First draft genome sequence data of TA4-1, the type strain of Gram-positive bacterium Streptomyces chiangmaiensis.

TA4-1 is the type strain of Streptomyces chiangmaiensis. The TA4-1 strain was isolated from a stingless bee (Tetragonilla collina). Here we present the draft genome sequence data of S. chiangmaiensis TA4-1. The Illumina NextSeq 550 sequencer was used to generate paired-end reads from the genomic DNA of the pure culture of S. chiangmaiensis TA4-1. The draft genome sequence of strain TA4-1 consists of 776 contigs with a total size of 9,707,984 base pairs, an N50 of 32,937 base pairs, and a GC content of 69.73 %. Digital DNA-DNA hybridisation (dDDH) and average nucleotide identity (ANI) analysis showed that S. yaanensis CGMCC 4.7035 had the highest dDDH value (32.7 %) and ANIm value (88.50 %) when compared with TA4-1. The presented data indicate the potential for a reference genome sequence in bacterial taxonomy, comparative genomics, and the investigation of bioactive compound biosynthesis in S. chiangmaiensis TA4-1. The draft genome sequence data have been deposited at NCBI under the Bioproject accession number PRJNA680432.

Draft genome sequence data of multidrug-resistant Pseudomonas aeruginosa WO7 from a hospital wastewater treatment plant in Thailand.

Multidrug-resistant Pseudomonas aeruginosa WO7 was isolated from an untreated water sample from a hospital wastewater treatment plant in Thailand. This report presents the draft genome sequence data of P. aeruginosa WO7. Genomic DNA was obtained from a pure culture of P. aeruginosa WO7, and paired-end reads were generated using an Illumina MiSeq sequencer. The draft genome consisted of 111 contigs with a total size of 6,784,206 base pairs, an N50 of 209,424 base pairs, and a GC content of 65.85%. The dDDH value between WO7 and Pseudomonas aeruginosa DSM 50071T was determined to be 90.7%, indicating that the strain is Pseudomonas aeruginosa. The data presented indicate the potential for bacterial classification, comparative genomics, comprehensive analysis of antimicrobial resistance, and assessment of bacterial virulence factors in P. aeruginosa. The draft genome sequence data have been deposited at the NCBI under Bioproject accession number PRJNA550309.

Streptomyces monashensis MSK03-mediated synthesis of gold nanoparticles: characterization and antibacterial activity.

Nanotechnology is a cutting-edge field with diverse applications, particularly in the utilization of gold nanoparticles (AuNPs) due to their stability and biocompatibility. AuNPs serve as pivotal components in medical applications, with a specific emphasis on their significant antibacterial efficacy. This study focuses on synthesizing AuNPs using the cell-free supernatant of Streptomyces monashensis MSK03, isolated from terrestrial soil in Thailand. The biosynthesis process involved utilizing the cell-free supernatant of S. monashensis MSK03 and hydrogen tetrachloroauric acid (HAuCl4) under controlled conditions of 37 °C and 200 rpm agitation. Characterization studies revealed spherical AuNPs with sizes ranging from 7.1 to 40.0 nm (average size: 23.2 ± 10.7 nm), as confirmed by TEM. UV-Vis spectroscopy indicated a localized surface plasmon resonance (LSPR) band at 545 nm, while XRD analysis confirmed a crystalline structure with characteristics of cubic lattice surfaces. The capping molecules on the surface of AuNPs carry a negative charge, indicated by a Zeta potential of -26.35 mV, and FTIR analysis identified functional groups involved in reduction and stabilization. XANES spectra further confirmed the successful reduction of Au3+ to Au0. Moreover, the synthesized AuNPs demonstrated antibacterial activity against drug-resistant strains of Pseudomonas aeruginosa and Acinetobacter baumannii. Interestingly, the AuNPs showed non-toxicity to Vero cell lines. These significant antibacterial properties of the produced nanoparticles mean they hold great promise as new antimicrobial treatments for tackling the increasing issue of antibiotic resistance.

Draft genome sequence data of the multidrug-resistant bacterium Staphylococcus haemolyticus 010503B isolated from an aerosol sample in a hospital waiting area in Thailand.

Staphylococcus haemolyticus 010503B is a multidrug-resistant bacterium isolated from an outpatient clinic in a hospital waiting area in Thailand. Here we present the draft genome sequence of S. haemolyticus 010503B. The paired-end reads were generated on the Illumina NextSeq 550 sequencer using genomic DNA from the pure culture of S. haemolyticus 010503B. The draft genome consisted of 114 contigs with a total size of 2,457,654 base pairs, an N50 of 57,312 base pairs and a GC content of 32.60%. The dDDH between 010503B and Staphylococcus haemolyticus SM 131T was 91.9%, identifying the strain as Staphylococcus haemolyticus. The data presented holds promise for bacterial classification, comparative genomics, analysing antimicrobial resistance comprehensively, and assessing bacterial virulence factors of S. haemolyticus. The draft genome sequence data has been deposited at NCBI under Bioproject accession number PRJNA550309.

Draft genome sequence data of the endophytic actinobacterium Streptomyces justiciae WPN32, a potential bioactive compounds producer.

Streptomyces justiciae WPN32 is an endophytic actinobacterium isolated from the rhizosphere of turmeric field soil at the Botanical Garden of Suranaree University of Technology (Nakhon Ratchasima Province, Thailand). Here we present the draft genome sequence of S. justiciae WPN32. It was sequenced on the Illumina NextSeq 550 sequencer. The draft genome consisted of 123 contigs with a total size of 9,832,147 base pairs, an N50 of 237,572 base pairs and a GC content of 70.87%. The dDDH between WPN32 and Streptomyces justiciae 3R004T was 80.1%, identifying the strain as Streptomyces justiciae. The data presented here may aid microbial taxonomy, comparative genomics and identification of gene clusters associated with the synthesis of bioactive compounds. The draft genome sequence data has been deposited at NCBI under Bioproject accession number PRJNA680432.

Draft genome sequence data of heavy metal-resistant Morganella morganii WA01/MUTU, a silver nanoparticle (AgNP) synthesising bacterium.

Morganella morganii WA01/MUTU is a heavy metal tolerant strain capable of producing silver nanoparticles (AgNPs) from AgNO3. Here we present the draft genome sequence of M. morganii WA01/MUTU isolated from a water sample collected in Nakhon Pathom province, Thailand. The draft genome was sequenced on the Illumina NextSeq 550 sequencer. The genome consisted of 34 contigs with a total size of 3,991,804 bp, an N50 value of 364,423 bp and a GC content of 50.93%. The digital DNA-DNA hybridisation (dDDH) between WA01/MUTU and Morganella morganii (NBRC 3848) was 83.9%, identifying the strain as Morganella morganii. The data presented here can be used in comparative genomics to identify gene clusters involved in AgNP biosynthesis and secondary metabolite production. The draft genome sequence data was deposited at NCBI under Bioproject accession number PRJNA493966.

Development of Duplex LAMP Technique for Detection of Porcine Epidemic Diarrhea Virus (PEDV) and Porcine Circovirus Type 2 (PCV 2).

Porcine epidemic diarrhea virus (PEDV) and porcine circovirus type 2 (PCV2) are both important global pathogenic viruses which have a significant impact on the swine industry. In this study, a duplex loop-mediated isothermal amplification (duplex LAMP) method was developed in combination with lateral flow dipstick (LFD) for simultaneous detection of PEDV and PCV2 using specific sets of primers and probes designed based on the conserved regions of a spike gene (KF272920) and an ORF gene (EF493839), respectively. The limit of detection (LOD) values of the duplex LAMP-LFD for the detection of PEDV and PCV2 were 0.1 ng/µL and 0.246 ng/µL, respectively. The LOD of duplex LAMP-LFD was 10-times more sensitive than conventional PCR and RT-PCR-agarose gel-electrophoresis (PCR-AGE and RT-PCR-AGE). No cross-reaction to each other and to other pathogenic viruses that can infect pigs were observed according to analytical specificity tests. The duplex LAMP-LFD method for the simultaneous detection of PEDV and PCV2 co-infection could be completed within approximately 1.5 h, and only a simple heating block was required for isothermal amplification. The preliminary validation using 50 swine clinical samples with positive and negative PEDV and/or PCV2 revealed that the sensitivity, specificity, and accuracy of duplex LAMP-LFD were all 100% in comparison to conventional PCR and RT-PCR. Hence, this study suggests that duplex LAMP-LFD is a promising tool for the early detection and initial screening of PEDV and PCV2, which could be beneficial for prevention, planning, and epidemiological surveys of these diseases.

Hybrid Plasticizers Enhance Specificity and Sensitivity of an Electrochemical-Based Sensor for Cadmium Detection.

In addition to their use as an additive to improve physical properties of solvent polymeric membranes, plasticizers have a considerable impact on the specificity and sensitivity of membrane-modified electrochemical sensors. In this work, we aim at the hybridization of two different plasticizers using the electropolymerization technique in the development of a cadmium(II)-selective electrochemical sensor based on screen-printed gold electrode along with cyclic voltammetric measurement. At this point, we first screen for the primary plasticizer yielding the highest signal using cyclic voltammetry followed by pairing it with the secondary plasticizers giving rise to the most sensitive current response. The results show that the hybridization of DOS and TOTM with 3:1 weight ratio (~137.7-µm-thick membrane) renders a signal that is >26% higher than that from the sensor plasticized by DOS per se in water. The solution of 0.1 mM hydrochloric acid (pH 4) is the optimal supporting electrolyte. In addition, hybrid plasticizers have adequate redox capacity to induce cadmium(II) transfer from bulk solution to the membrane/water interfaces. Conversion of voltammetric signals to semi-integral currents results in linearity with cadmium(II) concentration, indicating the irreversible cadmium(II) transfer to the membrane. The DOS:TOTM hybrid sensor also exhibits high sensitivity, with a limit of detection (LOD) and limit of quantitation (LOQ) of 95 ppb and 288 ppb, respectively, as well as greater specificity towards cadmium(II) than that obtained from the single plasticizer sensor. Furthermore, recovery rates of spiked cadmium(II) in water samples were higher than 97% using the hybrid plasticizer sensor. Unprecedentedly, our work reports that the hybridization of plasticizers serves as ion-to-electron transducer that can improve the sensor performance in cadmium(II) detection.

Metabolic engineering of Escherichia coli for biosynthesis of ß-nicotinamide mononucleotide from nicotinamide.

The ß-nicotinamide mononucleotide (NMN) is a key intermediate of an essential coenzyme for cellular redox reactions, NAD. Administration of NMN is reported to improve various symptoms, such as diabetes and age-related physiological decline. Thus, NMN is attracting much attention as a promising nutraceutical. Here, we engineered an Escherichia coli strain to produce NMN from cheap substrate nicotinamide (NAM) and glucose. The supply of in vivo precursor phosphoribosyl pyrophosphate (PRPP) and ATP was enhanced by strengthening the metabolic flux from glucose. A nicotinamide phosphoribosyltransferase with high activity was newly screened, which is the key enzyme for converting NAM to NMN with PRPP as cofactor. Notably, the E. coli endogenous protein YgcS, which function is primarily in the uptake of sugars, was firstly proven to be beneficial for NMN production in this study. Fine-tuning regulation of ygcS gene expression in the engineered E. coli strain increased NMN production. Combined with process optimization of whole-cell biocatalysts reaction, a final NMN titre of 496.2 mg l-1 was obtained.

Genome Sequence of Streptomyces cavourensis BUU135, Isolated from Soil from a Tropical Fruit Farm in Thailand.

Streptomyces cavourensis BUU135 is a bacterial species isolated from the soil of a tropical fruit farm. The genome of S. cavourensis BUU135 comprises a gene encoding nebramycin 5' synthase, which produces nebramycin 5' by catalyzing the O-carbamoylation reaction of tobramycin. The newly sequenced 7.66-Mb draft genome of S. cavourensis BUU135 may contribute to the discovery of novel natural products derived from this organism.

First Draft Genome Sequence of Thermophilic Laceyella tengchongensis BKK01, Isolated from Municipal Solid Waste in Thailand.

Laceyella tengchongensis BKK01 is a thermophilic bacterium isolated from municipal solid waste. The genome of L. tengchongensis BKK01 includes a gene putatively encoding gramicidin S synthase. Gramicidin S has antibiotic activity against some bacteria and fungi. The newly sequenced 3.44-Mb draft genome of L. tengchongensis BKK01 will shed some light on the biosynthesis of gramicidin S.

Draft Genome Sequence of Haloferax volcanii SS0101, Isolated from Salt Farms in Samut Sakhon, Thailand.

Haloferax volcanii SS0101 is a halophilic archaeon isolated from salt farms in Thailand. The genome sequence of H. volcanii SS0101 contains a gene encoding capreomycidine synthase, a key enzyme for capreomycidine biosynthesis. This 3.8-Mb draft genome sequence of H. volcanii SS0101 will provide the tools for investigating genes involved in capeomycidine production in haloarchaea.

A Novel Nucleic Lateral Flow Assay for Screening phaR-Containing Bacillus spp.

Polyhydroxyalkanoate (PHA) synthase is a key enzyme for PHA production in microorganisms. The class IV PHA synthase is composed of two subunits: PhaC and PhaR. The PhaR subunit, which encodes the phaR gene, is only present in class IV PHA synthases. Therefore, the phaR gene is used as a biomarker for bacteria that contain a class IV PHA synthase, such as some Bacillus spp. The phaR gene was developed to screen phaR-containing Bacillus spp. The phaR screening method involved two steps: phaR gene amplification by PCR and phaR amplicon detection using a DNA lateral flow assay. The screening method has a high specificity for phaR-containing Bacillus spp. The lowest amount of genomic DNA of B. thuringiensis ATCC 10792 that the phaR screening method could detect was 10 pg. This novel screening method improves the specificity and sensitivity of phaR gene screening and reduces the time and cost of the screening process, which could enhance the opportunity to discover good candidate PHA producers. Nevertheless, the screening method can certainly be used as a tool to screen phaR-containing Bacillus spp. from environmental samples.

Elevation of Cleaved p18 Bax Levels Associated with the Kinetics of Neuronal Cell Death during Japanese Encephalitis Virus Infection.

Japanese encephalitis virus (JEV) infection induces uncontrolled neuronal apoptosis, leading to irreversible brain damage. However, the mechanism of JEV-induced neuronal apoptosis has not been clearly elucidated. This study aimed to investigate both virus replication and neuronal cell apoptosis during JEV infection in human neuroblastoma SH-SY5Y cells. As a result, the kinetic productions of new viral progeny were time- and dose-dependent. The stimulation of SH-SY5Y cell apoptosis was dependent on the multiplicity of infections (MOIs) and infection periods, particularly during the late period of infection. Interestingly, we observed that of full-length Bax (p21 Bax) level started to decrease, which corresponded to the increased level of its cleaved form (p18 Bax). The formation of p18 Bax resulting in cytochrome c release into the cytosol appeared to correlate with JEV-induced apoptotic cell death together with the activation of caspase-3/7 activity, especially during the late stage of a robust viral infection. Therefore, our results suggest another possible mechanism of JEV-induced apoptotic cell death via the induction of the proteolysis of endogenous p21 Bax to generate p18 Bax. This finding could be a new avenue to facilitate novel drug discovery for the further development of therapeutic treatments that could relieve neuronal damage from JEV infection.

Draft Genome Sequence of Multidrug-Resistant Proteus mirabilis CKTH01, Isolated from Raw Chicken Meat.

Proteus mirabilis CKTH01 is a pathogenic bacterium isolated from raw chicken meat. The genome sequence of P. mirabilis CKTH01 contains genes encoding multidrug efflux pumps, which are the virulence factors of the antibiotic-resistant bacterium. This 3.98-Mb draft genome sequence of P. mirabilis CKTH01 will contribute to the understanding of the distribution of multidrug-resistant P. mirabilis in raw chicken meat at the open markets.

Dissemination of Carbapenem-Resistance among Multidrug Resistant Pseudomonas aeruginosa carrying Metallo-Beta-Lactamase Genes, including the Novel blaIMP-65 Gene in Thailand.

BACKGROUND: Pseudomonas aeruginosa is considered as one of the most emerging threats in this century. Serious infections caused by this pathogen are often treated by carbapenems which are the last resource of antibiotics. Metallo-beta-lactamases (MBLs) production is one of the most important carbepenem resistance mechanisms and is usually related with nosocomial infections caused by P. aeruginosa. This study was aimed to determine the prevalence of MBL genes and distribution pattern of MBLs producing P. aeruginosa strains in Thailand. MATERIALS AND METHODS: Specific primers were designed to detect MBL genes including IMP-, VIM-, and NDM-type MBL genes. Multilocus sequence typing method was used to determine the dissemination pattern of carbapenem-resistance among multidrug-resistant (CR-MDR) P. aeruginosa. RESULTS: A total of 153 P. aeruginosa clinical isolates were characterized as CR-MDR. Among those, 31 P. aeruginosa clinical isolates (20.3%) presented metallo-beta-lactamase genes which could be divided into VIM-type (14 strains) and IMP-type (17 strains). blaIMP-1, blaIMP-13, blaIMP-14a, and blaVIM-2 genes were detected. Moreover, a novel IMP-type MBL, blaIMP-65 was discovered and it was demonstrated to be the unique group of MBLs in Thailand. It was of interest that ST235 was the major ST type in Thailand followed by ST964 and ST111 and ST235 was detected in both MBL harboring and non-MBL harboring strains. CONCLUSION: This study reported the dissemination of MBL gene including novel MBL, blaIMP-65. This study was also demonstrated major ST of P. aeruginosa which was ST235, followed by ST964 and ST111. Moreover, it is also the first report on many P. aeruginosa STs in Thailand: ST273, ST292, ST621, ST1584, and ST1816 which emphasized the dissemination trait difference of MBLs harboring P. aeruginosa in Thailand.

Draft Genome Sequence of Thermophilic Halotolerant Aeribacillus pallidus TD1, Isolated from Tao Dam Hot Spring, Thailand.

Aeribacillus pallidus TD1 is a thermophilic bacterium isolated from a hot spring in Thailand. The genome sequence of A. pallidus TD1 contains a gene-encoded naphthalene dioxygenase, which is a key enzyme for naphthalene degradation. This 3.7-Mb draft genome sequence of A. pallidus TD1 will contribute to the understanding of polycyclic aromatic hydrocarbon (PAH) degradation in high-temperature environments.

Draft Genome Sequence of Bacillus salarius IM0101, Isolated from Hypersaline Soil in Inner Mongolia, China.

Bacillus salarius IM0101 is a halophilic bacterium that was isolated from soil in Inner Mongolia, China. The genome sequence of B. salarius IM0101 contains a biomarker gene for polyhydroxyalkanoate (PHA) synthesis. This 6.9-Mb draft genome sequence of B. salarius IM0101 will provide new insights into the organism's PHA production machinery.

Development of an immunoFET biosensor for the detection of biotinylated PCR product.

ImmunoFET (IMFET) biosensor is a simple platform for the detection of biotinylated products of polymerase chain reaction (PCR). Construction of the IMFET biosensor started with adsorption of 1.5 mg/mL of protein A (PA) onto the insulated gate surface of ISFET for 90 min. Next, the immobilized 1/500 dilution of anti-biotin antibody was adsorbed onto the PA layer for 60 min. The IMFET biosensor was subsequently ready for detection of the biotinylated amplicon. The IMFET biosensor showed highly specific binding to the biotinylated PCR product of the phaE gene of Haloquadratum walsbyi DSM 16854. The phaE gene is a biomarker of polyhydroxyalkanoate (PHA) producers that contain PHA synthase class III. The lowest amount of DNA template of H. walsbyi DSM 16854 that the IMFET biosensor could detect was 125 fg. The IMFET biosensor has a lower amount of detection compared with a DNA lateral flow biosensor from our previous study. The degree of linearity of the biosensor signal was influenced by the concentration of the biotinylated amplicon. The IMFET biosensor also has a short response time (approximately 30 times) to detect the phaE amplicon compared to an agarose gel electrophoresis. The IMFET biosensor is a promising tool for the detection of the biotinylated PCR product, and it can be integrated into a micro total analysis system (µTAS).

Development of molecular approach based on PCR assay for detection of histamine producing bacteria.

Histamine fish poisoning becomes highly concern not only in public health but also economic aspect. Histamine is produced from histidine in fish muscles by bacterial decarboxylase enzyme. Several techniques have been developed to determine the level of histamine in fish and their products but the effective method for detecting histamine producing bacteria is still required. This study was attempted to detect histamine producing bacteria by newly developed PCR condition. Histamine producing bacteria were isolated from scombroid fish and determined the ability to produce histamine of isolated bacteria by biochemical and TLC assays. PCR method was developed to target the histidine decarboxylase gene (hdc). The result showed that fifteen histamine producing bacterial isolates and three standard strains produced an amplicon at the expected size of 571 bp after amplified by PCR using Hdc_2F/2R primers. Fifteen isolates of histamine producing bacteria were classified as M. morganii, E. aerogenes, and A. baumannii. The lowest detection levels of M. morganii and E. aerogenes were 10(2) and 10(5) Cfu/mL in culture media and 10(3) and 10(6) Cfu/mL in fish homogenates, respectively. The limit of detection by this method was clearly shown to be sensitive because the primers could detect the presence of M. morganii and E. aerogenes before the histamine level reached the regulation level at 50 ppm. Therefore, this PCR method exhibited the potential efficiency for detecting the hdc gene from histamine producing bacteria and could be used to prevent the proliferation of histamine producing bacteria in fish and fish products.