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Matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry (MALDI‐TOF MS) is an excellent tool for bacterial identification. It allows high throughput, sensitive and specific applications in clinical diagnostics and environmental research. Currently, there is no optimal standardized protocol for sample preparation and culture conditions to profile bacteria. The performance of MALDI‐TOF MS is affected by several variables, such as sample preparation, culture media and culture conditions, incubation time/growth stage, incubation temperature, high salt content, blood in the culture media, and others. This review thus aims to clarify why a uniformed protocol is not plausible, to assess the effects these factors have on MALDI‐TOF MS identification score, and discuss possible optimizations for its methodology, in relation to specific bacterial representatives and strain requirements. [ABSTRACT FROM AUTHOR]

The aim of this study was to compare the effectiveness of gas chromatography (GC) and near‐infrared spectroscopy (NIR) spectroscopy in detecting butter adulteration. Butter samples with a known percentage of added margarine or pork fat were prepared. Chemometric techniques (principal component analysis and partial least squares regression) were applied on the NIR spectra and GC data, which enabled the qualitative separation of samples depending on the added proportion of adulterant, and the quantitative prediction of the proportion of added adulterant or certain fatty acid content. C12:0; C16:0; C18:0; C16:1; C18:1n9; C18:2n6 showed significant trend changes depending on the adulteration degree. NIR has proven to be a suitable technique for the rapid determination of butter adulteration. Novelty impact statement: Fast and nondestructive technique as near‐infra red (NIR) spectroscopy can be used in qualitative and quantitative detection of butter originality. Obtained models by chemometrics methods allow prediction of proportion of added adulterant in butter and identify specific fatty acids. NIR may be applied in screening control of the correctness of butter labeling to improve the system of quality control in food fraud. [ABSTRACT FROM AUTHOR]

Gallibacterium anatis is considered one of the most common bacterial causative agents of reproductive tract disorders in poultry. In this study, phylogenetic analysis of partial rpoB sequences and biotyping using MALDI-TOF MS was done in order to investigate the genetic diversity of Gallibacterium isolates from 13 farms with different biosecurity measures and management practices. Sampling was done as a part of regular monitoring, except for Farms 9–13 that were included in the study to represent extensive production systems with lowest biosecurity levels. Pharyngeal and cloacal swabs were taken from live birds, while swabs from trachea, liver, peritoneum and oviduct were taken during necropsies. After cultivation and identification, strains from each farm were randomly selected for sequencing and biotyping. Both results showed high level of heterogeneity among the isolates originating from farms with low biosecurity levels, unlike isolates from farms with higher biosecurity levels and proper management that were more closely related and clustered together. Such correlation was statistically significant. Low biosecurity levels enable horizontal transmission of the pathogens, as well as gene transfer. The results confirm the importance of adequate biosecurity measures and management on poultry farms as they greatly affect the genetic diversity of the pathogens. Therefore, implementation of basic biosecurity measures could help control the heterogeneity of Gallibacterium strains, which would alleviate control of the infection prevalence on farms through immunoprophylaxis, and consequently improve poultry production. Also, the genetic diversity of G. anatis on poultry farms could be a good bioindicator of management practices and biosecurity measures used. RESEARCH HIGHLIGHTS High correlation between low biosecurity and high diversity of Gallibacterium anatis. Diversity of Gallibacterium is a good bioindicator of management practices on farms. [ABSTRACT FROM AUTHOR]

Vibrio (Listonella) anguillarum is a pathogenic bacterium causing septicaemia in a wide range of marine organisms and inducing severe mortalities, thus it is crucial to conduct its accurate and rapid identification. The aim of this study was to assess MALDI-TOF MS as a method of choice for identification of clinical V. anguillarum isolates from affected marine fish. Since the method accuracy might be influenced by the type of the medium used, as well as by the incubation conditions, we tested V. anguillarum isolates grown on standard media with and without the addition of NaCl, cultured at three incubation temperatures, and at three incubation periods. The best scores were retrieved for V. anguillarum strains grown on NaCl-supplemented tryptone soy agar (TSA) at 22°C and incubated for 48h (100% identification to species level; overall score 2.232), followed by incubation at 37°C and 48h (100% to species level; score 2.192). The strains grown on non-supplemented TSA gave the best readings when incubated at 22°C for 72h (100% identification to species level; overall score 2.182), followed by incubation at 15°C for 72h (100% to species level; score 2.160). Unreliable identifications and no-identifications were growing with the incubation duration at 37°C, on both media, amounting to 88.89% for 7d incubation on supplemented TSA, and 92.60% for 7d incubation on non-supplemented TSA. The age of the cultured strains and use of media significantly impacted the mass spectra, demonstrating that for reliable identification, MALDI-TOF MS protein fingerprinting with the on-target extraction should be performed on strains grown on a NaCl-supplemented medium at temperatures between 15 and 22°C, incubated for 48–72 hours. [ABSTRACT FROM AUTHOR]

MALDI‐TOF MS was tested for the identification of Photobacterium damselae subsp. piscicida on isolates grown on two media, cultured at three incubation times and applied on the target plate by the direct sample spotting (DS), by the on‐target extraction (OTE) and by the full extraction (FE) method, in triplicates. The identification of samples grown on blood agar (BA) outperformed identification on tryptic soya agar (TSA) by 0.64% for DS and OTE. The OTE gave the highest scores in both culture media, all incubation times and replicates. Reliable 24‐hr species identification was 61.54%, 84.61% and 53.85% for samples grown on TSA and identified by DS, OTE and FE, respectively. For isolates grown on BA, they were 76.92%, 96.15% and 30.77%, respectively. When identified by OTE, the 48‐hr identification was 93.58%, but for 72 hr declined to 71.79%. The reliable identification with the highest score from the first measurement was 100% only for OTE from BA (24 hr), whereas OTE from TSA gave 84.61% (24 hr), 76.92% (48 hr) and 84.61% (72 hr). The reliable MALDI‐TOF MS identification of Ph. damselae subsp. piscicida is incubation time, media, target plate preparation and replicate‐dependent. [ABSTRACT FROM AUTHOR]

Abstract The aim of the study was to compare bacterial composition and load in waters and fish related to the wastewater treatment plant (WWTP), particularly waters and wild fish affected by sugarplant processing (sugar cane and sugar beet). Aeromonads were the most frequently isolated group from water and fish. A. hydrophila was a prevailing species in isolates from water, followed by A. veronii, Rheinheimera soli and Ochrobactrum anthropi. Of indicator bacteria for aquatic contamination from fish tissues, the most prominent were V. cholerae , Enterobacter cloacae and E. sakazakii. Sugar cane processing contributed to high viable cell counts at 37 °C while sugar beet processing contributed to high bacterial counts at 22 °C. Heterotrophs from gills of effluent fish were highest during sugar cane processing. Counts retrieved from fish skin were more uniform between effluent fish and fish from downstream waters. Antimicrobial resistance of bacteria isolated from water was high against amoxicillin, sulfamethoxazole, flumequine, norfloxacin and oxolinic acid in samples from the inflow of raw municipal wastewaters to WWTP, while resistance found in bacteria from the inflow of sugarplant mostly related to sulfamethoxazole and amoxicillin. The PCA analysis associated the occurrence of high heterotroph counts, P. aeruginosa , and intestinal enterococci on skin and gills with sugar cane, and yeasts and molds with sugar beet processing. Fish living in treated wastewaters and related water bodies could pose a microbial hazard if fished for human consumption, possibly causing infection when being handled and processed, as a risk of human pathogens penetrating fish tissues. Graphical abstract Image 1 Highlights • We evaluated microbiological impact of sugarplant effluent on wild fish. • Ubiquitous and pathogenic bacteria were retrieved from tissues; mostly aeromonads. • 63% of fish aeromonads showed resistance against tested antimicrobials. • Gill and kidney histopathology showed an increased number of bacterial cells. • Multivariate tools correlated effectively microbiological data, tissues and seasons. [ABSTRACT FROM AUTHOR]

Identification of bacteria in aquatic and environmental applications, for monitoring purposes and research, for health assessments and therapy considerations of farmed and free-living aquatic organisms, still relies on conventional phenotypic and biochemical protocols. Although molecular techniques based on DNA amplification and sequencing are finding ways into diagnostic laboratories, they are time-consuming, costly and difficult in the case of multiplex assays. Matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) is a rapid and accurate proteomic method reliable for identification of unknown bacteria to the genus and species level. Upon extension of databases, it will certainly find its position in environmental sciences. The paper presents an overview of the principle of the method, its effectiveness in comparison with conventional and molecular identification procedures, and applicability on environmental and aquatic isolates, discussing its advantages and shortcomings, as well as possible future implementations. [ABSTRACT FROM AUTHOR]

We demonstrate improved power of Iatroscan thin layer chromatography/flame ionization detection (TLC–FID) technique for analysis of complex marine lipid mixture by developing protocol for the separation and analysis of glycolipids including sulfoquinovosyldiacylglycerols (SQDG), monogalactosyldiacylglycerols (MGDG) and digalactosyldiacylglycerols (DGDG). We have modified the common protocol used so far for the analysis of lipid classes by replacing the elution step which uses pure acetone for the elution of acetone mobile polar lipids, with the elution step containing chloroform–acetone (72:28, v:v) for separation of MGDG and DGDG. To separate SQDG from the complex lipid matrix we introduced solvent mixture acetone–chloroform–methanol–formic acid (33:33:33:0.6, v:v:v:v). Quantification of glycolipid classes was performed after calibration with glycolipid standards for the masses between 0.2 and 2.7–5.0 μg. With this new protocol we have successfully separated three glycolipids from the complex particulate lipid mixture of the seawater samples. Such an approach extends the power of existing protocol for the analysis of lipids which altogether ensure detection and quantification of 18 lipid classes what was demonstrated on seawater samples. This enables to gain a very broad system overview of the particularly complex environments as are seas, oceans and freshwaters. [ABSTRACT FROM AUTHOR]

The reactivity of new biologically active thymine derivatives substituted with 2-(arylsulfonamidino)ethyl group at N1 and N3 position was investigated in the gas phase using CID experiments (ESI-MS/MS) and by density functional theory (DFT) calculations. Both derivatives show similar chemistry in the negative mode with a retro-Michael addition (Path A) being the most abundant reaction channel, which correlate well with the fluoride induced retro-Michael addition observed in solution. The difference in the fragmentation of N-3 substituted thymine 5 and N-1 substituted thymine 1 in the positive mode relates to the preferred cleavage of the sulfonyl group ( m/z 155, Path B) in N-3 isomer and the formation of the acryl sulfonamidine 3 ( m/z 309) via Path A in N-1 isomer. Mechanistic studies of the cleavage reaction conducted by DFT calculations give the trend of the calculated activation energies that agree well with the experimental observations. A mechanism of the retro-Michael reaction was interpreted as a McLafferty type of fragmentation, which includes H proton shift to one of the neighboring oxygen atoms in a 1,5-fashion inducing N1(N3)-C bond scission. This mechanism was found to be kinetically favorable over other tested mechanisms. Significant difference in the observed fragmentation pattern of N-1 and N-3 isomers proves the ESI-MS/MS technique as an excellent method for tracking the fate of similar sulfonamidine drugs. Also, the observed N-1 and/or N-3 thymine alkylation with in situ formed reactive acryl sulfonamidine 3 as a Michael acceptor may open interesting possibilities for the preparation of other N-3 substituted pyrimidines. [Figure not available: see fulltext.] [ABSTRACT FROM AUTHOR]

Abstract: The electronic structures of 3- and 5-hydroxy-flavones, naringenin and 3-bromochromone have been studied by UV photoelectron spectroscopy (UPS) and high-level Green''s function (GF) calculations. Our UPS data show that hydroxyl substitution strongly influences the electronic structures of flavones and flavanones which in turn influences their biological (radical scavenging or antioxidant) activity. In particular, the experimental antioxidant activity correlates with HOMO π-ionization energies. [Copyright &y& Elsevier]

In this study, the purposefulness of using the API20E biochemical identification system as a supportive tool for enhancing the discrimination of environmental bacteria by MALDI-TOF MS method was evaluated. The identification results of MALDI-TOF MS and API20E for 321 Gram-negative strains isolated from the riverine freshwater and its sediment, and from the tissues of fish from the same water body were compared. Of 190 isolates identified with probable to highly probable species-level identification, and secure genus to probable species identification, 14 isolates (7.37%) had identification score over 2.300, and from the same group 19 isolates (10%) had excellent or very good identification to the genus by API20E system. With regard to agreement at genus level, out of 231 strains with genus designation available by API20E at any level of identification reliability, MALDI-TOF MS genus identification agreed in 163 (70.6%) strains. Of these, 135 (82.8%) were Aeromonas species and the remaining isolates belonged to 7 different genera. Although API20E resulted in frequent misidentification due to a limited profile index, its individual biochemical reactions might contribute to overall characterization of isolates. For example, for all reliable A. hydrophila strain identifications with MALDI-TOF MS, ONPG, GLU and OX reactions were unarguably positive for all fish and water/sediment isolates, whereas only fish isolates yielded additional 100% positive TDA and VP reactions. Thus, after initial identification with MALDI-TOF MS, environmental isolates with lower identification scores should be further analyzed. Before commencing confirmatory testing with nucleic acid-based methods, biochemical API20E tests could be applied as a purposeful and inexpensive identification support in targeting better identification accuracy. In this study, this was particularly evident with A. hydrophila, Chryseobacterium sp. and Pseudomonas sp. This identification strategy could significantly resolve methodological and cost-related shortcomings frequently occurring with large number of environmental isolates. [ABSTRACT FROM AUTHOR]

The gas-phase reaction products of Cr+, Fe+ and Ni+ ions produced by laser desorption ionization (LDI) of a stainless steel sample with iodine molecules in a Fourier transform ion cyclotron resonance mass spectrometer (FT ICR MS) uncover differences in the behavior of individual metal ions in the ligation process as well as in the reactions with ubiquitous water: Fe being prone to ligation of two (but also to hydrolysis and disappearance), Ni+ to ligation of three and Cr+ preferring to ligate four iodine atoms. The surplus energy of metal ions produced in the excited states enables them to dissociate I2 and exchange charge with the I2 molecules and I atoms forming I2+ and I3+ ions. [ABSTRACT FROM AUTHOR]

Wastewaters from a treatment plant discharging into a canal harboring fish may present sources of microbiological hazard for wild fish. Such fish, inhabiting microbiologically polluted bodies of water, can be contaminated by human pathogens and, if used for human consumption, may pose a risk to public health. Hence, in this work the aim was to identify tested strains from tissues of wild fish living in the receiving water bodies, captured from locations up to 12 km from the point of discharge of treated water of town Virovitica in order to assess the bacterial threat of the WWTP on fish and potentially on public health. A rather rich diversity of bacterial genera was isolated from gill tissues and internal organs. The most frequent isolate was Aeromonas hydrophila which has gained public health recognition as an opportunistic pathogen. Vibrio cholerae, an indicator bacterium for aquatic contamination, was retrieved from all investigated tissues. Opportunistic human pathogens as well as some zoonotic agents were also retrieved from fish tissues (Enterobacter amnigenus, Acinetobacter spp., Ochrobactrum anthropi, Pseudomonas aeruginosa, Flavimonas (Pseudomonas) oryzihabitans, Shewanella putrefaciens and others). Public health hazard is particularly pronounced regarding local recreational fishermen who fish out, handle and consume fish from respective waters. [ABSTRACT FROM AUTHOR]

Two main parameters that structure the marine ecosystem by affecting species distribution, abundance, community structure, timing of major life cycle events, and trophic state of the ecosystem are temperature and nutrient availability. Faced with climate change, eukaryotic plankton cope at multiple levels through physiological accommodation, adaptive evolution, shift in time and/or space of habitat, and/or community responses. Thirty‐two years of our phytoplankton research indicate that Chaetoceros curvisetus/pseudocurvisetus adjust to climate change more successfully than the majority of the accompanying phytoplankton taxa in the mesotrophic part of the NW Adriatic Sea, the Mediterranean. While the abundance of the entire accompanying phytoplankton community has decreased significantly since 2003 (the period of the northern Adriatic warming and oligotrophication) compared to the previous period (1986–2003), the abundance of C. curvisetus/pseudocurvisetus remained unchanged, while its contribution to the community increased significantly. Accommodation strategies include a change in the timing of high abundance and blooms in the surface layer and successful blooming in the deeper layers during warm months. Apart from the observed in situ accommodation, physiological acclimation to warming may involve changes in photosynthesis, respiration, growth, and cell biochemistry. Here, we conducted laboratory experiments with C. pseudocurvisetus to investigate how warming affects its biochemical response through the fatty acid remodeling of phospholipid classes. Long‐term field observations and short‐term laboratory experiments suggest that marine diatoms C. curvisetus/pseudocurvisetus are potential global winners with the ability to acclimate/adapt to climate change. [ABSTRACT FROM AUTHOR]

Organic farming preserves biodiversity and organic products can be the source of many microbial species. The species diversity in organically grown wheat, spelt and rye was investigated in order to find strains suitable for sourdough fermentation. Colonies representing various morphological appearances were isolated and catalase-negative colonies were identified by mass spectrometer Microflex LT ™ MALDI-TOF. The fermentation products (lactic, acetic, formic and phenyllactic acid) were determined by high performance liquid chromatography, while the antifungal activity was determined using an overlay agar method. Wheat flours showed less microbial biodiversity than the rye and spelt flours. The most common genera in the tested flour were Lactobacillus, Pediococcus and Enterococcus. Isolated Lactobacillus farciminis, Pediococcus pentosaceus, Leuconostoc citreum and Lactobacillus brevis showed the best acidification activity. Lactobacillus brevis, Pediococcus pentosaceus, Weissella cibaria and Lactobacillus farciminis showed significant antifungal activity against A. niger 357 and Penicillium sp. 505. The prefermented medium of Lactobacillus farciminis were characterized by high content of lactic and phenyllactic acid. [ABSTRACT FROM AUTHOR]

Human milk not only provides a perfect balance of nutrients to meet all the needs of the infant in the first months of life but also contains a variety of bacteria that play a key role in tailoring the neonatal faecal microbiome. Microbiome analysis of human milk and infant faeces from mother-breastfed infant pairs was performed by sequencing the V1–V3 region of the 16S rRNA gene using the Illumina MiSeq platform. According to the results, there is a connection in the composition of the microbiome in each mother-breastfed infant pair, supporting the hypothesis that the infant's gut is colonised with bacteria from human milk. MiSeq sequencing also revealed high biodiversity of the human milk microbiome and the infant faecal microbiome, whose composition changes during lactation and infant development, respectively. A total of 28 genetically distinct strains were selected by hierarchical cluster analysis of RAPD-PCR (Random Amplified Polymorphic DNA-Polymerase Chain Reaction) electrophoresis profiles of 100 strains isolated from human milk and identified by 16S RNA sequencing. Since certain cellular molecules may support their use as probiotics, the next focus was to detect (S)-layer proteins, bacteriocins and exopolysaccharides (EPSs) that have potential as therapeutic biomolecules. SDS-PAGE (Sodium Dodecyl-Sulfate Polyacrylamide Gel Electrophoresis) coupled with LC-MS (liquid chromatography-mass spectrometry) analysis revealed that four Levilactobacillus brevis strains expressed S-layer proteins, which were identified for the first time in strains isolated from human milk. The potential biosynthesis of plantaricin was detected in six Lactiplantibacillus plantarum strains by PCR analysis and in vitro antibacterial studies. 1H NMR (Proton Nuclear Magnetic Resonance) analysis confirmed EPS production in only one strain, Limosilactobacillus fermentum MC1. The overall microbiome analysis suggests that human milk contributes to the establishment of the intestinal microbiota of infants. In addition, it is a promising source of novel Lactobacillus strains expressing specific functional biomolecules. [ABSTRACT FROM AUTHOR]

Due to high anthropogenic pressures, science-based coastal management required to ensure the sustainable use of coastal areas highly depends on environmental indicators used for decision-making. In this paper, we argue for the inclusion of Vibrio spp. abundance as a supplemental indicator of water quality for science-based coastal management by examining the environmental and bacterial indicators at a fish farm and a control site in Mali Ston Bay in the Adriatic Sea. Unexpectedly, heterotrophic bacteria, enterococci and Vibrio spp. were more abundant in the cold season, while E. coli and total coliforms, following a more traditional pattern, were more abundant in the warm season. Each of the currently used indicators has a specific purpose: heterotrophic bacteria indicate the presence of both nonpathogenic and pathogenic bacteria, while enterococci are pathogenic bacteria indicating fecal pollution. Vibrio spp. abundance additionally represents a non-fecal bacteria that can cause vibriosis in humans and aquatic organisms. Since vibriosis is the leading cause of disease-related fish mortality in aquaculture, pathogenic Vibrio spp. have large health and economic implications. These implications, as well as the added interpretative value when compared to other bacterial indicators, make Vibrio spp. abundance a good candidate as a water quality indicator. Significant dependence of the abundance on depth further differentiates Vibrio spp. from other indicators, thus bolstering the candidacy - especially in aquaculture areas. Before inclusion of any Vibrio spp. indicators into legislature, further research is needed particularly into (i) abundance thresholds characterizing water quality, and (ii) identification of species whose abundance should be monitored for best estimate of the disease risks. [Display omitted] • Analysis of environmental and bacterial indicators revealed no difference between fish farm and control. • Contrary to expectations, heterotrophic bacteria and Vibrio spp. thrived in the cold season. • Vibrio spp. abundance provides additional information on water quality and health risks. • Vibrio spp. could indicate poor water quality, even when other indicators suggest the quality is excellent. [ABSTRACT FROM AUTHOR]

• As-rich mixture of pharmacolite, hornesite, and talmessite from Crven Dol mine. • Stenotrophomonas sp., Microbacterium spp. and bacterial consortium were isolated. • Novel hyper-resistant bacteria survive 32 g/L arsenite and 176 g/L arsenate. • As-tolerance involve efflux systems including ArsB, Acr3(1) and Acr3(2) proteins. • SEM/EDS analyses confirmed intracellular accumulation of arsenic. Novel hyper-resistant bacteria were isolated from the Crven Dol mine (Allchar, North Macedonia), arsenic-rich extreme environment. Bacteria were recovered from a secondary mineral mixture, an alteration of hydrothermal realgar rich in arsenates (pharmacolite, hornesite, and talmessite). The sample was recovered from the dark part of the mine at 28 m depth. Three bacterial strains and a bacterial consortium were isolated for their capacity to survive exposure to 32 g/L (209 mM) of arsenite, and 176 g/L (564 mM) of arsenate. The 16S rRNA gene analysis identified bacterial isolates as Stenotrophomonas sp. and two Microbacterium spp. This analysis also revealed that bacterial consortium comprise two Bacteriodetes exhibiting similarity to Olivibacter ginsengisoli and to uncultured bacterium, and one γ-proteobacteria with similarity to Luteimonas sp. Among all isolates Stenotrophomonas sp. exhibited the highest tolerance to As compound as well as the capacity to accumulate As inside the cells. Analysis of genes involved in As-resistance showed that recovered isolates possess the genes encoding the ArsB, Acr3(1) and Acr3(2) proteins, indicating that at least a part of their resistance could be ascribed to As-efflux systems described in isolates obtained from human-polluted environments. [ABSTRACT FROM AUTHOR]

Due to the development of resistance to antimicrobial agents, bacterium Acinetobacter baumannii is nowadays a leading cause of nosocomial outbreaks. Clinically relevant A. baumannii outside hospital settings including natural soils affected by human waste represents a public-health risk for humans and animals. The aim of this study was to investigate the potential of metal-loaded zeolites to eliminate viable A. baumannii from artificially contaminated natural soils. A. baumannii isolate was subjected to the activity of natural zeolitised tuff (NZ) and Cu-modified (CuNZ) or Ag-modified zeolite (AgNZ) in wet, slightly acidic terra rossa and slightly alkaline red palaeosol. A. baumannii survived in terra rossa and red palaeosol supplemented with 1 wt% of NZ for seven days and four months, respectively. The addition of 1 wt% of CuNZ to terra rossa and red palaeosol shortened the survival of A. baumannii to three and 14 days, respectively. The addition of 0.1 wt% of AgNZ to both soils resulted in complete removal of viable A. baumannii within 1 h of contact, while the total native heterotrophic bacterial counts remained high. Since AgNZ is prepared with a simple modification of cost-effective and environmentally friendly natural zeolite, it is a promising material for the remediation of soils contaminated with pandrug-resistant A. baumannii. [ABSTRACT FROM AUTHOR]

The strain Pseudomonas aeruginosa san ai, isolated from an extreme environment (industrial mineral cutting oil, pH 10), is able to survive and persist in the presence of a variety of pollutants such as heavy metals and organic chemicals. The genome of P. aeruginosa san ai is 6.98 Mbp long with a GC content of 66.08% and 6485 protein encoding genes. A large number of genes associated with proteins, responsible for microbial resistance to heavy metal ions and involved in catabolism of toxic aromatic organic compounds were identified. P. aeruginosa san ai is a highly cadmium-resistant strain. Proteome analysis of biomass after cadmium exposal confirmed a high tolerance to sublethal concentrations of cadmium (100 mg/L), based on: extracellular biosorption, bioaccumulation, biofilm formation, controlled siderophore production and a pronounced metalloprotein synthesis. Proteins responsible for survival in osmostress conditions during exposure to elevated concentrations of cadmium (200 mg/L) demonstrate a strong genetic potential of P. aeruginosa san ai for survival and adaptation. Sequencing of P. aeruginosa san ai genome provides valuable insights into the evolution and adaptation of this microbe to environmental extremes at the whole-genome level, as well as how to optimally use the strain in bioremediation of chemically polluted sites. [ABSTRACT FROM AUTHOR]

The novel N-1-sulfonylcytosine-cyclam conjugates 1 and 2 conjugates are ionized by electrospray ionization mass spectrometry (ESI MS) in positive and negative modes (ES+ and ES-) as singly protonated/deprotonated species or as singly or doubly charged metal complexes. Their structure and fragmentation behavior is examined by collision induced experiments. It was observed that the structure of the conjugate dictated the mode of the ionization: 1 was analyzed in ES- mode while 2 in positive mode. Complexation with metal ions did not have the influence on the ionization mode. Zn2+ and Cu2+ complexes with ligand 1 followed the similar fragmentation pattern in negative ionization mode. The transformation from 2°-amine in 1 to 3°-amine of cyclam ring in 2 leads to the different fragmentation patterns due to the modification of the protonation priority which changed the fragmentation channels within the conjugate itself. Cu2+ ions formed complexes practically immediately, and the priority had the cyclam portion of the ligand 2. The structure of the formed Zn2+ complexes with ligand 2 depended on the number of 3° amines within the cyclam portion of the conjugate and the ratio of the metal:ligand used. The cleavage of the cyclam ring of metal complexes is driven by the formation of the fragment that suited the coordinating demand of the metal ions and the collision energy applied. Finally, it was shown that the structure of the cyclam conjugate dictates the fragmentation reactions and not the metal ions. [ABSTRACT FROM AUTHOR]

S-layers represent the simplest biological membranes developed during the evolution and are one of the most abundant biopolymers on Earth. Current fundamental and applied research aim to reveal the chemical structure, morphogenesis and function of S-layer proteins (Slps). This is the first paper that describes the Slps of certain Lactobacillus brevis strain isolated from sauerkraut. The whole genome sequence (WGS) analysis of the L. brevis SF9B strain uncovered three genes encoding the putative Slps, but merely one, identified as similar to the SlpB of L. brevis ATCC 14869, was expressed. Slp-expressing SF9B cells exhibited increased survival in simulated gastrointestinal (GI) conditions and during freeze-drying. Their survival in stress conditions was additionally enhanced by microencapsulation, especially when using alginate with gelatine as a matrix. Thus prepared cells were subjected to simulated GI conditions and their mortality was only 0.28 ± 0.45 log CFU/mL. Furthermore, a correlation between the high surface hydrophobicity and the remarkable aggregative capacity of SF9B strain was established. The results indicate a prominent role of Slps in adhesion to mucin, extracellular matrix (ECM) proteins, and particularly to Caco-2 cells, where the removal of Slps utterly abolished the adhesiveness of SF9B cells for 7.78 ± 0.25 log CFU/mL. [ABSTRACT FROM AUTHOR]

Obesity is composed of multifunctional interactions of eating habits, behaviors, microbiota, genetics, and other unknown factors. We hypothesize that correlations occur between the fat mass and obesity-associated single nucleotide polymorphisms ( FTO SNPs), the composition of microorganisms in the saliva, and life habits in obese women from Zagreb County. Our results of the analysis of 3 FTO SNPs showed a statistically significant positive correlation among the frequencies of the high-risk genotypes AA rs9939609 ( P = .0367), CC rs1421085 ( P = .0367), and GG rs17817449 ( P = .0065) of the FTO gene in obese cases. Interestingly, 39.13% of obese women were triple homozygous for all 3 risk alleles. Furthermore, the composition of the oral microbiota in the obese group showed a higher occurrence of a major human pathogen, bacterium Staphylococcus aureus , but a significantly low presence of bacteria Streptococcus oralis , Streptococcus mitis , and Serratia ureilytica compared with the control group. The investigation also revealed that obese women prefer to consume candy and snacks and more meat and meat-derived products, sleep less than 6 hours per day, and had higher hypertension in comparison with the control group. These results support the hypothesis that female obesity is strongly related to all 3 variants of the FTO gene and perhaps a specific composition of microbiota in saliva due to dietary habits. Considering the bimodal distribution of the SNPs and bacterial content of saliva in obese women taken together are factors to consider for risk of obesity. [ABSTRACT FROM AUTHOR]

The aim of this report is to present the electrospray ionization mass spectrometry results of the non-covalent interaction of two biologically active ligands, N-1-(p-toluenesulfonyl)cytosine, 1-TsC, 1 and N-1-methanesulfonylcytosine, 1-MsC, 2 and their Cu(II) complexes Cu(1-TsC-N3)2Cl2, 3 and Cu(1-MsC-N3)2Cl2 and 4 with biologically important cations: Na+, K+, Ca2+, Mg2+ and Zn2+. The formation of various complex metal ions was observed. The alkali metals Na+ and K+ formed clusters because of electrostatic interactions. Ca2+ and Mg2+ salts produced the tris ligand and mixed ligand complexes. The interaction of Zn2+ with 1–4 produced monometal and dimetal Zn2+ complexes as a result of the affinity of Zn2+ ions toward both O and N atoms. [ABSTRACT FROM AUTHOR]

Relating the treated wastewater quality and its impact on organismic biosensors (Prussian carp, Carassius gibelio and earthworm, Eisenia fetida ) was the main objective of the study. The impact on health status of fish living downstream, microbiological contamination and antimicrobial resistance, fish tissue structure, blood biochemistry, oxidative stress, genotoxic effects, as well as multixenobiotic resistance mechanism (MXR) was assessed. Treated wastewater discharged from the WWTP modified the environmental parameters and xenobiotic concentrations of the receiving surface waters. Potential bacterial pathogens from fish and respective waters were found in relatively low numbers, although they comprised aeromonads with a zoonotic potential. High resistance profiles were determined towards the tested antimicrobial compounds, mostly sulfamethoxazole and erythromycin. Histopathology primarily revealed gill lamellar fusion and reduction of interlamellar spaces of effluent fish. A significant increase in plasma values of urea, total proteins, albumins and triglycerides and a significant decrease in the activity of plasma superoxide dismutase were noted in carp from the effluent-receiving canal. Micronucleus test did not reveal significant differences between the examined groups, but a higher frequency of erythrocyte nuclear abnormalities was found in fish sampled from the effluent-receiving canal. Earthworms indicated to the presence of MXR inhibitors in water and sludge samples, thus proving as a sensitive sentinel organism for environmental pollutants. The integrative approach of this study could serve as a guiding principle in conducting evaluations of the aquatic habitat health in complex bio-monitoring studies. [ABSTRACT FROM AUTHOR]

Pathogenic oomycete Aphanomyces astaci , the causative agent of crayfish plague, is listed among the world's 100 worst invasive alien species due to its fast spread and detrimental impact on native European crayfish populations, as well as losses in crayfish aquaculture. During the infection process, A. astaci mycelium invades the host through its cuticle where it interacts with the resident epibiotic bacteria. Since host microbial communities may have an important role in the defense against pathogens, in this study we aimed to i) identify the epibiotic bacterial isolates from the cuticle of two commercially relevant crayfish – the signal crayfish Pacifastacus leniusculus and the narrow-clawed crayfish Pontastacus leptodactylus , and ii) distinguish those that can inhibit the growth of A. astaci mycelium. Results of 16S rRNA gene sequencing and MALDI–TOF mass spectrometry revealed that the majority of collected isolates belonged to the phylum Proteobacteria (69%), followed by Bacteroidetes (17%) and Firmicutes (8%). Using the in vitro plate assays, we tested the potential of the isolates to inhibit A. astaci mycelial growth and classified them as inhibitors (33%) or non-inhibitors (67%). More than half of the identified inhibitors (56%) belonged to the genus Pseudomonas. Our results represent the first step in the understanding of interactions between A. astaci and epibiotic bacteria on the crayfish cuticle and open the perspective for testing in vivo the application of selected bacterial inoculums for crayfish plague prevention and/or treatment. The development of such A. astaci biocontrol measures is urgently needed in astaciculture. • Bacteria were isolated from the cuticle of commercially relevant crayfish. • A third of the isolates inhibited the growth of crayfish pathogen A. astaci. • Pseudomonas isolates were the most potent A. astaci inhibitors. • Our results could start the development of new approaches in A. astaci control. [ABSTRACT FROM AUTHOR]