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1.
Int J Food Microbiol ; 351: 109264, 2021 Aug 02.
Article in English | MEDLINE | ID: mdl-34098468

ABSTRACT

Fresh meat is commonly packaged in modified atmosphere to decelerate spoilage processes. The applied gas mixture affects the growth of spoilage organisms and selectively shapes the spoilage community. In this study, we investigated the impact of O2 and CO2 on the growth of Photobacterium (P.) phosphoreum and P. carnosum strains in situ on chicken meat by packaging under different modified atmospheres (air, 70% O2/30% CO2, 70% N2/30% CO2, 100% N2). Combination of 70% O2 and 30% CO2 resulted in significant growth reduction of the analyzed strains, suggesting inhibitory effects of both gases in combination. In contrast, 30% CO2 alone had only a minor effect and photobacteria are supposed to have a growth advantage over other meat spoilers in this atmosphere. Additionally, single growth of the strains in the different atmospheres was compared when challenged with the presence of Pseudomonas (Ps.) fragi or Brochothrix (B.) thermosphacta as prominent co-contaminants in different ratios (10:1, 1:1, 1:10). Presence of co-contaminants resulted in increased cell numbers of P. carnosum TMW2.2149 but reduced or unchanged cell numbers of P. phosphoreum TMW2.2103 in most packaging atmospheres. The initial ratio of photobacteria and co-contaminants defined the relative abundance during storage but did not change the type of the interaction. Our results suggest either a commensalistic (P. carnosum) or competitive interaction (P. phosphoreum) of photobacteria and co-contaminants on modified atmosphere packaged chicken, respectively. Furthermore, in a mix comprising seven prominent spoilers, strains of both Photobacterium species prevailed as a constant part of the spoilage microbiome during 7 days of refrigerated storage on chicken meat packaged under O2/CO2 atmosphere.


Subject(s)
Atmosphere/chemistry , Food Packaging/methods , Photobacterium/growth & development , Poultry/microbiology , Animals , Bacteria/drug effects , Bacteria/growth & development , Carbon Dioxide/analysis , Carbon Dioxide/pharmacology , Chickens , Food Microbiology , Microbial Interactions , Microbiota/drug effects , Oxygen/analysis , Oxygen/pharmacology , Photobacterium/drug effects
2.
Food Microbiol ; 99: 103679, 2021 Oct.
Article in English | MEDLINE | ID: mdl-34119089

ABSTRACT

Photobacterium spp. occur frequently in marine environments but have been recently also found as common spoilers on chilled meats. The environmental conditions in these ecological niches differ especially regarding salinity and ambient pressure. Linking the occurrence of photobacteria in different niches may elucidate its ecology and bring insights for the food industry. We investigated tolerance of Photobacterium (P.) phosphoreum and P. carnosum strains to high hydrostatic pressure and salinity and aligned our observations with presence of relevant genes. The strains were isolated from packaged meats and salmon (or the sea) to identify adaptations to marine and terrestrial habitats. Growth of all P. carnosum strains was reduced by 40 MPa hydrostatic pressure and >3% sodium chloride, suggesting loss of traits associated with marine habitats. In contrast, P. phosphoreum strains were only slightly affected, suggesting general adaptation to marine habitats. In accordance, these strains had gene clusters associated with marine niches, e.g. flagellar and lux-operons, being incomplete in P. carnosum. Occurrence of P. carnosum strains on packaged salmon and P. phosphoreum strains on meats therefore likely results from cross-contamination in meat and fish processing. Still, these strains showed intermediate traits regarding pressure- and halotolerance, suggesting developing adaptation to their respective environment.


Subject(s)
Meat/microbiology , Photobacterium/metabolism , Salmon/microbiology , Sodium Chloride/metabolism , Animals , Cattle , Chickens , Food Microbiology , Hydrostatic Pressure , Photobacterium/chemistry , Photobacterium/growth & development , Photobacterium/isolation & purification , Seawater/microbiology , Sodium Chloride/analysis
3.
J Appl Microbiol ; 129(6): 1497-1510, 2020 Dec.
Article in English | MEDLINE | ID: mdl-32538525

ABSTRACT

AIMS: This study describes the effect of phage therapy on hatching of longfin yellowtail (Seriola rivoliana) eggs challenged with Photobacterium damselae subsp. damselae. METHODS AND RESULTS: A lytic phage (vB_Pd_PDCC-1) against P. damselae subsp. damselae was isolated and characterized. The use of phage vB_Pd_PDCC-1 increased the hatching rate of eggs, and reduced presumptive Vibrio species to non-detectable numbers, even in non-disinfected eggs. High-throughput 16S rRNA gene sequencing analysis revealed that phage vB_Pd_PDCC-1 caused significant changes in the composition and structure of the associated microbiota, allowing that members (e.g. those belonging to the family Vibrionaceae) of the class Gammaproteobacteria to be displaced by members of the class Alphaproteobacteria. CONCLUSIONS: To the best of our knowledge, this represents the first study evaluating phage therapy to control potential negative effects of P. damselae subsp. damselae during hatching of longfin yellowtail eggs. SIGNIFICANCE AND IMPACT OF THE STUDY: The Seriola genus includes several important commercial fish species due to its rapid growth and easy adaptability to confinement conditions. However, bacterial infections (especially those caused by Vibrio and Photobacterium species) are among the main limiting factors for the intensification of marine fish aquaculture, particularly during early development stages. Therefore, the use of phages, which are natural killers of bacteria, represents a promising strategy to reduce the mortality of farmed organisms caused by pathogenic bacteria.


Subject(s)
Bacteriophages/physiology , Biological Control Agents/pharmacology , Fish Diseases/therapy , Fishes/microbiology , Gram-Negative Bacterial Infections/veterinary , Photobacterium/drug effects , Animals , Aquaculture , Fish Diseases/microbiology , Fishes/physiology , Gram-Negative Bacterial Infections/microbiology , Gram-Negative Bacterial Infections/therapy , Microbiota/drug effects , Ovum/microbiology , Ovum/physiology , Phage Therapy , Photobacterium/growth & development
4.
J Food Prot ; 83(4): 621-627, 2020 Apr 01.
Article in English | MEDLINE | ID: mdl-32221566

ABSTRACT

ABSTRACT: The effects of high hydrostatic pressure (HHP) treatments on histamine-forming bacteria (HFB) Morganella morganii and Photobacterium phosphoreum in phosphate buffer and tuna meat slurry were investigated using viability counting and scanning electron microscopy. The first-order model fits the destruction kinetics of high pressure on M. morganii and P. phosphoreum during the pressure hold period. The D-values of M. morganii (200 to 600 MPa) and P. phosphoreum (100 to 400 MPa) in phosphate buffer ranged from 16.4 to 0.08 min and 26.4 to 0.19 min, respectively, whereas those in tuna meat slurry ranged from 51.0 to 0.09 min and 71.6 to 0.19 min, respectively. M. morganii had higher D-values than P. phosphoreum at the same pressure, indicating it was more resistant to HHP treatment. HFB had a higher D-value in tuna meat slurry compared with that in phosphate buffer, indicating that the HFB were more resistant to pressure in tuna meat slurry. The Zp values (pressure range that results in a 10-fold change in D-value) of M. morganii and P. phosphoreum were 162 and 140 MPa in phosphate buffer and 153 and 105 MPa in tuna meat slurry, respectively. Damage to the cell wall and cell membrane by HHP treatments can be observed by scanning electron microscopy. To our knowledge, this is the first report to demonstrate that HHP can be applied to inactivate the HFB M. morganii and P. phosphoreum by inducing morphological changes in the cells.


Subject(s)
Food Handling/methods , Food Preservation/methods , Morganella morganii , Photobacterium , Animals , Histamine , Morganella morganii/growth & development , Morganella morganii/metabolism , Photobacterium/growth & development , Photobacterium/metabolism , Pressure
5.
Food Microbiol ; 88: 103405, 2020 Jun.
Article in English | MEDLINE | ID: mdl-31997761

ABSTRACT

Sensory, chemical and microbial changes for Atlantic cod (Gadus morhua L.) filets from capture-based-aquaculture in Greenland were studied. The objective was to determine shelf-life and indices of spoilage for iced or superchilled fillets when stored in air, or modified atmosphere packed (MAP; 40% CO2 and 60% N2). MAP iced storage extended the sensory shelf-life from 15 days to 21 days compared to storage in air. With superchilling at -1.7 °C sensory shelf-life was above 32 days, and no formation of total volatile nitrogen (TVN) was observed irrespective of storage in air or MAP. pH of ≥7.0, TVN (≥35 mg-N/100 g) and trimethylamine (≥20 mg-N TMA/100 g) were promising indices of spoilage. Aerobic viable counts were less valuable indices of spoilage as the dominating microbiota of cod in air (Pseudomonas spp., Photobacterium spp., Shewanella spp., Acinetobacter spp.) changed to Photobacterium spp. in MAP cod. Spoilage activity determined as the yield factor for TVN formation was 6-200 folds higher for Photobacterium spp. compared to Shewanella spp. and Pseudomonas spp. Photobacterium carnosum was responsible for TVN formation in iced cod irrespective of storage in air or MAP, and it was identified at the specific spoilage organism that limited iced product shelf-life.


Subject(s)
Atmosphere , Bacteria/growth & development , Cold Temperature , Food Preservation/methods , Gadus morhua/microbiology , Seafood/microbiology , Animals , Aquaculture , Colony Count, Microbial , Food Microbiology/methods , Food Packaging/methods , Food Storage , Greenland , Ice , Photobacterium/growth & development , Pseudomonas/growth & development
6.
J Appl Microbiol ; 129(1): 37-50, 2020 Jul.
Article in English | MEDLINE | ID: mdl-31505106

ABSTRACT

AIMS: Several virulence factors of three new Photobacterium species: Photobacterium toruni, Photobacterium malacitanum and Photobacterium andalusiense associated with diseases of cultured redbanded seabream (Pagrus auriga) were studied. The exoenzymatic activities, adherence and cytotoxic capabilities, and iron-uptake mechanisms were determined both in bacterial extracellular products (ECP) and whole bacterial cells. The histopathology damages provoked on redbanded seabream by the ECP was also studied. METHODS AND RESULTS: The highest exoenzymatic activities of the ECP were alkaline- and acid-phosphatase, phosphohydrolase and lipase. The ECP were strongly lethal for fish at 4-96 h post-inoculation (p.i). Histological changes were evident at 96 hpi of ECP, affecting head kidney, splenic parenchyma and heart. Cytotoxicity assays, on three fish lines and one human cell line, were conducted using whole bacterial cells and their ECP. The new species tested were cytotoxic only for fish cell lines using whole bacterial cells. Bacterial adherence showed an adherence index moderate on CHSE-214 cell line. All strains showed variable haemolytic activity, and were able to grow under iron-limiting conditions, although the CAS reactivitiy was very low. However, all strains produced high amounts of extracelullar citrate that could be used as iron carrier, and use haem as iron source, except the P. toruni strains because a deletion in the genomic region encoding this ability in all Vibrionaceae members. CONCLUSIONS: The toxic activity of the bacterial ECPs was thermolabile, and not associated with their thermoresistant lipopolysaccharide content. The virulence of the strains tested could not be related to the haemolytic activity. Iron uptake could be based on the use of endogenous citrate as iron carrier and P. toruni lacks the ability to use haem as iron source. SIGNIFICANCE AND IMPACT OF THE STUDY: The study analyses for the first time the virulence properties of three new species of Photobacterium pathogenic for fish.


Subject(s)
Fish Diseases/microbiology , Gram-Negative Bacterial Infections/veterinary , Photobacterium/pathogenicity , Sea Bream/microbiology , Animals , Aquaculture , Cell Line , Fish Diseases/pathology , Gram-Negative Bacterial Infections/microbiology , Humans , Photobacterium/growth & development , Photobacterium/metabolism , Photobacterium/physiology , Virulence , Virulence Factors/metabolism
7.
Food Res Int ; 122: 506-516, 2019 08.
Article in English | MEDLINE | ID: mdl-31229106

ABSTRACT

Market globalization and changes in purchasing habits pose a challenge to the fishery industry because of the short shelf life of fish products. In view of this scenario, it would be very helpful if tools capable of predicting the shelf-life of fish could be developed. Thus, the objective of this study was to employ a modelling approach capable of predicting the evolution of the microbiota of hake fillets packaged under a modified atmosphere (MAP) rich in CO2 (50% CO2 / 50% N2) when stored at temperatures ranging between 1 and 10 °C. Growth curves of ten microbial groups were obtained at four different temperatures and fitted with the Baranyi model. Photobacterium showed high growth rates in hake fillets (0.99 days-1 at 4 °C), similar to those of Shewanella, lactic acid bacteria, and non-specific microbial groups investigated, and significantly higher than those of Pseudomonas. Furthermore, no lag phase was observed for Photobacterium regardless of the temperature investigated. On the other hand, Enterobacteriaceae and moulds and yeasts displayed low growth fitness, and their counts increased by <1.5-2 Log10 cycles along the incubation period regardless of storage temperature. The influence of storage temperature on growth parameters (λ, µmax and Yend) was subsequently studied, and secondary models were developed for the eight most relevant microbial groups. All of the final equations developed in this study showed R2 values ≥0.90, and RMSE values ≤0.50. In addition, results obtained in this investigation strongly suggest that Photobacterium would be the main responsible microorganism for the spoilage of hake fillets stored under MAP conditions (50% CO2/50% N2) along the entire range of temperatures investigated (1-10 °C).


Subject(s)
Gadiformes/microbiology , Seafood/microbiology , Animals , Colony Count, Microbial , Consumer Behavior , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Food Contamination , Food Microbiology , Food Packaging , Humans , Lactobacillales/growth & development , Lactobacillales/isolation & purification , Photobacterium/growth & development , Photobacterium/isolation & purification , RNA, Ribosomal, 16S/genetics , RNA, Ribosomal, 16S/isolation & purification , Sequence Analysis, DNA , Shewanella/growth & development , Shewanella/isolation & purification , Temperature
8.
Microbiol Res ; 222: 52-59, 2019 May.
Article in English | MEDLINE | ID: mdl-30928030

ABSTRACT

Modified atmosphere packaging (MAP) is widely used in food industry to extend the microbiological shelf life of meat. Common CO2-containing gas atmospheres for poultry meat packaging are either nearly O2-free or high O2 MAPs. In this work, we compared spoilage microbiota of skinless chicken breast in CO2/O2 (30/70%) and CO2/N2 (30/70%) MAP, which are culturable with conventional methods and identified isolates by MALDI-TOF MS. These data were compared to metatranscriptome sequencing enabling a culture-independent overview on the composition of microbiota at species level. While typical MAP meat spoilers were confirmed in the transcriptomic approach, we also found high numbers of transcripts mapping to Photobacterium spp. sequences in these samples. As photobacteria were recently shown to occur in different MAP and vacuum packaged meats, we used the respective part of the metatranscriptomic data for prediction of Photobacterium spp. major metabolic routes in situ, upon growth in MAP poultry meat. It is predicted that they employ similar metabolism in both atmospheres: In the lack of carbohydrates upon meat spoilage, the pyruvate pool is filled via glycerol originating from lipolysis and amino acid conversions. From the pyruvate pool, gluconeogenesis is fed enabling cell wall biosynthesis and growth as well as catabolism to lactate and other metabolites, or anaplerosis towards the citric acid cycle. Production is predicted of several biogenic amines including tyramine and cadaverine, enabling generation of proton motive force. Taken together, photobacteria express metabolic pathways upon growth on meat, which should lead to compounds overlapping with those of known potent meat spoilers.


Subject(s)
Atmosphere/chemistry , Food Microbiology , Food Packaging , Meat/microbiology , Microbiota/physiology , Photobacterium/metabolism , Animals , Carbon Dioxide/analysis , Colony Count, Microbial , Food Preservation , Food Storage , Gene Expression Regulation, Bacterial , Metabolic Networks and Pathways , Nitrogen/analysis , Oxygen/analysis , Photobacterium/genetics , Photobacterium/growth & development , Poultry , Sequence Analysis, RNA , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Time Factors , Transcriptome
9.
J Biosci Bioeng ; 128(1): 64-71, 2019 Jul.
Article in English | MEDLINE | ID: mdl-30709706

ABSTRACT

An efficient heterotrophic nitrifying/aerobic denitrifying strain, Photobacterium sp. NNA4 was isolated from a recirculating aquaculture system (RAS). NNA4 was capable of utilizing ammonia, nitrate or nitrite as sole N-source with maximal removal rates of 12.5 mg/L/h for NH4+N, 16.4 mg/L/h for NO3--N, and 4.5 mg/L/h for NO2--N, respectively. Optimal nitrification conditions were: sodium succinate as C-source, 30-37°C, NaCl 1-4%, pH 7.0-8.0, dissolved oxygen 5.89 mg/L, C/N > 10. Gas chromatography/mass spectrometry and gas chromatography/isotope ratio mass spectrometry analyses showed that N2 and N2O were aerobic denitrification products of nitrite and nitrate. NNA4 could tolerate high concentration of hydroxylamine and displayed efficient hydroxylamine-transforming capability. Hydroxylamine oxidoreductase activity using potassium ferricyanide as electron acceptor was 0.042 U. Results revealed that strain NNA4 could oxidize NH2OH directly to N2O at aerobic conditions. In view of its high removal ability of inorganic nitrogen pollutants and broad salinity tolerance range, NNA4 has great potential in denitrification treatment of types of wastewater with either low salinity (e.g., municipal facilities) or high salinity (e.g., aquaculture, seafood processing).


Subject(s)
Denitrification , Heterotrophic Processes , Hydroxylamine/metabolism , Nitrification , Photobacterium , Aerobiosis , Ammonia/isolation & purification , Animals , Aquaculture/methods , Equipment Reuse , Humans , Nitrates/metabolism , Nitrogen/isolation & purification , Nitrogen/metabolism , Oxidoreductases/genetics , Oxidoreductases/metabolism , Photobacterium/enzymology , Photobacterium/genetics , Photobacterium/growth & development , Photobacterium/metabolism , Wastewater/chemistry , Wastewater/microbiology , Water Pollutants, Chemical/isolation & purification , Water Purification/methods
10.
Microbiology (Reading) ; 164(6): 920-933, 2018 06.
Article in English | MEDLINE | ID: mdl-29757128

ABSTRACT

How DNA metabolism is adapted to survival of organisms such as the bacterium Photobacterium profundum SS9 at high pressure is unknown. Previously, a high pressure-sensitive P. profundum SS9 transposon mutant (FL31) was identified, with an insertion in a putative rctB gene. The Vibrio cholerae RctB protein is essential for replication initiation at the origin of chromosome II, oriCII. Using a plasmid-based system in E. coli we have identified the replication origin of chromosome II from P. profundum SS9 and have shown that the putative rctB gene, disrupted in FL31, is essential for oriCII function. Moreover, we found that a region corresponding to the V. cholerae oriCII incompatibility region (incII) exerts an inhibitory effect on P. profundum oriCII. The truncated rctB gene in FL31 confers insensitivity to incII inhibition, indicating that the C-terminus of RctB is important for the negative regulation of replication. The RctB proteins of V. cholerae and P. profundum are partially interchangeable, but full functionality is achieved only with the cognate origin. Our findings provide the first characterization of the replication origin of chromosome II in a deep-sea bacterium.


Subject(s)
Chromosomes, Bacterial/genetics , DNA Replication , Photobacterium/genetics , Replication Origin/genetics , Adaptation, Physiological/genetics , Atmospheric Pressure , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Chromosomes, Bacterial/metabolism , DNA Helicases/chemistry , DNA Helicases/genetics , DNA Helicases/metabolism , DNA, Bacterial/genetics , DNA, Bacterial/metabolism , Escherichia coli/genetics , Genes, Bacterial/genetics , Mutation , Photobacterium/growth & development , Photobacterium/metabolism , Plasmids/genetics , Plasmids/metabolism , Trans-Activators/chemistry , Trans-Activators/genetics , Trans-Activators/metabolism , Vibrio cholerae/genetics
11.
Biocontrol Sci ; 22(2): 89-96, 2017.
Article in English | MEDLINE | ID: mdl-28659560

ABSTRACT

 We compared the TBT-resistant ability of resting cells prepared from isolates that formed colonies on nutrient agar plates containing 100 µM tributyltin (TBT) chloride, such as Photobacterium sp. TKY1, Halomonas sp. TKY2, and Photobacterium sp. NGY1, with those from taxonomically similar type strains. Photobacterium sp. TKY1 showed the highest ability among those three isolates. The number of surviving Photobacterium sp. TKY1 cells was hardly decreased after 1 h of exposure to 100 µM TBTCl, regardless of the number of resting cells in the range from 109.4 to 104.2 CFU mL-1. In such an experimental condition, the maximum number of TBT molecules available to associate with a single cell was estimated to be approximately 6.0 x 1011.8. Resting cells prepared from type strains Photobacterium ganghwense JCM 12487T and P. halotolerans LMG 22194T, which have 16S rDNA sequences highly homologous with those of Photobacterium sp. TKY1, showed sensitivity to TBT, indicating that TBT-resistant marine bacterial species are not closely related in spite of their taxonomic similarity. We also estimated the impact of TBT-resistant bacterial species to indigenous microbial populations of TBT-polluted surface sediments. The number of surviving TBT-sensitive Vibrio natriegens ATCC 14048T cells, 106.2±0.3 CFU mL-1, was reduced to 104.4±0.4 CFU mL-1 when TBT-resistant Photobacterium sp. TKY1 cells, 109.1±0.2 CFU mL-1, coexisted with 109.4±0.2 CFU mL-1 of V. natriegens ATCC 14048T cells in the presence of 100 µM TBTCl. These results indicate that the toxicity of TBT to TBT-sensitive marine bacterial populations might be enhanced when a TBT-resistant marine bacterial species inhabits TBT-polluted surface sediments.


Subject(s)
Drug Resistance, Microbial , Geologic Sediments/microbiology , Photobacterium/drug effects , Trialkyltin Compounds/pharmacology , DNA, Ribosomal , Photobacterium/growth & development , Water Microbiology , Water Pollutants, Chemical/pharmacology
12.
Appl Microbiol Biotechnol ; 101(11): 4371-4385, 2017 Jun.
Article in English | MEDLINE | ID: mdl-28497204

ABSTRACT

Photobacterium species are Gram-negative coccobacilli which are distributed in marine habitats worldwide. Some species are unique because of their capability to produce luminescence. Taxonomically, about 23 species and 2 subspecies are validated to date. Genomes from a few Photobacterium spp. have been sequenced and studied. They are considered a special group of bacteria because some species are capable of producing essential polyunsaturated fatty acids, antibacterial compounds, lipases, esterases and asparaginases. They are also used as biosensors in food and environmental monitoring and detectors of drown victim, as well as an important symbiont.


Subject(s)
Bacterial Proteins/genetics , Photobacterium/genetics , Photobacterium/physiology , Biosensing Techniques , DNA, Bacterial , Ecosystem , Genome, Bacterial , Luminescence , Photobacterium/growth & development , Phylogeny , RNA, Ribosomal, 16S , Sequence Analysis, DNA , Symbiosis
13.
Mikrobiologiia ; 85(2): 177-86, 2016.
Article in Russian | MEDLINE | ID: mdl-27476206

ABSTRACT

A comparative analysis of the four commercially available and laboratory luminescent sensor strains to the toxic effect of 10 carbon-based nanomatherials (CBNs) and 10 metal nanoparticles (MNPs) was carried out in this study. The bioluminescence inhibition assays with marine Photobacterium phosphoreum and recombinant Escherichia coli strains were varied in minimal toxic concentrations and EC50 values but led to well correlated biotoxicity evaluation for the most active compounds were ranked as Cu > (MgO, CuO) > (fullerenol, graphene oxide). The novel sensor strain Bacillus subtilis EG 168-1 exhibited the highest sensitivity to CBNs and MNPs that increased significantly number of toxic compounds causing the bacterial bioluminescence inhibition effect.


Subject(s)
Anti-Bacterial Agents , Bacillus subtilis/growth & development , Carbon , Escherichia coli/growth & development , Metal Nanoparticles/chemistry , Photobacterium/growth & development , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Carbon/chemistry , Carbon/pharmacology
14.
J Basic Microbiol ; 56(11): 1234-1243, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27282981

ABSTRACT

Proteomic approaches were applied to investigate whether Photobacterium damselae subsp. piscicida (Phdp) can directly sense and respond to growth conditions under different salinities, 0.85% and 3.5% NaCl concentrations, mimicking the osmotic conditions in host and marine water bodies, respectively. Proteins significantly altered were analyzed by two-dimensional gel electrophoresis (2-DE), liquid chromatography-electrospray ionization-quadrupole-time-of-flight tandem mass spectrometry (LC-ESI-Q-TOF MS/MS) and bioinformatics analysis, thus resulting in 16 outer membrane proteins (OMPs), 12 inner membrane proteins (IMPs), and 20 cytoplasmic proteins (CPs). Quantitative real-time PCR was also applied to monitor the mRNA expression level of these target proteins. Cluster of orthologous groups of protein (COG) analysis revealed that when shifting from 3.5% to 0.85% salinity, the majority of the up-regulated proteins were involved in posttranslational modification, protein turnover, and chaperones, while the down-regulated proteins were mainly related to energy production and conversion, compatible solutes (carbohydrates, amino acids and their derivatives) biogenesis and transport. Differentially expressed proteins identified in the current study could be used to elucidate the salt adaptation mechanisms of Phdp during their transition between host cells and the marine habitats.


Subject(s)
Photobacterium/genetics , Photobacterium/physiology , Proteome , Salt Tolerance , Animals , Electrophoresis, Gel, Two-Dimensional , Fish Diseases/microbiology , Osmoregulation/genetics , Photobacterium/growth & development , Photobacterium/ultrastructure , Proteomics , Real-Time Polymerase Chain Reaction , Salt Tolerance/genetics , Sodium Chloride/pharmacology , Tandem Mass Spectrometry
15.
PLoS One ; 11(3): e0151351, 2016.
Article in English | MEDLINE | ID: mdl-26977596

ABSTRACT

Myeolchi-aekjeot (MA) in Korea is produced outdoors without temperature controls, which is a major obstacle to produce commercial MA products with uniform quality. To investigate the effects of temperature on MA fermentation, pH, bacterial abundance and community, and metabolites were monitored during fermentation at 15°C, 20°C, 25°C, and 30°C. Initial pH values were approximately 6.0, and pH values increased after approximately 42 days, with faster increases at higher temperatures. Bacterial abundances increased rapidly in all MA samples after quick initial decreases during early fermentation and then they again steadily decreased after reaching their maxima, which were significantly greater at higher temperatures. Bacterial community analysis revealed that Proteobacteria and Tenericutes were predominant in all initial MA samples, but they were rapidly displaced by Firmicutes as fermentation progressed. Photobacterium and Mycoplasma belonging to Proteobacteria and Tenericutes, respectively, which may include potentially pathogenic strains, were dominant in initial MA, but decreased with the growth of Chromohalobacter, which occurred faster at higher temperatures--they were dominant until 273 and 100 days at 15°C and 20°C, respectively, but not detected after 30 days at 25°C and 30°C. Chromohalobacter also decreased with the appearance of subsequent genera belonging to Firmicutes in all MA samples. Tetragenococcus, halophilic lactic acid bacteria, appeared predominantly at 20°C, 25°C, and 30°C; they were most abundant at 30°C, but not detected at 15°C. Alkalibacillus and Lentibacillus appeared as dominant genera with the decrease of Tetragenococcus at 25°C and 30°C, but only Lentibacillus was dominant at 15°C and 20°C. Metabolite analysis showed that amino acids related to tastes were major metabolites and their concentrations were relatively higher at high temperatures. This study suggests that high temperatures (approximately 30°C) may be appropriate in MA fermentation, in the light of faster disappearance of potentially pathogenic genera, higher amino acids, growth of Tetragenococcus, and faster fermentation.


Subject(s)
Fermentation , Food Microbiology , Food , Temperature , Bioreactors , Firmicutes/growth & development , Firmicutes/metabolism , Mycoplasma/growth & development , Mycoplasma/metabolism , Photobacterium/growth & development , Photobacterium/metabolism , Proteobacteria/growth & development , Proteobacteria/metabolism , Republic of Korea , Tenericutes/growth & development , Tenericutes/metabolism
16.
Int J Food Microbiol ; 215: 131-42, 2015 Dec 23.
Article in English | MEDLINE | ID: mdl-26433458

ABSTRACT

Proliferation of microbial population on fresh poultry meat over time elicits spoilage when reaching unacceptable levels, during which process slime production, microorganism colony formation, negative organoleptic impact and meat structure change are observed. Spoilage organisms in raw meat, especially Gram-negative bacteria can be difficult to combat due to their cell wall composition. In this study, the natural antimicrobial agents ε-poly-L-lysine (ε-PL) and isoeugenol were tested individually and in combinations for their activities against a selection of Gram-negative strains in vitro. All combinations resulted in additive interactions between ε-PL and isoeugenol towards the bacteria tested. The killing efficiency of different ratios of the two antimicrobial agents was further evaluated in vitro against Pseudomonas putida. Subsequently, the most efficient ratio was applied to a raw turkey meat model system which was incubated for 96 h at spoilage temperature. Half of the samples were challenged with P. putida, and the bacterial load and microbial community composition was followed over time. CFU counts revealed that the antimicrobial blend was able to lower the amount of viable Pseudomonas spp. by one log compared to untreated samples of challenged turkey meat, while the single compounds had no effect on the population. However, the compounds had no effect on Pseudomonas spp. CFU in unchallenged meat. Next-generation sequencing offered culture-independent insight into population diversity and changes in microbial composition of the meat during spoilage and in response to antimicrobial treatment. Spoilage of unchallenged turkey meat resulted in decreasing species diversity over time, regardless of whether the samples received antimicrobial treatment. The microbiota composition of untreated unchallenged meat progressed from a Pseudomonas spp. to a Pseudomonas spp., Photobacterium spp., and Brochothrix thermosphacta dominated food matrix on the expense of low abundance species. We observed a similar shift among the dominant species in meat treated with ε-PL or the antimicrobial blend, but the samples differed markedly in the composition of less abundant species. In contrast, the overall species diversity was constant during incubation of turkey meat challenged with P. putida although the microbiota composition did change over time. Untreated or ε-PL treated samples progressed from a Pseudomonas spp. to a Pseudomonas spp. and Enterobacteriaceae dominated food matrix, while treatment with the antimicrobial blend resulted in increased relative abundance of Hafnia spp., Enterococcaceae, and Photobacterium spp. We conclude that the blend delayed the onset of spoilage of challenged meat, and that all antimicrobial treatments of unchallenged or challenged meat affect the progression of the microbial community composition. Our study confirms that the antimicrobial effects observed in vitro can be extrapolated to a food matrix such as turkey meat. However, it also underlines the consequence of species-to-species variation in susceptibility to antimicrobials, namely that the microbial community change while the CFU remains the same. Addition of antimicrobials may thus prevent the growth of some microorganisms, allowing others to proliferate in their place.


Subject(s)
Eugenol/analogs & derivatives , Food Preservation/methods , Food Preservatives/pharmacology , Meat/microbiology , Polylysine/pharmacology , Pseudomonas putida/drug effects , Turkeys/microbiology , Animals , Bacterial Load , Brochothrix/drug effects , Brochothrix/growth & development , Enterobacteriaceae/drug effects , Enterobacteriaceae/growth & development , Eugenol/pharmacology , Food Microbiology , Hafnia/drug effects , Hafnia/growth & development , Microbial Sensitivity Tests , Microbiota/drug effects , Photobacterium/drug effects , Photobacterium/growth & development , Pseudomonas putida/growth & development
17.
Chemosphere ; 141: 290-6, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26298026

ABSTRACT

Anti-cancer drugs are discussed as high risk substances in regard to human health and considered as problematic for the environment. They are of potential environmental relevance due to their poor biodegradability and toxicological properties. Methotrexate (MTX) is an antimetabolite that was introduced in the pharmaceutical market in the 40's and still today is one of the most consumed cytotoxic compounds around the world. In the present study MTX was only partially biodegraded in the closed bottle test (CBT). Therefore, it was submitted to three different advanced oxidation processes (AOPs): UV/H2O2, UV/Fe(2+)/H2O2 and UV/TiO2. The irradiation was carried out with a Hg medium-pressure lamp during 256min whereas the analytical monitoring was done through LC-UV-MS/MS and DOC analysis. MTX was easily removed in all the irradiation experiments, while the highest mineralization values and rates were achieved by the UV/Fe(2+)/H2O2 treatment. The lowest resulted from the UV/H2O2 reactions. The UV/H2O2 treatment resulted in little biodegradable transformation products (TPs). However, the same treatment resulted in a reduction of the toxicity of MTX by forming less toxic TPs. Analysis by LC-UV-MS/MS revealed the existence of nine TPs formed during the photo-catalytic treatments. The pH of the solutions decreased from 6.4 (t 0min) to 5.15 in the UV/H2O2 and from 6.4 (t 0min) to 5.9 in the UV/TiO2 at the end of the experiments. The initial pH of the UV/Fe(2+)/H2O2 experiments was adjusted to 5 and after the addition of H2O2 the pH decreased to around 3 and remained in this range until the end of the treatments.


Subject(s)
Antineoplastic Agents/analysis , Methotrexate/analysis , Water Pollutants, Chemical/analysis , Water Purification/methods , Aerobiosis , Aliivibrio fischeri/drug effects , Aliivibrio fischeri/growth & development , Antineoplastic Agents/chemistry , Antineoplastic Agents/toxicity , Biodegradation, Environmental , Chromatography, Liquid , Hydrogen Peroxide/chemistry , Methotrexate/chemistry , Methotrexate/toxicity , Oxidation-Reduction , Photobacterium/drug effects , Photobacterium/growth & development , Photolysis , Tandem Mass Spectrometry , Titanium/chemistry , Toxicity Tests , Ultraviolet Rays , Water Pollutants, Chemical/chemistry , Water Pollutants, Chemical/toxicity
18.
Biofizika ; 60(2): 316-21, 2015.
Article in Russian | MEDLINE | ID: mdl-26016027

ABSTRACT

This contribution presents the results of analysis of the dynamics of the bioluminescence of luminous bacteria Photobacterium phosphoreum IMV B-7071 under optimal conditions of their growth. A quasi-harmonic nature of the bacterial bioluminescence dynamics was detected. The observed periods of these changes have similar values compared with those in the earlier defined periods of changes in physicochemical properties of water. The relationship between biorhythms and a quasi-harmonic nature of changes in physicochemical properties of water is discussed.


Subject(s)
Luminescence , Photobacterium/chemistry , Water/chemistry , Luminescent Measurements , Periodicity , Photobacterium/growth & development
19.
Food Microbiol ; 47: 111-5, 2015 May.
Article in English | MEDLINE | ID: mdl-25583344

ABSTRACT

Most existing models for the spoilage of modified atmosphere packed Atlantic salmon are based on the growth of the spoilage organism Photobacterium phosphoreum. However, there is evidence that this organism is not the specific spoilage organism on salmon produced and packaged in Australia. We developed a predictive model for the growth of bacteria in Australian-produced Atlantic salmon stored under modified atmosphere conditions (30-98% carbon dioxide in nitrogen) at refrigeration temperatures (0-10 °C). As expected, both higher levels of carbon dioxide and lower temperatures decreased the observed growth rates of the total population. A Belehrádek-type model for growth rate fitted the data best with an acceptably low root mean square error. At low temperatures (∼0 °C) the growth rates in this study were similar to those predicted by other models but at higher temperatures (∼10 °C) the growth rates were significantly lower in the current study.


Subject(s)
Bacteria/growth & development , Food Packaging , Food Preservation , Models, Statistical , Photobacterium/growth & development , Salmo salar/microbiology , Animals , Australia , Carbon Dioxide , Cold Temperature , Colony Count, Microbial , Food Microbiology , Nitrogen
20.
J Food Sci ; 79(8): S1568-74, 2014 Aug.
Article in English | MEDLINE | ID: mdl-25046980

ABSTRACT

A total of 18 farmed turbot (Scophthalmus maximus) were slaughtered over 4 successive weeks in November 2012 and stored in polystyrene boxes with ice until analyzed. The fish were stored between 1 and 22 d and presented to a taste panel and further analyzed for quality index method (QIM), microbiological analysis by real-time quantitative PCR (qPCR), taste, pH, color by computer imaging, protein denaturation with differential scanner calorimeter (DSC), texture hardness, and shear force. Results show small, but significant changes in physical and visual attributes such as texture and color. No gaping was observed. Only small changes in texture were observed explained by lack of myosin denaturation. The fillets became more white and yellow during storage, whereas the major changes occurred during the 1st week. A panel evaluating QIM and taste could not distinguish major differences in appearance and taste and over 15 d storage period, but were able to quantify the age by smell. Analysis of microorganisms on the epidermis displayed growth of Carnobacterium maltaromaticum, potentially inhibiting growth of other spoilage bacteria. Fish stored for 22 d were rejected by the taste panel caused by a stale smell and taste, but not bitter or rancid. It is concluded that turbot has a shelf life of at least 16 d.


Subject(s)
Carnobacterium/isolation & purification , Flatfishes/microbiology , Food Storage , Seafood/microbiology , Animals , Calorimetry, Differential Scanning , Carnobacterium/growth & development , Color , Cooking , DNA, Bacterial/genetics , Food Contamination/analysis , Food Microbiology , Hardness , Humans , Hydrogen-Ion Concentration , Lactobacillus/growth & development , Lactobacillus/isolation & purification , Linear Models , Muscle, Skeletal/chemistry , Photobacterium/growth & development , Photobacterium/isolation & purification , Sequence Analysis, DNA , Taste
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