Pseudomonas Lundensis Research Articles

Pseudomonas fragi and Pseudomonas lundensis drove the co-spoilage in chilled pork: Insights from the metabolome

Pseudomonas fragi and Pseudomonas lundensis are key spoilage microorganisms in aerobically stored chilled meat, which had co-spoilage effect on chilled pork. However, the mechanism of co-spoilage interaction of P. fragi and P. lundensis has not been elucidated. The metabolism of nutrients in pork by mixed bacteria may differ from that by single strains. This study applied metabolomics and in vitro metabolite utilization experiment to assess the mechanism of that co-metabolize raw pork during storage. Results showed that co-spoilage group had 104 differential metabolites (histamine, N-methylhydantoin and D-gluconic acid, etc.) and 78 differential metabolites (putrescine, uracil acid and uracil, etc.) compared to P. fragi and P. lundensis group, respectively. These differential metabolites were mainly related to histidine metabolism, arginine biosynthesis, and purine. The co-spoilage effect of P. fragi and P. lundensis was related to the promotion of deamination and decarboxylation of histidine to produce histamine, the promotion of α-ketoglutarate and glutamate-mediated transamination, and the full utilization of the arginine deiminase pathway by the co-culture bacteria in the degradation of arginine to produce putrescine. In vitro experiment, co-cultures of these strains resulted in greater consumption of glucose, higher utilization activity, and promoted deamination and decarboxylation of amino acids. These findings reveal the complex and competitive interactions of co-culture of P. fragi and P. lundensis, providing insight into microbial spoilage mechanisms in chilled pork.

Global biochemical profiling of fast-growing Antarctic bacteria isolated from meltwater ponds by high-throughput FTIR spectroscopy.

Fourier transform infrared (FTIR) spectroscopy is a biophysical technique used for non-destructive biochemical profiling of biological samples. It can provide comprehensive information about the total cellular biochemical profile of microbial cells. In this study, FTIR spectroscopy was used to perform biochemical characterization of twenty-nine bacterial strains isolated from the Antarctic meltwater ponds. The bacteria were grown on two forms of brain heart infusion (BHI) medium: agar at six different temperatures (4, 10, 18, 25, 30, and 37°C) and on broth at 18°C. Multivariate data analysis approaches such as principal component analysis (PCA) and correlation analysis were used to study the difference in biochemical profiles induced by the cultivation conditions. The observed results indicated a strong correlation between FTIR spectra and the phylogenetic relationships among the studied bacteria. The most accurate taxonomy-aligned clustering was achieved with bacteria cultivated on agar. Cultivation on two forms of BHI medium provided biochemically different bacterial biomass. The impact of temperature on the total cellular biochemical profile of the studied bacteria was species-specific, however, similarly for all bacteria, lipid spectral region was the least affected while polysaccharide region was the most affected by different temperatures. The biggest temperature-triggered changes of the cell chemistry were detected for bacteria with a wide temperature tolerance such Pseudomonas lundensis strains and Acinetobacter lwoffii BIM B-1558.

From disinfection to pathogenicity: Occurrence, resistome risks and assembly mechanism of biocide and metal resistance genes in hospital wastewaters

Hospital wastewaters (HWWs) represent critical reservoir for the accumulation and propagation of resistance genes. However, studies on biocide and metal resistance genes (BMRGs) and their associated resistome risks and driving mechanisms in HWWs are still in their infancy. Here, metagenomic assembly was firstly used to investigate host pathogenicity and transferability profiles of BMGRs in a typical HWWs system. As a result, genes conferring resistance to Ethidium Bromide, Benzylkonium Chloride, and Cetylpyridinium Chloride dominated biocide resistance genes (BRGs), whereas Cu resistance gene was the largest contributor of metal resistance genes (MRGs). Most BMRGs experienced significant reduction from anoxic-aerobic treatment to sedimentation stages but exhibited enrichment after chlorine disinfection. Network analysis indicated intense interactions between BMRGs and virulence factors (VFs). Polar_flagella, belonging to the adherence was identified to play important role in the network. Contig-based analysis further revealed noteworthy shifts in host associations along the treatment processes, with Pseudomonadota emerging as the primary carrier, hosting 91.1% and 85.3% of the BRGs and MRGs. A total of 199 opportunistic pathogens were identified to carry 285 BMRG subtypes, which mainly included Pseudomonas alcaligenes, Pseudomonas lundensis, and Escherichia coli. Notably, ruvB conferring resistance to Cr, Cetylpyridinium Chloride, and Dodine were characterized with the highest frequency carried by pathogens. Diverse co-occurrence patterns between BMRGs and mobile genetic elements (MGEs) were found from the raw influent to final effluent. Overall, 10.5% BRGs and 8.84% MRGs were mobile and among the 4 MGEs, transposase exhibited the greatest potential for the BMRGs dissemination. Furthermore, deterministic processes played a dominant role in bacterial communities and BMRGs assembly in HWWs. Bacterial communities contributed more than MGEs in shaping the resistome. Taken together, this work demonstrated widespread BMRGs pollution throughout the HWWs treatment system, emphasizing the potential for informing resistome risk and ecological mechanism in medical practice.

Bioaugmentation strategies based on bacterial and methanogenic cultures to relieve stress in anaerobic digestion of protein-rich substrates

Anaerobic co-digestion of protein-rich substrates is a prominent strategy for converting valuable feedstocks into methane, but it releases ammonia, which can inhibit the overall process. This study developed a cutting-edge combined culturomic and metagenomic approach to investigate the microbial composition of an ammonia-tolerant biogas plant. Newly-isolated microorganisms were used for bioaugmentation of stressed batch reactors fed with casein, maize silage and their combination. A co-culture enriched with proteolytic bacteria was isolated, selected and compared with the proteolytic collection strain Pseudomonas lundensis DSM6252. The co-culture and P. lundensis were combined with the ammonia-resistant archaeon Methanoculleus bourgensis MS2 to boost process stability. A microbial population pre-adapted to casein was also tested for evaluating the digestion of protein-rich feedstock. The promising results suggest combining proteolytic bacteria and M. bourgensis could exploit microbial co-cultures to improve anaerobic digestion stability and ensure stable productivity even under the harshest of ammonia conditions.

Growth performance and metabolic changes in lettuce inoculated with plant growth promoting bacteria in a hydroponic system

Crop cultivation in hydroponic systems is often exposed to both abiotic and biotic stresses, which impact growth. One possible approach to alleviating these stresses and improving plant growth is by utilizing plant growth-promoting bacteria (PGPB). Our previous study has shown the efficacy of two PGPB strains, Pseudomonas lundensis UB 53 and Pseudomonas migulae UB 54, in promoting growth and stimulating the generation of plant defense enzymes to mitigate biotic and abiotic stress in lettuce grown in a nutrient film technique (NFT) hydroponic system. Therefore, this study purposed to assess metabolic changes in lettuce in response to P. lundensis UB 53 and P. migulae UB 54 inoculation in a NFT hydroponic system. The results exhibited that these bacteria enhanced IAA accumulation and lettuce growth in a NFT hydroponic system. A total of 58 metabolites were detected in the root samples, including primary and secondary metabolites. It also influenced 12 metabolic pathways, with emphasis on five pathways, including the metabolism of alpha-linolenic acid, inositol phosphate, pyruvate, sulfur, and the phosphatidylinositol signaling system. These results highlight the significant roles of myo-inositol and acetic acid in metabolic pathways during the interaction between lettuce and PGPB.

Novel PVA/carboxylated cellulose antimicrobial hydrogel grafted with curcumin and ε-polylysine for chilled chicken preservation.

PVA/CC/CUR/PL composite films containing curcumin (CUR) and ε-polylysine (PL) were prepared by casting and chemical grafting methods to address the threat to food spoilage. Morphological analysis showed that the grafting of CUR and PL resulted in a rough cross-section of the polymer matrix. Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis confirmed the grafting of CUR and PL into the polymer matrix via esterification and amidation reactions, respectively. Thermal weight loss analysis showed that grafting process positively improved the thermal stability. The PVA/CC/CUR/PL films exhibited strong bactericidal activity, reaching 99.0% and 99.8% for Pseudomonas lundensis and Shewanella putrefaciens, respectively. After 8days of storage, the total number of colonies and the TVB-N content in the PVA/CC/CUR/PL group decreased by 1.51 lg CFU/g and 13.77mg/100g, respectively. Therefore, PVA/CC/CUR/PL films are considered as a promising bactericidal material with good mechanical properties, functionality, and other excellent characteristics.

Antibacterial efficacy of phenyllactic acid against Pseudomonas lundensis and Brochothrix thermosphacta and its synergistic application on modified atmosphere/air-packaged fresh pork loins

Microbial contamination is a crucial problem that is difficult to solve for the meat industry. Therefore, this study explored the antibacterial efficacy of phenyllactic acid (PLA) against Pseudomonas lundensis (PL) and Brochothrix thermosphacta (BT) solely and in combination (PL + BT). It also provided insights into its synergistic preservation effect during inoculation in chilled (4 °C) fresh pork loins under air (AP) and modified atmosphere packaging (MAP). The minimum inhibitory concentration (MIC) of PLA was 10 mg/mL. Growth kinetics, scanning electron microscopy (SEM), zeta potential, and cell viability investigations showed that PLA treatment exhibited reduced bacterial growth, aided morphological alterations, and leakage in cell membrane integrity in vitro. Nonetheless, PLA and MAP (70 %N2/30 %CO2) showed an excellent synergistic antibacterial ability against spoilage indicators(total glucose, pH, TVB-N, and TBARS), bacterial counts than AP, without impairing organoleptic acceptability. These results demonstrate the broad antibacterial efficacy of PLA as a biopreservative for the meat industry.

Photodynamic inactivation of spoilers Pseudomonas lundensis and Brochothrix thermosphacta by food-grade curcumin and its application on ground beef

This study evaluated the bactericidal effects of curcumin-mediated photodynamic inactivation (PDI) combined with citric acid on meat spoilage bacteria, and their preservative effects of refrigerated beef. After treatment of PDI (60 μM curcumin and 33.01 J/cm2) combined with 0.5 mg/mL citric acid, the populations of Pseudomonas lundensis and Brochothrix thermosphacta in PBS solution were reduced by 3.07 and 6.14 log CFU/mL, respectively, compared to 0.31 and 1.39 log CFU/mL for PDI. Combined treatment enhanced reactive oxygen species (ROS) activity in the two strains, and damaged their cell morphology, especially B. thermosphacta. With increasing curcumin concentration, the growth rate of spoilage bacteria was significantly inhibited (P < 0.05), and the changes of dominant spoilage species in bacterial community were retarded in aerobic-packaged and vacuum-packaged ground beef at 4 °C. Compared to the control, trichloroacetic acid-soluble peptide and total volatile basic nitrogen production were greatly restrained, and changes in pH values and brown color were apparently reduced for the combined treatment, which could extend the shelf life by 2 days. These results suggested the synergistic potency of PDI combined with citric acid against spoilage bacteria in meat products.Industrial relevance: The proliferation and spread of some psychrotrophs are major causes of quality deterioration and cross-contamination in fresh meat during cold chain. This study revealed that photosensitized curcumin combined with citric acid significantly decreased the survival of two psychrotrophic spoilage bacteria and extended the shelf-life of ground beef. Thus, curcumin-based PDI provides a new insight for the development of efficient and economic preservation in the meat processing industry.

Bacterial Skin Microbiota of Seabass from Aegean Fish Farms and Antibiotic Susceptibility of Psychrotrophic Pseudomonas.

Farming seabass (Dicentrarchus labrax) is an essential activity in the Mediterranean basin including the Aegean Sea. The main seabass producer is Turkey accounting for 155,151 tons of production in 2021. In this study, skin swabs of seabass farmed in the Aegean Sea were analysed with regard to the isolation and identification of Pseudomonas. Bacterial microbiota of skin samples (n = 96) from 12 fish farms were investigated using next-generation sequencing (NGS) and metabarcoding analysis. The results demonstrated that Proteobacteria was the dominant bacterial phylum in all samples. At the species level, Pseudomonas lundensis was identified in all samples. Pseudomonas, Shewanella, and Flavobacterium were identified using conventional methods and a total of 46 viable (48% of all NGS+) Pseudomonas were isolated in seabass swab samples. Additionally, antibiotic susceptibility was determined according to standards of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) and Clinical and Laboratory Standards Institute (CLSI) in psychrotrophic Pseudomonas. Pseudomonas strains were tested for susceptibility to 11 antibiotics (piperacillin-tazobactam, gentamicin, tobramycin, amikacin, doripenem, meropenem, imipenem, levofloxacin, ciprofloxacin, norfloxacin, and tetracycline) from five different groups of antibiotics (penicillins, aminoglycosides, carbapenems, fluoroquinolones, and tetracyclines). The antibiotics chosen were not specifically linked to usage by the aquaculture industry. According to the EUCAST and CLSI, three and two Pseudomonas strains were found to be resistant to doripenem and imipenem (E-test), respectively. All strains were susceptible to piperacillin-tazobactam, amikacin, levofloxacin, and tetracycline. Our data provide insight into different bacteria that are prevalent in the skin microbiota of seabass sampled from the Aegean Sea in Turkey, and into the antibiotic resistance of psychrotrophic Pseudomonas spp.

Unraveling the Antibacterial Mechanism of Plasma-Activated Lactic Acid against Pseudomonas ludensis by Untargeted Metabolomics.

Plasma-activated liquid is a novel non-thermal antibacterial agent against a wide spectrum of foodborne bacteria, yet fewer studies focused on its disinfection of meat spoilage bacteria. In this study, the antibacterial properties of plasma-activated lactic acid (PALA) on Pseudomonas lundensis, isolated and identified from spoilage beef, were investigated. A plasma jet was used to treat lactic acid (0.05-0.20%) for 60-120 s. The results presented that the 0.2% LA solution treated with plasma for 120 s caused a 5.64 log reduction. Additionally, the surface morphology, membrane integrity and permeability were altered slightly and verified by scanning electron microscopy, double staining of SYTO-9 and propidium iodide, and a K+ test kit. The intracellular organization of the cells, observed by transmission electron microscopy, was damaged significantly. Increased intracellular reactive oxygen species (ROS) levels exceeded the antioxidant ability of glutathione (GSH), leading to a reduction in the activity of malate dehydrogenase (MDH), succinic dehydrogenase (SDH) and intracellular ATP levels. Metabolomics analysis indicated that the energy and synthesis of essential components, such as DNA and amino acid-related metabolic pathways, were disturbed. In conclusion, this research established a theoretical basis for the use of PALA in refrigerated beef preservation by shedding light on the bacteriostatic effect of PALA against Pseudomonas lundensis.

Respiratory depression driven by membrane damage as a mechanism for linalool to inhibit Pseudomonas lundensis and its preservation potential for beef.

This study aimed to investigate the mechanism of linalool against Pseudomonas lundensis and its application on beef. Field emission scanning electron microscopy found that linalool exerted antibacterial activity with a minimum inhibitory concentration (MIC) of 1.5ml l-1 by disrupting cell structure. Loss of cell membrane integrity was monitored due to leakage of nucleic acids and K+. In addition, respiratory depression appeared in Ps. lundensis based on inhibition of enzyme activities including hexokinase (HK), glucose 6-phosphate dehydrogenase (G6PDH), phosphofructokinase (PFK), pyruvate kinase (PK), pyruvate dehydrogenase (PDH), citrate synthase (CS), succinate dehydrogenase (SDH), and malate dehydrogenase (MDH). Subsequently, energy limitation also occurred according to the decrease in ATP content and ATPase activity. Molecular docking confirmed that linalool can combine with enzymes in cell wall (ddlB) and energy synthesis (AtpD) pathways to exert antibacterial effect. Of note, linalool has advantages for beef preservation by delaying quality changes including pH, total volatile basic nitrogen (TVB-N) and total viable count (TVC). Linalool has significant inhibitory effect on Ps. lundensis, and respiratory depression driven by membrane damage is the main inhibitory mechanism.

In Vitro and In Situ Characterization of Psychrotrophic Spoilage Bacteria Recovered from Chilled Chicken

Spoilage bacteria play a remarkable role in the spoilage of chilled chicken. In this paper, a total of 42 isolates belonging to 16 species of four genera were isolated from chilled chicken and displayed different characterizations of psychrotrophic spoilage. Six isolates of J7, J8, Q20, Q23, R1, and R9 with differences in proteolytic capabilities were further characterized for in situ spoilage potential evaluation. Pseudomonas lundensis J8 exhibited the strongest spoilage potential in situ, displaying a fast growth rate, increased pH velocity, high total volatile basic nitrogen, and high peptide content in the chicken samples. The volatile flavor analysis of chicken samples via electronic nose indicated that the content of characteristic odors representing spoilage, including sulfides, organic sulfide, and hydride, increased during storage. Additionally, the principle component and correlation analyses revealed that the spoilage odors produced by different species of bacteria were significantly different and positively correlated with the results of protease activity in vitro. The characteristics of spoilage bacteria in chilled chicken provided a comprehensive insight into microbial assessment during storage.

Mathematical Modeling of the Effects of Temperature and Modified Atmosphere Packaging on the Growth Kinetics of Pseudomonas Lundensis and Shewanella Putrefaciens in Chilled Chicken.

The effects of modified atmosphere packaging (MAP) on the growth and spoilage characteristics of Pseudomonas lundensis LD1 and Shewanella putrefaciens SP1 in chilled chicken at 0–10 °C were studied. MAP inhibited microbial growth, TVB-N synthesis, and lipid oxidation. The inhibitory effect of MAP became more significant as the temperature decreased. The kinetic models to describe the growth of P. lundensis LD1 and S. putrefaciens SP1 at 0–10 °C were also established to fit the primary model Gompertz and the secondary model Ratkowsky. The models had a high degree of fit to describe the growth of dominant spoilage bacteria in chilled chicken. The observed numbers of P. lundensis LD1 and S. putrefaciens SP1 at 2 °C were compared with the predicted numbers, and the accuracy factor and bias factor ranged from 0.93 to 1.14. These results indicated that the two models could help predict the growth of P. lundensis and S. putrefaciens in chilled chicken at 0–10 °C. The analyzed models provide fast and cost-effective alternatives to replace traditional culturing methods to assess the influence of temperature and MAP on the shelf life of meat.

Antibiotic Resistance Properties among Pseudomonas spp. Associated with Salmon Processing Environments.

Continuous monitoring of antimicrobial resistance in bacteria along the food chain is crucial for the assessment of human health risks. Uncritical use of antibiotics in farming over years can be one of the main reasons for increased antibiotic resistance in bacteria. In this study, we aimed to classify 222 presumptive Pseudomonas isolates originating from a salmon processing environment, and to examine the phenotypic and genotypic antibiotic resistance profiles of these isolates. Of all the analyzed isolates 68% belonged to Pseudomonas, and the most abundant species were Pseudomonas fluorescens, Pseudomonas azotoformans, Pseudomonas gessardii, Pseudomonas libanesis, Pseudomonas lundensis, Pseudomonas cedrina and Pseudomonas extremaustralis based on sequencing of the rpoD gene. As many as 27% of Pseudomonas isolates could not be classified to species level. Phenotypic susceptibility analysis by disc diffusion method revealed a high level of resistance towards the antibiotics ampicillin, amoxicillin, cefotaxime, ceftriaxone, imipenem, and the fish farming relevant antibiotics florfenicol and oxolinic acid among the Pseudomonas isolates. Whole genome sequencing and subsequent analysis of AMR determinants by ResFinder and CARD revealed that no isolates harbored any acquired resistance determinants, but all isolates carried variants of genes known from P. aeruginosa to be involved in multidrug efflux pump systems.

Synergism With ε-Polylysine Hydrochloride and Cinnamon Essential Oil Against Dual-Species Biofilms of Listeria monocytogenes and Pseudomonas lundensis.

Various pathogenic and spoilage bacteria frequently coexist in meat processing environments and can form multispecies biofilms, causing significant health and economic issues. Despite the prevalence and coexistence, only less is known about possible interactions between Listeria monocytogenes (LM) and spoilers like Pseudomonas species, and their community-wide resistance against natural preservatives. This study evaluates the interactions between mono- or dual-species biofilms formed by LM and Pseudomonas lundensis (PL), as well as the sensitivity of these bacteria in dual-species biofilms to ε-polylysine hydrochloride (ε-PLH) alone or combined with cinnamon essential oil (CEO). The results showed that the biofilm cell density of P. lundensis in dual species was higher (p < 0.05) than LM, constituting about 85% of the total population. More biofilms and exopolysaccharide both in mono- or dual species of the two psychrotrophic strains were greatly produced at 15°C than at 30°C. The biomass, biovolume, and thickness of dual-species biofilms were significantly lower than single PL biofilm when tested using crystal violet staining, confocal laser scanning microscopy, and scanning electron microscopy, indicating the competitive interactions between them prevail. Additionally, ε-PLH significantly reduced the biofilm development as mono- and dual species in a concentration-dependent manner, especially single LM biofilm, which was consistent with the decrease in autoinducer-2 (AI-2) activity. LM as dual-species biofilms exhibited lower sensitivity to ε-PLH than its mono-biofilm probably due to protective effect conferred by PL. ε-PLH in combination with CEO, at the maximum sublethal concentrations (MSCs), showed enhanced inhibitory activity against dual-species biofilm formation, as evidenced by thin spare spatial structures and reduced AI-2 activity. In addition, the preformed dual biofilms were dramatically eradicated following treatment with ε-PLH combined with CEO at higher than minimum inhibitory concentration in comparison with either of the compounds used alone, indicating the synergistic antibiofilm of the two preservatives. This study reveals the competitive interactions between the two strains in dual-species biofilms, in which the dominant PL significantly contributed toward the tolerance of LM to ε-PLH, and the use of combined preservatives shows it is an effective strategy to control the multispecies biofilms in meat processing.

Pseudomonas species prevalence, protein analysis, and antibiotic resistance: an evolving public health challenge

Psychrotrophic Pseudomonas is one of the significant microbes that lead to putrefaction in chilled meat. One of the biggest problems in the detection of Pseudomonas is that several species are seemingly identical. Currently, antibiotic resistance is one of the most significant challenges facing the world's health and food security. Therefore, this study was designed to apply an accurate technique for eliminating the identification discrepancy of Pseudomonas species and to study their resistance against various antimicrobials. A total of 320 chicken meat specimens were cultivated, and the isolated bacteria’ were phenotypically recognized. Protein analysis was carried out for cultured isolates via Microflex LT. The resistance of Pseudomonas isolates was recorded through Vitek® 2 AST-GN83 cards. Overall, 69 samples were identified as Pseudomonas spp. and included 18 Pseudomonas lundensis (P. lundensis), 16 Pseudomonas fragi (P. fragi), 13 Pseudomonas oryzihabitans (P. oryzihabitans), 10 Pseudomonas stutzeri (P. stutzeri), 5 Pseudomonas fluorescens (P. fluorescens), 4 Pseudomonas putida (P. putida), and 3 Pseudomonas aeruginosa (P. aeruginosa) isolates. Microflex LT identified all Pseudomonas isolates (100%) correctly with a score value ≥ 2.00. PCA positively discriminated the identified isolates into various groups. The antimicrobial resistance levels against Pseudomonas isolates were 81.16% for nitrofurantoin, 71% for ampicillin and ampicillin/sulbactam, 65.22% for cefuroxime and ceftriaxone, 55% for aztreonam, and 49.28% for ciprofloxacin. The susceptibilities were 100% for cefotaxime, 98.55% for ceftazidime, 94.20% for each piperacillin/tazobactam and cefepime, 91.3% for cefazolin. In conclusion, chicken meat was found to be contaminated with different Pseudomonas spp., with high incidence rates of P. lundensis. Microflex LT is a potent tool for distinguishing Pseudomonads at the species level.

Investigation of Pseudomonas Species in Chicken Drumstick Samples

This study aimed to detect the contamination of refrigerated poultry meat with psychrotrophic bacteria, such as Pseudomonas species. Pseudomonas spp. can grow well in the skin and muscle of poultry meat by using carbonhydrates and amino acids at refrigeration temperature (4 °C). They are mainly responsible for the spoilage of poultry meat with their enzymatic activiy. For this purpose, a total of 107 chicken drumstick samples were analyzed for the presence of Pseudomonas species (Pseudomonas fragi, Pseudomonas fluorescens, Pseudomonas putida, Pseudomonas lundensis, Pseudomonas aeruginosa). Of the samples, 92.5% found to be contaminated with Pseudomonas spp. A total of 99 isolates were confirmed as Pseudomonas spp. by PCR, targeting the 16S rDNA gene. Among the 99 isolates, 78.7% were identified as P. fluorescens, whereas P. fragi, P. putida, P. lundensis, and P. aeruginosa were not detected in the study. The findings show that P. fluorescens is the most prevalent species in refrigerated poultry meat, thus posing a potential risk for the spoilage of poultry meat.

Metabolomics reveals spoilage characteristics and interaction of Pseudomonas lundensis and Brochothrix thermosphacta in refrigerated beef

Pseudomonas lundensis and Brochothrix thermosphacta are key spoilage microorganisms in aerobically stored chilled meat. The present study aimed to investigate the physicochemical and metabolomic profiles of refrigerated ground beef inoculated P. lundensis (PL) and B. thermosphacta (BT) as mono- or co-culture (BP). P. lundensis was the dominant spoilage strain in the co-culture of ground beef. A large amount of TCA-soluble peptide, TVB-N and TBA were formed in the PL and BP, while acetion was mainly produced in the BT, as accompanied by the different sensory and color changes. Meat metabolome indicated that 95, 396, and 409 metabolites with significant differences, were identified in ground beef inoculated BT, PL, and BP, respectively. These differential metabolites covered 58 metabolic pathways, in which histidine metabolism was identified as an important pathway related to spoilage in the three groups. Specifically, creatine, inosine, anserine, uracil, alanine, glutamine, 3-methylhistidine and 3-hydroxycapric acid were enriched as potential spoilage biomarkers. Taken together, those findings reveal the complex and competitive interactions of their co-culture of B. thermosphacta and P. lundensis, which provided a comprehensive insight into microbial spoilage mechanism in chilled beef.

The Psychrotrophic Pseudomonas lundensis, a Non-aeruginosa Pseudomonad, Has a Type III Secretion System of the Ysc Family, Which Is Transcriptionally Active at 37°C.

ABSTRACTThe type III secretion system (T3SS) is a needle-like structure found in Gram-negative pathogens that directly delivers virulence factors like toxins and effector molecules into eukaryotic cells. The T3SS is classified into different families according to the type of effector and host. Of these, the Ysc family T3SS, found in Yersinia species and Pseudomonas aeruginosa, confers high virulence to bacteria against eukaryotic hosts. Here, we present the first identification and transcriptional analyses of a Ysc T3SS in a non-aeruginosa Pseudomonas species, Pseudomonas lundensis, an environmental psychrotrophic bacterium and important agent of frozen food spoilage. We have identified and sequenced isolates of P. lundensis from three very distinct ecological niches (Antarctic temporary meltwater pond, U.S. supermarket 1% pasteurized milk, and cystic fibrosis lungs) and compared these to previously reported food spoilage isolates in Europe. In this paper, we show that strains of P. lundensis isolated from these diverse environments with ambient temperatures ranging from below freezing to 37°C all possess a Ysc family T3SS secretion system and a T3S effector, ExoU. Using in vitro and in vivo transcriptomics, we show that the T3SS in P. lundensis is transcriptionally active, is expressed more highly at mammalian body temperature (37°C) than 4°C, and has even higher expression levels when colonizing a host environment (mouse intestine). Thus, this Ysc T3SS-expressing psychrotrophic Pseudomonad has an even greater range of growth niches than previously appreciated, including diseased human airways.

Antibacterial Mechanism of 3-Carene against the Meat Spoilage Bacterium Pseudomonas lundensis and Its Application in Pork during Refrigerated Storage.

Pseudomonas lundensis is the main bacterium responsible for meat spoilage and its control is of great significance. 3-Carene, a natural monoterpene, has been proved to possess antimicrobial activities. This study aimed to investigate the antibacterial activity and mechanism of 3-carene against the meat spoilage bacterium P. lundensis, and explore its application on pork. After 3-carene treatment, cellular structural changes were observed. Cell walls and membranes were destroyed, resulting in the leakage of alkaline phosphatase and cellular contents. The decreased activity of Ca2+-Mg2+-ATPase and Na+-K+-ATPase showed the imbalance of intracellular ions. Subsequently, adenosine triphosphate (ATP) content and oxidative respiratory metabolism characteristics indicated that 3-carene inhibited the metabolism of the tricarboxylic acid cycle in P. lundensis. The results of binding 3-carene with the vital proteins (MurA, OmpW, and AtpD) related to the formation of the cell wall, the composition of the cell membrane, and the synthesis of ATP further suggested that 3-carene possibly affected the normal function of those proteins. In addition, the growth of P. lundensis and increase in pH were inhibited in pork during the 5 days of cold storage after the samples were pre-treated with 3-carene. These results show the anti-P. lundensis activity and mechanism of 3-carene, and its potential use in meat preservation under refrigerated conditions.