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1.
Article in English | MEDLINE | ID: mdl-38683659

ABSTRACT

A strain belonging to the genus Psychrobacter, named PraFG1T, was isolated from the peritoneal effusion of a stray dog during necropsy procedures. The strain was characterized by the phylogenetic analyses based on the nucleotide sequences of 16S and 23S rRNA genes and of gyrB, which placed the strain in the genus Psychrobacter. The nucleotide sequence of the chromosome confirmed the placement, showing an average nucleotide identity of 72.1, 77.7, and 77.5 % with the closest related species, namely Psychrobacter sanguinis, Psychrobacter piechaudii, and Psychrobacter phenylpyruvicus, respectively, thus indicating a novel species. The polyphasic characterization by biochemical and fatty acid profiling as well as MALDI-TOF supported those findings. The strain was halotolerant, capable of growing within a temperature range between 4 and 37 °C, it was positive for catalase and oxidase, indole producing, nitrate reducing, and not able to use 5-keto-d-gluconic acid as a carbon source. Taken together, the data suggest that strain PraFG1T could be considered as representing a novel species, with the name Psychrobacter raelei sp. nov. (type strain PraFG1T=CIP 111873T=LMG 32233T).


Subject(s)
Bacterial Typing Techniques , DNA, Bacterial , Fatty Acids , Peritonitis , Phylogeny , Psychrobacter , RNA, Ribosomal, 16S , RNA, Ribosomal, 23S , Sequence Analysis, DNA , Animals , Psychrobacter/genetics , Psychrobacter/isolation & purification , Psychrobacter/classification , RNA, Ribosomal, 16S/genetics , DNA, Bacterial/genetics , Peritonitis/microbiology , Dogs , RNA, Ribosomal, 23S/genetics , Dog Diseases/microbiology , Gram-Positive Bacterial Infections/microbiology
2.
Article in English | MEDLINE | ID: mdl-35037845

ABSTRACT

In this study, two bacterial strains designated F2608T and F1192T, isolated from marine sediment sampled in Weihai, PR China, were characterized using a polyphasic approach. Strains were aerobic, Gram-stain-negative and motile. According to the results of phylogenetic analyses based on their 16S rRNA genes, these two strains should be classified under the genus Psychrobacter and they both show <98.5% sequence similarity to their closest relative, Psychrobacter celer JCM 12601T. Moreover, strain F2608T showed 97.5% sequence similarity to strain F1192T. Strain F2608T grew at 4-37 °C (optimum, 30-33 °C) and at pH 6.0-9.0 (optimum, pH 6.5-7.0) in the presence of 0-12% (w/v) NaCl (optimum, 4.0-5.0%). Strain F1192T grew at 4-37 °C (optimum, 30 °C) and at pH 5.5-9.0 (optimum, pH 7.0-7.5) in the presence of 0.5-12% (w/v) NaCl (optimum, 3.0-4.0%). The genomic DNA G+C contents of strain F2608T and strain F1192T were 47.4 and 44.9 %, respectively. Genomic characteristics including average nucleotide identity and digital DNA-DNA hybridization values clearly separated strain F2608T from strain F1192T. The sole isoprenoid quinone in these two strains was ubiquinone 8 and the major cellular fatty acids (>10.0%) were C18:1 ω9c and C17:1 ω8c. The major polar lipids of these two strains were phosphatidylglycerol, phosphatidylethanolamine and diphosphatidylglycerol. Based on the results of polyphasic analysis, the two strains represent two novel species of the genus Psychrobacter, for which the names Psychrobacter halodurans sp. nov. and Psychrobacter coccoides sp. nov. are proposed. The type strains are F2608T (=MCCC 1K05774T=KCTC 82766T) and F1192T (=MCCC 1K05775T=KCTC 82765T), respectively.


Subject(s)
Geologic Sediments/microbiology , Phylogeny , Psychrobacter , Seawater/microbiology , Bacterial Typing Techniques , Base Composition , China , DNA, Bacterial/genetics , Fatty Acids/chemistry , Phospholipids/chemistry , Psychrobacter/classification , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
3.
Int J Food Microbiol ; 328: 108668, 2020 Sep 02.
Article in English | MEDLINE | ID: mdl-32474228

ABSTRACT

Biofilms are comprised of microorganisms embedded in a self-produced matrix that normally adhere to a surface. In the food processing environment they are suggested to be a source of contamination leading to food spoilage or the transmission of food-borne pathogens. To date, research has mainly focused on the presence of (biofilm-forming) bacteria within food processing environments, without measuring the associated biofilm matrix components. Here, we assessed the presence of biofilms within a meat processing environment, processing pork, poultry and beef, by the detection of microorganisms and at least two biofilm matrix components. Sampling included 47 food contact surfaces and 61 non-food contact surfaces from eleven rooms within an Austrian meat processing plant, either during operation or after cleaning and disinfection. The 108 samples were analysed for the presence of microorganisms by cultivation and targeted quantitative real-time PCR based on 16S rRNA. Furthermore, the presence of the major matrix components carbohydrates, extracellular DNA and proteins was evaluated. Overall, we identified ten biofilm hotspots, among them seven of which were sampled during operation and three after cleaning and disinfection. Five biofilms were detected on food contact surfaces (cutters and associated equipment and a screw conveyor) and five on non-food contact surfaces (drains and water hoses) resulting in 9.3 % of the sites being classified as biofilm positive. From these biofilm positive samples, we cultivated bacteria of 29 different genera. The most prevalent bacteria belonged to the genera Brochothrix (present in 80 % of biofilms), Pseudomonas and Psychrobacter (isolated from 70 % biofilms). From each biofilm we isolated bacteria from four to twelve different genera, indicating the presence of multi-species biofilms. This work ultimately determined the presence of multi-species biofilms within the meat processing environment, thereby identifying various sources of potential contamination. Especially the identification of biofilms in water hoses and associated parts highlights the need of a frequent monitoring at these sites. The knowledge gained about the presence and composition of biofilms (i.e. chemical and microbiological) will help to prevent and reduce biofilm formation within food processing environments.


Subject(s)
Brochothrix/isolation & purification , Food Handling , Meat/microbiology , Pseudomonas/isolation & purification , Psychrobacter/isolation & purification , Animals , Austria , Biofilms/classification , Biofilms/growth & development , Cattle , Disinfection/methods , Food Microbiology , Foodborne Diseases/microbiology , Poultry/microbiology , RNA, Ribosomal, 16S/analysis
4.
Int J Syst Evol Microbiol ; 70(1): 211-219, 2020 Jan.
Article in English | MEDLINE | ID: mdl-31617840

ABSTRACT

One slightly beige-white pigmented, Gram-stain-negative, rod-shaped bacterium, strain I-STPP5bT, was isolated from the trachea of a Gentoo penguin chick individual (Pygoscelin papua) investigated in Fildes Bay, Chilean Antarctic (62° 12' S, 58° 57' W). I-STPP5bT consists of a 3.4 Mb chromosome with a DNA G+C content of 44.4 mol%. Of the 3056 predicted genes, 1206 were annotated as hypothetical proteins and 51 were tRNAs. Phylogenetic analysis based on nearly full-length 16S rRNA gene sequences showed that the isolate shared a 16S rRNA gene sequence identity to the type strains of Psychrobacter phenylpyruvicus (98.8 %), Psychrobacter arenosus and Psychrobacter pasteurii (both 98.3 %), Psychrobacter piechaudii (98.2 %) and Psychrobacter sanguinis (98.1 %), but 16S rRNA gene sequence similarities to all other Psychrobacter species were ≤98.0 %. Partial gyrB nucleotide and amino acid sequence similarities among strain STPP5bT and the next related type strains were all below 81.8 and 92.9%, respectively. DNA-DNA hybridisation (DDH) with P. phenylpyruvicus LMG 5372T, P. arenosus DSM 15389T and P. sanguinis DSM 23635T also showed low values (all below 30 %). The main cellular fatty acids of the strain were C18 : 1ω9c and C16 : 1ω7c and/or C16 : 1ω6c. Based on phylogenetic, chemotaxonomic, genomic and phenotypic analyses we propose a new species of the genus Psychrobacter, with the name Psychrobacter pygoscelis sp. nov. and strain I-STPP5bT (=CIP 111410T= CCM 8799T=LMG 30301T) as type strain.


Subject(s)
Phylogeny , Psychrobacter/classification , Spheniscidae/microbiology , Trachea/microbiology , Animals , Antarctic Regions , Bacterial Typing Techniques , Base Composition , Chile , DNA, Bacterial/genetics , Fatty Acids/chemistry , Nucleic Acid Hybridization , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
5.
Molecules ; 24(12)2019 Jun 14.
Article in English | MEDLINE | ID: mdl-31207974

ABSTRACT

A novel RNase R, psrnr, was cloned from the Antarctic bacterium Psychrobacter sp. ANT206 and expressed in Escherichia coli (E. coli). A bioinformatics analysis of the psrnr gene revealed that it contained an open reading frame of 2313 bp and encoded a protein (PsRNR) of 770 amino acids. Homology modeling indicated that PsRNR had reduced hydrogen bonds and salt bridges, which might be the main reason for the catalytic efficiency at low temperatures. A site directed mutation exhibited that His 667 in the active site was absolutely crucial for the enzyme catalysis. The recombinant PsRNR (rPsRNR) showed maximum activity at 30 °C and had thermal instability, suggesting that rPsRNR was a cold-adapted enzyme. Interestingly, rPsRNR displayed remarkable salt tolerance, remaining stable at 0.5-3.0 M NaCl. Furthermore, rPsRNR had a higher kcat value, contributing to its efficient catalytic activity at a low temperature. Overall, cold-adapted RNase R in this study was an excellent candidate for antimicrobial treatment.


Subject(s)
Adaptation, Biological , Cold Temperature , Environmental Microbiology , Ice Cover/microbiology , Psychrobacter/physiology , Ribonucleases/metabolism , Salt Tolerance , Amino Acid Sequence , Antarctic Regions , Bacterial Proteins/chemistry , Bacterial Proteins/genetics , Enzyme Activation , Kinetics , Models, Biological , Molecular Conformation , Molecular Structure , Psychrobacter/isolation & purification , Ribonucleases/genetics
6.
Int J Food Microbiol ; 293: 102-113, 2019 Mar 16.
Article in English | MEDLINE | ID: mdl-30677559

ABSTRACT

Thawed hake (Merluccius capensis and M. paradoxus) and plaice (Pleuronectes platessa) fillets were used as a model to evaluate the effect of storage temperature (0 or 10 °C) and biological variability (fish species, lot to lot) on bacterial growth kinetics and microbial successions. Both culture dependent methods (plate counts on non-selective and selective media) and culture independent methods (qPCR and 16S rRNA gene metabarcoding) were used. Bacterial counts exceeded 107 cfu/g within 2-3 days at 10 °C and 7-8 days at 0 °C. Plate counts on three media (Plate Count Agar +0.5% NaCl, Iron Agar Lyngby and Pseudomonas Selective medium) and 16S rRNA gene counts estimated by qPCR were highly correlated. Growth was modelled using the D-model and specific growth rate ranged between 0.97 and 1.24 d-1 and 3.54 and 5.90 d-1 at 0 and 10 °C, respectively. The initial composition of the microbiota showed lot-to-lot variation, but significant differences between the two fish species were detected. Alpha diversity significantly decreased during storage. When bacterial counts exceeded 107 cfu/g, the microbiota was dominated by members of the genera Pseudomonas, Psychrobacter, Acinetobacter, Serratia, Flavobacterium, Acinetobacter, Carnobacterium, Brochothrix and Vagococcus. However, Photobacterium and Shewanella, two genera frequently associated with fish spoilage, were either absent or minor components of the microbiota. As expected, storage temperature significantly affected the abundance of several species. The inference of microbial association networks with three different approaches (an ensemble approach using the CoNet app, Sparse Correlations for Compositional data, and SParse InversE Covariance Estimation for Ecological Association Inference) allowed the detection of both a core microbiota, which was present throughout storage, and a number of taxa, which became dominant at the end of spoilage and were characterized by a disproportionate amount of negative interactions.


Subject(s)
Food Contamination/analysis , Food Storage , RNA, Ribosomal, 16S/isolation & purification , Seafood/microbiology , Acinetobacter/genetics , Acinetobacter/isolation & purification , Animals , Bacterial Load , Brochothrix/genetics , Brochothrix/isolation & purification , Carnobacterium/genetics , Carnobacterium/isolation & purification , Cold Temperature , Colony Count, Microbial , DNA, Bacterial/genetics , DNA, Bacterial/isolation & purification , Fishes , Food Microbiology , Microbial Consortia , Photobacterium/genetics , Photobacterium/isolation & purification , Pseudomonas/genetics , Pseudomonas/isolation & purification , Psychrobacter/genetics , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Shewanella/genetics , Shewanella/isolation & purification
7.
Int J Food Microbiol ; 293: 87-93, 2019 Mar 16.
Article in English | MEDLINE | ID: mdl-30677560

ABSTRACT

The effect of air packaging (AP), vacuum packaging (VP) and modified atmosphere packaging (MAP, 75% CO2/25% N2) on the composition of the microbial community of lightly salted grass carp (Ctenopharyngodon idellus) fillets stored at 4 °C was studied. Physicochemical characteristics (total volatile basic nitrogen (TVB-N) value, pH value and biogenic amines (BA)) were also monitored. Based on sensory evaluation, the shelf life of fillets under AP, VP, and MAP was 8, 16 and 24 days, respectively. High-throughput sequencing showed that Acinetobacter and Pseudomonas dominated the microflora of fresh grass carp. At the sensory rejection time, Pseudomonas and Psychrobacter dominated the AP products. Lactococcus became the predominant genus of both VP and MAP fillets, and there was no obvious difference in the composition of the dominant microbiota between these two treatments at the end of the shelf life.


Subject(s)
Carps/microbiology , Food Packaging , Food Preservation , Vacuum , Acinetobacter/isolation & purification , Adult , Animals , Atmosphere , Biogenic Amines/analysis , DNA, Bacterial/isolation & purification , Female , Food Storage , High-Throughput Nucleotide Sequencing , Humans , Hydrogen-Ion Concentration , Lactococcus/isolation & purification , Male , Microbiota/drug effects , Pseudomonas/isolation & purification , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/isolation & purification , Sodium Chloride , Taste , Young Adult
8.
Microb Biotechnol ; 12(5): 1049-1063, 2019 09.
Article in English | MEDLINE | ID: mdl-29105344

ABSTRACT

In recent years, the marine environment has been the subject of increasing attention from biotechnological and pharmaceutical industries. A combination of unique physicochemical properties and spatial niche-specific substrates, in wide-ranging and extreme habitats, underscores the potential of the marine environment to deliver on functionally novel bioactivities. One such area of ongoing research is the discovery of compounds that interfere with the cell-cell signalling process called quorum sensing (QS). Described as the next generation of antimicrobials, these compounds can target virulence and persistence of clinically relevant pathogens, independent of any growth-limiting effects. Marine sponges are a rich source of microbial diversity, with dynamic populations in a symbiotic relationship. In this study, we have harnessed the QS inhibition (QSI) potential of marine sponge microbiota and through culture-based discovery have uncovered small molecule signal mimics that neutralize virulence phenotypes in clinical pathogens. This study describes for the first time a marine sponge Psychrobacter sp. isolate B98C22 that blocks QS signalling, while also reporting dual QS/QSI activity in the Pseudoalteromonas sp. J10 and ParacoccusJM45. Isolation of novel QSI activities has significant potential for future therapeutic development, of particular relevance in the light of the pending perfect storm of antibiotic resistance meeting antibiotic drug discovery decline.


Subject(s)
Acyl-Butyrolactones/metabolism , Biological Products/metabolism , Paracoccus/drug effects , Porifera/microbiology , Pseudoalteromonas/drug effects , Psychrobacter/metabolism , Quorum Sensing/drug effects , Animals , Psychrobacter/isolation & purification , Virulence/drug effects
9.
Food Microbiol ; 76: 543-552, 2018 Dec.
Article in English | MEDLINE | ID: mdl-30166186

ABSTRACT

The rinds of surface-ripened cheeses have expected aesthetic properties, including distinct colors, that contribute to overall quality and consumer acceptance. Atypical rind pigments are frequently reported in small-scale cheese production, but the causes of these color defects are largely unknown. We provide a potential microbial explanation for a striking purple rind defect in a surface-ripened cheese. A cheese producer in the United States reported to us several batches of a raw-milk washed-rind cheese with a distinctly purple rind. We isolated a Proteus species from samples with purple rind defect, but not from samples with typical rind pigments, suggesting that this strain of Proteus could be causing the defect. When provided tryptophan, a precursor in the indigo and indirubin biosynthesis pathway, the isolated strain of Proteus secreted purple-red pigments. A Psychrobacter species isolated from both purple and normal rinds also secreted purple-red pigments. Using thin-layer chromatography and liquid chromatography-mass spectrometry, we confirmed that these bacteria produced indigo and indirubin from tryptophan just as closely related bacteria make these compounds in purple urine bag syndrome in medical settings. Experimental cheese communities with or without Proteus and Psychrobacter confirmed that these Proteobacteria cause purple pigmentation of cheese rinds. Reports of purple rinds in two other cheeses from Europe and the observation of pigment production by Proteus and Psychrobacter strains isolated from other cheese rinds suggest that purple rind defect has the potential to be widespread in surface-ripened cheeses.


Subject(s)
Cheese/microbiology , Indigo Carmine/metabolism , Proteus/isolation & purification , Psychrobacter/isolation & purification , Animals , Cattle , Cheese/analysis , Color , Indoles/metabolism , Milk/metabolism , Milk/microbiology , Pigments, Biological/metabolism , Proteus/genetics , Proteus/metabolism , Psychrobacter/genetics , Psychrobacter/metabolism , Tryptophan/metabolism
10.
Int J Food Microbiol ; 278: 36-43, 2018 Aug 02.
Article in English | MEDLINE | ID: mdl-29698856

ABSTRACT

Bacterial diversity of whole gilt-head sea bream (Sparus aurata L. 1758) originating from Ionian and Aegean Sea aquaculture farms and stored at 0 (ice), 4 and 8 °C was determined by 16S rRNA gene amplicon sequencing method using the Illumina's MiSeq platform. The composition of Aerobic Plate Counts (APC) was also monitored by 16S rRNA gene sequencing. The rejection time point of sea bream from either area, as determined by sensory evaluation, was about 14, 6 and 3 days at 0, 4 and 8 °C, respectively. APC was approximately 4.5 log cfu/g at day 0 and ranged from 7.5 to 8.5 log cfu/g at sensory rejection. Culture-depended analysis showed that Pseudomonas and Shewanella were the most abundant microorganisms grown on plates for both seas. Moreover, culture-independent analysis of DNA extracted directly from fish flesh showed that sea bream originating from different geographical areas exhibited different bacterial diversity. Pseudomonas and Psychrobacter were the dominant microorganisms of chill-stored fish from Ionian (apart from 8 °C, where Carnobacterium dominated) and Aegean Sea, respectively. In addition, small changes of storage temperature greatly affected bacterial microbiota of stored fish. Various bacterial species, not detected by conventional microbiological methods, were also revealed through 16S amplicon sequencing. In conclusion, the use of NGS approach is a promising methodology for assessing bacterial diversity of sea bream originating from different geographical areas and stored at various temperatures.


Subject(s)
Food Microbiology , Pseudomonas/isolation & purification , Psychrobacter/isolation & purification , Sea Bream/microbiology , Shewanella/isolation & purification , Animals , Aquaculture , Female , Fisheries , Food Preservation/methods , Greece , Microbiota/genetics , Oceans and Seas , Pseudomonas/genetics , Psychrobacter/genetics , RNA, Ribosomal, 16S/genetics , Shewanella/genetics , Temperature
11.
Int J Syst Evol Microbiol ; 67(9): 3192-3197, 2017 Sep.
Article in English | MEDLINE | ID: mdl-28840795

ABSTRACT

Six Gram-negative, non-motile, non-spore-forming, non-pigmented, oxidase- and catalase-positive bacterial strains were deposited in 1972, in the Collection of the Institut Pasteur (CIP), Paris, France. The strains, previously identified as members of the genus Moraxella on the basis of their phenotypic and biochemical characteristics, were placed within the genus Psychrobacter based on the results from comparative 16S rRNA gene sequence studies. Their closest phylogenetic relatives were Psychrobacter sanguinis CIP 110993T, Psychrobacter phenylpyruvicus CIP 82.27T and Psychrobacter lutiphocae CIP 110018T. The DNA G+C contents were between 42.1 and 42.7 mol%. The predominant fatty acids were C18 : 1ω9c, C16 : 0, C12 : 0 3-OH, and C18 : 0. Average nucleotide identity between the six strains and their closest phylogenetic relatives, as well as their phenotypic characteristics, supported the assignment of these strains to two novel species within the genus Psychrobacter. The proposed names for these strains are Psychrobacter pasteurii sp. nov., for which the type strain is A1019T (=CIP 110853T=CECT 9184T), and Psychrobacter piechaudii sp. nov., for which the type strain is 1232T (=CIP110854T=CECT 9185T).


Subject(s)
Phylogeny , Psychrobacter/classification , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , DNA, Ribosomal/genetics , Fatty Acids/chemistry , France , Psychrobacter/genetics , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
12.
Extremophiles ; 21(3): 581-589, 2017 May.
Article in English | MEDLINE | ID: mdl-28314921

ABSTRACT

Psychrobacter has been regarded as an important genus for bacterial cold adaptation studies. However, members of this genus are highly varied in terms of both cold adaptability and genome content. To get an understanding of the diversity of members of this genus, five Psychrobacter strains (G, K5, 273-4, PAMC21119 and PRwf-1), with publicly available complete/draft genome, were selected and comprehensive comparative genomics analyses were performed among them. The closest phylogenetic relationship, highest average nucleotide identity (96.78%) and best sequence synteny were identified between strains G and K5. These findings suggest they belong to the same species, despite the long geographic distance between them (Antarctic and Siberia). 4542 gene clusters in total were identified from the five genomes, and of which 1424 were shared by all of them. The number of genes unique to strains G, K5, 273-4, PAMC21119 and PRwf-1 are 183, 188, 300, 637 and 665, respectively. COG assignment revealed their differences in gene content related to stress response. The extensive sequence rearrangements and the large number of genes unique to strain PAMC21119 and PRwf-1 suggest they may have experienced a high level of gene exchanges in the permafrost soil and the surface of fish skin.


Subject(s)
Genome, Bacterial , Polymorphism, Genetic , Psychrobacter/genetics , Adaptation, Physiological , Ecosystem , Permafrost/microbiology , Phylogeny , Psychrobacter/classification , Psychrobacter/isolation & purification , Sequence Homology , Synteny
13.
Int J Syst Evol Microbiol ; 66(12): 5091-5098, 2016 Dec.
Article in English | MEDLINE | ID: mdl-27609593

ABSTRACT

Two closely related aerobic, Gram-stain-negative, rod-shaped bacteria (S6-60T and S6-67) were isolated from the mucus of the coral, Pocilloporaeydouxi, from the Andaman Sea, India. Heterotrophic growth on marine agar was observed at 4-37 °C and at pH 6.5-10.0; optimum growth occurred at 25-30 °C and at pH 7-9. 16S rRNA gene sequence analysis confirmed that the isolates belong to the genus Psychrobacter; the two isolates shared more than 99.5 % pairwise sequence similarity. Strain S6-60T showed a maximum 16S rRNA similarity of 98.92 % with Psychrobacter pacificensis DSM 23406T. DNA-DNA homology between the two isolates, S6-60T and S6-67, was above 90 %, whereas strain S6-60T showed less than 70 % homology with closely related type species. The DNA G+C content was 47.7 mol%. It contained phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and phospholipid as the major polar lipids. C10 : 0, C12 : 0 3OH, C16 : 0, C18 : 1ω9c, C17 : 1ω8c and C16 : 1ω7c were found to be the predominant fatty acids. Based on a polyphasic analysis, the isolates (S6-60T and S6-67) represent a novel species of the genus Psychrobacter for which the name Psychrobacter pocilloporae sp. nov. is proposed with S6-60T(=JCM 31058T=LMG 29157T) as the type strain.


Subject(s)
Anthozoa/microbiology , Phylogeny , Psychrobacter/classification , Animals , Bacterial Typing Techniques , Base Composition , DNA, Bacterial/genetics , Fatty Acids/chemistry , India , Nucleic Acid Hybridization , Phospholipids/chemistry , Psychrobacter/genetics , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA
14.
Extremophiles ; 20(3): 337-49, 2016 May.
Article in English | MEDLINE | ID: mdl-27033516

ABSTRACT

Permafrost on the Qinghai-Tibet Plateau is one of the most sensitive regions to climate warming, thus characterizing its microbial diversity and community composition may be important for understanding their potential responses to climate changes. Here, we investigated the prokaryotic diversity in a 10-m-long permafrost core from the Qinghai-Tibet Plateau by restriction fragment length polymorphism analysis targeting the 16S rRNA gene. We detected 191 and 17 bacterial and archaeal phylotypes representing 14 and 2 distinct phyla, respectively. Proteobacteria was the dominant bacterial phylum, while archaeal communities were characterized by a preponderance of Thaumarchaeota. Some of prokaryotic phylotypes were closely related to characterized species involved in carbon and nitrogen cycles, including nitrogen fixation, methane oxidation and nitrification. However, the majority of the phylotypes were only distantly related to known taxa at order or species level, suggesting the potential of novel diversity. Additionally, both bacterial α diversity and community composition changed significantly with sampling depth, where these communities mainly distributed according to core horizons. Arthrobacter-related phylotypes presented at high relative abundance in two active layer soils, while the deeper permafrost soils were dominated by Psychrobacter-related clones. Changes in bacterial community composition were correlated with most measured soil variables, such as carbon and nitrogen contents, pH, and conductivity.


Subject(s)
Microbiota , Permafrost/microbiology , Archaea/genetics , Archaea/isolation & purification , Archaea/metabolism , Carbon/analysis , Carbon/metabolism , Nitrogen/analysis , Nitrogen/metabolism , Permafrost/chemistry , Psychrobacter/genetics , Psychrobacter/isolation & purification , Psychrobacter/metabolism , RNA, Ribosomal, 16S/genetics , Tibet
15.
BMC Genomics ; 17: 117, 2016 Feb 16.
Article in English | MEDLINE | ID: mdl-26879123

ABSTRACT

BACKGROUND: Marine cold-temperature environments are an invaluable source of psychrophilic microbial life for new biodiscoveries. An Arctic marine bacterial strain collection was established consisting of 1448 individual isolates originating from biota, water and sediment samples taken at a various depth in the Barents Sea, North of mainland Norway, with an all year round seawater temperature of 4 °C. The entire collection was subjected to high-throughput screening for detection of extracellular laccase activity with guaiacol as a substrate. RESULTS: In total, 13 laccase-positive isolates were identified, all belonging to the Psychrobacter genus. From the most diverse four strains, based on 16S rRNA gene sequence analysis, all originating from the same Botryllus sp. colonial ascidian tunicate sample, genomic DNA was isolated and genome sequenced using a combined approach of whole genome shotgun and 8 kb mate-pair library sequencing on an Illumina MiSeq platform. The genomes were assembled and revealed genome sizes between 3.29 and 3.52 Mbp with an average G + C content of around 42%, with one to seven plasmids present in the four strains. Bioinformatics based genome mining was performed to describe the metabolic potential of these four strains and to identify gene candidates potentially responsible for the observed laccase-positive phenotype. Up to two different laccase-like multicopper oxidase (LMCO) encoding gene candidates were identified in each of the four strains. Heterologous expression of P11F6-LMCO and P11G5-LMCO2 in Escherichia coli BL21 (DE3) resulted in recombinant proteins exhibiting 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulphonic acid (ABTS) and guaiacol oxidizing activity. CONCLUSIONS: Thirteen Psychrobacter species with laccase-positive phenotype were isolated from a collection of Arctic marine bacteria. Four of the isolates were genome sequenced. The overall genome features were similar to other publicly available Psychrobacter genome sequences except for P11G5 harboring seven plasmids. However, there were differences at the pathway level as genes associated with degradation of phenolic compounds, nicotine, phenylalanine, styrene, ethylbenzene, and ethanolamine were detected only in the Psychrobacter strains reported in this study while they were absent among the other publicly available Psychrobacter genomes. In addition, six gene candidates were identified by genome mining and shown to possess T1, T2 and T3 copper binding sites as the main signature of the three-domain laccases. P11F6-LMCO and P11G5-LMCO2 were recombinantly expressed and shown to be active when ABTS and guaiacol were used as substrates.


Subject(s)
Genome, Bacterial , Oxidoreductases/metabolism , Phylogeny , Psychrobacter/classification , Arctic Regions , Bacterial Typing Techniques , Base Composition , Base Sequence , Cold Temperature , DNA, Bacterial/genetics , Molecular Sequence Data , Norway , Psychrobacter/enzymology , Psychrobacter/genetics , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Seawater/microbiology , Sequence Analysis, DNA
16.
Arch Microbiol ; 198(2): 129-35, 2016 Mar.
Article in English | MEDLINE | ID: mdl-26546315

ABSTRACT

The genus Psychrobacter contains environmental, psychrophilic and halotolerant gram-negative bacteria considered rare opportunistic pathogens in humans. Metagenomics was performed on the cerebrospinal fluid (CSF) of a pediatric patient with meningitis. Nucleic acids were extracted, randomly amplified, and sequenced with the 454 GS FLX Titanium next-generation sequencing (NGS) system. Sequencing reads were assembled, and potential virulence genes were predicted. Phylogenomic and phylogenetic studies were performed. Psychrobacter sp. 310 was identified, and several virulence genes characteristic of pathogenic bacteria were found. The phylogenomic study and 16S rRNA gene phylogenetic analysis showed that the closest relative of Psychrobacter sp. 310 was Psychrobacter sanguinis. To our knowledge, this is the first report of a meningitis case associated with Psychrobacter sp. identified by NGS metagenomics in CSF from a pediatric patient. The metagenomic strategy based on NGS was a powerful tool to identify a rare unknown pathogen in a clinical case.


Subject(s)
Cerebrospinal Fluid/microbiology , Meningitis/microbiology , Metagenomics , Moraxellaceae Infections/microbiology , Psychrobacter/genetics , Adolescent , Base Sequence , Fatal Outcome , Genome, Bacterial/genetics , Humans , Male , Meningitis/cerebrospinal fluid , Mexico , Molecular Sequence Data , Moraxellaceae Infections/cerebrospinal fluid , Phylogeny , Psychrobacter/classification , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Virulence Factors/genetics
17.
Antonie Van Leeuwenhoek ; 108(6): 1283-1292, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26362329

ABSTRACT

A Gram-negative, non-motile, non-spore-forming, psychrotolerant and halotolerant bacterium designated BSw21516B(T), was obtained from seawater in Kongsfjorden, a glacial fjord in the Arctic Svalbard and subjected to taxonomic analysis using a polyphasic approach. This bacterium was observed to optimally grow at 25-29 °C; between at 4 and 34 °C, but not at >35 °C; and in the presence of 0-8 % (w/v) NaCl at an optimum concentration of 2-5 % (w/v) NaCl. Strain BSw21516B(T) was found to contain Ubiquinone-8 (Q-8) as a predominant respiratory lipoquinone and C18:1 ω9c and summed feature 3 (C16:1 ω7c and/or iso-C15:0 2-OH) as predominant cellular fatty acids. Phylogenetic analysis of 16S rRNA and gyrB gene sequences showed that this isolate belongs to the genus Psychrobacter and is closely related to Psychrobacter fozii LMG 21280(T), which was isolated from a sediment sample in Antarctica. DNA hybridization experiments revealed a low level of DNA-DNA relatedness (less than 58.6 %) between strain BSw21516B(T) and its closest relatives. Based on these results a new species Psychrobacter fjordensis sp. nov. is proposed (type strain BSw21516B(T) = KCTC 42279(T) = CCTCC AB 2014020(T)).


Subject(s)
Psychrobacter/classification , Psychrobacter/isolation & purification , Seawater/microbiology , Arctic Regions , Bacterial Typing Techniques , Cluster Analysis , Cytosol/chemistry , DNA Gyrase/genetics , DNA, Bacterial/chemistry , DNA, Bacterial/genetics , DNA, Ribosomal/chemistry , DNA, Ribosomal/genetics , Estuaries , Fatty Acids/analysis , Molecular Sequence Data , Nucleic Acid Hybridization , Phylogeny , Psychrobacter/genetics , Psychrobacter/physiology , Quinones/analysis , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Sodium Chloride/metabolism , Svalbard , Temperature
18.
Microbiol Res ; 179: 38-44, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26411893

ABSTRACT

Food production and processing industry holds a perpetual relationship with microorganisms and their by-products. In the present study, we aimed to identify beneficial cold-adapted bacteria devoid of any food spoilage properties and study their antagonism against common food-borne pathogens at low temperature conditions. Ten isolates were obtained on selective isolation at 5 °C, which were spread across genera Pseudomonas, Sphingomonas, Psychrobacter, Leuconostoc, Rhodococcus, and Arthrobacter. Methanol extracts of strains were found to contain several bioactive metabolites. Among the studied isolates, methanol extracts of S. faeni ISY and Rhodococcus fascians CS4 were found to show antagonism against growth of Escherichia coli, Proteus mirabilis, Enterobacter aerogenes, Listeria monocytogenes and Vibrio fischeri at refrigeration temperatures. Characterization of the abundant yellow pigment in methanol extracts of S. faeni ISY through UV-Vis spectrophotometry, high performance liquid chromatography (HPLC) and mass spectrometry (LC-MS) revealed the presence of astaxanthin, which, owing to its presence in very large amounts and evidenced to be responsible for antagonistic activity of the solvent extract.


Subject(s)
Anti-Bacterial Agents/pharmacology , Cold Temperature , Sphingomonas/metabolism , Arthrobacter/drug effects , Arthrobacter/isolation & purification , Colony Count, Microbial , Food Microbiology , Leuconostoc/drug effects , Leuconostoc/isolation & purification , Listeria monocytogenes/drug effects , Listeria monocytogenes/isolation & purification , Methanol/chemistry , Microbial Sensitivity Tests , Pseudomonas/drug effects , Pseudomonas/isolation & purification , Psychrobacter/drug effects , Psychrobacter/isolation & purification , Rhodococcus/drug effects , Rhodococcus/isolation & purification , Sphingomonas/isolation & purification , Xanthophylls/pharmacology
19.
Int J Syst Evol Microbiol ; 65(9): 3046-3052, 2015 Sep.
Article in English | MEDLINE | ID: mdl-26065735

ABSTRACT

A novel, aerobic, psychrotolerant, Gram-stain-positive, endospore-forming strain, NHI-2(T), was isolated from oil-contaminated soil near a gas station in Mongolia. This strain was characterized by motile rods and grew over a wide range of temperatures ( -2 to 40 °C) with optimal growth at 28-30 °C. It tolerated salt concentrations of up to 7% over a five-day incubation period. Analysis of 16S rRNA gene sequence indicated that strain NHI-2(T) belongs to the genus Psychrobacillus. Sequence similarity between NHI-2(T) and members of the genus Psychrobacillus with validly published names ranged from 97.83 to 98.18%. DNA-DNA hybridization indicated less than 70% relatedness to reference strains within the genus. The G+C content of the genomic DNA was 36 mol%. This strain contained MK-8 as a predominant isoprenoid menaquinone. NHI-2(T) had ornithine in the cell wall similar to reference strains of the genus Psychrobacillus. The major fatty acids present in NHI-2(T )were anteiso-C15 : 0 (51.0%), iso-C15 : 0 (9.1%) and anteiso-C17 : 0 (8.0%). The major polar lipids were phosphatidylethanolamine, diphosphatidylglycerol and phosphatidylglycerol. These data highlight that the phenotype of strain NHI-2(T) differs from that of related species in terms of chemotaxonomic properties and genotype characteristics. Therefore, this strain is proposed as a representative of a novel species, named Psychrobacillus soli. The type strain is NHI-2(T) ( = KEMB 9005-135(T) = KACC 18243(T) = NBRC 110600(T)).


Subject(s)
Psychrobacter , Soil Microbiology , Bacillaceae/classification , Base Composition , DNA, Bacterial , Environmental Pollution , Fatty Acids/analysis , Molecular Sequence Data , Mongolia , Nucleic Acid Hybridization , Phylogeny , Psychrobacter/isolation & purification , Psychrobacter/metabolism , RNA, Ribosomal, 16S/genetics , Sequence Analysis, DNA , Soil
20.
PLoS One ; 10(5): e0125844, 2015.
Article in English | MEDLINE | ID: mdl-25970606

ABSTRACT

A survey was carried out on the microbial community of 20 groundwater samples (4 low and 16 high arsenic groundwater) and 19 sediments from three boreholes (two high arsenic and one low arsenic boreholes) in a high arsenic groundwater system located in Hetao Basin, Inner Mongolia, using the 454 pyrosequencing approach. A total of 233,704 sequence reads were obtained and classified into 12-267 operational taxonomic units (OTUs). Groundwater and sediment samples were divided into low and high arsenic groups based on measured geochemical parameters and microbial communities, by hierarchical clustering and principal coordinates analysis. Richness and diversity of the microbial communities in high arsenic sediments are higher than those in high arsenic groundwater. Microbial community structure was significantly different either between low and high arsenic samples or between groundwater and sediments. Acinetobacter, Pseudomonas, Psychrobacter and Alishewanella were the top four genera in high arsenic groundwater, while Thiobacillus, Pseudomonas, Hydrogenophaga, Enterobacteriaceae, Sulfuricurvum and Arthrobacter dominated high arsenic sediments. Archaeal sequences in high arsenic groundwater were mostly related to methanogens. Biota-environment matching and co-inertia analyses showed that arsenic, total organic carbon, SO4(2-), SO4(2-)/total sulfur ratio, and Fe(2+) were important environmental factors shaping the observed microbial communities. The results of this study expand our current understanding of microbial ecology in high arsenic groundwater aquifers and emphasize the potential importance of microbes in arsenic transformation in the Hetao Basin, Inner Mongolia.


Subject(s)
Arsenic/chemistry , Groundwater/microbiology , Soil Microbiology , Water Microbiology , Water Pollutants, Chemical/chemistry , Acinetobacter/genetics , Acinetobacter/isolation & purification , Arthrobacter/genetics , Arthrobacter/isolation & purification , China , Comamonadaceae/genetics , Comamonadaceae/isolation & purification , Enterobacteriaceae/genetics , Enterobacteriaceae/isolation & purification , Groundwater/chemistry , Microbiota , Molecular Typing , Pseudomonas , Psychrobacter/genetics , Psychrobacter/isolation & purification , RNA, Ribosomal, 16S/genetics , Shewanella/genetics , Shewanella/isolation & purification , Thiobacillus/genetics , Thiobacillus/isolation & purification
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