Alteromonas Putrefaciens Research Articles

Storage life of croaker(<i>Pseudotholitus senegalensis</i>)in ice and ambient temperature

Storage life of croaker (Pseudotolithus senegalensis) was 20 days in ice and 12 hrs at ambient temperature. Rejection of the raw fish by a five-member taste panel was for strong fishy to sour odours and soft texture. Cooked samples were rejected based on sour and ammoniacal faecal odours with mushy texture. The predominant flavour was bitterness. The initial bacterial load we. Predominantly masophilic in nature and included Micrococcus, Bacillus, Pseudomonas, Coryneforms, Flavobacterium, Alteromonas putrefaciens and Enterobacteria. Bacterial counts were within the range 106 - 110 on rejection and the principal spoilage organisms were Pseudomonas sp and Alteromonas putrefaciens during iced storage while Bacillus sp, Pseudomonas sp, Alteromonas putrefaciens and Proteus dominated the spoilage flora at ambient temperature. TMA and TVN proved to be reliable quality indices during ambient storage while their reliability as quality indices during ice storage was questioned

Three automated feeding strategies of natural complex nutrients utilizing on-line turbidity values in fed-batch culture: A case study on the cultivation of a marine microorganism

A comparison of three automated feeding strategies was made utilizing on-line turbidity values in fed-batch cultures. Control indices in these feeding strategies were (i) total cell mass ( VX), (ii) total cell mass ( VX) and estimated cell yield on the basis of the total amount of the substrate supplied ( Y ̂ x/lms ), and (iii) growth rate (d( VX)/d t). A marine microorganism, Alteromonas putrefaciens, was cultivated by feeding a solution composed of peptone (220 g/ l) and yeast extract (110 g/ l). When the automatic feeding strategy (i) was applied, a considerable amount of peptide accumulated in the later phase, indicating ‘overfeeding’ of the natural complex nutrients. The improved feeding strategies (ii) and (iii) enabled the overfeeding to be avoided, resulting in more effective utilization of the nutrients fed. Overall, the maximum cell concentrations were increased more than 1.5 times, while the total amounts of nutrients fed were reduced by a half.

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-galactosamine, galactose, and melibiose. The hemagglutinin showed binding ability to heparin and LPS, as demonstrated by heparin-Sepharose chromatography and centrifugation experiments, respectively. It was also found that the hemagglutinin can bind to various bacteria such as Escherichia coli, Bacillus subtilis, Vibrio anguillarum, Pseudomonas perfectomarinus, Achromobacter aquamarinus, and Alteromonas putrefaciens, and can agglutinate all of them.

Heterotrophic Bacterial Flora Associated with European Eel Anguilla anguilla Reared in Freshwater.

The heterotrophic bacterial microflora of European eel reared in freshwater and its surround-ing aqueous environment is reported. Quality of eel culture water is also discussed with respect to its physicochemical and bacteriological characteristics. The presence of bacterial species pathogenic for eel and its serological classification was investigated in fishery water and eels. Overall 675 isolates from fishery water and healthy or diseased were eel identified at genus level. Pseudo-monas was the most frequently encountered genus. Moraxella sp. and Acinetobacter sp. were abundant in fishery water, and Aeromonas sp. in elver and internal organs of silver eel. Plesiomonas sp., Escherichia sp., Enterobacter sp., Serratia sp., Citrobacter sp., and Streptococcus sp. were minority genera that were only recovered from eel. The species Aeromonas hydrophila, A. sobria, and Alteromonas putrefaciens were mostly isolated from diseased eel whereas other potential fish pathogens such as P. fluorescens, V. anguillarum, and Plesiomonas shigelloides were generally observed in fishery water or healthy eel. Following the typing scheme of Guinee and Jensen, most of the isolates of A. hydrophila from diseased European eels, as well as the strain A. hydrophila Y-62 pathogenic for Japanese eel, belonged to O19 serogroup. A relationship between origin and serogroup for A. sobria was not observed. None Vibrio anguillarum isolates agglutinated with O1 and O2 antisera of Sorensen and Larsen.

Release of 226Ra from uranium mill tailings by microbial Fe(III) reduction

Uranium mill tailings were anaerobically incubated in the presence of H 2 with Alteromonas putrefaciens, a bacterium known to couple the oxidation of H 2 and organic compounds to the reduction of Fe(III) oxides. There was a direct correlation between the extent of Fe(III) reduction and the accumulation of dissolved 226Ra. In sterile tailings in which Fe(III) was not reduced, there was negligible leaching of 226Ra. The behavior of Ba was similar to that of Ra in inoculated and sterile systems. These results demonstrate that under anaerobic conditions, microbial reduction of Fe(III) may result in the release of dissolved 226Ra from uranium mill tailings.

Microbial sensor system for nondestructive evaluation of fish meat quality

A microbial sensor system consisting of the bacterium Alteromonas putrefaciens) immobilized within membranes, a flow cell, an oxygen electrode, peristaltic pumps, a buffer tank, a thermostatically controlled bath and a recorder, was constructed for the nondestructive quality evaluation of bluefin tuna. The chemical compounds on fish meat surfaces which are the indicators of fish meat quality were rapidly determined by using the proposed sensor system. Fish meat quality was determined from the rate of current decrease of the sensor. Good correlations were obtained between fish meat quality and sensor response. One assay could be completed within one minute.

Volatile compounds associated with microbial growth on normal and high pH beef stored at chill temperatures.

Volatile compounds produced by Pseudomonas fragi and mixed, natural floras on beef of normal pH (5.5-5.8; glucose greater than 1500 micrograms/g) and high pH (6.3-6.8; glucose less than 10 micrograms/g) included a range of alkyl esters and a number of sulphur-containing compounds including dimethylsulphide but not hydrogen sulphide. Production of the last was a property common to the other Gram-negative organisms tested viz. Hafnia alvei, Enterobacter agglomerans, Serratia liquefaciens, Alteromonas putrefaciens and Aeromonas hydrophila, all of which produced similar off-odours and, with the exception of E. agglomerans, 'greening' on high pH meat. Serratia liquefaciens also produced greening of normal pH meat. Acetoin and diacetyl were major end products of Brochothrix thermosphacta but the related 2,3-butanediol was formed only on normal pH meat. The Enterobacteriaceae produced the same compounds but only on normal pH meat and together with Br. thermosphacta were probable sources of these compounds and of the free and esterified branched-chain alcohols detected in the naturally contaminated samples.

Hydrogen and Formate Oxidation Coupled to Dissimilatory Reduction of Iron or Manganese by Alteromonas putrefaciens

The ability of Alteromonas putrefaciens to obtain energy for growth by coupling the oxidation of various electron donors to dissimilatory Fe(III) or Mn(IV) reduction was investigated. A. putrefaciens grew with hydrogen, formate, lactate, or pyruvate as the sole electron donor and Fe(III) as the sole electron acceptor. Lactate and pyruvate were oxidized to acetate, which was not metabolized further. With Fe(III) as the electron acceptor, A. putrefaciens had a high affinity for hydrogen and formate and metabolized hydrogen at partial pressures that were 25-fold lower than those of hydrogen that can be metabolized by pure cultures of sulfate reducers or methanogens. The electron donors for Fe(III) reduction also supported Mn(IV) reduction. The electron donors for Fe(III) and Mn(IV) reduction and the inability of A. putrefaciens to completely oxidize multicarbon substrates to carbon dioxide distinguish A. putrefaciens from GS-15, the only other organism that is known to obtain energy for growth by coupling the oxidation of organic compounds to the reduction of Fe(III) or Mn(IV). The ability of A. putrefaciens to reduce large quantities of Fe(III) and to grow in a defined medium distinguishes it from a Pseudomonas sp., which is the only other known hydrogen-oxidizing, Fe(III)-reducing microorganism. Furthermore, A. putrefaciens is the first organism that is known to grow with hydrogen as the electron donor and Mn(IV) as the electron acceptor and is the first organism that is known to couple the oxidation of formate to the reduction of Fe(III) or Mn(IV). Thus, A. putrefaciens provides a much needed microbial model for key reactions in the oxidation of sediment organic matter coupled to Fe(III) and Mn(IV) reduction.

Changes in Polyamine Contents of Common Squid and Scallop during Storage

Changes in polyamine contents and viable counts of total bacteria were examined in the meats of common squid and scallop with and without the addition of chloramphenicol during storage at 10°C. In the common squid and scallop meats without chloramphenicol, the levels of agmatine, putrescine and cadaverine increased remarkably as decomposition proceeded. These polyamines appear to be useful as potential indicators of the freshness and decomposition of common squid and scallop. In the meats added with chloramphenicol, viable counts increased slowly and polyamines were produced in very small amounts. From these results it was concluded that polyamines were produced from amino acids by bacterial decarboxylases and not by endogenous muscle enzymes. When viable counts of total bacteria reached the level of 103/g in common squid and 103/g to 104/g in scallop, the formation of polyamines was observed. Alteromonas putrefaciens produced putrescine, cadaverine, agmatine and tryptamine.

Composition of Lipopolysaccharides from Alteromonas putrefaciens (Shewanella putrefaciens)

Lipopolysaccharides from eight strains of Alteromonas putrefaciens (Shewanella putrefaciens) representing four groups defined by DNA-DNA homology studies have been compared. All products were of the R-type, lacking a polymeric side chain. In each case lipid A was based on glucosamine and contained a complex range of fatty acids. In general, the major non-hydroxy acids were 11-methyldodecanoic acid and tridecanoic acid, and the major hydroxy acids were 3-hydroxyundecanoic acid, 3-hydroxydodecanoic acid, 3-hydroxy-11-methyldodecanoic acid, 3-hydroxytridecanoic acid and 3-hydroxytetradecanoic acid. All lipopolysaccharides contained a 3-deoxy-2-octulosonic acid, only detectable after dephosphorylation. All core oligosaccharides contained d-galactose and d-glycero-d-manno-heptose, and all except those of group I strains also contained d-galactosamine. Group I strains (including NCIB 10471, the type strain) were distinguished from others by the presence in the core of 3-amino-3,6-dideoxyglucose, d-glucose, and acid-stable phosphate residues. Quinovosamine and d-fructose were found only in group IV strains. Core oligosaccharides from the group II and group III strains, as isolated, were deficient in heptose and seemed to have the same structure. During mild acid hydrolysis of the lipopolysaccharides, various monosaccharides and phosphates were released. The monosaccharides were d-galactose (group I), d-fructose (group IV), and d-galacturonic acid (mainly groups II and III). Phosphates identified were ethanolamine pyrophosphate, ethanolamine phosphate, pyrophosphate, and orthophosphate. The chemotaxonomic implications of the results are discussed.

The effect of potassium sorbate on the structural integrity of Alteromonas putrefaciens

Treatment of Alteromonas putrefaciens with potassium sorbate at pH 7.0 resulted in increased hydrophobicity of the cell wall and lysis on exposure to lysozyme. These effects could be overcome to some extent by the addition of magnesium ions to the incubation medium. Transmission and scanning electron microscopy provided evidence of outer membrane damage in sorbate‐treated cells. Dissociated sorbate ion was shown to have an inhibitory effect on A. putrefaciens.

Microbial reduction of manganese oxides: Interactions with iron and sulfur

Alteromonas putrefaciens (strain MR-1) is capable of rapid Mn(IV) reduction under conditions of neutral pH and temperatures characteristic of the Oneida Lake, New York, sediments from which it was isolated. MR-1 also reduces Fe 3+ to Fe 2+, and disproportionates thiosulfate to sulfide and sulfite; independently, the Fe 2+ and sulfide act as rapid reductants of Mn. The addition of Fe 3+ or thiosulfate to cultures of MR-1 in the presence of oxidized Mn increases the rate and extent of Mn reduction relative to that observed in the absence of Fe 3+ or thiosulfate. Furthermore, when Fe 3+ and Mn oxides are present conjointly, Fe 2+ does not appear until reduction of the oxidized Mn is complete. These results demonstrate that observed rates of Fe 2+ and sulfide production may underestimate the total rates of Fe and sulfate reduction in those environments containing oxidized Mn. These results also demonstrate the potential impact that a single microbe can exert on sediment geochemistry, and provide the basis for preliminary models of the complexity of microbial and geochemical interactions that occur.

Dissimilative Iron Reduction by the Marine Eubacterium Alteromonas putrefaciens Strain 200

The ability of microorganisms to mediate the transformation of iron coupled with the participation of iron as a reactive redox species in natural waters and sediments has prompted recent speculation that such bacteria play an important role in such activities as global iron cycling, biologically induced mineral formation, and localized competition for available organic substrates. Microaerophilic soil bacteria dominate the list of microorganisms capable of iron reduction. However, screening experiments have shown that the marine eubacterium Alteromonas putrefaciens strain 200 is one of the most efficient iron reducers yet isolated. Respiratory inhibitor studies have suggested that a branched respiratory chain and two ferri-reductases (one constitutive and one induced by growth under microaerophilic conditions (2 µM O2)) are involved in iron reduction. Further complementary biochemical and genetic studies have concentrated on characterizing the components of the induced iron reduction system. Ethyl methane sulfonate (EMS) has been used to develop mutagenesis procedures for A.putrefaciens 200; under optimal conditions, the frequency of total auxotrophy for EMS-treated cultures is approximately 0.4%. Tests for specific auxotrophic requirements have indicated the presence of a complex distribution of loci required for biosynthetic pathways. Mutants in both high-rate and low-rate ferri-reductase activity have been identified via a newly developed technique for screening EMS-treated A.putrefaciens 200 cultures. Ferri-reductase-deficient mutants generated by these techniques have been shown to contain reduced cytochrome content. Preliminary results indicate that the broad host range cloning vector pVK100 (Inc P) is mobilizable into A.putrefaciens 200. Current work is aimed at construction of an A.putrefaciens 200 clone bank, and complementation of the ferri-reductase mutants.

Bacterial Manganese Reduction and Growth with Manganese Oxide as the Sole Electron Acceptor

Microbes that couple growth to the reduction of manganese could play an important role in the biogeochemistry of certain anaerobic environments. Such a bacterium, Alteromonas putrefaciens MR-1, couples its growth to the reduction of manganese oxides only under anaerobic conditions. The characteristics of this reduction are consistent with a biological, and not an indirect chemical, reduction of manganese, which suggest that this bacterium uses manganic oxide as a terminal electron acceptor. It can also utilize a large number of other compounds as terminal electron acceptors; this versatility could provide a distinct advantage in environments where electron-acceptor concentrations may vary.

Polar Lipids, Fatty Acids, and Isoprenoid Quinones of Alteromonas putrefaciens ( Shewanella putrefaciens)

The polar lipids, fatty acids, and isoprenoid quinones of eight strains of Alteromonas putrefaciens (Shewanella putrefaciens) representing different phenotypes and genotypes have been analysed. The polar lipids consisted almost entirely of phosphatidylethanolamine and phosphatidylglycerol, with variable proportions of their lyso derivatives (such derivatives were most prominent in extracts from strains of relatively high mol % G+C). In contrast to the results of previous studies, ornithine amide lipids and glycolipids were not encountered. Fatty acid profiles were complex, usually with 13-methyltetradecanoic acid, pentadecanoic acid, cis-hexadec-9-enoic acid, and ds-heptadec-9-enoic acid as the major components. All strains contained ubiquinone-8 and ubiquinone-7, with the former preponderant in all strains but one. In addition all strains contained menaquinone-7 (a trace only in one strain) and its methyl derivative thermoplasmaquinone-7 (traces only in two strains). Thus, rather diverse strains of A. putrefaciens share an overall lipid profile highly unusual for a Gram-negative bacterium, and distinctively different from that of recognized Alteromonas species. The classification of the organism as Shewanella putrefaciens within Vibrionaceae does not eliminate this chemotaxonomic problem.

Characterization of iron-reducing Alteromonas putrefaciens strains from oil field fluids

Gram-negative, aerobic bacteria capable of facultative growth using ferric ions or thiosulfate or sulfite as electron acceptors were readily isolated from oil field fluids. Morphological and biochemical data are presented which support the classification of these isolates as being strains of Alteromonas putrefaciens (mol% G + C in the range 42–56%). The isolates expressed a range of halotolerance and temperature tolerance as some would grow in the presence of 7.5% NaCl and some grew well at 4 °C. These physiological characteristics, together with the ability to produce ferrous ions and sulfide and their ubiquitous presence in oil field fluids, indicate that these bacteria could contribute to the corrosion of metal in the oil field environment.

Sulfur isotope fractionation by strains of Alteromonas putrefaciens isolated from oil field fluids

Strains of Alteromonas putrefaciens which reduce sulfite and thiosulfate to H2S are readily isolated from oil field fluids in north central Alberta. Data are presented on H2S production during anaerobic sulfite reduction by four strains of A. putrefaciens. Analysis of the sulfur isotope composition (δ34S) of the evolved sulfide shows normal and "inverse" isotope fractionation patterns which are diagnostic of dissimilatory sulfite reduction.

Microbial Sensors for the Detection of Fish Freshness

ABSTRACTA microbial sensor system consisting of immobilized living spoilage‐causing bacterium, Alteromonas putrefaciens, and an oxygen electrode was prepared and applied to the continuous determination of fish freshness. Fish freshness was expressed as a ratio of A/B, where A is the current decrease of the microbial sensor obtained by the application of l0–50 μL fish extract and B is the current decrease by the equivalent volume of culture medium. Freshness of bluefin tuna and yellowtail meats stored in ice for up to 2 weeks was determined by both the proposed and the conventional K value enzyme sensor method. Good comparative results were obtained. One assay could be completed within 13 min.

Spoilage Pattern of Atlantic Cod Fillets Treated with Glucose Oxidase/Gluconic Acid

Atlantic cod fillets were subjected to two experimental treatments as potential means of prolonging freshness: (1) 30 sec dip in glucose oxidase (1 unit/ml) plus glucose (4% w/v); and (2) 30 sec dip in gluconic acid (0.125% w/v). A control group was dipped in tap water. The fillets were packed in 10-lb commercial containers and stored at 1-2°C for a 12d period. Quality of the fillets was assessed microbiologically (total plate count, Alteromonas putrefaciens, and fluorescent pseudomonads), chemically (trimethylamine content) and organoleptically (raw quality grading and sensory assessment). Contrary to previous claims, both the enzyme and acid treatments showed insignificant inhibition of the microbial indicators, and few if any benefits were apparent in chemical, raw quality, or sensory assesment of the fillets. A glucose oxidase enzyme system for extending the shelf life of cod fillets was therefore considered to have little potential.

Identification and decarboxylase activities of bacteria isolated from decomposed mahimahi ( Coryphaena hippurus) after incubation at 0 and 32°C

Identification and amine-forming ability of bacterial isolates from decomposed mahimahi ( Coryphaena hippurus) were studied in order to identify spoilage microflora and determine the potential for a chemical index of spoilage based on the metabolites produced. Mesophilic bacteria isolated from a fish incubated at 32°C for 24 h were essentially all Gram-negative rods; 89% of these were Vibrio alginolyticus. Strong histamine-forming (> 100 mg/100 ml) mesophiles consisted of eight cultures of Morganella morganii and one of Proteus mirabilis. Weak histamine-forming (< 10 mg/100 ml) mesophiles were all V. alginolyticus, and these comprised 15% of the isolates assigned to that species. Decarboxylation of ornithine and lysine occurred in 38 and 92%, respectively, of the mesophilic isolates. Psychrotrophic isolates obtained from a fish incubated 14 days at 0°C were predominantly Gram-negative genera. Of these, 9% were histamine-forming cultures of Alteromonas putrefaciens, a weak histamine former that produced < 1 mg/100 ml at 5 and 20°C. Decarboxylation of ornithine and lysine occurred in 13 and 15%, respectively, of the psychrotrophic isolates.