Zoospore encystment was triggered within minutes after treatment with CaCl₂. Subsequently, high rates of germ tube formation occurred in a synchronized manner. This method was employed to compare development-related gene expression in strains of different virulence. Soon after germination, spores showed strong up-regulation of two effector genes, PvRxLR18 and PvRxLR28, particularly in the high virulence strain. On infected grapevine leaf-discs of cultivars with different susceptibility, a similar up-regulation was found at 6 hours post inoculation (hpi). This effect was much more evident in the high virulence than in the low virulence strain and was significantly higher on leaves of the tolerant cultivar Regent than on Müller-Thurgau. In addition, PvRxLR67 was up-regulated 24 hpi in the high virulence strain indicating that different effectors are active in later infection stages. Differences in the expression pattern of RxLR effector genes between the two strains corroborated with infection symptoms visible by sporulation.     

A Sub-population of Group A Streptococcus Elicits a Population-wide Production of Bacteriocins to Establish Dominance in the Host

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Graphene oxide/silver nanohybrid: Optimization,antibacterial activity and its impregnation on bacterial cellulose as a potential wound dressing based on GO‐Ag nanocomposite‐coated BC

Recently, bacterial cellulose (BC) based wound dressing have raised significant interests in medical fields. However, to our best knowledge, it is apparent that the BC itself has no antibacterial activity. In this study, we optimized graphene oxide‐silver (GO‐Ag) nanohybrid synthesis using Response Surface Methodology and impregnate it to BC and carefully investigate their antibacterial activities against both the Gram‐negative bacteria Escherichia coli and the Gram‐positive bacteria Staphylococcus aureus. We discover that, compared to silver nanoparticles, GO‐Ag nanohybrid with an optimal GO suspension's pH and ratio is much more effective and shows synergistically enhanced, strong antibacterial activities at rather low dose. The GO‐Ag nanohybrid is more toxic to E. coli than that to S. aureus. The antibacterial and mechanical properties of BC/GO‐Ag composite are further investigated.     

Pseudomonas aeruginosa: A risk factor for fertility in male mice

The present study was designed to evaluate the effect of P. aeruginosa on reproductive potential of male mice via a series of in vitro and in vivo experiments. In vitro studies involved sperm parameters, Mg²⁺ATPase activity and acrosome status. In vivo study employed male mice which in the right vas deferens received 20?μl of either PBS (Group I) or 10⁴ cfu of P. aeruginosa (Group II). The animals were sacrificed on day 3, 7 and 14 and various parameters viz. body weight, TSI (%), bacterial load, spermiogram {i.e. sperm count, motility (%), viability (%) and morphology}, lipid peroxidation and tissue histopathology were evaluated. The results revealed that cell free supernatant of P. aeruginosa resulted in reduced motility, viability, Mg²⁺dependent ATPase activity and premature acrosomal loss of mouse spermatozoa in vitro. In vivo study showed that in comparison to group I, group II revealed significant alterations in all the parameters on all the days of sacrifice. Further, when reproductive organs of right and left side of mice in group II were compared, the right side demonstrated more devastating effects in terms of altered TSI (%) of testis and cauda epididymis, higher bacterial counts, azoospermia, increased malondialdehyde levels and severe inflammation in tissue histopathology in comparison to left side where bacteria disseminated in reduced numbers, thereby, resulting in insignificant changes in TSI (%), spermiogram, malondialdehyde levels and tissue histology. This study demonstrates that the colonization of P. aeruginosa in male genital tract could be a risk factor for fertility.     

Solution structure analysis of the periplasmic region of bacterial flagellar motor stators by small angle X-ray scattering

The bacterial flagellar motor drives the rotation of helical flagellar filaments to propel bacteria through viscous media. It consists of a dynamic population of mechanosensitive stators that are embedded in the inner membrane and activate in response to external load. This entails assembly around the rotor, anchoring to the peptidoglycan layer to counteract torque from the rotor and opening of a cation channel to facilitate an influx of cations, which is converted into mechanical rotation. Stator complexes are comprised of four copies of an integral membrane A subunit and two copies of a B subunit. Each B subunit includes a C-terminal OmpA-like peptidoglycan-binding (PGB) domain. This is thought to be linked to a single N-terminal transmembrane helix by a long unstructured peptide, which allows the PGB domain to bind to the peptidoglycan layer during stator anchoring. The high-resolution crystal structures of flagellar motor PGB domains from Salmonella enterica (MotB_C2) and Vibrio alginolyticus (PomB_C5) have previously been elucidated. Here, we use small-angle X-ray scattering (SAXS). We show that unlike MotB_C2, the dimeric conformation of the PomB_C5 in solution differs to its crystal structure, and explore the functional relevance by characterising gain-of-function mutants as well as wild-type constructs of various lengths. These provide new insight into the conformational diversity of flagellar motor PGB domains and experimental verification of their overall topology.     

Roles of bacteria in the bark beetle holobiont – how do they shape this forest pest?

Bark beetles are well‐known forest pests, some species inducing massive attacks on trees, resulting in the devastation of entire woodlands. Bark beetles are associated with microorganisms, forming an entity known as ‘holobiont’. Beetles and fungi are the best‐studied members of this multipartite symbiosis. However, recent studies have shown that bacteria may play important roles in the bark beetle holobiont, such as providing certain nutrients, promoting the growth of beneficial fungi, detoxifying the environment by lowering the levels of phenolic compounds synthesised by the host tree or by inhibiting the growth of antagonistic fungi whereas some bacterial symbionts have the potential to kill beetles under certain conditions. Therefore, bacteria probably greatly affect the life cycle of bark beetles; hence, more research is needed to clarify the extent to which a bacterial associate is implicated in a bacterial bark beetle symbiosis and how much it determines host's performance. This review summarises all of the known activities of bacteria present in the bark beetle holobiont, indicates some important gaps in the knowledge of this symbiosis and provides some guidance for overcoming the difficulties in investigating this relationship in future studies.     

Paracoccidioides restrepiensis B339 (PS3) and P. lutzii LDR2 yeast cells and soluble components display in vitro hemolytic and hemagglutinating activities on human erythrocytes

Paracoccidioidomycosis (PCM) is a systemic mycosis caused by thermally dimorphic fungi of the Paracoccidioides species complex. Several pathogenic fungi produce hemagglutinins and hemolysins, which are virulence factors involved in adhesion of pathogens to host tissues or cells and in destruction of erythrocytes. The present research investigated hemolytic and hemagglutinating activities of yeast cells and soluble components from P. restrepiensis (PS3; formerly P. brasiliensis B339) and P. lutzii (LDR2). Different concentrations of live and heat‐killed yeast cells and soluble components from a cell free antigen preparation (native or heated, 56°C or 100°C, 30 min) were mixed with 1% human erythrocyte suspensions. Yeast cells from both species caused hemolysis, P. lutzii LDR2 being more strongly hemolytic than P. restrepiensis B339, whereas the opposite phenomena occurred with soluble components in most conditions. Live or heat‐killed yeast cells of both fungi agglutinated erythrocytes, but only heated soluble components from P. restrepiensis B339 showed hemagglutinating activity. In conclusion, yeast cells of P. restrepiensis B339 and P. lutzii LDR2 produce hemolysins and hemagglutinins, which are most likely predominantly restricted to yeast cells in P. lutzii LDR2 and predominantly released in soluble form by P. restrepiensis B339, requiring further study.