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[studies on the morphology of blood of the european eel (anguilla agnuilla). iii. studies on monocytic and lymphoid cells].investigations concerning the monocytic and lymphocytic cell series were carried out on the peripheral blood and dabbings of the kidney, the spleen and intestines of healthy eels (anguilla anguilla), and eels taken ill with vibrio anguillarum. by means of histological, cytochemical, phaseoptical, and electronmicroscopical examinations independent development lines of the monocytes and lymphocytes were established. the place of origination for the monocytes was found to be the kidney, for the lym ...197772711
an aeromonas species implicated in ulcer-disease of the cod (gadus morhua).vibrio anguillarum is the most frequently isolated bacterial species involved in ulcer disease in salt water fish. however, an aeromonas species was very often incriminated in ulcers and septicaemia in cods (gadus morhua) in danish coastal areas. these aeromonas strains were very uniform in their biochemical activities only differing in growth on simmons citrate agar, and the mean value for the guanine + cytosine content of dna was 59 per cent with a standard deviation of 0.4. primary serologica ...1977325501
antitumor activity of marine bacteria, vibrio anguillarum, in mice.antitumor activity of marine bacteria, vibrio anguillarum p-b-1, against ehrlich carcinoma cells in ddy mice was investigated. ehrlich carcinoma cells were inoculated intraperitoneally (ip) into mice and v. anguillarum (1.0 mg/mouse) was administered ip twice before and 4 times after the tumor inoculation. groups of mice administered v. anguillarum survived 80 approximately 90% and their mean survival was 54.0 approximately 56.0 days (range, 24.0 approximately 60 days) at day 60 after inoculatio ...1979510844
rainbow trout complement fixation used for titration of antibodies against several pathogens.since it is impossible to fix guinea pig complement (c) with rainbow trout (salmo gairdneri) antibodies, whereas it is possible with brown trout (salmo trutta) antibodies, a c fixation test has been designed which uses haemolysin and c from rainbow trout. immunization of trout against lysed sheep red blood cells (srbc) elicited production of haemolytic igm. normal trout serum (nts) can be used as a source of c at a dilution at which its "natural" haemolytic activity (against various homeotherms ...1979547824
[biochemical properties of the fish pathogen vibrio anguillarum (author's transl)]. 1977600705
[host specific pathogenecity in vibrio anguillarum strains isolated from salmonids and coalfish (author's transl)]. 1977600710
vibrio anguillarum and larval mortality in a california coastal shellfish hatchery.vibrio anguillarum was isolated as a pathogen in the commercial culture of oyster spat at pigeon point, calif. a water-soluble, heat-stable exotoxin extracted from cultures of the vibrio inhibited larval swimming and contributed to larval mortality. although the vibrio was insensitive to penicillin in standard plate testing, this antibiotic proved useful in preventing mass larval mortalities in the hatchery.1978623467
evidence for plasmid contribution to the virulence of fish pathogen vibrio anguillarum.analysis of the plasmid deoxyribonucleic acid complement of high- and low- virulent strains of the fish pathogen vibrio anguillarum showed a correlation between enhanced virulence and the presence of a 50-megadalton plasmid class. all 50-megadalton plasmids isolated from different high-virulent v. anguillarum strains were homologous as judged by the analysis of plasmid deoxyribonucleic acid-deoxyribonucleic acid hybridization. the 50-megadalton plasmid class did not have polynucleotide sequences ...1977924679
failure of vaccines to protect salmon from vibriosis enzootic in puget sound, washington.juvenile chinook salmon in fresh water were vaccinated either orally or parenterally with heat- or formalin-killed bacterins prepared with vibrio anguillarum. subsequent exposure to naturally occuring vibriosis in the marine waters of puget sound (washington state) indicated that no protection resulted. it was demonstrated, however, that protection could be achieved with the passive transfer of immune serum.1976937795
study of genetic relationships among marine species of the genera beneckea and photobacterium by means of in vitro dna/dna hybridization.strains representative of species of the marine genera beneckea and photobacterium were used as reference standards in in vitro dna/dna competition experiments. within a given species, strains were found to be related by over 80% competition. (competition was defined as the amount of radioactive dna displaced by heterologous dna relative to the amount displaced by homologous dna.) on the basis of interspecies competition values (expressed as averages), the following groupings could be made: 1. " ...19761015934
aeromonas proteolytica--a halophilic aeromonad?aeromonas proteolytica, a halophilic bacterium originally isolated from the intestine of a marine isopod, was compared with 93 strains of aeromonas hydrophila and representative strains of five similar genera in a computer and serological analysis. aeromonas proteolytica was peritrichate and non-reactive with six a. hydrohila antisera, and possessed a percentage guanine + cytosine ratio (49.5) outside the range for other aeromonads (55-62%). moreover, in the computer analysis it shared a lower l ...19751148936
characterization of the factors responsible for death of fish infected with vibrio anguillarum.vibrio anguillarum produced substances toxic for goldfish (crassius auretus) that are released from living bacteria and associated with heat-killed bacteria. heating (100 degrees c) enhances the potency of the extracellular toxin.19751164698
serological responses and immunity produced in salmonids by vaccination with australian strains of vibrio anguillarum.the australian populations of rainbow trout (onchorhynchus mykiss) and atlantic salmon (salmo salar) were found to have similar immunological responses to local strains of vibrio anguillarum as those reported for the more genetically diverse populations of these fish and strains of v. anguillarum found in the northern hemisphere. in addition, our studies more precisely defined the respective responses of rainbow trout and atlantic salmon to immersion vaccination by the bath and dip methods.19921289240
highly preferred site of insertion of tn7 into the chromosome of vibrio anguillarum.the possible usefulness of tn7 as a tool for genetic studies in vibrio anguillarum was examined. using the plasmid prk2073 as the transposon donor, tn7 transposes at high frequency into the chromosome of v. anguillarum. however, hybridization analysis of the mutants dna digested with different enzymes revealed that all isolates have the insertions in the same site. this indicates that like in many other gram-negative bacteria, tn7 shows a specificity of transposition in the chromosome of v. angu ...19921319594
virulence plasmid pjm1 prevents the conjugal entry of plasmid dna into the marine fish pathogen vibrio anguillarum 775.studies involving the introduction of cloned homologous genes into vibrio anguillarum revealed that several plasmids could not be conjugally introduced into v. anguillarum 775(pjm1), but were transmissible to the pjm1-cured derivative h775-3. recombinant pbr322 plasmids containing v. anguillarum genomic dna inserts were mobilized from escherichia coli donors, using prk2013, into v. anguillarum h775-3 recipients at frequencies of 10(-6) to 10(-5) per recipient. when identical matings were perform ...19921336793
towards a phylogeny of the genus vibrio based on 16s rrna sequences.the inter- and intrageneric relationships of the genus vibrio were investigated by performing a comparative analysis of the 16s rrnas of 10 species, including four pathogenic representatives. the results of immunological and 5s rrna studies were confirmed in that the genus is a neighboring taxon of the family enterobacteriaceae. with regard to the intrageneric structure, vibrio alginolyticus, vibrio campbellii, vibrio natriegens, vibrio harveyi, vibrio proteolyticus, vibrio parahaemolyticus, and ...19921371064
occurrence of 2,4-dihydroxy-3,3,4-trimethylpyroglutamic acid as an n-acyl substituent in the o-polysaccharide chain of the lipopolysaccharide of vibrio anguillarum v-123.a new and highly branched amino acid was found as an n-acyl substituent of the o-polysaccharide chain obtained from the lipopolysaccharide of vibrio anguillarum v-123 (serogroup jo-2) and evidence is presented to support the structure as 2,4-dihydroxy-3,3,4-trimethylpyroglutamic acid. acid hydrolysis of the o-polysaccharide gave the lactone of 2,4-dihydroxy-3,3,4-trimethylglutamic acid, together with 2-amino-2-deoxy-d-galacturonic acid, 2-amino-2,6-dideoxy-d-glucose (d-quinovosamine), and 4-amin ...19921394311
structure of the o-polysaccharide chain of the lipopolysaccharide of vibrio anguillarum v-123.the o-polysaccharide chain (ps-1), released by mild acidic treatment of the lps of v. anguillarum v-123 (serogroup jo-2), a pathogenic bacterium of marine and estuarine fish, consists of 2-amino-2-deoxy-d-galacturonic acid, 2-amino-2,6-dideoxy-d-glucose (d-quinovosamine), and 4-amino-4,6-dideoxy-d-glucose (d-viosamine) n-acylated with 2,4-dihydroxy-3,3,4-trimethylpyroglutamic acid. strong-acid hydrolysis of ps-1 afforded alpha-galna-(1----4)-alpha-galna-(1----3)-quin (a1) and alpha-galna-(1----3 ...19921394312
effectiveness of an oral enteric coated vibrio vaccine for use in salmonid fish.an oral enteric-coated vibrio anguillarum vaccine was developed by initially coating lyophilized bacteria onto 0.9 mm diameter dextrose sugar beads followed by an eudragit l-30d coat to serve as enteric protection. vaccine efficacy was determined by both an in vivo challenge with live pathogen and measurements of serum and mucus antibody levels by elisa. survival rates after challenge were 80.3%, 83.3% and 70.3% among the vaccine group, positive and negative controls respectively. serum and mucu ...19921398784
cloning and nucleotide sequence analysis of a chloramphenicol acetyltransferase gene from vibrio anguillarum.the chloramphenicol resistant gene (cat) encoding chloramphenicol acetyltransferase (cat) in a transferable r plasmid (pja7324) isolated from the fish pathogen vibrio anguillarum strain pt24 was cloned into the plasmid vector puc19. the nucleotide sequence analysis of 1,348 base pair dna identified an open reading frame encoding a protein of 216 amino acid residues with a calculated molecular mass of 25,471 daltons. the predicted amino acid sequences for this cat gene are 37-69% homologous with ...19921406372
temperature-dependent in vitro antimicrobial activity of four 4-quinolones and oxytetracycline against bacteria pathogenic to fish.the in vitro antimicrobial activities of oxolinic acid, flumequine, sarafloxacin, enrofloxacin, and oxytetracycline against strains of bacteria pathogenic to fish (aeromonas salmonicida subsp. salmonicida, atypical a. salmonicida, vibrio salmonicida, vibrio anguillarum, and yersinia ruckeri) were determined at two different incubation temperatures, 4 and 15 degrees c, by a drug microdilution method. the main objective of the study was to examine the effect of incubation temperature on the in vit ...19921416857
cloning of a metalloprotease gene involved in the virulence mechanism of vibrio anguillarum.genetic evidence has previously suggested that a zinc metalloprotease is involved in the invasive mechanism of the fish pathogen vibrio anguillarum nb10. in this study, the metalloprotease gene was cloned and sequenced. the sequence encodes a polypeptide (611 amino acids) that contains a putative signal sequence followed by a large leader sequence and the mature protein (44.6 kda). since the purified protein has a molecular mass of 36 kda instead of the predicted 44.6 kda, the mature protein is ...19921429449
application of ozone disinfection to remove enterococcus seriolicida, pasteurella piscicida, and vibrio anguillarum from seawater.survival of bacterial fish pathogens, including enterococcus seriolicida, vibrio anguillarum, and pasteurella piscicida, in ozonated seawater was determined in a batch system. bacterial counts of all fish pathogens decreased at more than 0.040 to 0.060 mg of total residual oxidants (tros) per liter, whereas no decrease in viable counts was observed at less than 0.018 to 0.028 mg of tros per liter. the 99% inactivation point was achieved at concentrations of 0.111 mg/liter for e. seriolicida, 0.0 ...19921476447
nucleotide sequence analysis of the class g tetracycline resistance determinant from vibrio anguillarum.the nucleotide sequence of the class g tetracycline resistance determinant previously isolated from vibrio anguillarum has been determined. two open reading frames of divergent polarity were identified. a resistance gene (tet a) encodes a protein of 393 amino acid residues (deduced molecular mass of 40.9 kda), and a repressor gene (tet r) encodes a protein consisting of 210 amino acids with a calculated molecular mass of 23.6 kda. based on the deduced amino acid sequences, the proteins of tet a( ...19921479961
agglutinin activity in pacific oyster (crassostrea gigas) hemolymph following in vivo vibrio anguillarum challenge.hemolymph from the pacific oyster (crassostrea gigas) contains lectins that agglutinate horse (gigalin e) and human (gigalin h) erythrocytes. the gigalins also agglutinate bacteria, including vibrio anguillarum, and were adsorbed from oyster hemolymph at different temperatures by living, heat-killed, and freeze-dried v. anguillarum cells. baseline activities of the two gigalins were established by measuring their activities in oyster hemolymph over a period of 4 years. a normal distribution of g ...19921499833
characterization of the reca gene of vibrio anguillarum.we have cloned and sequenced the reca gene from two strains, 775 and 531a, of the fish pathogen, vibrio anguillarum. although both strains showed different sensitivities to methyl methanesulfonate (mms), the reca genes were identical. in vitro expression of the v. anguillarum reca gene produced a polypeptide of about 40 kda, in agreement with the value obtained from the nucleotide sequence. we identified the transcription start point by primer extension. the promoter for the reca gene mapped to ...19921544576
expression of the b subunit of escherichia coli heat-labile enterotoxin in a marine vibrio and in a mutant that is pleiotropically defective in the secretion of extracellular proteins.a marine vibrio (designated vibrio sp. 60) that is related to vibrio anguillarum was used as a host for a plasmid that encodes the non-toxic b subunit (etxb) of escherichia coli heat-labile enterotoxin. expression of etxb in vibrio sp. 60 resulted in the efficient and selective secretion of the b subunit into the extracellular growth medium. this indicated that vibrio sp. 60, which does not normally produce cholera-like enterotoxins, nonetheless possesses a secretory machinery that permits these ...19921588306
point mutations in two conserved glycine residues within the integral membrane protein fhub affect iron(iii) hydroxamate transport.a region of substantial homology, comprising 32 amino acids around a highly conserved glycine residue, is located near the c-terminal ends of the hydrophobic fhu, fec, fep, fat, and btu transport proteins involved in the uptake of ferrisiderophores and vitamin b12 into escherichia coli and vibrio anguillarum. furthermore, a region similar in location and sequence containing an invariant glycine at an equivalent position was identified in the hydrophobic component of all other periplasmic binding ...19921588908
intestinal colonization potential of turbot (scophthalmus maximus)- and dab (limanda limanda)-associated bacteria with inhibitory effects against vibrio anguillarum.of more than 400 bacteria isolated from turbot (scophthalmus maximus), 89 have previously been shown to inhibit the in vitro growth of the fish pathogen vibrio anguillarum. the aim of the present study was to investigate the potential of seven of these strains, as well as of intestinal isolates (four strains) from a closely related fish, dab (limanda limanda), for colonizing farmed turbot as a means of protecting the host from infection by v. anguillarum. in addition, the inhibitory effect of th ...19921610180
development of a gene transfer system for curing of plasmids in the marine fish pathogen vibrio salmonicida.all reported natural isolates of the marine fish pathogen vibrio salmonicida contain plasmids, and in another marine fish pathogen, vibrio anguillarum, it has been shown that a plasmid is important for expression of virulence by the organism. to study the function of the plasmids in v. salmonicida, we developed a gene transfer system based on the plasmid rsf1010 replicon. the gene transfer system was used to construct a plasmid-free strain, and this strain was found to behave similarly to the wi ...19921622274
molecular characterization of the iron transport system mediated by the pjm1 plasmid in vibrio anguillarum 775.complementation of insertion mutants showed that the polypeptides fatd, fatc, fatb, and fata are essential for the iron-transport process encoded by pjm1. sequence analysis followed by homology studies indicated that transport of ferric anguibactin into vibrio anguillarum 775 follows the same mechanism as reported for transport of fe(3+)-hydroxamates, fe(3+)-catecholates, ferric dicitrate, and vitamin b12 into escherichia coli. homology of fata, part of the receptor complex, to seven e. coli rec ...19911748657
occurrence of plasmids in danish isolates of vibrio anguillarum serovars o1 and o2 and association of plasmids with phenotypic characteristics.two hundred and twenty-eight isolates of vibrio anguillarum serovar o1 (125 isolates) and serovar o2 (103 isolates) have been characterized with regard to plasmid contents, biochemical properties, and in vitro hemagglutination and hydrophobic properties. among 74 v. anguillarum isolates from diseased fish, 63 carried only a 67-kb plasmid (pjm1), 9 carried an additional 98-kb plasmid, and 1 isolate carried only the 98-kb plasmid. only one isolate was without plasmids. in v. anguillarum serovar o1 ...19911768088
isolation and characterization of turbot (scophtalmus maximus)-associated bacteria with inhibitory effects against vibrio anguillarum.more than 400 isolates from the intestine and the external surface of farmed scophtalmus maximus as well as from fish food and hatchery water were screened for inhibitory effects against the fish pathogen vibrio anguillarum hi 11345 and seven other fish pathogens. the bacteria with inhibitory effects were then characterized with regard to their sites of colonization, especially the intestinal regions and sites within each region. of the total number of bacterial isolates from the intestine, 28% ...19911768092
characterization of vibrio anguillarum and closely related species isolated from farmed fish in norway.a total of 264 bacterial strains tentatively or definitely classified as vibrio anguillarum were examined. the strains were isolated from diseased or healthy norwegian fish after routine autopsy. with the exception of five isolates from wild saithe (pollachius virens), the strains originated from nine different species of farmed fish. the bacteria were subjected to morphological, physiological, and biochemical studies, numerical taxonomical analyses, serotyping by slide agglutination and enzyme- ...19911768147
distribution of plasmid- and chromosome-mediated iron uptake systems in vibrio anguillarum strains of different origins.we investigated the incidence of plasmid-mediated and chromosome-mediated iron uptake systems in strains of vibrio anguillarum that belong to serotypes o1 and o2 and were isolated from different fish species and in different geographic areas. all of the strains gave positive reactions in cas agar medium and in the arnow test, which indicated that catechol types of siderophores were produced. the majority of v. anguillarum serotype o1 strains harbored a 65-kb plasmid similar to plasmid pjm1 from ...19911836126
polypeptides p40, pom2, and pangr are required for iron uptake and for virulence of the marine fish pathogen vibrio anguillarum 775.insertions were created in three iron uptake genes in plasmid pjm1 of vibrio anguillarum 775 to assess their in vivo effects on virulence in fish. insertions that blocked p40, pom2, and pangr expression resulted in iron uptake-negative strains and in 4.2 x 10(5)-, 8.8 x 10(5)-, and 2.5 x 10(5)-fold attenuations in virulence, respectively. a strain with an insertion in the pangr coding region still synthesized significant constitutive levels of the outer membrane protein pom2 and persisted in fis ...19911846864
regulation of plasmid-mediated iron transport and virulence in vibrio anguillarum.iron is essential for bacterial growth and metabolism. in vertebrates this metal is complexed by high-affinity iron-binding proteins, such as transferrin in serum. the fish pathogen vibrio anguillarum possesses a very efficient iron-uptake system which is encoded in the virulence plasmid pjm1. this allows the bacterium to utilize the otherwise unavailable iron in the fish host, resulting in the septicemic disease vibriosis. this system includes the siderophore anguibactin and transport component ...19911854586
activation of rainbow trout complement by c-reactive protein.the activation of the complement system of rainbow trout by trout c-reactive protein (crp) was investigated. complement fixation tests were performed by using rabbit hemolysin-sensitized sheep erythrocytes and rainbow trout complement. purified crp increased the consumption of complement in the presence of streptococcus pneumoniae c-polysaccharide (cps), indicating the activation of the complement system. in contrast to this, acute phase serum activated the complement in the absence of cps. cons ...19911903618
a novel lipopolysaccharide-binding hemagglutinin isolated from hemocytes of the solitary ascidian, halocynthia roretzi: it can agglutinate bacteria.a hemagglutinin was isolated from hemocytes of the ascidian, halocynthia roretzi, by a procedure including extraction and ion-exchange chromatography on cm-cellulose. the molecular weight of the hemagglutinin was estimated to be 120,000 by gel filtration. it was resistant to acid treatment but sensitive to alkali or heat treatment. the hemagglutinating activity was inhibited by heparin, chondroitin sulfate, and lipopolysaccharide (lps), but not by mono- and disaccharides such as n-acetyl-galacto ...19911904830
restriction fragment length polymorphism of the vibrio anguillarum serovar o1 virulence plasmid.seventy-eight strains of vibrio anguillarum serovar o1, all harboring one 65- to 70-kilobase plasmid, were typed according to restriction fragment length polymorphism of the plasmid. six types, three of which comprised 96% of the strains examined, were produced with the restriction endonuclease bamhi. the fragment length polymorphism type did not correlate to any of 12 different phenotypic properties tested.19901981001
purification and characterization of a secreted protease from the pathogenic marine bacterium vibrio anguillarum.vibrio anguillarum is a pathogenic marine bacterium which causes the disease vibriosis in salmonid fish, which is characterized by a fatal hemorrhagic septicemia accompanied by massive tissue destruction. in this paper, the purification of the major caseinolytic extracellular protease from v. anguillarum is presented. the purification steps include ammonium sulfate precipitation, deae-sepharose chromatography, sephacryl s-200 chromatography, and deae high-pressure liquid chromatography. the puri ...19912012804
monoclonal antibodies to salmonid immunoglobulin: characterization and applicability in immunoassays.monoclonal antibodies to rainbow trout (oncorhynchus mykiss) igm were prepared and characterized for use in immunoassays. antibodies produced by the five clones reacted with the heavy chain of the immunoglobulin. no indication of different heavy chain isotype specificity was observed for the mabs. one clone discerned igm from rainbow trout while the other four clones cross-reacted with igm from atlantic salmon (salmo salar) and from brown trout (salmo trutta). the monoclonal antibodies identifie ...19902086271
inhibitory effect of halocyamine, an antimicrobial substance from ascidian hemocytes, on the growth of fish viruses and marine bacteria.halocyamine a, an antimicrobial substance isolated from hemocytes of the solitary ascidian halocynthia roretzi, inhibited in vitro the growth of fish rna viruses (infectious hematopoietic necrosis virus and infectious pancreatic necrosis virus). pretreatment of rna virus with halocyamine a reduced the infectivity of the virus toward host cells. the growth of marine bacteria, achromobacter aquamarinus and pseudomonas perfectomarinus, was also inhibited by halocyamine a but that of alteromonas put ...19902121517
iron-binding compounds and related outer membrane proteins in vibrio cholerae non-o1 strains from aquatic environments.a total of 156 strains of vibrio cholerae non-o1 from aquatic origins were examined for the presence of iron uptake mechanisms and compared with o1 strains and other vibrio species. all non-o1 strains were able to grow in iron-limiting conditions, with mics of ethylenediaminedi (o-hydroxyphenylacetic acid) ranging from 20 microm to 2 mm. the production of siderophores was demonstrated by growth in chrome azurol s agar and cross-feeding assays. all strains produced phenolate-type compounds, as as ...19902144956
identification and characterization of a zinc metalloprotease associated with invasion by the fish pathogen vibrio anguillarum.an invasiveness-defective mutant of the fish-pathogenic bacterium vibrio anguillarum was isolated. compared with the wild type, this mutant had a 1,000-fold higher 50% lethal dose after immersion infection of rainbow trout, oncorhynchus mykiss, while after intraperitoneal infection, the mutant had only a 10-fold higher 50% lethal dose. in addition, the mutant showed a lower level of protease activity. two forms of the protease (pa and pb) were found after sodium dodecyl sulfate-polyacrylamide ge ...19902228244
vibrios associated with red tides caused by mesodinium rubrum.vibrios were isolated from red tides caused by mesodinium rubrum and also throughout the year in the ria de pontevedra, spain. the isolates were grouped into 14 phena by numerical toxonomy. strains associated with red tides were restricted to four phena: phena i and ii were vibrio alginolyticus, and phena iii and iv were vibrio tubiashii and vibrio anguillarum, respectively. v. anguillarum-like strains (phena v through xi) predominated throughout the year outside the red tide areas. cytotoxicity ...19902268167
a regulatory gene, angr, of the iron uptake system of vibrio anguillarum: similarity with phage p22 cro and regulation by iron.the angr locus in vibrio anguillarum encodes a trans-acting transcriptional activator which modulates several fe2(+)-regulated loci in the anguibactin biosynthesis gene cluster. in this paper, the complete nucleotide (nt) sequence of the angr gene and deduced amino acid (aa) sequence of the angr protein are presented. a region upstream from the angr gene is shown to have similarity with fe2(+)-regulated operators in escherichia coli which bind the fur protein. the involvement of a fur-like regul ...19902311935
isolation and characterization of rainbow trout c-reactive protein.an acute phase serum component, c-reactive protein (crp), was isolated from the sera of rainbow trout (salmo gairdneri). the isolation was based on its calcium-dependent binding affinity for pneumococcal c-polysaccharide (cps) according to the isolation procedure of human c-reactive protein. in sds-page, the nonreduced crp showed two subunits with molecular weights of 43,700 and 26,600, respectively, at a molar ratio of 1:1. the reduced crp showed a single subunit of 26,600. the molecular weight ...19902338157
detection of vibrio anguillarum antigen by the dot blot assay.the dot blot assay, modified and adapted for detection of antigens from vibrio anguillarum in fish tissues, was specific for v. anguillarum and did not react with antigens of v. ordalii, pseudomonas sp., or yersinia ruckeri. the blot assay enabled detection of as little as 2.3 ng of a mixture of protein antigens obtained from cell-free extracts of v. anguillarum; it was about 100 times more sensitive than either the indirect fluorescent antibody technique or bacterial isolation for detecting v. ...19852411956
survival of vibrio anguillarum and vibrio salmonicida at different salinities.the fish pathogenic bacteria vibrio anguillarum and v. salmonicida showed the capacity to survive for more than 50 and 14 months, respectively, in seawater microcosms. a salinity of 5% proved lethal to v. anguillarum harvested in the late-exponential growth phase, whereas a salinity of 9% was lethal to the bacterium after it had been starved at a salinity of 30% for 67 days. the lethal salinity for v. salmonicida harvested in the late-exponential growth phase was probably in the vicinity of 10%. ...19892475069
monoclonal antibodies against surface antigens of vibrio anguillarum serogroup 02. 19882476007
genetics and molecular biology of siderophore-mediated iron transport in bacteria.the possession of specialized iron transport systems may be crucial for bacteria to override the iron limitation imposed by the host or the environment. one of the most commonly found strategies evolved by microorganisms is the production of siderophores, low-molecular-weight iron chelators that have very high constants of association for their complexes with iron. thus, siderophores act as extracellular solubilizing agents for iron from minerals or organic compounds, such as transferrin and lac ...19892531838
regulation of the iron uptake system in vibrio anguillarum: evidence for a cooperative effect between two transcriptional activators.we have identified a 110-kda polypeptide that has a trans-acting regulatory activity on the expression of the pjm1 plasmid iron-uptake genes in vibrio anguillarum. this protein is encoded by the angr gene and maps in a 3.6-kilobase-pair pjm1 dna region located downstream of the iron transport genes. full expression of this gene occurs under iron-limiting conditions and requires a 2.9-kilobase-pair upstream region in cis that maps within the coding region of the om2 outer membrane protein, essent ...19892542936
protection of rainbow trout against vibriosis and furunculosis by the use of attenuated strains of vibrio anguillarum.the fish pathogen vibrio anguillarum causes a lethal infection in rainbow trout (salmo gairdneri). three different avirulent mutants, constructed by transposon insertion mutagenesis (van20 and van70) or as antibiotic-resistant mutants (van1000), were isolated by screening 200 individual isolated mutants for avirulence. when used as live vaccines, all three avirulent mutants were able to induce protective immunity against the homologous as well as a heterologous strain of v. anguillarum. when van ...19892548446
[chemical properties of lipopolysaccharide (lps) isolated from vibrio anguillarum pt514].vibrio anguillarum, one of the causative agents of fish vibriosis, is serologically and biochemically divided into three groups (a, b and c). the chemical composition and molecular architecture of lipopolysaccharide (lps) isolated from v. anguillarum pt 514, which belongs to serogroup b, were investigated. the lps contained glucose (glc), fructose (fru), l-glycero-d-mannoheptose (l-d hep), glucosamine (glcn) and 4-amino-4,6-dideoxyglucose as sugar constituents in molar ratios of 8.9:0.7:3.0:1.1: ...19892621792
identification of polypeptides encoded by cloned pjm1 iron uptake dna isolated from vibrio anguillarum 775.the xhoi fragment containing much of the iron uptake region of plasmid pjm1 was isolated from vibrio anguillarum 775 and cloned into plasmid pbr322. plasmid-encoded polypeptides were examined in maxicells of escherichia coli, and transposon mutagenesis was used to map insertion mutations in the structural dna encoding the om2 polypeptide. tn1000 insertions that mapped within om2 and blocked maxicell expression of om2 resulted in the loss of ferric iron-anguibactin receptor function when plasmids ...19892651396
bacterial action of fertilization envelope extract from eggs of the fish cyprinus carpio and plecoglossus altivelis.the vitelline envelope (ve) and fertilization envelope (fe) in eggs of the fish cyprinus carpio and plecoglossus altivelis were purified by homogenization of eggs or embryos in 5 mm tris-hcl buffer, ph 7.0, containing 2 mm ethylenediamine tetraacetic acid disodium salt (edta), except for processing of ves in plecoglossus eggs, and by repeated washing wih the same buffer. to extract the outermost layer material, the purified ves and fes were processed overnight at 4 degrees c in 5 mm tris-hcl buf ...19892661718
molecular cloning of the reca analog from the marine fish pathogen vibrio anguillarum 775.the reca analog from vibrio anguillarum 775 was isolated by complementation of reca mutations in escherichia coli, and its protein product was identified. the reca analog promoted recombination between two partially deleted lactose operons, stimulated both spontaneous and mitomycin c-induced phage production in reca- lambda lysogens, and restored near wild-type levels of resistance to uv radiation and methyl methanesulfonate.19892681167
relationships between plasmids and phenotypes of presumptive strains of vibrio anguillarum isolated from different fish species.on the basis of plasmid composition as well as serological and biochemical properties, 26 strains identified as vibrio anguillarum isolated from diseased fish could be assigned to two different groups. except for three reference strains, these++ strains were isolated from norwegian fish. the four strains isolated from rainbow trout (salmo gairdneri), the only strain isolated from char (salvelinus alpinus), and three of six strains isolated from atlantic salmon (salmo salar) harbored a plasmid of ...19892729984
activation of trout macrophages and production of crp after immunization with vibrio anguillarum.to examine the mechanism of the protection of rainbow trout (salmo gairdneri) against vibrio anguillarum in the early stage of immunization, the activation of macrophages and production of c-reactive protein (crp) were investigated. fish immunized with formalin-killed bacteria emulsified in freund's complete adjuvant (fca) resisted intraperitoneal challenge with living bacteria seven and ten days after immunization. the activation of macrophages was demonstrated by a significant increase of the ...19892776933
identification of vibrio anguillarum in fish by using partial 16s rrna sequences and a specific 16s rrna oligonucleotide probe.16s rrna from seven different vibrio anguillarum strains was partially sequenced and compared. from this sequence information we could design a 25-base-long oligonucleotide and use it as a specific probe for identification of v. anguillarum. this was determined by rna-dna colony hybridization and slot-blot hybridization. strong, specific hybridization to the probe was observed for all v. anguillarum strains tested. furthermore, no cross-hybridization could be seen against five other bacterial sp ...19892782871
new tetracycline resistance determinant on r plasmids from vibrio anguillarum.two classes of tetracycline resistance determinants on r plasmids were detected in vibrio anguillarum strains isolated from ayu (sweat fish; plecoglossus altivelis) farms in japan. tetracycline resistance genes categorized as class b were prevalent from 1973 to 1977; however, a new tetracycline resistance gene, which was not classified into tetracycline resistance determinant class a, b, c, or d, has been prevalent since 1981.19872823701
genetic and molecular characterization of essential components of the vibrio anguillarum plasmid-mediated iron-transport system.the iron-transport genes from the pjm1 plasmid of vibrio anguillarum have been cloned and sequenced. five open-reading frames have been identified, one of which encodes the outer membrane receptor for ferric anguibactin, om2. this coding region corresponds to a protein of 726 amino acids with a mr of 78,777. the protein has a hydrophobic signal sequence of 35 amino acids and a potential membrane-associated hydrophobic region at the carboxyl terminus. a 2.3-kilobase iron-regulated mrna was transc ...19882830268
genetic analysis of the iron uptake region of the vibrio anguillarum plasmid pjm1: molecular cloning of genetic determinants encoding a novel trans activator of siderophore biosynthesis.clones carrying the iron uptake region of the vibrio anguillarum plasmid pjm1 were subjected to insertion mutagenesis, using transposon tn3::hoho1 which carries a promoterless lacz gene and can thus generate lacz transcriptional fusions if inserted downstream from an indigenous promoter. four classes of insertion mutants were obtained based on the level of expression of components of the iron uptake system, and six genetic units were defined according to the phenotype of the mutants. five of the ...19882832388
stress alters immune function and disease resistance in chinook salmon (oncorhynchus tshawytscha).we examined the effects of acute stress on the immune system and disease resistance of juvenile chinook salmon (oncorhynchus tshawytscha) in laboratory and clinical trials. immune function, as measured by the ability of lymphocytes from the anterior kidney to generate specific antibody-producing cells (apc) in vitro, was depressed 4 h after stress, when plasma cortisol levels were highest. at the same time, resistance to the fish pathogen, vibrio anguillarum, was also depressed. compared with co ...19892918264
synthesis of glycoconjugates derived from various lipopolysaccharides of the vibrionaceae family.conjugation of simple ketoses (such as 3-deoxy-d-manno-2-octulosonic acid and n-acetylneuraminic acid) and of various o-specific polysaccharides (from aeromonas hydrophila and aeromonas salmonicida) to the bifunctional spacer 1,6-hexanediamine, was achieved by reductive amination. the saccharide--1-(6-amino)-hexane alkyamines obtained were converted into the corresponding isothiocyanate derivatives and coupled to the free epsilon-amino group of lysine residues of the protein carrier bovine serum ...19892920731
evidence for the role of a siderophore in promoting vibrio anguillarum infections.vibrio anguillarum strain 775 harbouring the virulence plasmid pjm1 produces a plasmid-mediated siderophore that can crossfeed siderophore-deficient, receptor-proficient mutants of v. anguillarum in vitro. experimental infections of salmonid fishes with mixtures consisting of the wild-type strain and a siderophore-deficient, receptor-proficient mutant resulted in recovery of both the wild-type and the mutant strain, while in infections with mixtures consisting of the wild-type strain and a sider ...19862957462
chromosome-mediated iron uptake system in pathogenic strains of vibrio anguillarum.we describe in this work a new iron uptake system encoded by chromosomal genes in pathogenic strains of vibrio anguillarum. this iron uptake system differs from the plasmid-encoded anguibactin-mediated system present in certain strains of v. anguillarum in several properties. the siderophore anguibactin is not utilized as an external siderophore, and although characteristic outer membrane proteins are synthesized under iron-limiting conditions, these are not related to the plasmid-mediated outer ...19882965144
plasmid-mediated iron sequestering systems in pathogenic strains of vibrio anguillarum and escherichia coli. 19852990437
plasmids mediating iron uptake in vibrio anguillarum strains isolated from turbot in spain.vibrio strains isolated from diseased turbot in an experimental fish farm on the atlantic coast of northwest spain were identified as vibrio anguillarum. the isolates shared many biochemical characteristics with v. anguillarum strains obtained from other sources, and harboured a plasmid species that showed extensive homology with plasmid pjm1, carried by v. anguillarum strain 775 isolated from an epizootic in north america. restriction endonuclease analysis showed that the two plasmids were very ...19852997366
characterization of anguibactin, a novel siderophore from vibrio anguillarum 775(pjm1).anguibactin, a siderophore produced by cells of vibrio anguillarum 775 harboring the pjm1 plasmid, has now been isolated from the supernatants of iron-deficient cultures. this iron-reactive material was purified by adsorption onto an xad-7 resin and subsequent gel filtration on a sephadex lh-20 column. the resulting neutral compound produced an ion at m/z 348 in mass spectrometry and contained one sulfur, four oxygen, and four nitrogen atoms as determined by elemental analysis. its strong uv abs ...19863013839
drug-resistant plasmids from fish pathogens.the epidemiological surveillance of drug-resistant strains of the fish pathogenic bacteria vibrio anguillarum and pasteurella piscicida carrying transferable r plasmids in fish farms is described. the dna structure of r plasmids, and the drug-resistant determinants of r plasmids from the fish pathogens aeromonas hydrophila,aeromonas salmonicida, edwardsiella tarda, v. anguillarum, and p. piscicida are discussed.19883079184
effects of the antimicrobic tiamulin on seven gram-negative bacterial fish pathogens.in vitro and in vivo test were carried out with tiamulin and gram-negative bacterial pathogens of fish. determination of minimum inhibitory concentration for 51 strains of seven species of gram-negative bacterial pathogens showed that only strains of vibrio anguillarum were sensitive at 1.6-6.25 ppm, while the rest of test strains required 25- greater than 100 ppm. control of infection was not achieved when tiamulin was fed for 14 days at 5 or 50 mg/kg to rainbow trout (salmo gairdneri) experime ...19883127604
purification and some properties of a chloramphenicol acetyltransferase mediated by plasmids from vibrio anguillarum.vibrio anguillarum strains were isolated from chloramphenicol-resistant bacteria in diseased fish. plasmid rms418, which confers chloramphenicol resistance, was transferred from v. anguillarum gn11379 to escherichia coli k12 by conjugation. the rms418-encoded chloramphenicol acetyltransferase (cat) [ec 2.3.1.99] was isolated and purified to homogeneity using affinity chromatography on immobilized p-amino-chloramphenicol or atp. the general cat could be adsorbed by a matrix with a chloramphenicol ...19883146569
study of vibrio anguillarum strains from different sources with emphasis on ecological and pathobiological properties.a total of 317 vibrio anguillarum strains were isolated from water, sediment, and diseased as well as healthy rainbow trout at a danish mariculture farm and from feral fish caught close to the farm. all strains were examined serologically. ten sera permitted determination of the o group in 66.7% of the strains from diseased rainbow trout. furthermore, the o group could be determined in 45.1 to 65.4% of the strains from mucus, gills, and intestinal contents of healthy rainbow trout, while only 22 ...19883190226
vibrio anguillarum antigen stimulates mitogenesis and polyclonal activation of salmonid lymphocytes.an antigen preparation of vibrio anguillarum, a salmonid pathogen, acts as a potent in vitro mitogenic stimulator of splenic and pronephric (anterior kidney) lymphocytes from coho salmon (oncorhynchus kisutch), chinook salmon (o. tshawytscha) and rainbow trout (salmo gairdneri). this antigen (va) is comparable in its mitogenic activity to concanavalin a (con a), escherichia coli lipopolysaccharide (lps), and phytohemagglutinin (pha). va gives peak mitogenic responses in coho five days after init ...19873315760
increased production of the siderophore anguibactin mediated by pjm1-like plasmids in vibrio anguillarum.the virulence of the fish pathogen vibrio anguillarum 775 is mediated by the pjm1 plasmid-specified iron uptake system which is expressed under conditions of iron limitation. other v. anguillarum strains isolated from various geographical locations harbor plasmids that are highly related to pjm1 and that are also associated with the high-virulence phenotype of these strains. in this work, we found that a pjm1-like plasmid, pjhc1, from one of these virulent strains encoded an iron uptake system t ...19883372022
purification and properties of agglutinins from conger eel, conger myriaster (brevoort), skin mucus.the skin mucus of the conger eel, conger myriaster, contains galactose-specific agglutinins. hemagglutinating activity is independent of divalent cations and is destroyed by heating at 50 degrees c for 15 min. the mucus agglutinins, named congerins, are a mixture of proteins with different electrical charges. three of these molecules were isolated by affinity chromatography on acid-treated sepharose 4b and by ion-exchange chromatography. they are simple proteins with the same molecular weight of ...19883384160
quantitative measurement of endotoxin in rainbow trout (salmo gairdneri) serum by the chromogenic substrate method.the amount of endotoxin in serum collected from normal rainbow trout (salmo gairdneri) and trout inoculated with viable vibrio anguillarum or lipopolysaccharide (lps) extracted from bacteria was determined by the chromogenic substrate method. the mean values of endotoxin in four different groups of normal rainbow trout sera ranged from 31.9 to 65.3 pg/ml. when fish were inoculated with viable bacteria (1 x 10(8], they became septicaemic and a large amount of endotoxin ng/ml) was detected in the ...19873429359
salmonid b lymphocytes demonstrate organ dependent functional heterogeneity.the passive hemolytic plaque assay was used to examine the functional heterogeneity of antibody producing cells in salmonid immune organs. in this study, the antibody response to vibrio anguillarum antigens was induced by the injection of a somatic antigen extract. this antigen was also coated onto sheep red blood cells (srbc) for plaque forming cell (pfc) determination. previous studies have demonstrated that this response is antibody dependent and antigen specific (kaattari and irwin, 1985). t ...19863490046
antibacterial activity and mechanism of action of 3'-azido-3'-deoxythymidine (bw a509u).the thymidine analog 3'-azido-3'-deoxythymidine (bw a509u; azidothymidine [azt]) had potent bactericidal activity against many members of the family enterobacteriaceae, including strains of escherichia coli, salmonella typhimurium, klebsiella pneumoniae, shigella flexneri, and enterobacter aerogenes. azt also had bactericidal activity against vibrio cholerae and the fish pathogen vibrio anguillarum. azt had no activity against pseudomonas aeruginosa, gram-positive bacteria, anaerobic bacteria, m ...19873551832
structural elucidation of the o-specific polysaccharide of the phenol-phase soluble lipopolysaccharide of vibrio anguillarum.the structure of the o-specific polysaccharide of the phenol-soluble cellular lipopolysaccharide of vibrio anguillarum has been investigated. the studies involved the use of methylation analysis, partial hydrolysis with 48% hydrogen fluoride, smith degradation, oxidation with chromium trioxide, and comprehensive proton and carbon-13 nuclear magnetic resonance studies, in which one- and two-dimensional experiments were carried out. as a result of these studies it is proposed that the o-specific p ...19873566967
oxytetracycline residues in rainbow trout analysed by a rapid hplc method.a rapid and sensitive hplc method was developed for the determination of oxytetracycline in fish tissues (muscle and liver) based on a clean-up and concentration procedure on sep-pak c18. at a coastal fishfarm rainbow trout (salmo gairdneri) suffering from vibrio anguillarum were treated with 75 mg oxytetracycline per kg fish and day for ten days. oxytetracycline residues above the limit of determination (0.005 micrograms/g) were found in fish 82 days after treatment. the recoveries from spiked ...19873653453
virulence, cytotoxic and inflammatory activities of vibrio anguillarum and aeromonas salmonicida isolated from cultivated salmonid fish in sweden. 19873687646
phagocytic activity of macrophages of rainbow trout against vibrio anguillarum and the opsonising effect of antibody and complement.the phagocytosis of vibrio anguillarum by peritoneal macrophages from normal rainbow trout was enhanced by antibody and complement. treatment of either macrophages or the bacteria by antibody also enhanced opsonisation. five weeks after immunisation with v anguillarum, the phagocytic activity of macrophages from rainbow trout was increased significantly compared with the activity of those from unvaccinated fish. although agglutinin titres did not increase until three weeks after immunisation, se ...19863738229
effects of metals on the chemiluminescent response of rainbow trout (salmo gairdneri) phagocytes.quantification of an induced chemiluminescent (cl) response in phagocytes is currently being evaluated as an indicator system for determining those environmental pollutants that may predispose fish to disease. a cl assay was developed using phagocytes from the pronephros of rainbow trout (salmo gairdneri). the cl response of phagocytes to phorbol myristate acetate, a chemical inducer of cl, was shown to be dose-dependent. the response to five species of bacteria was also evaluated. staphylococcu ...19863765345
immunization of carp (cyprinus carpio) with a vibrio anguillarum bacterin: indications for a common mucosal immune system.uptake and transport of formalin-killed vibrio anguillarum bacteria were studied in the second gut segment of carp and the resulting reaction of the immune system was investigated. within a few hours after anal administration antigenic determinants of bacteria were present in intraepithelial macrophages of the second gut segment. in gut and skin mucus and bile, immunoglobulins (ig's) were detected, but the amount was much lower than found in serum. in mucus of the second gut segment, 4 times mor ...19863770268
vibrio anguillarum: prevalence of typical and atypical strains in marine recipients with special reference to carbohydrate pollution. 19853836565
cloning and expression in escherichia coli of vibrio parahaemolyticus thermostable direct hemolysin and thermolabile hemolysin genes.two hemolysin genes of vibrio parahaemolyticus wp1, a thermostable direct (tsd) hemolysin gene and a thermolabile hemolysin gene, were cloned into the pbr322 vector in escherichia coli k-12 c600. a large amount of the tsd hemolysin produced in e. coli k-12 accumulated in the periplasmic space. the tsd hemolysin gene was localized on a 0.9-kilobase hindiii-bamhi fragment by identifying qualitatively the production of the tsd hemolysin by a reverse passive hemagglutination assay in the osmotic sho ...19853886626
salmonid spleen and anterior kidney harbor populations of lymphocytes with different b cell repertoires.an o-antigen extract of the fish pathogen vibrio anguillarum was used to stimulate plaque-forming cells (pfc) in coho salmon. the kinetics of the response demonstrates peak pfc per organ on day 16 post immunization for the spleen and anterior kidney. significant pfc were also seen in thymic tissue. pfc responses were shown to be immunoglobulin mediated and antigen specific. inhibition profiles of responding pfc populations demonstrate that the cells of the anterior kidney are more restricted in ...19853899761
serotyping of vibrio anguillarum.a serotyping scheme based on the detection of o antigens by slide agglutination in fish-pathogenic strains of vibrio anguillarum is presented. over a period of 5 years 270 vibrio strains from feral and cultured fish, 189 strains from the environment, and 36 strains from invertebrates were collected. the strains were divided into 10 distinct serotypes (o1 through o10). more than 90% of the fish-pathogenic strains, but only 40% of the environmental strains, were typable; 71% of the strains isolate ...19863963811
iron-regulated outer membrane protein om2 of vibrio anguillarum is encoded by virulence plasmid pjm1.vibrio anguillarum 775 harboring the virulence plasmid pjm1 synthesized an outer membrane protein of 86 kilodaltons, om2, that was inducible under conditions of iron limitation. pjm1 dna fragments obtained by digestion with restriction endonucleases were cloned into cosmid vectors and transferred into escherichia coli. the om2 protein was synthesized in e. coli, demonstrating that it is actually encoded by the pjm1 plasmid. mobilization of the recombinant plasmids to v. anguillarum was accomplis ...19853968037
characterization of extracellular substance of vibrio anguillarum toxic for rainbow trout and mice.an extracellular toxic substance was separated from the cell-free culture filtrate of vibrio anguillarum (strain ncmb571). two fractions (gi and gii + iii) obtained by sephadex g-200 chromatography following deae-cellulose chromatography were lethal to rainbow trout and mice. material separated from the gi fraction by sepharose 4b affinity chromatography (gi-a fraction) was lethal to these animals. by sodium dodecylsulfate polyacrylamide gel electrophoresis, the gi and gi-a fractions were found ...19854079843
vibrio anguillarum isolated from a nasal abscess of the cod fish (gadus morhua). 19725049109
vibrio anguillarum as a cause of disease in winter flounder (pseudopleuronectes americanus). 19725081124
characterization and relatedness of marine vibrios pathogenic to fish: physiology, serology, and epidemiology.cultural characteristics and serological relationships of pathogenic marine vibrios isolated from fish in the pacific northwest were studied. these organisms were compared with cultures of vibrio anguillarum, a known fish pathogen. on the basis of morphological and cultural characteristics, the pacific northwest strains of vibrio were found to be closely related to v. anguillarum. serological analyses of thermostable antigens served to distinguish three serotypes among the vibrios. serotype 1 wa ...19695391230
vibriosis (vibrio anguillarum) as an epizootic disease in rainbow trout (salmo gairdneri). 19705532773
vibrio anguillarum from an epizootic in rainbow trout (salmo gairdneri) in the u.s.a. 19685752922
molecular cloning and expression of genetic determinants for the iron uptake system mediated by the vibrio anguillarum plasmid pjm1.plasmid pjm1 from an invasive strain of vibrio anguillarum encodes an iron uptake system which mediates the biosynthesis of a siderophore and a membrane receptor for the iron-siderophore complex. this system has been associated with the ability of v. anguillarum to cause hemorrhagic septicemic disease in marine fish. recombinant derivatives containing essential regions of the pjm1-mediated iron uptake system cloned into cosmid vector pvk102 were introduced into low-virulence iron uptake-deficien ...19846094511
characterization of the surface antigens of the marine fish pathogens vibrio anguillarum and vibrio ordalii.the technique of immunoblotting was used to identify the surface antigens of the marine vibrios pathogenic for fish, vibrio anguillarum and vibrio ordalii. polyclonal antisera raised in rabbits to strains representing the two most common serotypes causing vibriosis in fish in north america were used. the results demonstrated that antigenic specificity was conferred by the lipopolysaccharides, with three serotypes being displayed among the strains examined. the lipopolysaccharides of strains chos ...19846204733
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