Genomic and vaccine preclinical studies reveal a novel mouse-adapted Helicobacter pylori model for the hpEastAsia genotype in Southeast Asia.

Introduction. Helicobacter pylori infection is a major global health concern, linked to the development of various gastrointestinal diseases, including gastric cancer. To study the pathogenesis of H. pylori and develop effective intervention strategies, appropriate animal pathogen models that closely mimic human infection are essential.Gap statement. This study focuses on the understudied hpEastAsia genotype in Southeast Asia, a region marked by a high H. pylori infection rate. No mouse-adapted model strains has been reported previously. Moreover, it recognizes the urgent requirement for vaccines in developing countries, where overuse of antimicrobials is fuelling the emergence of resistance.Aim. This study aims to establish a novel mouse-adapted H. pylori model specific to the hpEastAsia genotype prevalent in Southeast Asia, focusing on comparative genomic and histopathological analysis of pathogens coupled with vaccine preclinical studies.Methodology. We collected and sequenced the whole genome of clinical strains of H. pylori from infected patients in Vietnam and performed comparative genomic analyses of H. pylori strains in Southeast Asia. In parallel, we conducted preclinical studies to assess the pathogenicity of the mouse-adapted H. pylori strain and the protective effect of a new spore-vectored vaccine candidate on male Mlac:ICR mice and the host immune response in a female C57BL/6 mouse model.Results. Genome sequencing and comparison revealed unique and common genetic signatures, antimicrobial resistance genes and virulence factors in strains HP22 and HP34; and supported clarithromycin-resistant HP34 as a representation of the hpEastAsia genotype in Vietnam and Southeast Asia. HP34-infected mice exhibited gastric inflammation, epithelial erosion and dysplastic changes that closely resembled the pathology observed in human H. pylori infection. Furthermore, comprehensive immunological characterization demonstrated a robust host immune response, including both mucosal and systemic immune responses. Oral vaccination with candidate vaccine formulations elicited a significant reduction in bacterial colonization in the model.Conclusion. Our findings demonstrate the successful development of a novel mouse-adapted H. pylori model for the hpEastAsia genotype in Vietnam and Southeast Asia. Our research highlights the distinctive genotype and pathogenicity of clinical H. pylori strains in the region, laying the foundation for targeted interventions to address this global health burden.

Synthesis of Biodegradable and Antimicrobial Nanocomposite Films Reinforced for Coffee and Agri-Food Product Preservation.

The antimicrobial activity of silver nanoparticles is widely known. However, their application to biodegradable polymeric materials is still limited. In this work, we report a strategy involving the green synthesis of nanocomposite films based on a natural biodegradable matrix. Nanometer-sized silver nanoparticles (C-AgNPs) were synthesized with the aid of ultrasound waves between the silver nitrate solution and the nanocurcumin solution. The green synthesized C-AgNPs were found to have particle sizes in the range of 5-25 nm and demonstrated good antimicrobial activity against Clostridium perfringens, Staphylococcus aureus, Bacillus subtilis, Macrophoma theicola, and Aspergillus flavus. Owing to their physical-chemical and mechanical properties and the excellent antimicrobial activities, the obtained AgNPs were used together with chitosan, cassava starch, and poly(vinyl alcohol) (PVA) to make nanocomposite films, which are suitable for the packaging requirements of various key agricultural and food products such as coffee beans, bamboo straws, and fruits. The nanocomposite films lost up to 85% of their weight after being buried in the soil for 120 days. This indicates that the films made with natural biodegradable materials are environmentally friendly.

Prophylactic immunization to Helicobacter pylori infection using spore vectored vaccines.

BACKGROUND: Helicobacter pylori infection remains a major public health threat leading to gastrointestinal illness and increased risk of gastric cancer. Mostly affecting populations in developing countries no vaccines are yet available and the disease is controlled by antimicrobials which, in turn, are driving the emergence of AMR. MATERIALS AND METHODS: We have engineered spores of Bacillus subtilis to display putative H. pylori protective antigens, urease subunit A (UreA) and subunit B (UreB) on the spore surface. Following oral dosing of mice with these spores, we evaluated immunity and colonization in animals challenged with H. pylori. RESULTS: Oral immunization with spores expressing either UreA or UreB showed antigen-specific mucosal responses (fecal sIgA) including seroconversion and hyperimmunity. Following challenge, colonization by H. pylori was significantly reduced by up to 1-log. CONCLUSIONS: This study demonstrates the utility of bacterial spores for mucosal vaccination to H. pylori infection. The heat stability and robustness of Bacillus spores coupled with their existing use as probiotics make them an attractive solution for either protection against H. pylori infection or potentially for therapy and control of active infection.

High-resolution genomic analysis of four local Vietnamese chicken breeds.

In Vietnam, local chicken breeds account for over 70% of the national poultry population. Although these breeds are abundant, their productivity is low and their use is threatened by the extensive importation of foreign productive breeds. In this context, conservation programmes targeting several emblematic breeds have been established. The goal of these programmes was to characterize endangered breeds and maintain a pool of characteristic birds for preserving their genetic heritage. To contribute to these programmes, we comprehensively characterized four Vietnamese local chicken breeds (Dong Tao, Ho, Mia and Mong) at the genomic level using high-density single-nucleotide polymorphism (SNP) genotyping. Despite originating in geographically close areas, Dong Tao and Ho were evidently different from each other as well as from Mong and Mia, which shared a more recent common ancestor. The genomic inbreeding coefficient revealed high homozygosity amongst the four breeds (10%-20%). The observation of clear differentiation at the genomic level supported the presence of distinct breeds; nonetheless, the occurrence of crossbred birds in a presumably purebred sample demonstrated the need to apply genomic tools to unambiguously assign the birds to the correct breed. Moreover, the occurrence of substantial inbreeding and the presence of subgroups in certain breeds warranted attention to create future nuclei for use in the conservation of these local breeds.

Penicillium vietnamense sp. nov., the First Novel Marine Fungi Species Described from Vietnam with a Unique Conidiophore Structure and Molecular Phylogeny of Penicillium Section Charlesia.

Penicillium vietnamense sp. nov. was isolated from Nha Trang Bay, Vietnam in June 2017. It is phylogenetically distinct from the sister species of Penicillium section Charlesia series Indica based on multi-locus sequence typing results using internal transcribed spacer, large subunit ribosomal RNA, ß-tubulin, calmodulin, and RNA polymerase II second largest subunit regions. It showed strong growth on Czapek yeast autolysate agar at 37 °C, a strong acid production on Creatine sucrose agar, and produced short stipes, small vesicles, and subglobose to globose conidia delicately roughened with very short ridges. As the first novel marine fungi species described from Vietnam and discovered in a unique environment, the data could be significant for understanding the taxonomy and geographical distribution of marine fungi in tropical coastal systems such as Vietnam.

Convergence of Bar and Cry1Ac Mutant Genes in Soybean Confers Synergistic Resistance to Herbicide and Lepidopteran Insects.

Soybean is a globally important crop species, which is subject to pressure by insects and weeds causing severe substantially reduce yield and quality. Despite the success of transgenic soybean in terms of Bacillus thuringiensis (Bt) and herbicide tolerance, unforeseen mitigated performances have still been inspected due to climate changes that favor the emergence of insect resistance. Therefore, there is a need to develop a biotech soybean with elaborated gene stacking to improve insect and herbicide tolerance in the field. In this study, new gene stacking soybean events, such as bialaphos resistance (bar) and pesticidal crystal protein (cry)1Ac mutant 2 (M#2), are being developed in Vietnamese soybean under field condition. Five transgenic plants were extensively studied in the herbicide effects, gene expression patterns, and insect mortality across generations. The increase in the expression of the bar gene by 100% in the leaves of putative transgenic plants was a determinant of herbicide tolerance. In an insect bioassay, the cry1Ac-M#2 protein tested yielded higher than expected larval mortality (86%), reflecting larval weight gain and weight of leaf consumed were less in the T1 generation. Similarly, in the field tests, the expression of cry1Ac-M#2 in the transgenic soybean lines was relatively stable from T0 to T3 generations that corresponded to a large reduction in the rate of leaves and pods damage caused by Lamprosema indicata and Helicoverpa armigera. The transgenic lines converged two genes, producing a soybean phenotype that was resistant to herbicide and lepidopteran insects. Furthermore, the expression of cry1Ac-M#2 was dominant in the T1 generation leading to the exhibit of better phenotypic traits. These results underscored the great potential of combining bar and cry1Ac mutation genes in transgenic soybean as pursuant of ensuring resistance to herbicide and lepidopteran insects.

Marine and brackish Luticola D.G.Mann (Bacillariophyta) species from the Java Sea and South China Sea coasts with the description of three new species.

In this study, samples were collected from the Java Sea coasts, from the South China Sea in Hainan Island coasts and Quang Yên region and Rú Chá mangrove near Hue in Central Vietnam. In studied samples a total of eight Luticola species have been observed. Three of the taxa studied are described herein as species new to science - Luticolaorientalis sp. nov., L.cribriareolata sp. nov. and L.halongiana sp. nov. Under light microscopy (LM) L.orientalis sp. nov. and L.cribriareolata sp. nov. are similar with rhombic-lanceolate to rhombic/ elliptic-lanceolate to elliptic valve shapes and narrowly rounded apices. Both species can be easily distinguished by stria density (higher density in L.orientalis). Under SEML.cribriareolata is characterized by cribrate areola occlusions, a character thus far observed only in three established species. The remaining species of the whole genus known thus far are characterized by hymenate areola. Similar morphology Luticola species have been observed from tropical mangrove forests from Madagascar but they all can be easily distinguished based on the lack of grooves in the central area. The third species - L.halongiana sp. nov. has rhombic-elliptic to rhombic-lanceolate valves with broadly rounded to slightly protracted apices in larger specimens. This species has a relatively broad central area. Also unique among brackish-water Luticola is the small, rounded stigma positioned almost midway between the valve center and valve margin. In the habitats from which the new species are described we also identified five established Luticola taxa including, L.belawanensis, L.celebesica, L.inserata, L.seposita and L.tropica. For those species we provide detailed SEM characteristics of valve ultrastructure, as well as the range of environmental conditions and geographic distribution within the study area.

Biodiversity and Enzyme Activity of Marine Fungi with 28 New Records from the Tropical Coastal Ecosystems in Vietnam.

The coastal marine ecosystems of Vietnam are one of the global biodiversity hotspots, but the biodiversity of marine fungi is not well known. To fill this major gap of knowledge, we assessed the genetic diversity (ITS sequence) of 75 fungal strains isolated from 11 surface coastal marine and deeper waters in Nha Trang Bay and Van Phong Bay using a culture-dependent approach and 5 OTUs (Operational Taxonomic Units) of fungi in three representative sampling sites using next-generation sequencing. The results from both approaches shared similar fungal taxonomy to the most abundant phylum (Ascomycota), genera (Candida and Aspergillus) and species (Candida blankii) but were different at less common taxa. Culturable fungal strains in this study belong to 3 phyla, 5 subdivisions, 7 classes, 12 orders, 17 families, 22 genera and at least 40 species, of which 29 species have been identified and several species are likely novel. Among identified species, 12 and 28 are new records in global and Vietnamese marine areas, respectively. The analysis of enzyme activity and the checklist of trophic mode and guild assignment provided valuable additional biological information and suggested the ecological function of planktonic fungi in the marine food web. This is the largest dataset of marine fungal biodiversity on morphology, phylogeny and enzyme activity in the tropical coastal ecosystems of Vietnam and Southeast Asia. Biogeographic aspects, ecological factors and human impact may structure mycoplankton communities in such aquatic habitats.

Productive performance and egg and meat quality of two indigenous poultry breeds in Vietnam, Ho and Dong Tao, fed on commercial feed.

The objective of this work was the evaluation of the meat production and laying performances, and the meat and egg quality of two breeds of Vietnamese broiler chickens, Ho and Dong Tao, fed on a commercial diet. In a survey, we continuously recorded for 28 weeks, the data on the production performance and meat quality of 250 chicks from each breed. We investigated egg laying and egg quality using 36 Ho and 32 Dong Tao hens during 52 weeks of laying. The growth patterns were similar for the two breeds. Feed conversion ratios were also similar, and demonstrated the low efficiency of these two breeds when compared to commercial broilers. Slaughter age proved to affect several carcass yield characteristics, showing that slaughtering between 16 and 20 weeks might be better than at the usual age of 28 weeks. Yield, carcass composition and meat quality differed between the two studied breeds. The eggs production and number of embryonated eggs were low for the two breeds when compared to other breeds, with a lower hatching performance in Ho than in Dong Tao. In summary, the production performances of Ho and Dong Tao chickens were low, even when birds were fed a commercial diet. The study demonstrates the need to find ways to improve the production and reproduction performances of these animals, in order to contribute to the program of conservation and exploitation of these two breeds.

Molecular screening and genetic diversity analysis of anticancer Azurin-encoding and Azurin-like genes in human gut microbiome deduced through cultivation-dependent and cultivation-independent studies

Azurin, a bacteriocin produced by a human gut bacterium Pseudomonas aeruginosa, can reveal selectively cytotoxic and induce apoptosis in cancer cells. After overcoming two phase I trials, a functional region of Azurin called p28 has been approved as a drug for the treatment of brain tumor glioma by FDA. The present study aims to improve a screening procedure and assess genetic diversity of Azurin genes in P. aeruginosa and Azurin-like genes in the gut microbiome of a specific population in Vietnam and global populations. Firstly, both cultivation-dependent and cultivation-independent techniques based on genomic and metagenomic DNAs extracted from fecal samples of the healthy specific population were performed and optimized to detect Azurin genes. Secondly, the Azurin gene sequences were analyzed and compared with global populations by using bioinformatics tools. Finally, the screening procedure improved from the first step was applied for screening Azurin-like genes, followed by the protein synthesis and NCI in vitro screening for anticancer activity. As a result, this study has successfully optimized the annealing temperatures to amplify DNAs for screening Azurin genes and applying to Azurin-like genes from human gut microbiota. The novelty of this study is the first of its kind to classify Azurin genes into five different genotypes at a global scale and confirm the potential anticancer activity of three Azurin-like synthetic proteins (Cnazu1, Dlazu11, and Ruazu12). The results contribute to the procedure development applied for screening anticancer proteins from human microbiome and a comprehensive understanding of their therapeutic response at a genetic level

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Molecular screening and genetic diversity analysis of anticancer Azurin-encoding and Azurin-like genes in human gut microbiome deduced through cultivation-dependent and cultivation-independent studies.

Azurin, a bacteriocin produced by a human gut bacterium Pseudomonas aeruginosa, can reveal selectively cytotoxic and induce apoptosis in cancer cells. After overcoming two phase I trials, a functional region of Azurin called p28 has been approved as a drug for the treatment of brain tumor glioma by FDA. The present study aims to improve a screening procedure and assess genetic diversity of Azurin genes in P. aeruginosa and Azurin-like genes in the gut microbiome of a specific population in Vietnam and global populations. Firstly, both cultivation-dependent and cultivation-independent techniques based on genomic and metagenomic DNAs extracted from fecal samples of the healthy specific population were performed and optimized to detect Azurin genes. Secondly, the Azurin gene sequences were analyzed and compared with global populations by using bioinformatics tools. Finally, the screening procedure improved from the first step was applied for screening Azurin-like genes, followed by the protein synthesis and NCI in vitro screening for anticancer activity. As a result, this study has successfully optimized the annealing temperatures to amplify DNAs for screening Azurin genes and applying to Azurin-like genes from human gut microbiota. The novelty of this study is the first of its kind to classify Azurin genes into five different genotypes at a global scale and confirm the potential anticancer activity of three Azurin-like synthetic proteins (Cnazu1, Dlazu11, and Ruazu12). The results contribute to the procedure development applied for screening anticancer proteins from human microbiome and a comprehensive understanding of their therapeutic response at a genetic level.

The complete mitochondrial DNA of the tropical oyster Crassostrea belcheri from the Can Giò' mangrove in Vietnam.

The complete mitochondrial genome of the oyster Crassostrea belcheri from the Can Giò' mangrove in Vietnam has been sequenced. It consists of a circular DNA molecule of 21020 base pairs (bp), coding for 12 proteins, 20 transfer RNAs, and two ribosomal RNAs. Like the mitogenomes of Crassostrea iredalei and Crassostrea sp. DB1, it contains a non-coding region and two ORFs. The C. belcheri mitogenome provides information that could improve the molecular phylogeny of Asian oysters and be useful to the development of oyster aquaculture in South East Asia.

Complete mitogenome of Cerithidea obtusa, the red chut-chut snail from the Can Gio Mangrove in Vietnam.

We sequenced the complete mitogenome of the red chut-chut snail Cerithidea obtusa, from the Can Gio mangrove in Vietnam. The mitogenome is 15,708 bp long. It is colinear with the mitogenomes of other members of the superfamily Cerithioidea, and the maximum-likelihood phylogeny obtained with the cox1, cox2 and cox3 genes of several Caenogastropoda associated all Cerithioidea together inside a strongly supported clade.

Discovery of Azurin-Like Anticancer Bacteriocins from Human Gut Microbiome through Homology Modeling and Molecular Docking against the Tumor Suppressor p53.

Azurin from Pseudomonas aeruginosa is known anticancer bacteriocin, which can specifically penetrate human cancer cells and induce apoptosis. We hypothesized that pathogenic and commensal bacteria with long term residence in human body can produce azurin-like bacteriocins as a weapon against the invasion of cancers. In our previous work, putative bacteriocins have been screened from complete genomes of 66 dominant bacteria species in human gut microbiota and subsequently characterized by subjecting them as functional annotation algorithms with azurin as control. We have qualitatively predicted 14 putative bacteriocins that possessed functional properties very similar to those of azurin. In this work, we perform a number of quantitative and structure-based analyses including hydrophobic percentage calculation, structural modeling, and molecular docking study of bacteriocins of interest against protein p53, a cancer target. Finally, we have identified 8 putative bacteriocins that bind p53 in a same manner as p28-azurin and azurin, in which 3 peptides (p1seq16, p2seq20, and p3seq24) shared with our previous study and 5 novel ones (p1seq09, p2seq05, p2seq08, p3seq02, and p3seq17) discovered in the first time. These bacteriocins are suggested for further in vitro tests in different neoplastic line cells.

Chlorine Gas Sensing Performance of On-Chip Grown ZnO, WO3, and SnO2 Nanowire Sensors.

Monitoring toxic chlorine (Cl2) at the parts-per-billion (ppb) level is crucial for safe usage of this gas. Herein, ZnO, WO3, and SnO2 nanowire sensors were fabricated using an on-chip growth technique with chemical vapor deposition. The Cl2 gas-sensing characteristics of the fabricated sensors were systematically investigated. Results demonstrated that SnO2 nanowires exhibited higher sensitivity to Cl2 gas than ZnO and WO3 nanowires. The response (RCl2/Rair) of the SnO2 nanowire sensor to 50 ppb Cl2 at 50 °C was about 57. Hence, SnO2 nanowires can be an excellent sensing material for detecting Cl2 gas at the ppb level under low temperatures. Abnormal sensing characteristics were observed in the WO3 and SnO2 nanowire sensors at certain temperatures; in particular, the response level of these sensors to 5 ppm of Cl2 was lower than that to 2.5 ppm of Cl2. The sensing mechanism of the SnO2 nanowire sensor was also elucidated by determining Cl2 responses under N2 and dry air as carrier gases. We proved that the Cl2 molecule was first directly adsorbed on the metal oxide surface and was then substituted for pre-adsorbed oxygen, followed by lattice oxygen.

Fabrication of highly sensitive and selective H2 gas sensor based on SnO2 thin film sensitized with microsized Pd islands.

Ultrasensitive and selective hydrogen gas sensor is vital component in safe use of hydrogen that requires a detection and alarm of leakage. Herein, we fabricated a H2 sensing devices by adopting a simple design of planar-type structure sensor in which the heater, electrode, and sensing layer were patterned on the front side of a silicon wafer. The SnO2 thin film-based sensors that were sensitized with microsized Pd islands were fabricated at a wafer-scale by using a sputtering system combined with micro-electronic techniques. The thicknesses of SnO2 thin film and microsized Pd islands were optimized to maximize the sensing performance of the devices. The optimized sensor could be used for monitoring hydrogen gas at low concentrations of 25-250 ppm, with a linear dependence to H2 concentration and a fast response and recovery time. The sensor also showed excellent selectivity for monitoring H2 among other gases, such as CO, NH3, and LPG, and satisfactory characteristics for ensuring safety in handling hydrogen. The hydrogen sensing characteristics of the sensors sensitized with Pt and Au islands were also studied to clarify the sensing mechanisms.

Screening of marine bacteria with bacteriocin-like activities and probiotic potential for ornate spiny lobster (Panulirus ornatus) juveniles.

Bacteriocins are ribosomally synthesized antimicrobial peptides, which have been found in diverse bacterial species of terrestrial origins and some from the sea. New bacteriocins with new characteristics, new origins and new applications are likely still awaiting discovery. The present study screened bacteria isolated from marine animals of interest to the aquaculture industry for antimicrobial and bacteriocin-like activities in order to uncover biodiversity of bacteriocin producers, and explore the potential application in aquaculture. In total, 24 of 100 screened isolates showed antimicrobial activities and 7 of these exerted bacteriocin-like activities. Sequencing of 16S rRNA genes identified the isolates as members of the six genera Proteus, Providencia, Klebsiella, Alcaligenes, Bacillus and Enterococcus. In some cases, further analysis of housekeeping genes, rpoB for Proteus and recA for Klebsiella, as well as biochemical tests was necessary for identification to species level, and some of the Proteus isolates may represent novel species. The seven bacteriocinogenic isolates showed a wide antimicrobial spectrum against foodborne and animal pathogens, which opens the way to their potential use as marine drugs and probiotics in food, aquaculture, livestock and clinical settings. As a case study, the protective effect of shortlisted bacteriocinogenic isolates were tested in aquaculture-raised spiny lobster (Panulirus ornatus) juveniles. A single-strain (Bacillus pumilus B3.10.2B) and a three-strain (B. pumilus B3.10.2B, Bacillus cereus D9, Lactobacillus plantarum T13) probiotic preparation were added to the feed of Panulirus ornatus juveniles, which were subsequently challenged with the pathogen Vibrio owensii DY05. Juveniles in the probiotic treatments displayed increased growth and reduced feed conversion rates after 60 days, and increased survival rate after pathogen challenge relative to the control. This study represents the first evidence of bacteriocin production by bacteria associated with lobster, tiger shrimp, snubnose pompano and cobia and the first description of V. owensii as a pathogen in P. ornatus juveniles.

Transcriptome and proteome analyses in response to 2-methylhydroquinone and 6-brom-2-vinyl-chroman-4-on reveal different degradation systems involved in the catabolism of aromatic compounds in Bacillus subtilis.

Bacillus subtilis is exposed to a variety of antimicrobial compounds in the soil. In this paper, we report on the response of B. subtilis to the fungal-related antimicrobials 6-brom-2-vinyl-chroman-4-on (chromanon) and 2-methylhydroquinone (2-MHQ) using proteome and transcriptome analyses. Chromanon, a derivative of aposphaerins from Aposphaeria species caused predominant protein damage in B. subtilis as indicated by the induction of the HrcA, CtsR, and Spx regulons. The expression profile of the ganomycin-related substance 2-MHQ was similar to that of catechol as reflected by the common induction of the thiol-specific oxidative stress response. Several putative ring-cleavage dioxygenases and oxidoreductases were differentially up-regulated by 2-MHQ, catechol, and chromanon including yfiDE, ydfNOP, yodED, ycnDE, yodC, and ykcA. The nitroreductase encoding yodC gene is induced in response to catechol, 2-MHQ, and chromanon, which depend on the MarR-type repressor YodB. The yfiDE (catDE) operon encodes a catechol-2,3-dioxygenase which is most strongly induced by catechol. The yodED (mhqED), ydfNOP (mhqNOP) operons, and ykcA (mhqA) respond most strongly to 2-MHQ and encode putative hydroquinone-specific extradiol dioxygenases. The ycnDE operon was most strongly induced by chromanon. Mutational analyses revealed that the putative hydroquinone-specific dioxygenases MhqO and MhqA confer resistance to 2-MHQ in B. subtilis.