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molecular cloning, nucleotide sequence, and expression in escherichia coli of a hemolytic toxin (aerolysin) gene from aeromonas trota.aeromonas trota ak2, which was derived from atcc 49659 and produces the extracellular pore-forming hemolytic toxin aerolysin, was mutagenized with the transposon mini-tn5km1 to generate a hemolysin-deficient mutant, designated strain ak253. southern blotting data indicated that an 8.7-kb noti fragment of the genomic dna of strain ak253 contained the kanamycin resistance gene of mini-tn5km1. the 8.7-kb noti dna fragment was cloned into the vector pgem5zf(-) by selecting for kanamycin resistance, ...19989647817
aeromonas as a cause of segmental colitis. 19989647047
[ocular infection caused by aeromonas spp.: considerations on its treatment and importance of resistance to antimicrobial agents]. 19989646574
effects of mercuric chloride and sodium selenite on some immune responses of blue gourami, trichogaster trichopterus (pallus).the immunotoxicological effects of mercuric chloride and sodium selenite on blue gourami were studied. some immune responses ranging from non-specific to specific were investigated. these include tissue lysozyme activity, kidney lymphocyte proliferation and plasma agglutinating antibody titre against bacteria. after 2 weeks of chronic exposure, 0.09 mg/l of hg2+ alone induced a significant increase of kidney lysozyme activity of 4196.3 +/- 1171.0 u/g, but it decreased to 1577.4 +/- 902.4 u/g whe ...19989646523
antimicrobial activity in the skin of the channel catfish ictalurus punctatus: characterization of broad-spectrum histone-like antimicrobial proteins.three antibacterial proteins were isolated from acid extracts of channel catfish (ictalurus punctatus) skin by cation exchange chromatography and reverse-phase high-pressure liquid chromatography. the molecular masses of the proteins were 15.5, 15.5 and 30 kd as determined by sds-polyacrylamide gel electrophoresis. mass spectrometry, amino acid composition and amino acid sequence data suggest that the most abundant protein is closely related to histone h2b. the h2b-like protein was inhibitory to ...19989645227
the effect of suspending solution supplemented with marine cations on the oxidation of biolog gn microplate substrates by vibrionaceae bacteria.bacteria belonging to the family vibrionaceae were suspended using saline and a solution prepared from a marine-cations supplement. the effect of this on the profile of oxidized substrates obtained when using biolog gn microplates was investigated. thirty-nine species belonging to the genera aeromonas, listonella, photobacterium, and vibrio were studied. of the strains studied, species of listonella, photobacterium, and vibrio could be expected to benefit from a marine-cations supplement that co ...19989643966
aeromonas hydrophila adenylyl cyclase 2: a new class of adenylyl cyclases with thermophilic properties and sequence similarities to proteins from hyperthermophilic archaebacteria.complementation of an escherichia coli cya mutant with a genomic library from aeromonas hydrophila allowed isolation of clones containing two different cya genes. whereas one of these genes (cyaa) coded for an adenylyl cyclase (ac1) belonging to the previously described class i adenylyl cyclases (acs), the second one (cyab) coded for a protein (ac2) that did not match any previously characterized protein when compared to protein sequence databases. in particular, it did not align with any of mem ...19989642185
macromolecular assembly and secretion across the bacterial cell envelope: type ii protein secretion systems.a decade ago, pugsley and colleagues reported the existence of a large region of klebsiella dna, distinct from the klebsiella gene encoding pullulanase, which was necessary for secretion of this enzyme to the cell surface in escherichia coli (d'enfert et al., 1987a,b). the pul genes it contained proved to be the tip of an iceberg. the sequences reported before 1992 (d'enfert et al., 1987a,b; d'enfert & pugsley, 1989; pugsley & reyss, 1990; reyss & pugsley, 1990) included only one gene (puld) tha ...19989641973
sialic acid-binding lectin with antibacterial activity from the horse mussel: further characterization and immunolocalization.a heterogeneous sialic acid-binding lectin with affinity for bacterial lps was isolated and partially characterized from hemolymph of the horse mussel modiolus modiolus.(1) using two-dimensional electrophoresis with immobilized ph gradients, the lectin revealed three subunits with different molecular weight and isoelectric points (pi); mr14 (pi approximately 5.1 and approximately 5.5), 17.5 (pi approximately 5.5) and 20 (pi approximately 4.9) kda. the affinity purified lectin existed in its nati ...19989639084
the isolation of pseudomonas and other gram(-) psychrotrophic bacteria in raw milks.in this research, totally 200 raw milk samples in different areas of ankara were collected from various dairy plant. the isolated psychotrophic bacteria from the raw milk samples are the species of pseudomonas spp., acinetobacter spp., alcaligenes and aeromonas. isolation of pseudomonas and other gram(-) psychrotrophic bacteria types are determined as p. aeruginosa 11 (5.5%), p. putida 11 (5.5%), p. fluorescens biotype i 10 (5.0%), p. fluorescens biotype ii 4 (2.0%), p. fluorescens biotype iii 6 ...19989637013
occurrence of gram-negative bacteria in drinking water undergoing softening treatment.a study was carried out on the presence of gram-negative bacteria in the municipal waters of bologna (italy) undergoing softening using domestic ion exchangers with an automatic disinfection mechanism. the softening process was seen to cause a 15 fold increase in 22 degrees c and 36 degrees c heterotrophic plate counts. there was a 30 fold increase in gram-negative bacteria and their number correlated directly with temperature and inversely with active residual chlorine. organic matter had no ef ...19979636986
colonization of vibrio pelagius and aeromonas caviae in early developing turbot (scophthalmus maximus l.) larvae.polyclonal antisera made in rabbits against whole washed cells of vibrio pelagius and aeromonas caviae were used for detection of these bacterial species in the rearing water and gastrointestinal tract of healthy turbot (scophthalmus maximus) larvae exposed to v. pelagius and/or aer. caviae. the results demonstrated that this method is suitable for detection of v. pelagius and aer. caviae in water samples and larvae at population levels higher than 10(3) ml-1 and 10(3) larva-1. populations of ae ...19989633637
the effect of essential oils of basil on the growth of aeromonas hydrophila and pseudomonas fluorescens.basil essential oils, including basil sweet linalool (bsl) and basil methyl chavicol (bmc), were screened for antimicrobial activity against a range of gram-positive and gram-negative bacteria, yeasts and moulds using an agar well diffusion method. both essential oils showed antimicrobial activity against most of the micro-organisms examined except clostridium sporogenes, flavimonas oryzihabitans, and three species of pseudomonas. the minimum inhibitory concentration (mic) of bmc against aeromon ...19989633630
men 10700, a new penem antibiotic: in-vitro activity and its correlation with beta-lactamase stability, pbp affinity and diffusion through the bacterial cell wall.the in-vitro activity of men 10700, a novel penem, was compared with that of imipenem, ritipenem, ampicillin/sulbactam, cefotaxime, ciprofloxacin and amikacin against 1088 strains taken from 21 genera, including gram-negative, gram-positive and anaerobic bacteria. mic data showed that men 10700 was very active against staphylococci and streptococci (mic90 < or = 0.5 mg/l) and against most members of the enterobacteriaceae (mic90 < or = 2 mg/l), with reduced activity only against providencia stua ...19989630405
aeromonas hydrophila soft-tissue infection as a complication of snake bite: report of three cases.aeromonas hydrophila soft-tissue infection has been associated with fish and reptile bites. there have bee three recent cases from brazil of abscesses complicating snake bites in which a. hydrophila was isolated from the purulent exudates. one of the snakes responsible for the bites was a specimen of bothrops moojeni, and the others were most probably also lance-headed vipers. these snakes have a local necrotizing, myotoxic, oedema-inducing venom that must have favoured the multiplication in the ...19989625918
purification and characterization of an arylamine n-acetyltransferase from the bacteria aeromonas hydrophilia.n-acetyltransferase from aeromonas hydrophilia was purified by ultrafiltration, deae-sephacel, gel filtration chromatography on sephadex g-100, and deae-5pw on high performance liquid chromatography, as judged by sodium dodecyl sulfate-polyacrylamine gel electrophoresis (sds-page) on a 12.% (wt/vol) slab gel. the enzyme had a molecular mass 44.9 kda. the purified enzyme was thermostable at 37 degrees c for 1 h with a half-life 28 min at 37 degrees c, and displayed optimum activity at 37 degrees ...19989625795
incidence of non-01 vibrio cholerae and aeromonas spp. in fresh water in araraquara, brazil.the occurrence of aeromonas spp., vibrio cholerae, and plesiomonas shigelloides in fresh water from various sources in araraquara, state of são paulo, brazil was determined. samples from ten distinct irrigation systems used in vegetable cultivation, from five distinct streams, from two reservoirs, from one artificial lake, and from three distinct springs were analyzed. all isolates were serotyped and tested for hemolysin, cytotoxin, heat-stable (st) and heat-labile (lt) enterotoxins production; ...19989625786
diverse and related 16s rrna-encoding dna sequences in prostate tissues of men with chronic prostatitis.treatment of chronic prostatitis/chronic pelvic pain syndrome is often empirical because clinical culture methods fail to detect prostate-associated pathogens in >90% of patients. previously, we tested a variety of specific-microorganism pcrs and began a dna sequence study after we found that 77% of prostatitis patients were pcr positive for prokaryotic rrna-encoding dna sequences (rdnas) despite negative cultures using optimal techniques. in the present study, 36 rdna clones from 23 rdna-positi ...19989620394
characterization of a 14 kda plant-related serine protease inhibitor and regulation of cytotoxic activity in earthworm coelomic fluid.we have purified and characterized the serine protease inhibitor activity contained in the coelomic fluid of the earthworms, eisenia. serine protease inhibitor activity was stable between ph3 and 9.5, not flocculable by ph 3.0 and resistant to 100 degrees c for 15 min. or to 4 degrees c for 24 h. ten microl of coelomic fluid was sufficient to inhibit in vitro the protease activity of 0.12 microgram of trypsin. injection of living bacteria into earthworms resulted in increased serine protease act ...19989617579
immunostimulatory effects of dimerized lysozyme (klp-602) on the nonspecific defense mechanisms and protection against furunculosis in salmonids.utilization of natural immunostimulants in fish culture offers a wide range of attractive methods for inducing and building protection against diseases. lysozyme is an enzyme with bacteriolytic properties and is ubiquitous in its distribution among living organisms. this enzyme has antiviral, antibacterial and anti-inflammatory properties. in nature, lysozyme is found as a monomer. lysozyme dimer is significantly less toxic than its monomer, and its high biological activity has been ascertained ...19989613448
selenite bioremediation potential of indigenous microorganisms from industrial activated sludge.ten bacterial strains were isolated from the activated sludge waste treatment system (biox) at the exxon refinery in benicia, california. half of these isolates could be grown in minimal medium. when tested for selenite detoxification capability, these five isolates (members of the genera bacillus, pseudomonas, enterobacter and aeromonas), were capable of detoxifying selenite with kinetics similar to those of a well characterized bacillus subtilis strain (168 trp+) studied previously. the seleni ...19979608517
bacterial density and survey of cultivable heterotrophs in the surface water of a freshwater marsh habitat of anopheles quadrimaculatus larvae (diptera: culicidae).we examined surface water samples collected in september and october 1994 from a freshwater marsh habitat containing larval anopheles quadrimaculatus mosquitoes. bacterial densities in direct microscopic counts ranged from 9.7 x 10(5) to 1.3 x 10(7) cells/ml. densities of cultivable bacteria on trypticase soy agar medium ranged from 1.0 to 1.5 x 10(5) cells/ml. the majority of 888 isolates were gram-positive rods (41%) followed by gram-negative rods (28%). analysis of the cellular fatty acid pro ...19989599327
aeromonas hydrophila septicaemia in a grey seal (halichoerus grypus) from the baltic sea: a case study.aeromonas hydrophila septicaemia in a grey seal (halichoerus grypus) from the swedish part of the baltic sea is reported. the pathogen was isolated from both lung and spleen specimens. all of the a. hydrophila isolates produced haemolysin and vero active cytotoxin. the aerolysin gene was found in all tested isolates as evidenced by the polymerase chain reaction (pcr) technique. also, all isolates tested showed identical patterns of biochemical and antibiotic resistance. as aeromonas spp. commonl ...19989597306
interactions of bacteria with contact lenses: the effect of soluble protein and carbohydrate on bacterial adhesion to contact lenses.during contact lens wear, components in the tear film adsorb to the contact lens surface to form a protein-rich coating. in this study, we investigated the importance of these components in bacterial adhesion to contact lenses.19989586751
[selective screening of enterohemorrhagic escherichia coli depending on hemolysis on beutin's washed sheep blood agar plates].recently, beutin et al have studied hemolysin production in a large number of serologically diverse verotoxin (vt) producing escherichia coli (vtec) strains and found a close association between hemolysin production and vt production. this study was to examine whether hemolysin production on sheep blood agar can be used as an epidemiological marker for microbiological screening of enterohemorrhagic escherichia coli (ehec). in comparison of hemolysis on washed blood agar plates prepared in our la ...19989585695
efficient production of polyhydroxyalkanoates from plant oils by alcaligenes eutrophus and its recombinant strain.the ability of alcaligenes eutrophus to grow and produce polyhydroxyalkanoates (pha) on plant oils was evaluated. when olive oil, corn oil, or palm oil was fed as a sole carbon source, the wild-type strain of a. eutrophus grew well and accumulated poly(3-hydroxybutyrate) homopolymer up to approximately 80% (w/w) of the cell dry weight during its stationary growth phase. in addition, a recombinant strain of a. eutrophus phb-4 (a pha-negative mutant), harboring a pha synthase gene from aeromonas c ...19989581296
fructose and mannose metabolism in aeromonas hydrophila: identification of transport systems and catabolic pathways.aeromonas hydrophila was examined for fructose and mannose transport systems. a. hydrophila was shown to possess a phosphoenolpyruvate (pep): fructose phosphotransferase system (fructose-pts) and a mannose-specific pts, both induced by fructose and mannose. the mannose-pts of a. hydrophila exhibited cross-reactivity with escherichia coli mannose-pts proteins. the fructose-pts proteins exhibited cross-reactivities with e. coli and xanthomonas campestris fructose-pts proteins. in a. hydrophila gro ...19989579084
characterization and sequence of the chryseobacterium (flavobacterium) meningosepticum carbapenemase: a new molecular class b beta-lactamase showing a broad substrate profile.the metallo-beta-lactamase produced by chryseobacterium (formerly flavobacterium) meningosepticum, which is the flavobacterial species of greatest clinical relevance, was purified and characterized. the enzyme, named blab, contains a polypeptide with an apparent mr of 26000, and has a pi of 8.5. it hydrolyses penicillins, cephalosporins (including cefoxitin), carbapenems and 6-beta-iodopenicillanate, a mechanism-based inactivator of active-site serine beta-lactamases. the enzyme was inhibited by ...19989576862
infected stingray injury. 19989576395
phenotypic identification of aeromonas genomospecies from clinical and environmental sources.a collection of 983 aeromonas isolates from environmental and clinical sources have been identified to the genomospecies level. a phenotypic method identified 93% of the strains. the use of citrate and the production of acid from sorbitol enabled the members of the aeromonas hydrophila complex to be separated. the most common genomospecies from intestinal sources were aeromonas veronii biotype sobria and aeromonas caviae. the former, together with a. hydrophila, was the most frequently isolated ...19989575026
defined deletion mutants demonstrate that the major secreted toxins are not essential for the virulence of aeromonas salmonicida.the importance of the two major extracellular enzymes of aeromonas salmonicida, glycerophospholipid: cholesterol acyltransferase (gcat) and a serine protease (aspa), to the pathology and mortality of salmonid fish with furunculosis had been indicated in toxicity studies. in this study, the genes encoding gcat (sata) and aspa (aspa) have been cloned and mutagenized by marker replacement of internal deletions, and the constructs have been used for the creation of isogenic sata and aspa mutants of ...19989573081
molecular characterization of the aeromonas hydrophila aroa gene and potential use of an auxotrophic aroa mutant as a live attenuated vaccine.the aroa gene of aeromonas hydrophila so2/2, encoding 5-enolpyruvylshikimate 3-phosphate synthase, was cloned by complementation of the aroa mutation in escherichia coli k-12 strain ab2829, and the nucleotide sequence was determined. the nucleotide sequence of the a. hydrophila aroa gene encoded a protein of 440 amino acids which showed a high degree of homology to other bacterial aroa proteins. to obtain an effective attenuated live vaccine against a. hydrophila infections in fish, the aroa gen ...19989573055
[aeromonas infections]. 19979569900
analysis of the interaction of aeromonas caviae, a. hydrophila and a. sobria with mucins.aeromonas species are known to be involved in human gastrointestinal diseases. these organisms colonize the gastrointestinal tract. aeromonas hydrophila, a. caviae, and a. sobria have been demonstrated microscopically to adhere to animal cell lines that express mucous receptors, but quantitative studies of adherence to mucosal components such as mucin have not been published to date. purified bovine submaxillary gland, hog gastric mucin, and fish skin mucin were used as a model to study mucin-bi ...19989566493
study of the microbial ecology of cold-smoked salmon during storage at 8 degrees c.microbiological, chemical and sensory changes in cold-smoked salmon were studied during 5 weeks of vacuum storage at 8 degrees c. the aerobic 20 degrees c viable count reached its maximum level after 6 days (3 x 10(6) cfu g-1) however, the shelf-life of the product was estimated to be 2 or 3 weeks by the panellists, confirming that there is no correlation between those two factors. acid, pungent, sour and rancid odours and flavours and pasty texture were the main spoilage characteristics. trimet ...19989562883
a further review of the microbiological quality of bottled water sold in canada: 1992-1997 survey results. 19989553800
evaluation of the role of carnobacterium piscicola in spoilage of vacuum- and modified-atmosphere-packed cold-smoked salmon stored at 5 degrees c.the microflora on spoiled cold-smoked salmon often consists of a mixture of lactic acid bacteria (lab) and gram-negative bacteria. to elucidate the role of the different groups, a storage trial was carried out in which nisin and co2 were used for the selective inhibition of the two bacterial groups. the shelf-life of vacuum-packed cold-smoked salmon, recorded by sensory evaluation, was four weeks at 5 degrees c and the microflora was composed of lab (10(6)-10(7) cfu/g) with an associate gram-neg ...19989553794
misidentification of unusual aeromonas species as members of the genus vibrio: a continuing problem.two unusual cases of aeromonas infection are described, one associated with bacteremia (aeromonas schubertii) and another in which the organism was recovered from an infected gall bladder (aeromonas veronii biotype veronii). these strains were initially identified as vibrio damsela and vibrio cholerae by the vitek and api 20e systems, respectively. use of appropriate screening tests and familiarity with the newer aeromonas species could prevent initial misidentifications and potential public hea ...19989542946
identification of lactococcus garvieae by pcr.lactococcus garvieae junior synonym enterococcus seriolicida) is an emerging zoonotic agent isolated from economically important fish (rainbow trout and yellowtail), from cattle, and from humans. clindamycin susceptibility is the only phenotypic test which can differentiate l. garvieae from lactococcus lactis, another emerging agent in humans. a pcr assay for the identification of l. garvieae was developed and resulted in an amplified fragment of 1,100 bp in size. the pcr assay was shown to be s ...19989542921
activity of bacterial phosphomonoesterases in batch culture.the phosphomonoesterases catalyse the hydrolysis of primary esters of phosphoric acid which help the bacteria to survive in phosphate stressed environment. ninety-five bacterial isolates were obtained from domestic sewage and industrial effluents of gelatine and soap factories at jabalpur on a medium enriched with phosphate and were screened for phosphatase production. the phosphatase producers were tentatively identified as escherichia coli, vibrio vulnificus, aeromonas hydrophila, staphylococc ...19989536656
food borne microbial pathogens of cultured aquatic species.this article provides a broad overview of microbial pathogens associated with marine and fresh water aquatic animals, including vibrio species, escherichia coli, streptococcus iniae, salmonella species, and edwardsiella tarda. historically, cultured fish were not considered important vectors of human pathogens. this situation is changing, partly due to increasing animal densities as a consequence of a rapidly growing industry and partly due to increasing awareness by health care providers of pat ...19989532670
detection of psychotrophic aeromonads in drinking water.in the context of evaluating their pathogenic relevance, culture on solid media is the only approach presently suitable for culture of psychotrophic aeromonads from drinking water. in this respect, a check must be effected to ensure that the culture medium cannot engender selective culture losses for individual species. for this reason, media to which e.g. ampicillin has been added are unsuitable.19989531729
nucleotide and amino acid sequences of the metallo-beta-lactamase, imis, from aeromonas veronii bv. sobria.the aeromonas veronii bv. sobria metallo-beta-lactamase gene, imis, was cloned. the imis open reading frame extends for 762 bp and encodes a protein of 254 amino acids with a secreted modified protein of 227 amino acids and a predicted pi of 8.1. to confirm the predicted sequence, purified imis was digested and the resulting peptides were identified, yielding an identical sequence for imis, with 98% identity to cpha. both possessed the putative active-site sequence asn-tyr-his-thr-asp at positio ...19989527802
indwelling device-related and recurrent infections due to aeromonas species.from october 1995 to february 1997, 13 isolates of aeromonas species were recovered from four patients treated at national taiwan university hospital (taipei). one of the patients, a diabetic, had simultaneous aeromonas veronii biotype veronii bacteremia and a. veronii biotype sobria urinary tract infection. seven weeks after the episode, the patient had necrotizing fasciitis due to a. veronii biotype veronii. the other three patients all had underlying hepatobiliary malignancies complicated by ...19989524839
rflp-pcr analysis of the aroa gene as a taxonomic tool for the genus aeromonas.the aroa gene has been identified as a target in screening for the presence of most aeromonas species so far described by pcr. synthetic oligonucleotide primers of 24 and 25 nucleotides were used by pcr assay to amplify a sequence of the aroa gene, which encodes 3-phosphoshikimate-1-carboxyvinyltransferase, a key enzyme of aromatic amino acids and folate biosynthetic pathway. a 1236-bp dna fragment, representing most of the aroa gene, according to the nucleotide sequence of a. salmonicida, was a ...19979513265
isolation and identification of aeromonas species from human stools.one thousand and three diarrhoeal stool samples were processed in our laboratory during the period 1996/1997 for the presence of enteric pathogens especially aeromonas spp., which has emerged as a new agent causing diarrhoea. ampicillin sheep blood agar was found to be the best medium for the isolation of aeromonas spp. from stool specimens. enteric pathogens were found in 200 (20%) stools, of which aeromonas spp. was the second commonest pathogen isolated amounting to 21% of isolates. this stud ...19979510643
effects of temperature, medium composition, ph, salt and dissolved oxygen on haemolysin and cytotoxin production by aeromonas hydrophila isolated from oyster.the effects of temperature, medium composition, ph, salt content and dissolved oxygen (do) on the production of haemolysin and cytotoxin by one strain of aeromonas hydrophila isolated from oyster were investigated. four media were tested: brain heart infusion broth (bhib), casamino acid-yeast extract broth (cayeb), nutrient broth (nb), and trypticase soy broth (tsb). bhib was the best for toxin production even though the growth rates for aeromonas hydrophila in all of these media were quite simi ...19979506276
enzyme characteristics of beta-d-galactosidase- and beta-d-glucuronidase-positive bacteria and their interference in rapid methods for detection of waterborne coliforms and escherichia coli.bacteria which were beta-d-galactosidase and beta-d-glucuronidase positive or expressed only one of these enzymes were isolated from environmental water samples. the enzymatic activity of these bacteria was measured in 25-min assays by using the fluorogenic substrates 4-methylumbelliferyl-beta-d-galactoside and 4-methylumbelliferyl-beta-d-glucuronide. the enzyme activity, enzyme induction, and enzyme temperature characteristics of target and nontarget bacteria in assays aimed at detecting colifo ...19989501441
the role of seafood in foodborne diseases in the united states of america.in the united states of america, seafood ranked third on the list of products which caused foodborne disease between 1983 and 1992. outbreaks connected with fish vectors were caused by scombroid, ciguatoxin, bacteria and unknown agents; in shellfish, unknown agents, paralytic shellfish poisoning, vibrio spp. and other bacteria, followed by hepatitis a virus, were responsible for the outbreaks. at least ten genera of bacterial pathogens have been implicated in seafood-borne diseases. over the pas ...19979501377
meat from dairy cows: possible microbiological hazards and risks.the authors provide an overview of the circumstances associated with culling of dairy cattle in the united states of america (usa) and focus on the possible significant microbiological hazards associated with meat from cull dairy cows. cull dairy cows are an important source of food in the usa, accounting for at least approximately 17% of ground beef. the potential microbiological hazards for foodborne illness from cull dairy cows discussed here include salmonella (with special attention to s. t ...19979501354
acute rhabdomyolysis and myonecrosis complicating aeromonas bacteremia in neutropenic patients with hematologic malignancies: report of two cases.infections by aeromonas spp. are a rare cause of systemic infection in normal and immunocompromised hosts. we report the cases of two patients with acute non-lymphoblastic leukemia who developed septic shock by aeromonas species with unusual soft-tissue complications. one patient who was undergoing consolidation chemotherapy developed septic shock by aeromonas hydrophila with rhabdomyolysis and subsequent soft-tissue destruction consistent with myonecrosis. she recovered with combination antibio ...19979499669
virulence factors in aeromonas spp and their association with gastrointestinal disease.culture filtrates of eight aeromonas strains isolated from the feces of 487 subjects (292 diarrhoeic patients and 195 asymptomatic subjects) were tested for toxin production in cho and mccoy cells, and adhesion and invasive ability in caco-2 cells. among these isolates, three aeromonas sobria and one aeromonas caviae strains possessed virulence-associated properties. toxin production was the most common of the three virulence properties. two a. caviae were associated, in the absence of other dia ...19989497926
antibiotic resistance of bacterial litter isolates.use of antibiotics in subtherapeutic doses as growth-promoting feed additives for animal production is widespread in the u.s. and throughout the world. previous studies by our research group concluded that size fractionation of poultry (broiler) litter followed by storage facilitated reutilization of litter as a soil amendment or bedding supplement. however, litter microbial contamination, including antibiotic-resistant populations, and accumulation of metals and other elements may limit litter ...19989495488
internalization of aeromonas hydrophila by fish epithelial cells can be inhibited with a tyrosine kinase inhibitor.aeromonas hydrophila is a gram-negative bacterium that is pathogenic in fish, causing motile aeromonad septicaemia. it can enter (invade) fish cells, and survive as an intracellular parasite. the host-pathogen interaction and signal transduction pathway were studied by screening signal transduction inhibitors using carp epithelial cells and a virulent strain of the bacterium, ppd134/91. genistein, a tyrosine kinase inhibitor, postponed internalization of a. hydrophila into host cells, suggesting ...19989493367
inactivation of two haemolytic toxin genes in aeromonas hydrophila attenuates virulence in a suckling mouse model.the contribution of two unrelated aeromonas hydrophila beta-haemolytic toxins to virulence was assessed in a suckling mouse model. the first haemolysin gene, isolated from an a. hydrophila a6 cosmid bank, encoded a potential gene product of 621 amino acids and a predicted molecular size of 69.0 kda. the inferred amino acid sequence showed 89% identity to the ahh1 haemolysin of a. hydrophila atcc 7966, and 51% identity to the hlya haemolysin of vibrio cholerae ei tor strain o17. the second haemol ...19989493366
further characterization of renibacterium salmoninarum extracellular products.renibacterium salmoninarum, the agent of bacterial kidney disease in salmonids, releases high concentrations of extracellular protein in tissues of infected fish. the extracellular protein consists almost entirely of a 57-kda protein and derivatives of degradation and aggregation of the same molecule. the 57-kda protein and its derivatives were fractionated into defined ranges of molecular mass. separated fractions continued to produce degradation and aggregation products. one-dimensional electr ...19979480644
reduction of diverse electron acceptors by aeromonas hydrophilaaeromonas hydrophila atcc 7966 grew anaerobically on glycerol with nitrate, fumarate, fe(iii), co(iii), or se(vi) as the sole terminal electron acceptor, but did not ferment glycerol. final cell yields were directly proportional to the amount of terminal electron acceptor provided. twenty-four estuarine mesophilic aeromonads were isolated; all reduced nitrate, fe(iii), or co(iii), and five strains reduced se(vi). dissimilatory fe(iii) reduction by a. hydrophila may involve cytochromes. differenc ...19989477259
gene cloning, nucleotide sequencing, and purification and characterization of the low-specificity l-threonine aldolase from pseudomonas sp. strain ncimb 10558.a low-specificity l-threonine aldolase (l-ta) gene from pseudomonas sp. strain ncimb 10558 was cloned and sequenced. the gene contains an open reading frame consisting of 1,041 nucleotides corresponding to 346 amino acid residues. the gene was overexpressed in escherichia coli cells, and the recombinant enzyme was purified and characterized. the enzyme, requiring pyridoxal 5'-phosphate as a coenzyme, is strictly l specific at the alpha position, whereas it cannot distinguish between threo and er ...19989464392
expression and characterization of (r)-specific enoyl coenzyme a hydratase involved in polyhydroxyalkanoate biosynthesis by aeromonas caviae.complementation analysis of a polyhydroxyalkanoate (pha)-negative mutant of aeromonas caviae proved that orf3 in the pha locus (a 402-bp gene located downstream of the pha synthase gene) participates in pha biosynthesis on alkanoic acids, and the orf3 gene is here referred to as phaj(ac). escherichia coli bl21(de3) carrying phaj(ac). under the control of the t7 promoter overexpressed enoyl coenzyme a (enoyl-coa) hydratase, which was purified by one-step anion-exchange chromatography. the n-termi ...19989457873
a pore-forming toxin interacts with a gpi-anchored protein and causes vacuolation of the endoplasmic reticulum.in this paper, we have investigated the effects of the pore-forming toxin aerolysin, produced by aeromonas hydrophila, on mammalian cells. our data indicate that the protoxin binds to an 80-kd glycosyl-phosphatidylinositol (gpi)-anchored protein on bhk cells, and that the bound toxin is associated with specialized plasma membrane domains, described as detergent-insoluble microdomains, or cholesterol-glycolipid "rafts." we show that the protoxin is then processed to its mature form by host cell p ...19989456314
[detection of the thermostable lipase gene (lipah) in clinical isolates of aeromonas hydrophila belonging to the major o serogroups o11, o16 and o34].on the basis of dna hybridization data and phenotypes, most pathogenic strains of aeromonas hydrophila are grouped into hybridization group 1 (hg1). these pathogenic strains secret the theromostable lipase, and it's gene (lipah) has been cloned and sequenced. the present study was performed to identify the pathogenic strains of a. hydrophila by the polymerase chain reaction (pcr) technique to detect the lipah. synthetic oligonucleotide primers (lipah-ns-1 and -na-1) were used in the pcr. the pcr ...19979455059
virulence factors-pathogenicity relationships for aeromonas species from clinical and food isolates.the presence of virulence factors in 96 aeromonas strains isolated from food and clinical samples was studied. neither cytotoxic activity and hydrophobicity, not the presence of pili or an extra surface layer made it possible to establish differences between food and clinical strains. statistical studies showed that cytotoxin production was associated with a positive voges-proskauer reaction, inability to ferment arabinose and a positive lysine decarboxylation. therefore, when comparing cytotoxi ...19979449784
small bowel and fecal microbiology in children suffering from persistent diarrhea in bangladesh.the etiology of persistent diarrhea in children is multifactorial. the objective of the current study was to ascertain the role of microorganisms in the etiology and pathogenesis of persistent diarrhea in a group of children in bangladesh.19989443113
effect of amino acids on the production and activities of cephalosporin c acylase and penicillin v acylase from aeromonas species acy 95.amino acids altered the production and activities of cephalosporin c acylase and penicillin v acylase from aeromonas species acy 95 to a varying degree. dl-tryptophan enhanced the cephalosporin c acylase formation by 222% while suppressed the penicillin v acylase formation by 68%.19979440283
detection and speciation of bacteria through pcr using universal major cold-shock protein primer oligomers.the detection of bacteria using pcr is a well-established diagnostic technique. however, conventional pcr requires the use of dna primer oligomers that are specific to the target organism and, as a consequence, a sample can only be tested for the presence of that specific target. a significant advantage would be to probe a sample for the presence of any bacteria, followed by identification. to achieve this it is necessary to identify a dna sequence common to all bacteria. here we demonstrate tha ...19979439003
characterization of the bacterial flora isolated from a pilot-scale lagoon processing swine manure.the bacterial flora of an experimental plant that processes liquid swine manure by an aerated compartmented (multistage) lagoon system has been studied. the total flora is characterized by a larger number of oligotrophic bacteria than eutrophic ones. each compartment displays a specific flora, different from the flora in the manure, and consisting of a complex assembly of gram-negative and gram-positive ubiquitous species, such as aeromonas spp. and pseudomonas spp., and specialized species, suc ...19979436310
[prevalence of aeromonas spp. in surface water].some aeromonas strains are well recognized enteropathogens according to microbiological, clinical, immunological and epidemiological evidence. the main source of infection seems to be untreated water, these microorganisms can be found in virtually all aquatic environments. additionally, some aeromonas, which include enterotoxigenic strains, are capable of rapid growth at 5 degrees c and even of producing toxins. vegetable products irrigated with contaminated water may reach critical aeromonas le ...19979429640
movement of a loop in domain 3 of aerolysin is required for channel formation.aerolysin is a channel-forming toxin that must oligomerize in order to become insertion-competent. modeling based on the crystal structure of the proaerolysin dimer and electron microscopic images of the oligomer indicated that a loop in domain 3 must move away from the beta-sheet that forms the main body of the protein before oligomerization can proceed. in order to determine if movement actually occurs, strategically located amino acids in the loop and in the sheet were replaced with cysteines ...19989425098
enzyme-linked immunosorbent assay for detection of aeromonas hydrophila serogroup o:19.an enzyme-linked immunosorbent assay has been developed for the detection of aeromonas hydrophila serogroup o:19 isolated from epizootics in eels. the enzyme-linked immunosorbent assay specificity was confirmed after testing a. hydrophila o:19 and non-o:19 strains from different origins, as well as other aeromonas species and other fish pathogens such as vibrio vulnificus biotype 2, v. furnisii, v. damsela, yersinia ruckerii and edwardsiella tarda. the detection limits for a. hydrophila o:19 cel ...19979418247
reappraisal of the effect of temperature on the growth kinetics of aeromonas salmonicida.the effect of temperature (1-34 degrees c) on the maximum specific growth rate of aeromonas salmonicida could not be described by the classical growth models; for some strains, two optimal temperatures at 23 degrees c and 30 degrees c were observed, as well as an unexpected increase in the pseudolag time above 27 degrees c. this could be explained by the presence of two subsets, notably s-layer+ and s-layer- sub-populations. the a- cells had higher growth parameters (topt and mu opt) than the a+ ...19979418073
antimicrobial activity among pseudomonas and related strains of mineral water origin.siderophore production in 382 pseudomonas and related strains of mineral water origin were screened and the antimicrobial activities of 158 of these tested against nine target organisms of health significance. presence of siderophores could be detected in 54.4% and the majority of strains tested (91.2%) inhibited at least one of the nine target strains. staphylococcus, escherichia coli and aeromonas hydrophila were particularly sensitive. addition of iron eliminated the inhibitory activity in 96 ...19979418027
pathogenicity of atypical aeromonas salmonicida in atlantic salmon compared with protease production.the pathogenicity of extracellular products (ecps) from 24 atypical aeromonas salmonicida strains was studied with respect to: lethality in atlantic salmon, pathogenic effect in muscle, haemolytic activity, cytotoxicity in two fish cell lines and proteolytic activities. furthermore, the relationship between lethality of ecps and mortality caused by bacterial challenge was examined. correlation was demonstrated between the pathogenic properties and proteolytic activities of the ecps. cytolytic (g ...19979418019
[meropenem: microbiologic perspective].meropenem is a beta-lactamic carbapenem derived from thienamycin and is structurally characterized by the presence of a beta-methyl group in position c1 which confers stability to the molecule versus renal dehydropeptidase 1 (dhp-1), thereby making the coadministration of an enzyme inhibitor unnecessary. its esterochemical configuration of the lateral chain in c2 (dimethyl carbomoilpyrrolidenethium) increases the activity versus gram negative bacteria (enterobacteria and pseudomonas) and moreove ...19979410064
experimental pathogenicity of aeromonas spp. for the zebra mussel, dreissena polymorphaexperiments were conducted to determine whether species of aeromonas were pathogenic to the zebra mussel dreissena polymorpha. a. jandaei, a. veronii, and a. media, identified with biolog, were originally isolated from dead zebra mussels. when inoculated into living mussels, these bacteria resulted in the mortality of the bivalves. two additional species, a. salmonicida salmonicida (atcc 33678) and a. hydrophila (atcc 7966), were also demonstrated to be pathogenic to the mussels. in addition to ...19989405741
antimicrobial activity of 12 broad-spectrum agents tested against 270 nosocomial blood stream infection isolates caused by non-enteric gram-negative bacilli: occurrence of resistance, molecular epidemiology, and screening for metallo-enzymes.a total of 270 recent nosocomial blood stream isolates of non-enterobacteriaceae gram-negative bacilli representing nearly 50 u.s. medical centers were characterized. the numbers of isolates of individual organisms were: pseudomonas aeruginosa (n = 204), acinetobacter spp. (n = 48), and stenotrophomonas maltophilia (n = 18). mics were determined using the broth microdilution susceptibility method with 12 antimicrobial agents: piperacillin, piperacillin/tazobactam, ceftriaxone, ceftazidime, cefep ...19979401811
inhibition of the metallo-beta-lactamase produced from serratia marcescens by thiol compounds.low molecular weight thiol compounds have been found to be strong inhibitors of metallo-beta-lactamase (imp-1) produced by serratia marcescens tn9106, which was expressed by echerichia coli jm109 cells. mercaptoacetic acid and 2-mercaptopropionic acid strongly and competitively inhibited imp-1 with ki of 0.23 and 0.19 microm, respectively. 2-mercaptoethanol reversibly inhibited imp-1 but did not show simple competitive inhibition.19979401719
aeromonas sobria endophthalmitis.aeromonas sobria causes a rare gram-negative bacterial water-borne infection. it has been found in waters of north queensland and south-east asia. of all aeromonas species, a. sobria is the most virulent and invasive and has been reported to cause soft tissue infection and corneal ulcer.19979395834
species, biotype and serogroup of campylobacter spp. isolated from children with diarrhoea over a ten-year period.from 1981 to 1990, stool samples from 6403 gastroenteritis cases were examined for the presence of campylobacters as well as salmonella, shigella, aeromonas species and yersinia enterocolitica. the percentages of isolation were the following: campylobacters 10.8 (86.1% of isolates were c. jejuni and 13.9% were c. coli), salmonella spp. 8.4, aeromonas spp 1.4, yersinia enterocolitica 0.3. shigella spp. were isolated only occasionally. predominant biotypes of campylobacters were c. jejuni i (69.5% ...19979385599
bacterial pathogens associated with diarrhoea on the island of crete.stool specimens from 3,600 diarrhoeal patients from the island of crete, greece, were examined for bacterial pathogens, during a three-year period (1992-1994). one or more pathogens were identified in 826 patients (22.9%), more often from children. salmonella spp. were the most frequently isolated organisms in 13.6% of the patients, followed by campylobacter in 4.7%, and enteropathogenic escherichia coli (epec) in 3.9%. yersinia enterocolitica was found in 0.7%, shigella spp. in 0.7% and aeromon ...19979384274
[bacteria of the genus aeromonas as the causative agents of saprophytic infection].the bacteriological study of feces from 458 patients with acute enteric diseases revealed that in 3.2% of cases (in summer in 8.1% of cases) the disease was caused by aeromonas. in aeromonas strains isolated from river water in the volga delta, from fish and from raw meat the same pathogenicity factors occurred as in strains isolated from patients (hemolysin, dna-ase, protease, lecithinase, amylase, adhesins, capacity of binding congo red).19959381884
[the prevalence of aeromonas spp. in the diarrheal feces of children under the age of 5 years in the city of goiânia, goiás in the 1995-1996 biennium].from 1995 through 1996, 163 fecal specimens of children aged under 5 years were analysed, 91 being from diarrhea feces and 72 without diarrhea. the material was collected in transport medium and submitted to the enrichment procedure at 4 degrees c for 7 days. for the primary isolation starch ampicillin agar was used and incubated at 35 degrees c for 18 to 24 hours. twenty (20.9%) from the following specimens were isolated: aeromonas (a.) caviae (7.7%), a. salmonicida salmonicida (6.6%), a. sobri ...19979380898
[ascitic fluid infection and enteritis by aeromonas hydrophila]. 19979379726
[transient bacteremia caused by aeromonas jandaei]. 19979376410
general secretion pathway (eps) genes required for toxin secretion and outer membrane biogenesis in vibrio cholerae.the general secretion pathway (gsp) of vibrio cholerae is required for secretion of proteins including chitinase, enterotoxin, and protease through the outer membrane. in this study, we report the cloning and sequencing of a dna fragment from v. cholerae, containing 12 open reading frames, epsc to -n, which are similar to gsp genes of aeromonas, erwinia, klebsiella, pseudomonas, and xanthomonas spp. in addition to the two previously described genes, epse and epsm (m. sandkvist, v. morales, and m ...19979371445
determination of the amino acid sequence of the plant cytolysin enterolobin.the cytolytic seed protein enterolobin from seeds of enterolobium contortisiliquum was purified by using fplc on a mono q column giving a single peak in capillary electrophoresis. the complete amino acid sequence of the plant cytolysin was determined by an automated method, yielding a molecular mass of 54,806 da. databank searches and sequence alignment demonstrated a high degree of sequence identity and similarity between enterolobin and bacterial aerolysins from aeromonas hydrophila and a. sob ...19979367525
influence of iron, growth temperature and plasmids on siderophore production in aeromonas hydrophila.aeromonas hydrophila strains obtained from diarrhoeal samples of human patients (19 isolates) and freshwater ponds (11 isolates) were analysed for siderophore production. both clinical and environmental isolates showed significantly increased siderophore production under iron-limiting conditions both at 28 degrees c and at 37 degrees c. clinical isolates consistently produced higher levels of siderophores than did the environmental isolates. the role of plasmids in moderating siderophore product ...19979364139
incidence and clinical symptoms of aeromonas-associated travellers' diarrhoea in tokyo.in a survey examining the causes of travellers' diarrhoea treated in tokyo between july 1986 and december 1995, aeromonas species were isolated from 1265 (5.5%) of 23,215 travellers returning from developing countries. aeromonas species were the fourth most frequent enteropathogen isolated, following enterotoxigenic e. coli (8.5%), salmonella spp. (7.6%) and plesiomonas shigelloides (5.6%). aeromonas species were found in 1191 (5.6%) of 21,257 patients with diarrhoea and in 74 (3.8%) of 1958 hea ...19979363009
aeromonas sobria-associated left-sided segmental colitis.aeromonas species, once thought pathogenic only in immunocompromised hosts, have more recently been found to be associated with many different infections in previously healthy individuals. the most common site of these infections is the gastrointestinal tract. we describe a 67-yr-old woman who presented with abdominal pain and bloody diarrhea and was diagnosed with left-sided, segmental colitis due to aeromonas sobria, which was proven by stool culture. extensive work-up for ischemic colitis was ...19979362202
development and application of monoclonal antibodies for in situ detection of indigenous bacterial strains in aquatic ecosystems.strain-specific monoclonal antibodies (mabs) were developed for three different bacterial isolates obtained from a freshwater environment (lake plusssee) in the spring of 1990. the three isolates, which were identified by molecular methods, were as follows: cytophaga johnsonae px62, comamonas acidovorans px54, and aeromonas hydrophila pu7718. these strains represented three species that were detected in high abundance during a set of mesocosm experiments in lake plusssee by the direct analysis o ...19979361440
expression and characterization of the recombinant gene encoding chitinase from aeromonas caviae.the gene encoding a chitinase from aeromonas caviae was cloned by pcr techniques. its recombinant gene expression was performed using pet20b(+) in escherichia coli bl21 (de3). the recombinant chitinase with the extra 33 and 13 amino acids in its n- and c-termini, respectively, was purified to near homogeneity using his-tag affinity chromatography. the recombinant chitinase was found to be present in both the culture medium and the cytoplasm. a single protein band on the native polyacrylamide gel ...19979358657
cloning and sequence analysis of the gene (epra1) encoding an extracellular protease from aeromonas hydrophila.a gene (epra1) encoding the extracellular protease of aeromonas hydrophila ah1 has been cloned and sequenced. nucleotide sequence analysis of epra1 predicted a single open reading frame (orf) of 1038 bp encoding a 346 amino acid (aa) polypeptide, with a potential 21-aa signal peptide. when the epra1 gene was expressed in minicells, one major band of approx. 37 kda was identified, while protease activity staining experiments identified a caseinolytic band of approx. 29 kda determined by sds-page ...19979358060
cloning and expression of the cdna encoding prolyl oligopeptidase (prolyl endopeptidase) from bovine brain.prolyl oligopeptidase (ec 3.4.21.26, prolyl endopeptidase) cdna from bovine brain was cloned by pcr, and the amplified fragment was used as a probe to screen the cdna library from bovine brain. the obtained clone contained a 2.7 kb dna fragment with an open reading frame of 2130 nucleotides, and encoded a protein of 710 amino acids with a deduced molecular weight of 80640 da. the deduced amino acid sequence is 95, 94, 51 and 48% homologous to those of human t-cell, porcine brain, aeromonas hydro ...19979353562
comparison of bgb-mug and lstb-mug in microbiological surveillance of recreational waters.recreational water surveillance is an important tool to prevent health hazards for the population. therefore distinct guide and imperative values for fecal indicators are listed in the ec directive about water quality control. the detection methods, however, give laboratories some room to choose their own method, which has led to difficulties in the comparability of results. in 1989 an ad-hoc working group of the coastal countries of germany established detection methods, which by now are obliga ...19959353533
toxicity of crude extracellular products of aeromonas hydrophila in tilapia, tilapia nilotica.extracellular products (ecp) secreted from aeromonas hydrophila with haemolytic and proteolytic activity were studied with respect to temperature and time of incubation as well as the lethal toxicity on tilapia, tilapia nilotica. the highest production of the haemolysin product was achieved when aer. hydrophila was grown at 35 degrees c for 30 h. tilapia erythrocyte was found to be more susceptible than sheep erythrocyte for determining the haemolytic activity. the haemolytic activity against ti ...19979351277
enteropathogenic bacteria in the la paz river of bolivia.diarrheal diseases often result from ingestion of contaminated water or food. the population of la paz, bolivia is directly or indirectly exposed to the sewage-contaminated la paz river. we conducted a bacteriologic survey of the la paz river to quantify the level of bacterial contamination, with particular reference to enteropathogens. a total bacterial count exceeding 10(6) colony-forming units (cfu)/ml, including lactose fermenting and nonfermenting, gram-negative bacilli of approximately 10( ...19979347960
aeromonas spp. possess at least two distinct type iv pilus families.type iv pili have been purified from strains of most of the aeromonas species associated with gastroenteritis (a. veronii biovar sobria, a. hydrophila, a. trota and a. caviae). they appear to be a related family (molecular mass of pilin 19 to 23 kda) with a tendency to bundle-formation. hence, we have designated them 'bundle-forming pili' (bfp). a type iv pilus biogenesis gene cluster (tapabcd) recently cloned from a strain of a. hydrophila, however, encoded a 17 kda pilin which differed signifi ...19979344785
aeromonas popoffii sp. nov., a mesophilic bacterium isolated from drinking water production plants and reservoirs.we examined the taxonomic position of seven aeromonas isolates, recovered from flemish and scottish drinking water production plants and reservoirs, which were previously recognized by numerical analysis of genomic aflp fingerprints as members of an unknown aeromonas taxon that most closely resembled the species aeromonas bestiarum (dna hybridization group [hg] 2). the new phenotypic and dna-dna hybridization data obtained in this study show that the a. bestiarum-like strains constitute a separa ...19979336924
inclusion of aeromonas dna hybridization group 11 in aeromonas encheleia and extended descriptions of the species aeromonas eucrenophila and a. encheleia.the recently reported chemotaxonomic and genotypic description of two well-separated subgroups (i and ii) in aeromonas eucrenophila and their affiliation to aeromonas encheleia and the unnamed aeromonas dna hybridization group (hg) 11 (g. huys, m. altwegg, m.-l. hänninen, m. vancanneyt, l. vauterin, r. coopman, u. torck, j. lüthy-hottenstein, p. janssen, and k. kersters, syst. appl. microbiol. 19:616-623, 1996) has questioned the original species descriptions of a. eucrenophila and a. encheleia. ...19979336923
[post-traumatic endophthalmitis with multiple water pathogens].a 37-y-old worker for a recycling company suffered from a corneal perforation by a wire which had been in contact with dirt and moisture.19979333666
differences in eicosanoid and cytokine production between injury/hemorrhage and bacteremic shock in the pig.plasma concentrations of the eicosanoids leukotriene (lt)b4, ltc4d4e4, thromboxane (tx)a2 and prostaglandin (pg)i2, and tumor necrosis factor (tnf) were measured during acute bacteremic shock and injury/hemorrhage in two porcine models. as txa2 and pgi2 are rapidly metabolized, we measured their stable metabolites txb2 and 6-keto-pgf1 alpha. bacteremic shock was induced by a graded infusion of aeromonas hydrophila over 4 h. injury/hemorrhage was produced by a 30 min, 30% total blood volume hemor ...19979329129
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