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Chemistry (Weinheim An Der Bergstrasse,... Aug 2023Herein, we describe the efficient gram-scale synthesis of α2,3- and α2,6-sialyllactose oligosaccharides as well as mimetics from N-acyl mannosamines and lactose in...
Herein, we describe the efficient gram-scale synthesis of α2,3- and α2,6-sialyllactose oligosaccharides as well as mimetics from N-acyl mannosamines and lactose in metabolically engineered bacterial cells grown at high cell density. We designed new Escherichia coli strains co-expressing sialic acid synthase and N-acylneuraminate cytidylyltransferase from Campylobacter jejuni together with the α2,3-sialyltransferase from Neisseria meningitidis or the α2,6-sialyltransferase from Photobacterium sp. JT-ISH-224. Using their mannose transporter, these new strains actively internalized N-acetylmannosamine (ManNAc) and its N-propanoyl (N-Prop), N-butanoyl (N-But) and N-phenylacetyl (N-PhAc) analogs and converted them into the corresponding sialylated oligosaccharides, with overall yields between 10 % and 39 % (200-700 mg.L of culture). The three α2,6-sialyllactose analogs showed similar binding affinity for Sambucus nigra SNA-I lectin as for the natural oligosaccharide. They also proved to be stable competitive inhibitors of Vibrio cholerae neuraminidase. These N-acyl sialosides therefore hold promise for the development of anti-adhesion therapy against influenza viral infections.
Topics: Lactose; Neuraminidase; Escherichia coli; Sialyltransferases; Oligosaccharides
PubMed: 37294058
DOI: 10.1002/chem.202301555 -
Effects of plant-based proteins and handling stress on intestinal mucus microbiota in rainbow trout.Scientific Reports Dec 2023Via 16S rRNA gene amplicon sequencing, this study explores whether the gut mucus microbiota of rainbow trout is affected by the interaction of a plant-protein-based diet...
Via 16S rRNA gene amplicon sequencing, this study explores whether the gut mucus microbiota of rainbow trout is affected by the interaction of a plant-protein-based diet and a daily handling stressor (chasing with a fishing net) across two genetic lines (A, B). Initial body weights of fish from lines A and B were 124.7 g and 147.2 g, respectively. Fish were fed 1.5% of body weight per day for 59 days either of two experimental diets, differing in their fish meal [fishmeal-based diet (F): 35%, plant-based diet (V): 7%] and plant-based protein content (diet F: 47%, diet V: 73%). No diet- or stress-related effect on fish performance was observed at the end of the trial. However, we found significantly increased observed ASVs in the intestinal mucus of fish fed diet F compared to diet V. No significant differences in Shannon diversity could be observed between treatments. The autochthonous microbiota in fish fed with diet V was dominated by representatives of the genera Mycoplasma, Cetobacterium, and Ruminococcaceae, whereas Enterobacteriaceae and Photobacterium were significantly associated with diet F. The mucus bacteria in both genetic lines were significantly separated by diet, but neither by stress nor an interaction, as obtained via PERMANOVA. However, pairwise comparisons revealed that the diet effect was only significant in stressed fish. Therefore, our findings indicate that the mucus-associated microbiota is primarily modulated by the protein source, but this modulation is mediated by the stress status of the fish.
Topics: Animals; Oncorhynchus mykiss; Plant Proteins; RNA, Ribosomal, 16S; Gastrointestinal Microbiome; Microbiota; Diet; Animal Feed
PubMed: 38110473
DOI: 10.1038/s41598-023-50071-x -
Food Microbiology May 2024We aimed to evaluate the bacterial growth and diversity in vacuum-packed beef bags stored at different temperatures and to monitor blown-pack spoilage. We used...
We aimed to evaluate the bacterial growth and diversity in vacuum-packed beef bags stored at different temperatures and to monitor blown-pack spoilage. We used culture-based methods and high-throughput sequencing to study the development of the main bacterial groups naturally present in beef stored at 4 and 15 °C for 28 days. The growth of sulfite-reducing clostridium (SRC) was impaired in beef bags stored at 4 °C; significant differences among SRC counts were observed in beef bags stored at 4 and 15 °C on days 14, 21, and 28 (P = 0.001). Blown pack was observed in most beef bags stored at 15 °C, from day 14 to day 28, but not in beef bags stored at 4 °C. A storage temperature of 4 °C was able to maintain a stable bacterial microbiota (most prevalent: Photobacterium, Hafnia-Obesumbacterium, and Lactococcus). Remarkable changes in microbial abundance occurred at 15 °C from day 14 to day 28, with a predominance of strict anaerobes (Bacteroides) and the presence of Clostridium spp. The relative frequencies of strict anaerobes and Clostridium were statistically higher in the beef bags stored at 15 °C (P < 0.001 and P = 0.004, respectively). The temperature influenced the microbial counts and relative abundance of spoilage bacteria, leading to blown pack spoilage.
Topics: Animals; Cattle; Food Packaging; Meat; Temperature; Vacuum; Bacteria; Clostridium; Microbiota; Food Microbiology
PubMed: 38225050
DOI: 10.1016/j.fm.2023.104448 -
World Journal of Microbiology &... Sep 2023The present study evaluates the antibacterial properties of alkaloids and the crude extracts (ethanol, n-hexane and ethyl acetate) from seaweed Sargassum fusiforme...
The present study evaluates the antibacterial properties of alkaloids and the crude extracts (ethanol, n-hexane and ethyl acetate) from seaweed Sargassum fusiforme against coral pathogens (Photobacterium galatheae, Vibrio harveyi, Bordetella trematum, and Ochrobactrum pseudogrignonese) isolated from coral Porites lutea. To our knowledge, this is the first in vitro assay for such extracts on Porites lutea coral pathogens. Bacterial pathogens have been identified using 16S RNA and BankIt into gene bank and given the accession numbers (OR401000; OR401001; OR401336, and OR400998 respectively). GC-Mass profiling conducted for n-hexane compounds confirmed the presence of thirty-eight molecules, twelve of which have been previously reported for their bioactivity. The results revealed that alkaloids and n-hexane extract demonstrated eminent antibacterial activity compared to the other extracts against the tested coral pathogenic bacteria. Molecular docking was conducted to evaluate the twelve previously mentioned bioactive molecules to get a full understanding of the interaction of those bioactive molecules on vital bacterial proteins (Hemolysin protein (PDB ID: 1XEZ) and Cytoplasmic proteins (PDB ID: 3TZC)). Docked twelve molecules against hemolysin protein (PDB ID: 1XEZ) came exactly in line with the docked result of the same molecules with cytoplasmic proteins (PDB ID: 3TZC), proving the bioactivity of 6-O-Palmitoyl-L-ascorbic acid, 3TMS derivative; Glycerol monostearate, 2TMS derivative and Eicosanoic acid complexes in antibacterial activity action and score higher than reference ligand. Those three compounds will be investigated separately in future in vitro assay soon. Our conclusions align with the study's antibacterial in vitro assay results. The present study reports the novelty of different extracts of S. fusiforme as an antibacterial agent against coral pathogenic bacteria that trigger diseases in Porites lutea.
Topics: Animals; Molecular Docking Simulation; Hemolysin Proteins; Anthozoa; Anti-Bacterial Agents
PubMed: 37743438
DOI: 10.1007/s11274-023-03752-8 -
International Journal of Biological... Dec 2023As digestive and immune organs of animals, the gut was frequently used to evaluate the health status of aquatic animals. In previous oil source alternatives study, corn...
Integrated analysis of microbiome and host transcriptome reveals the damage/protective mechanism of corn oil and olive oil on the gut health of grouper (♀ Epinephelus fuscoguttatus × ♂ E. lanceolatu).
As digestive and immune organs of animals, the gut was frequently used to evaluate the health status of aquatic animals. In previous oil source alternatives study, corn oil (CO) had been found to induce gut inflammation, while olive oil (OO) had been found to be effective in protecting intestinal health. Three diets with different oil sources (fish oil, CO, OO) were formulated for an 8-week culture experiment, and it was proposed to combine 16S sequencing and transcriptome sequencing analysis to preliminarily elucidate the damage/protection mechanism of CO and OO on the gut health of grouper (♀ Epinephelus fuscoguttatus × ♂ E. lanceolatu). We found that CO indeed damaged to gut health and destroyed the gut structure, while OO had a positive outcome in protecting the gut structure, promoting digestibility and relieving enteritis. Photobacterium, Romboutsia and Epulopiscium were significantly enriched in OO group and Staphylococcus were significantly enriched in CO group. Transcriptome sequencing further revealed CO could activated Complement and coagulation cascades, Staphylococcus aureus infection, Systemic lupus erythematosus, and Tuberculosis pathways; conversely, OO activated B-cell signaling receptors, promoted B-cell proliferation and apoptosis, and thus activated B-cell signaling pathways to enhance immunity, whereas OO can regulate IL17 signaling pathway and TNF signaling pathway to inhibit NF-κB signaling pathway to reduce pro-inflammatory response. By integrating the microbiome and transcriptome, further identified all differential microorganisms were directly and significantly correlated with differential genes, and Clostridium_sensu_stricto_1, Romboutsia, Staphylococcus might as the core regulates the expression of differential gene in the organism. These results reveal that different oil sources alter gut gene expression mainly by modulating the composition and abundance of gut microbiota, further regulating the health status of the gut. Gut microbiota could be used as biomarkers to provide reference and solutions for the mitigation of inflammation in aquatic animals.
Topics: Animals; Bass; Olive Oil; Corn Oil; Transcriptome; Inflammation; Microbiota
PubMed: 37865354
DOI: 10.1016/j.ijbiomac.2023.127550 -
Fish and Shellfish Immunology Reports Dec 2023The effects of ssp (Phdp) on immune responses and intestinal ultrastructure of following infection and their amelioration by the probiotic bacteria and were...
The effects of ssp (Phdp) on immune responses and intestinal ultrastructure of following infection and their amelioration by the probiotic bacteria and were evaluated. Pathogen growth inhibition in coculture with each probiotic and its virulence against were confirmed with an LC of 10 CFU mL. Phdp administration to at sublethal levels resulted in depletion of superoxide dismutase, glutathione reductase, glutathione transferase and phenoloxidase activities, extensive lipid peroxidation and reduced survival. Following a combined administration of each probiotic and the pathogen, enzyme activities and survival were significantly higher, while lipid peroxidation was reduced, compared to the infected group with no probiotic treatment ( < 0.05). The transmission electron microscopy study revealed that pathogen infection resulted in disarranged and fragmented microvilli, formation of empty or pathogen containing cytoplasmic vacuoles and damaged mitochondria. In the probiotic-treated and Phdp-infected series, intestinal cells showed normal appearance, except for the presence of pathogen-containing vacuoles and highly ordered but laterally stacked microvilli. The results of the present study indicate that Phdp induces cell death through an oxidative stress response and probiotics enhance immune responses to protect it against the Phdp induced damage.
PubMed: 37671319
DOI: 10.1016/j.fsirep.2023.100113 -
Animal Nutrition (Zhongguo Xu Mu Shou... Sep 2023Several reports have revealed the vital role that probiotics play in fish growth and health. However, few works are available for host gut-derived probiotics on the...
Host gut-derived probiotics supplementation improves growth performance, serum and liver immunity, gut health, and resistive capacity against infection in hybrid grouper ( × ♂).
Several reports have revealed the vital role that probiotics play in fish growth and health. However, few works are available for host gut-derived probiotics on the growth, immunity, and gut microbiota of fish, especially in hybrid grouper (♀ × ♂) due to their isolation difficulty and functional verification. This study aimed at assessing 3 host gut-derived species' effects on the growth, immune and antioxidant-biochemical responses, haematological parameters, intestinal morphology, immune-related gene expression, gut microbiota, and disease resistance against in hybrid grouper. A total of 480 hybrid grouper (initial weight = 9.03 ± 0.02 g) were randomly allotted into 4 groups, namely, the group fed a basal diet without probiotic inclusion (control, B0), the group fed the basal diet with GPSAK4 (BV), the group fed the basal diet with GPSAK9 (BS), and the group fed the basal diet with GPSAK2 (BT) strains at 1.0 × 10 CFU/g. After a 6-week feeding trial, the results revealed significant improvements ( < 0.05) in the growth performance, whole fish-body proximate composition, blood haematological parameters, serum, liver, and intestinal biochemical indexes, intestinal morphology, and protection against . pathogen in the probiotic-treated groups compared with the untreated. Additionally, the expressions of intestinal tight junction genes (occludin and ), pro- and anti-inflammatory genes, including , , , , , , and , were upregulated ( < 0.05) after species administration. Host gut-derived supplementation shaped the gut microbiota by significantly increasing ( < 0.05) the relative abundance of Proteobacteria, Bacteroidetes, Actinobacteria (except the BS group), Acidobacteria (except the BT group), Cyanobacteria (except the BV and BT groups), and Verrucomicrobia phyla, as well as known beneficial genera (, , , 1 and 13, , and ), but significantly decreased ( < 0.05) the abundance of Firmicutes, Chloroflexi, and Fusobacteria phyla, and purported pathogenic genera ( and ) compared with the control group. Collectively, the results suggest that . GPSAK4, . GPSAK9 (especially this strain), . GPSAK2 dietary supplementation at 1.0 × 10 CFU/g has positive effects on the intestinal health of hybrid grouper via microbial composition modulation, thus enhancing the assimilation and absorption of nutrients to boost fish growth, immunity, and disease resistance.
PubMed: 37448647
DOI: 10.1016/j.aninu.2023.05.005 -
Microbiology Spectrum Aug 2023Scombrotoxin (histamine) fish poisoning is a common seafood-borne illness attributed to toxin production by histamine-producing bacteria (HPB) in fish tissues during...
Scombrotoxin (histamine) fish poisoning is a common seafood-borne illness attributed to toxin production by histamine-producing bacteria (HPB) in fish tissues during decomposition. In laboratory studies, growth of HPB and other bacterial species is affected by physical and chemical attributes, but natural communities of HPB are not well understood. To determine how environmental and water quality variables may affect density of HPB in the natural aquatic environment, we compared presence and abundance of HPB to ambient temperature, salinity, dissolved oxygen, fecal coliforms, male-specific coliphage, nutrient concentrations, carbon and nitrogen stable isotope ratios, and C:N in water samples collected from July 2017 to February 2018 along a natural salinity gradient in a tidal river on the coast of northern Gulf of Mexico. HPB in water samples were quantified using a real-time PCR, most probable number method. HPB species were identified via 16S rRNA gene sequences. Temperature and salinity were determined to be the main factors driving HPB presence and concentration. Canonical correspondence analysis revealed that different HPB were associated with different environmental conditions. Photobacterium damselae was found under warmer, higher-salinity conditions; Raoultella planticola was found at colder, lower-salinity conditions; Enterobacter aerogenes was found at warmer, lower-salinity conditions; and Morganella morganii was found at most sites, independent of environmental conditions. These results showed that naturally occurring HPB abundance and species composition can be affected by environmental conditions, which could manifest in various potentials for histamine formation and scombrotoxin fish poisoning risk based on environmental factors. This study determined the effects of environmental conditions on presence and abundance of naturally occurring histamine-producing bacteria in the northern Gulf of Mexico. Here, we show that HPB abundance and species composition are related to ambient temperature and salinity, with the magnitude of this effect dependent on the particular HPB species. This finding suggests that environmental conditions at fishing sites could affect the risk of human illness from scombrotoxin (histamine) fish poisoning.
Topics: Animals; Male; Humans; Histamine; Gulf of Mexico; RNA, Ribosomal, 16S; Water Quality; Bacteria; Foodborne Diseases
PubMed: 37310253
DOI: 10.1128/spectrum.04720-22 -
MSystems Jun 2023Facultative marine bacterial pathogens sense environmental signals so that the expression of virulence factors is upregulated on entry into hosts and downregulated...
Facultative marine bacterial pathogens sense environmental signals so that the expression of virulence factors is upregulated on entry into hosts and downregulated during the free-living lifestyle in the environment. In this study, we utilized transcriptome sequencing to compare the transcriptional profiles of subsp. , a generalist pathogen that causes disease in diverse marine animals and fatal infections in humans at NaCl concentrations that mimic the free-living lifestyle or host internal milieu, respectively. We here show that NaCl concentration constitutes a major regulatory signal that shapes the transcriptome and uncover 1,808 differentially expressed genes (888 upregulated and 920 downregulated in response to low-salt conditions). Growth at 3% NaCl, a salinity that mimics the free-living lifestyle, upregulated genes involved in energy production, nitrogen metabolism, transport of compatible solutes, utilization of trehalose and fructose, and carbohydrate and amino acid metabolism with strong upregulation of the arginine deiminase system (ADS). In addition, we observed a marked increase in resistance to antibiotics at 3% NaCl. On the contrary, the low salinity conditions (1% NaCl) that mimic those encountered in the host triggered a virulence gene expression profile that maximized the production of the type 2 secretion system (T2SS)-dependent cytotoxins damselysin, phobalysin P, and a putative PirAB-like toxin, observations that were corroborated by the analysis of the secretome. Low salinity also upregulated the expression of iron-acquisition systems, efflux pumps, and other functions related to stress response and virulence. The results of this study greatly expand our knowledge of the salinity-responsive adaptations of a generalist and versatile marine pathogen. IMPORTANCE Pathogenic species experience continuous shifts of NaCl concentration in their life cycles. However, the impact of salinity changes in gene regulation has been studied in a small number of species. In this study, we analyzed the transcriptional response of subsp. (), a generalist and facultative pathogen, to changes in salinity, and demonstrate that growth at 1% NaCl in comparison to 3% NaCl triggers a virulence program of gene expression, with a major impact in the T2SS-dependent secretome. The decrease in NaCl concentration encountered by bacteria on entry into a host is proposed to constitute a regulatory signal that upregulates a genetic program involved in host invasion and tissue damage, nutrient scavenging (notably iron), and stress responses. This study will surely inspire new research on pathobiology, as well as on other important pathogens of the family and related taxa whose salinity regulons still await investigation.
Topics: Humans; Animals; Virulence; Sodium Chloride; Salinity; Photobacterium; Iron
PubMed: 37288979
DOI: 10.1128/msystems.01253-22 -
Applied and Environmental Microbiology Jun 2024Marine bacteria experience fluctuations in osmolarity that they must adapt to, and most bacteria respond to high osmolarity by accumulating compatible solutes also known...
Marine bacteria experience fluctuations in osmolarity that they must adapt to, and most bacteria respond to high osmolarity by accumulating compatible solutes also known as osmolytes. The osmotic stress response and compatible solutes used by the coral and oyster pathogen were unknown. In this study, we showed that to alleviate osmotic stress biosynthesized glycine betaine (GB) and transported into the cell choline, GB, ectoine, dimethylglycine, and dimethylsulfoniopropionate, but not -inositol. -inositol is a stress protectant and a signaling molecule that is biosynthesized and used by algae. Bioinformatics identified -inositol () catabolism clusters in and other , , , and species. Growth pattern analysis demonstrated that utilized -inositol as a sole carbon source, with a short lag time of 3 h. An deletion mutant, which encodes an inositol dehydrogenase, was unable to grow on -inositol. Within the clusters were an MFS-type () and an ABC-type () transporter and analyses showed that both transported -inositol. IolG and IolA phylogeny among species showed different evolutionary histories indicating multiple acquisition events. Outside of , IolG was most closely related to IolG from a small group of fish and human pathogens and species. However, IolG from hypervirulent strains clustered with IolG from and divergently from , , and plant pathogens. The cluster was also present within , , , , , , , , , and , of which many species were associated with marine flora and fauna.IMPORTANCEHost associated bacteria such as encounter competition for nutrients and have evolved metabolic strategies to better compete for food. Emerging studies show that -inositol is exchanged in the coral-algae symbiosis, is likely involved in signaling, but is also an osmolyte in algae. The bacterial consumption of -inositol could contribute to a breakdown of the coral-algae symbiosis during thermal stress or disrupt the coral microbiome. Phylogenetic analyses showed that the evolutionary history of -inositol metabolism is complex, acquired multiple times in , but acquired once in many bacterial plant pathogens. Further analysis also showed that a conserved cluster is prevalent among many marine species (commensals, mutualists, and pathogens) associated with marine flora and fauna, algae, sponges, corals, molluscs, crustaceans, and fish.
PubMed: 38874337
DOI: 10.1128/aem.00920-24