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Phytochemical analysis and antibacterial activities of Eleutherine bulbosa(Mill.) Urb. extract against vibrio parahaemolyticus

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Asian Pacific Journal of Tropical Biomedicine 2019年第9期9卷 397-404页

作者： Waode Munaeni Widanarni Munti Yuhana Mia Setiawati Aris T.Wahyudi Department of Aquaculture Faculty of Fisheries and Marine ScienceIPB University(Bogor Agricultural University)Bogor 16680Indonesia Department of Aquaculture Faculty of Fisheries and Marine ScienceHalu Oleo UniversityKendari 93232Indonesia Department of Biology Faculty of Mathematics and Natural ScienceIPB University(Bogor Agricultural University)Bogor 16680Indonesia

Objective: To analyze compounds in Eleutherine bulbosa(E. bulbosa)(Mill.) Urb. extract and to determine its antibacterial capability against vibrio parahaemolyticus(V. parahaemolyticus).Methods: E. bulbosa bulb extract was preprared using 96% ethanol by the maceration method. Phytochemical investigation of E. bulbosa extract was analyzed using GC-MS, spectrophotometry and titrimetry methods. The zone of inhibition was identified by the diffusion agar method. The minimum inhibitory concentration and minimum bactericidal concentration were determined using the plate count method. The inhibitory rate against V. parahaemolyticus was determined by the microdilution method. Cellular leakage was evaluated by spectrophotometry and cellular damage was observed by scanning electron microscopy. Results: GC-MS analysis showed the high compound of the E. bulbosa extract was securixanthone E(7-hydroxy-1,2-dimethoxyxanthone). The compound groups also included fatty acid esters, isoquinolines, naphthalenes, and phenolics. The total phenolic content was(2.50 ± 0.00) mg/g, flavonoid(6.61 ± 0.00) mg/g, and tannin(0.03 ± 0.00)%. The greatest zone of inhibition and inhibitory rate were(11.83 ± 0.06) mm and(91.32 ± 2.76)%, respectively, at 10 mg/m L. The minimum inhibitory concentration was 0.156 mg/m L, while the minimum bactericidal concentration was 10 mg/m L. The E. bulbosa extract caused leakage and cellular damage to V. parahaemolyticus. Conclusions: The E. bulbosa extract possesses inhibitory activities against V. parahaemolyticus and causes cellular leakage and damage.

关键词： Phytochemical Eleutherine bulbosa Antibacterial vibrio parahaemolyticus

Molecular analyses of bacterioplankton communities with highly abundant vibrio clades: a case study in Bohai Sea coastal waters

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Journal of Oceanology and Limnology 2019年第5期37卷 1638-1648页

作者： WANG Shuping ZHANG Yuan HE Jia JIA Xiaobo LIN Jianing LI Meng WANG Qinglin State Key Laboratory of Environmental Criteria and Risk Assessment Chinese Research Academy of Environmental SciencesBeijing 100012China Institute for Advanced Study Shenzhen UniversityShenzhen 518000China College of Animal Science and Technology Hebei Normal University of Science and TechnologyQinhuangdao 066000China

We investigated the bacterioplankton abundance, community composition and the associated vibrio clades of natural seawater in Bohai Sea coastal waters. Seawater samples (10 L in triplicate) were collected at 0.5, 3, and 5 m depths near the coastal aquaculture zone of the Bohai Sea on May 12, 2016. Real-time PCR and 16S rRNA gene amplicon high-throughput sequencing methods were employed by which 485 operational taxonomic units (OTUs) at a 97% sequence similarity level were generated. Gammaproteobacteria, Alphaproteobacteria, and Bacteroidetes were the most abundant groups, accounting for 49.5%, 23.5%, and 18.9% of the total assemblage, respectively. Obvious variations in Pseudoalteromonas, vibrio, and Octadecabacter , which were the most abundant genera, could be observed among diff erent samples. Notably, the results of vibrio -specifi c real-time PCR indicated that vibrio had extremely high 16S rRNA gene copy numbers. The 16S rRNA gene sequencing results across all the samples also indicated that they occupied a large proportion of the total assemblage. Both the alpha diversity and major bacterioplankton group Pseudoalteromonas had significant correlations with the concentration of PO4^3-. Overall, studies on bacterioplankton communities with highly abundant vibrio clades can provide interesting insight into the microbial function and health assessment of the Bohai Sea coastal ecosystem.

关键词： Bohai Sea bacterioplankton vibrio 16S rRNA Illumina MiSeq

Electrochemical detection of a vibrio parahaemolyticus sequence-specific gene based on a gold electrode modified with a single stranded probe

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Science China Chemistry 2010年第6期53卷 1366-1370页

作者： LAN MinBo1,ZHOU Qin1,ZHAO YanHui1,TENG YuanJie1,CHEN Chen1,ZHAO HongLi1 & YUAN HuiHui2 1Key Laboratory for Advanced Materials and Analysis Test Research Center,East China University of Science and Technology,Shanghai 200237,China 2Key Laboratory for Ultrafine Materials,Ministry of Education,East China University of Science and Technology,Shanghai 200237,China Key Laboratory for Advanced Materials and Analysis Test Research Center East China University of Science and Technology Shanghai China Key Laboratory for Ultrafine Materials Ministry of Education East China University of Science and Technology Shanghai China

An electrochemical DNA biosensor for specific-sequences detection of vibrio parahaemolyticus (VP) was fabricated.A singlestranded 20-mer oligonucleotide (ssDNA) and 6-mercapto-1-hexanol (MCH) were immobilized via a thiol linker on gold disk electrodes by self-assembling.The ssDNA underwent hybridization in a hybridization solution containing complementary or non-complementary or single base pair mismatched DNA sequences of VP.Examination of changes in response to these three target DNAs showed that the developed biosensor had a high selectivity and sensitivity.

关键词： vibrio parahaemolyticus electrochemical DNA sensor self-assembling gold electrode

Purification,characterization and production optimization of a vibriocin produced by mangrove associated vibrio parahaemolyticus

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Asian Pacific Journal of Tropical Biomedicine 2014年第4期4卷 253-261页

作者： Baskar Balakrishnan Jayappriyan Kothilmozhian Ranishree Sathish Thadikamala Prabakaran Panchatcharam Center for Bioenergy Cooperative researchLincoln University Marine Biotechnology Andaman and Nicobar Centre for Ocean Science and TechnologyNational Institute of Ocean Technology Department of Microbiology PRIST University

Objective:To identify a potential bacterium which produces antimicrobial peptide(vibriocin),and its purification,characterization and production optimization.The bacteria subjected in the study were isolated from a highly competitive ecological niche of mangrove ecosystem.Methods:The bacterium was characterized by phenotype besides 16S rRNA gene sequence analysis.The antibacterial activity was recognised by using agar well diffusion method.The vibriocin was purified using ammonium sulphate precipitation,butanol extraction,gel filtration chromatography,ion-exchange chromatography and subsequently,by HPLC.Molecular weight of the substance identified in SDS-PAGE.Production optimization performed according to Taguchi's mathematical model using 6 different nutritional parameters as variables.Results:The objective bacterium was identified as vibrio parahaemolyticm.The vibriocin showed 18 KDa of molecular mass with mono peptide in nature and highest activity against pathogenic vibrio harveyi.The peptide act stable in a wide range of pH,temperature.UV radiation,solvents and chemicals utilized.An overall^20%of vibriocin production was improved,and was noticed that NaCl and agitation speed played a vital role in secretion of vibriocin.Conclusion:The vibriocin identified here would he an effective alternative for chemically synthesized drugs for the management of vibrio infections in mariculture industry.

关键词： vibriocin vibrio parahaemolyticus vibrio harveyi Mangrove rhizosphere Antimicrobial peptide

Agglutinin activity in hemolymph of Chinese shrimp and induction effect of vibrio

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Acta Oceanologica Sinica 1998年第3期17卷 357-361页

作者： Luo Rixiang(Received May 30, 1997 accepted July 20, 1997) Institute of Oceanology Chinese Academy of Sciences Qingdao 266071 China

Experiments showed the agglutinin activity in the hemolymph of both spring wild parents and autumncultured individuals of the shrimp Penaeus chinensis Osbeck with the body length of 12 - 16 cm as all 32. Thehemolymph of the above shrimps showed agglutinant reaction to all A-, B- and O-type human erythrocytes, with thereaction titer equal to that of rabbit blood cells. However, the agglutinin activity level was enhanced after being in-duced by Vibro, which was 128 after 24 h of induction and reached the peak value 256 after 48 h of induction, thendecreased gradually.

关键词： Agglutinin activity hemolymph Chinese shrimp induction effect vibrio

Emerging Food-borne Pathogens

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Biomedical and Environmental Sciences 2001年第1期14卷 44-52页

作者： JRGEN SCHLUNDT Programme of Food Safety

The epidemiology of foodborne diseases is changing. While in many developing nations the efficient treatment of diarrhoeal diseases through oral rehydration has probably led to the prevention of many food related deaths, the underlying problems have not been solved. In these countries, the disease incidence of food_and waterborne disease is still increasing, and now data from other parts of the world indicate that new foodborne pathogens have emerged as important public health problems. Over the last two decades, bacterial infections caused by Campylobacter and enterohaemorrhagic Escherichia coli have emerged, well_recognized pathogens, such as Salmonella enteritidis, have increased dramatically in disease incidence and important foodborne pathogens have become increasingly resistant to antimicrobial agents. The future strategy for prevention of foodborne disease should be founded in scientifically based evaluations of the whole food production chain ‘from farm to table’, including a description of the most important (risk) factors. Epidemiological studies of outbreaks as well as sporadic cases should be aimed at a better understanding of these factors. In terms of public health importance of the problem, the focus should be on the size of the problem, and the potential for improvement. The indications are that both in developed and developing countries there is significant potential for an improvement of the situation. With this aim in mind, international effort should focus on scientific assessments of the potential for risk reduction under different regional conditions.

关键词： FOODBORNE DISEASES EPIDEMIOLOGY SURVEILLANCE vibrio

Involvement of ayu NOD2 in NF-κB and MAPK signaling pathways: Insights into functional conservation of NOD2 in antibacterial innate immunity

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Zoological Research 2019年第2期40卷 77-88页

作者： Yi Ren Shui-Fang Liu Li Nie Shi-Yu Cai Jiong Chen Laboratory of Biochemistry and Molecular Biology School of Marine Sciences Ningbo University Key Laboratory of Applied Marine Biotechnology of Ministry of Education Ningbo University

Nucleotide oligomerization domain 2(NOD2) is a major cytoplasmic sensor for pathogens and is critical for the clearance of cytosolic bacteria in mammals.However, studies regarding NOD2, especially the initiated signaling pathways, are scarce in teleost species. In this study, we identified a NOD2 molecule(PaNOD2) from ayu(Plecoglossus altivelis).Bioinformatics analysis showed the structure of NOD2 to be highly conserved during vertebrate evolution. Dual-luciferase reporter assays examined the activation of NF-κB signaling and Western blotting analysis detected the phosphorylation of three MAP kinases(p-38, Erk1/2, and JNK1/2).Functional study revealed that, like its mammalian counterparts, PaNOD2 was the receptor of the bacterial cell wall component muramyl dipeptide(MDP), and the leucine-rich repeat motif was responsible for the recognition and binding of Pa NOD2 with the ligand. Overexpression of PaNOD2 activated the NF-κB signaling pathway, leading to the upregulation of inflammatory cytokines, including TNF-α and IL-1β in HEK293 T cells and ayu head kidney-derived monocytes/macrophages(MO/MΦ).Particularly, we found that PaNOD2 activated the MAPK signaling pathways, as indicated by the increased phosphorylation of p-38, Erk1/2, and JNK1/2, which have not been characterized in any teleost species previously. Our findings proved that the NOD2 molecule and initiated pathways are conserved between mammals and ayu. Therefore, ayu could be used as an animal model to investigate NOD2-based diseases and therapeutic applications.

关键词： Ayu NOD2 NF-κB signaling MAPK signaling Inflammatory cytokines vibrio anguillarum infection

Zonula occludin toxin,a microtubule binding protein

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World Journal of Gastroenterology 2000年第3期6卷 330-334页

作者： MariaRosaria DiPierro Alessio Fasano Center for Vaccine Development Division of Gastroenterology University of Baltimore at MarylandBaltimoreMD 21201USA

AIM To investigate the interaction of Zot withmicrotubule.METHODS Zot affinity column was applied topurify Zot-binding protein（s）from crudeintestinal cell lysates.After incubation at roomtemperature,the column was washed and theproteins bound to the Zot affinity column wereeluted by step gradient with NaCl(0.3 mol·L-1-0.5mol·L-1).The fractions were subjected to6.0%-15.0%（w/v）gradient SDS-PAGE andthen transferred to PVDF membrane for N-terminal sequencing.Purified Zot and tauprotein were blotted by using anti-Zot or anti-tauantibodies.Finally,purified Zot was tested in anin vitro tubulin binding assay.RESULTS Fractions from Zot affinity columnyielded two protein bands with a Mr of 60 kU and45kU respectively.The N-terminal sequence ofthe 60 kU band resulted identical to β-tubulin.Zot also cross-reacts with anti-tau antibodies.Inthe in vitro tubulin binding assay,Zot co-precipitate with Mt,further suggesting that Zotpossesses tubulin-binding properties.CONCLUSION Taken together,these resultssuggest that Zot regulates the permeability ofintestinal tight junctions by binding tointracellular Mt,with the subsequent activationof the intracellular signaling leading to thepermeabilization of intercellular tight junctions.

关键词： microtubules microtubule proteins microtubule-associated proteins intercellular junctions vibrio cholerae cholera toxin zonula occluding toxin

Cholera:a great global concern

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Asian Pacific Journal of Tropical Medicine 2011年第7期4卷 573-580页

作者： Shyamapada Mandal Manisha Deb Mandal Nishith Kumar Pal Department of Zoology Gurudas CollegeNarkeldangaKolkata-700 054India Department of Physiology and Biophysics KPC Medical College and Hospital1F Raja S C Mallick RoadJadavpurKolkata-700032India Department of Microbiology Institute of Post Graduate Medical Education and Research244B A J C Bose RoadKolkata-700 020India

Cholera,caused by the infection of toxigenic vibrio cholerae(V.cholerae) to humans,is a life threatening diarrheal disease with epidemic and pandemic potential.The V.cholerae,both 01 and 0139 serogroups,produce a potent enterotoxin(cholera toxin) responsible for the lethal symptoms of the disease.The O1 serogroup has two biotypes(phenotypes),classical and El Tor;each of which has two major serotypes(based on antigenic responses),Ogawa and Inaba and the extremely rare Hikojima.V.cholerae O1 strains interconvert and switch between the Ogawa and Inaba serotypes.Fluid and electrolyte replacement is the mainstay of treatment of cholera patients;the severe cases require antibiotic treatment to reduce the duration of illness and replacement of fluid intake.The antibiotic therapy cunenlly has faced difficulties due to the rapid emergence and spread o(multidrug resistant V.cholerae causing several outbreaks in the globe.Currently,cholera has been becoming endemic in an increasing number of geographical areas,reflecting a failure in implementation of control measures.However,the current safe oral vaccines lower the number of resistant infections and could thus represent an effective intervention measure to control antibiotic resistance in cholera.Overall,the priorities for cholera control remain public health interventions through improved drinking water,sanitation, surveillance and access to health care facilities,and lurther development of safe,effective and appropriate vaccines.Thus,this review describes the facts and phenomena related to the disease cholera,which is still a great threat mainly to the developing countries,and hence a grave global concern too.

关键词： Cholera Toxigenic vibrio cholerae Pandemics-epidemics-outbreaks Cholera vaccine Multidrug resistance