16S rRNA Gene Sequence Analysis Research Articles

Achromobacter sp. Strain BUKˍBCHˍTQ1: A Potential Paraquat-Degrading Bacterium Isolated from Pesticide Contaminated Agricultural Soil

Paraquat (1, 1′-dimethyl-4, 4′-bipyridinium dichloride) is one of the most frequently used herbicide in agriculture. It is a cationic non-systematic, non-selective contact compound that instantaneously interferes with the photosynthetic processes of plants. It has an immediate effect, once the compound comes into contact with the plants’ leaves, where the reaction occurs. However, the contamination of paraquat residue in soil can harm soil microbes, flora and fauna, farmer health and also soil ecology, which affects the soil fertility. The objective of this study was to isolate and characterize bacteria with the ability to break down and utilize paraquat as the primary carbon source. The isolation process involved the enrichment of mineral salt media (MSM) using serial dilution. The isolated bacterium underwent morphological, biochemical, and molecular identification following characterization. Results showed the isolate was identified as Achromobacter sp. with the accession number OQ372943 based on partial 16S rRNA gene sequence and phylogenetic analysis. The growth and degradation of paraquat by this isolate were optimum at a pH of 6.5, 276 mgL-1 of the substrate (paraquat), temperature of 35 °C, 200 µL of biomass size, and 48 h of incubation. The degradation efficiency of the isolate after 120 hours of incubation under optimal conditions was 91.01%. Hence, these results demonstrated a very high efficiency of paraquat degradation. Consequently, this isolate holds significant promise for paraquat degradation and could serve as a viable contender for remediating paraquat-contaminated environments.

Astaxanthin alleviates chronic prostatitis/chronic pelvic pain syndrome by increasing colonization of Akkermansia muciniphila in the intestine

BackgroundAstaxanthin (AST) is a natural compound with anti-inflammatory/immunomodulatory properties that has been found to have probiotic properties. However, the role and mechanism of AST in chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) are still not fully understood. PurposeThe aim of this study was to evaluate the effect of AST on CP/CPPS and elucidate the mediating role of the gut microbiota. Materials and methodsAn experimental autoimmune prostatitis (EAP) mouse model was utilized to test the potential role of AST on CP/CPPS. Antibiotic cocktail (ABX) treatment and fecal microbiota transplantation (FMT) were used to elucidate the gut microbiota-mediated effects on AST. In addition, 16S rRNA gene sequencing and qRT-PCR analyses were used to analyze changes in the gut microbiota of EAP mice and CP/CPPS patients. Finally, the mechanism by which AST exerts a protective effect on CP/CPPS was explored by untargeted metabolomics and gut barrier function assays. ResultsOral administration of AST reduced prostate inflammation scores, alleviated tactile sensitization of the pelvic region in EAP mice, reduced CD4+ T cell and CD68+ macrophage infiltration in the prostatic interstitium, and inhibited the up-regulation of systemic and localized pain/pro-inflammatory mediators in the prostate. After ABX, the protective effect of AST against CP/CPPS was attenuated, whereas colonization with fecal bacteria from AST-treated EAP mice alleviated CP/CPPS. 16S rRNA gene sequencing and qRT-PCR analyses showed that Akkermansia muciniphila in the feces of EAP mice and CP/CPPS patients showed a trend toward a decrease, which was associated with poor progression of CP/CPPS. In contrast, oral administration of AST increased the relative abundance of A. muciniphila, and oral supplementation with A. muciniphila also alleviated inflammation and pain in EAP mice. Finally, we demonstrated that both AST and A. muciniphila interventions increased serum levels of SCFAs acetate, up-regulated expression of colonic tight junction markers, and decreased serum lipopolysaccharide levels in EAP mice. ConclusionOur results showed that AST improved CP/CPPS by up-regulating A. muciniphila, which provides new potentially effective strategies and ideas for CP/CPPS management.

Gordonia metallireducens sp. nov., a tellurite- and selenite-resistant bacterium isolated from the sediment of an acid mine drainage stream.

A polyphasic taxonomic study was carried out on strain TSed Te1T, isolated from sediment of a stream contaminated with acid drainage from a coal mine. The bacterium forms pink-pigmented colonies and has a rod-coccus growth cycle, which also includes some coryneform arrangements. This bacterium is capable of growing in the presence of up to 750 μg ml-1 tellurite and 5000 μg ml-1 selenite, reducing each to elemental form. Nearly complete 16S rRNA gene sequence analysis associated the strain with Gordonia, with 99.5 and 99.3 % similarity to Gordonia namibiensis and Gordonia rubripertincta, respectively. Computation of the average nucleotide identity and digital DNA-DNA hybridization comparisons with the closest phylogenetic neighbour of TSed Te1T revealed genetic differences at the species level, which were further substantiated by differences in several physiological characteristics. The dominant fatty acids were C16 : 0, C18 : 1, C16 : 1 and tuberculostearic acid. The DNA G+C content was 67.6 mol%. Major polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside, while MK-9(H2) was the only menaquinone found. Mycolic acids of C56-C60 were present. Whole-cell hydrolysates contained meso-diaminopimelic acid along with arabinose and galactose as the major cell-wall sugars. On the basis of the results obtained in this study, the bacterium was assigned to the genus Gordonia and represents a new species with the name Gordonia metallireducens sp. nov. The type strain is TSed Te1T (=NRRL B-65678T=DSM 114093T).

Tessaracoccus caeni sp. nov., a novel member of Propionibacteriaceae isolated from activated sludge in Hefei, PR China.

A floc-forming bacterial strain, designated HF-7T, was isolated from the activated sludge of an industrial wastewater treatment plant in Hefei, PR China. Cells of this strain were Gram-stain-positive, catalase- and oxidase-negative, facultatively anaerobic, and rod-shaped. Growth occurred at 20-42 °C (optimum, 28 °C), at pH 5.5-10.5 (optimum, pH 7.5) and with 0-8.0 % (w/v) NaCl (optimum, 1 %). The major fatty acid was anteiso-C15 : 0. The polar lipid profile contained phosphatidylglycerol, diphosphatidylglycerol and phosphatidylinositol. The DNA G+C content was 67 mol% from whole genomic sequence analysis. Based on the results of 16S rRNA gene sequence analysis, this strain should be assigned to the genus Tessaracoccus and is closely related to Tessaracoccus arenae CAU 1319T (95.87 % similarity), Tessaracoccus lapidicaptus IPBSL-7T (95.19 %) and Tessaracoccus bendigoensis Ben 106T (94.63 %) but separated from them by large distances in different phylogenetic trees. Based on whole genome analysis, the orthologous average nucleotide identity and in silico DNA-DNA hybridization values against two of the closest relatives were 75.21-76.50 % and 14.2-24.4 %, respectively. The phylogenetic, genotypic, phenotypic and chemotaxonomic data demonstrated that strain HF-7T could be distinguished from its phylogenetically related species and represents a novel species within the genus Tessaracoccus, for which the name Tessaracoccus caeni sp. nov. is proposed. The type strain is HF-7T (=KCTC 49959T=CCTCC AB 2023019T).

Genomic Resource Insights into Strain WZ-12: An Antimicrobial-Producing Streptomyces Isolate from Star Anise Rhizosphere Soil

Strain WZ-12, isolated from the rhizosphere soil of star anise trees, exhibits significant antifungal efficacy against a variety of fungi, including Colletotrichum horii, the pathogenic fungus responsible for anthracnose in star anise. Based on 16S rRNA gene sequencing and phylogenetic analysis, the strain was identified as a member of the Streptomyces genus. The genome of WZ-12 comprises a linear chromosome of 9,169,400 bp and a plasmid of 378,325 bp, possessing a G + C content of 71.12%. Of note, the genome encompasses 8,172 protein-coding genes, 21 rRNA genes, 74 tRNA genes, and 89 other non-coding RNA genes. The COG, Swiss-Prot, GO, KEGG, and NR databases annotated up to 79.17%, 50.50%, 13.17%, 37.51%, and 90.43% of potential genes, respectively. Remarkably, the genome hosts 175 antimicrobial resistance genes and 33 secondary metabolite gene clusters that likely contribute to the biosynthesis of diverse bioactive compounds such as antibiotics and agents with antitumor, antioxidant, immunosuppressive, anti-inflammatory, or stress tolerance properties. These findings highlight the potential of WZ-12 as a promising source of novel bioactive molecules and suggest its significant value as a valuable resource for developing innovative antimicrobial agents.

Paenibacillus caseinilyticus sp. nov., isolated forest soil.

A milky-white-coloured, aerobic, Gram-stain-positive, rod-shaped and motile bacterial strain (GW78T) was isolated from forest soil. GW78T was catalase-positive and oxidase-negative. The strain was able to grow optimally at 37 °C and at pH 7.0 in Reasoner's 2A media. The phylogenetic and 16S rRNA gene sequence analysis of GW78T showed its affiliation with the genus Paenibacillus. The 16S rRNA gene sequence of GW78T revealed 98.3 % similarity to its nearest neighbour Paenibacillus mucilaginosus VKPM B-7519T. Its chemotaxonomic properties included MK-7 as the sole menaquinone, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylmonomethylethanolamine and phosphatidylethanolamine as major polar lipids, and anteiso-C15 : 0, C16 : 1 ω11c and anteiso-C17 : 0 as predominant fatty acids. Digital DNA-DNA hybridization and average nucleotide identity results with its closest relatives were <74.0 % and <14.0 %, respectively. Overall, 16S rRNA gene sequence comparisons, phylogenetic and genomic evidence, and phenotypic and chemotaxonomic data allow the differentiation of GW78T from other members of the genus Paenibacillus. Thus, we propose that strain GW78T represents a novel species of the genus Paenibacillus, with the name Paenibacillus caseinilyticus sp. nov. The type strain is GW78T (=KCTC 43430T=NBRC 116023T).

Arsenic stress management through arsenite and arsenate-tolerant growth-promoting bacteria in rice.

Arsenic (As) contamination is a major problem affecting soil and groundwater in India, harming agricultural crops and human health. Plant growth-promoting rhizobacteria (PGPR) have emerged as promising agents for reducing As toxicity in plants. This study aimed to isolate and characterize As-tolerant PGPR from rice field soils with varying As levels in five districts of West Bengal, India. A total of 663 bacterial isolates were obtained from the different soil samples, and 10 bacterial strains were selected based on their arsenite (As-III) and arsenate (As-V) tolerant ability and multiple PGP traits, including phosphate solubilization, production of siderophore, indole acetic acid, biofilm formation, alginate, and exopolysaccharide. These isolates were identified by 16S rRNA gene sequencing analysis as Staphylococcus sp. (4), Niallia sp. (2), Priestia sp. (1), Bacillus sp. (1), Pseudomonas sp. (1), and Citricoccus sp. (1). Among the selected bacterial strains, Priestia flexa NBRI4As1 and Pseudomonas chengduensis NBRI12As1 demonstrated significant improvement in rice growth by alleviating arsenic stress under greenhouse conditions. Both strains were also able to modulate photosynthetic pigments, soluble sugar content, proline concentration, and defense enzyme activity. Reduction in As-V accumulation inoculated with NBRI4As1 was recorded highest by 53.02% and 31.48%, while As-III by NBRI12As1 38.84% and 35.98% in the roots and shoots of rice plants, respectively. Overall, this study can lead to developing efficient As-tolerant bacterial strains-based bioinoculant application packages for arsenic stress management in rice.

Seasonal Variation of Gut Microbial Composition and Metabolism in Tibetan antelopes in Hoh Xil National Nature Reserve.

The Tibetan antelope is an endangered species suffering from poaching and habitat fragmentation. The intestinal flora and metabolites play a crucial role in the physiological homeostasis of hosts, which are influenced by various environmental factors like seasonal variation. In this particular research, our main goal was to explore the alterations in the metabolism and gut microbiota of Tibetan antelopes between the cold season (XB) and warm season (DA), using untargeted metabolomics and 16S rRNA gene-sequencing analyses. The findings indicated that Tibetan antelopes had a higher alpha-diversity of intestinal microbes during the cold season than during the warm season. Principal co-ordinate analysis revealed notable seasonal discrepancies in the function and structure of intestinal microbes in Tibetan antelopes. The relative abundance of Firmicutes was significantly increased during the cold season compared to during the warm season. Furthermore, the Tibetan antelope's primary metabolic functions of the intestinal micro-organisms were significantly higher during the cold season. The untargeted metabolomics analysis results showed a total of 532 metabolites that were significantly different between the cold season and warm season groups. These metabolites were found to be enriched in a total of 62 metabolic pathways. Among the most significant pathways of enrichment were the purine metabolism and pyrimidine metabolism. The levels of related metabolites in those pathways were remarkably higher in the warm season compared to the cold season. The comprehensive analysis of 16S rRNA and the metabolome reveals there is a significant correlation between differential microbiota and differential metabolites. Therefore, the gut microbiota changes caused by seasonal changes influenced the metabolites as well. This research reveals the function of seasonal changes in the intestinal flora and metabolites in the adaptation of Tibetan antelopes to environmental fluctuations and supplies a theoretical basis for instructing the protection management of Tibetan antelopes.

Multi-Antibiotic Resistant Citrobacter freundii in Eggs: A Silent Public Health Threat

Background: Improper handling of poultry, particularly in developing countries like Bangladesh, can lead to foodborne illnesses. Citrobacter freundii, a Gram-negative bacteria, is a silent emerging threat causing diseases and spreading antibiotic resistance. This study aimed to identify the multidrug resistant (MDR) C. freundii from poultry egg samples through biochemical and molecular characterization. Materials and Methods: A total of 50 poultry egg samples (both outer and inner) were collected from different farms located in Dhaka, Bangladesh within six months. Isolation of pathogens with selective bacteriological media such as McConkey agar, Xylose Lysine Deoxycholate agar, Salmonella-Shigella agar, and specific biochemical tests were performed to identify the Citrobacter isolates. The antibiotic sensitivity pattern of the isolates were determined by the disk diffusion method. Multidrug-resistant isolates were further identified by 16S rRNA gene sequencing and BLASTn analysis. Results: Out of 50 samples, 60% (30) were found positive for Gram negative Enterobacteriaceae. Among them, 50% of the samples were found to be contaminated with Salmonella enterica (15), 20% with E. coli (6), 6.67% with Pseudomonas spp. (2) and 23.33% with Citrobacter spp. They were highly resistant to erythromycin (83%), cefixime (82%), and amoxicillin (79%) antibiotics. On the other hand, they were sensitive against imipenem (93%) and intermediate for azithromycin (46%). Among all isolates, the most potent multidrug-resistant isolate was identified based on its 16S rRNA gene sequence, with one having the highest similarity (100%) to Citrobacter freundii strain A3.Conclusion: Poultry eggs can be a source of transmission of multidrug-resistant Citrobacter freundii from animal to human. It could be a serious health concern for developing countries since very little effort has been put into this issue.

Multi-Antibiotic Resistant Citrobacter freundii in Eggs: A Silent Public Health Threat

Background: Improper handling of poultry, particularly in developing countries like Bangladesh, can lead to foodborne illnesses. Citrobacter freundii, a Gram-negative bacteria, is a silent emerging threat causing diseases and spreading antibiotic resistance. This study aimed to identify the multidrug resistant (MDR) C. freundii from poultry egg samples through biochemical and molecular characterization. Materials and Methods: A total of 50 poultry egg samples (both outer and inner) were collected from different farms located in Dhaka, Bangladesh within six months. Isolation of pathogens with selective bacteriological media such as McConkey agar, Xylose Lysine Deoxycholate agar, Salmonella-Shigella agar, and specific biochemical tests were performed to identify the Citrobacter isolates. The antibiotic sensitivity pattern of the isolates were determined by the disk diffusion method. Multidrug-resistant isolates were further identified by 16S rRNA gene sequencing and BLASTn analysis. Results: Out of 50 samples, 60% (30) were found positive for Gram negative Enterobacteriaceae. Among them, 50% of the samples were found to be contaminated with Salmonella enterica (15), 20% with E. coli (6), 6.67% with Pseudomonas spp. (2) and 23.33% with Citrobacter spp. They were highly resistant to erythromycin (83%), cefixime (82%), and amoxicillin (79%) antibiotics. On the other hand, they were sensitive against imipenem (93%) and intermediate for azithromycin (46%). Among all isolates, the most potent multidrug-resistant isolate was identified based on its 16S rRNA gene sequence, with one having the highest similarity (100%) to Citrobacter freundii strain A3.Conclusion: Poultry eggs can be a source of transmission of multidrug-resistant Citrobacter freundii from animal to human. It could be a serious health concern for developing countries since very little effort has been put into this issue.

Association between gut microbiota and its functional metabolites with prenatal depression in women

BackgroundThe gut microbiota may affect mood through the microbiota-gut-brain axis. The purpose of this study was to examine the effect of the gut microbiota and its metabolites, such as short-chain fatty acids (SCFAs), on prenatal depression and to determine the role of 5-hydroxytryptamine (5-HT) on prenatal depression in association with the gut microbiota and its metabolites (i.e. SCFAs). MethodsEighty-six pregnant women in the third trimester were recruited. Prenatal depression was determined by a score of 10 via the Edinburgh Postpartum Depression Scale. Demographic data, stool, and blood samples were collected. The gut microbiota and its metabolites SCFAs were determined by 16S rRNA gene sequencing and liquid chromatography-mass spectrometry analysis. Plasma 5-HT was determined by gas chromatography-mass spectrometry analysis. ResultsAfter controlling relevant covariates, our results found the higher the abundance of Candidatus_Soleaferrea, the lower the risk of prenatal depression; the higher the concentration of propanoic acid, the higher risk of prenatal depression. Our results also found the lower the plasma 5-HT, the higher the risk of prenatal depression, and 5-HT was related to unclassified_c_Clostridia and NK4A214_group. However, results of this study did not support the moderating effect of plasma 5-HT on the association of Candidatus_Soleaferrea or propionic acid with prenatal depression. ConclusionsResults of this study supported that changes in certain gut microbiota, SCFAs, and plasma 5-HT during pregnancy were associated with prenatal depression. This finding provides new ideas for interventions based on diet or probiotics to regulate mood during pregnancy.

Thermobacterium salinum gen. nov., sp. nov., a novel member of the family Flavobacteriaceae isolated from a hydrothermal zone.

A Gram-stain-negative, aerobic, non-motile and rod-shaped bacterial strain, designated as strain TK19130T, was isolated from the Lonqi hydrothermal zone in the Southwest Indian Ridge. Growth occurred with 1-12 % (w/v) NaCl (optimum, 2-4 %), at 10-40 °C (optimum, 30-35 °C) and at pH 6.0-9.0 (optimum, pH 7.0-8.0). The genome of strain TK19130T was 3.15 Mb, with a DNA G+C content of 41.35 %. Based on the results of 16S rRNA gene sequence analysis, strain TK19130T was affiliated with the family Flavobacteriaceae, in which the highest similarity was 90.54 % to Aureisphaera salina A6D-50T, under the genus demarcation boundary (94.50 %). Average nucleotide identity values between strain TK19130T and adjacent strains were 67.17-72.00 %, lower than the recommended threshold of 73.98 % for genus delineation. The predominant respiratory quinone of strain TK19130T was menaquinone 6. Major polar lipids were phosphatidylethanolamine, three aminolipids and one unidentified polar lipid. Major fatty acids were detected as iso-C15 : 1 G, iso-C15 : 0 and iso-C17 : 0 3-OH. Based on the polyphasic taxonomic evidence presented above, strain TK19130T formed an independent branch representing a new species of a novel genus within the family Flavobacteriaceae, for which the name Thermobacterium salinum gen. nov., sp. nov. is proposed. The type strain is TK19130T (=CGMCC 1.18993T=JCM 35842T=MCCC M28200T).

Isolation and Phenotypic Microarray Profiling of Different Pseudomonas Strains Isolated from the Rhizosphere of Curcuma longa L.

In the present study, different Pseudomonas strains were isolated from the rhizospheric soil of Curcuma longa (turmeric) and further identified and characterized based on morphological, biochemical, and molecular characteristics through the 16S rRNA gene sequencing analysis. The identified bacterial strains belong to the Pseudomonas genus viz. Pseudomonas sp. CL10, Pseudomonas sp. CL11, and P. fluorescence CLI4. However, the isolated strains tested positive for IAA production, siderophore production, and the solubilization of tricalcium phosphate during plant growth promoting traits analysis. Further phenotype microArray (PM) technology was used to evaluate the antibiotic and chemical sensitivity of the isolated bacterial strains. The antibiotics phleomycin, oxacillin, vancomycin, novobiocin, spiramycin, and rifampicin, as well as chemicals like, 5-7 dichloro-8-hydroxy quanaldine, 5-7 dichloro-8-hydroxyquinoline, domophenbrobide, and 3-5 dimethoxy benzyl alcohol, showed resistance in all the rhizobacterial strains. However, upon further detailed study, Pseudomonas sp. CL10 exhibited resistance to thirteen antibiotics, CL11 to fourteen, and CL14 showed resistance against seventeen antibiotics and chemical classes. The results of the study indicate that some of these strains can be used as bioinoculum to enhance the plant growth and health.

Robiginitalea aurantiaca sp. nov. and Algoriphagus sediminis sp. nov., isolated from coastal sediment.

Two novel rod-shaped, Gram-stain-negative, aerobic and non-motile bacterial strains, designated M39T and C2-7T, were isolated from the coastal sediment of Xiaoshi Island, Weihai, PR China. Growth of strain M39T occurred at 15-37 °C, at pH 6.0-9.0 and in the presence of 1.0-9.0 % (w/v) NaCl. Strain C2-7T grew at 15-40 °C, at pH 6.0-8.0 and in the presence of 0.5-8.0 % (w/v) NaCl. Phylogenetic analysis based 16S rRNA gene sequences revealed that strains M39T and C2-7T belong to the phylum Bacteroidota. Based on the results of 16S rRNA gene sequence analysis, the closest relative of strain M39T was Robiginitalea marina KCTC 92035T (95.4 %), and the closest relative of strain C2-7T was Algoriphagus namhaensis DPG-3T (97.0 %). The percentage of conserved protein and average nucleotide identity values between strain M39T and some species of the genus Robiginitalea were 66.9-77.6% and 69.3-71.0 %, respectively, while those between strain C2-7T and some species of the genus Algoriphagus were 68.0-70.1% and 56.1-72.6 %, respectively. The major cellular fatty acids (>10 %) of strain M39T consisted of iso-C15 : 1 F, iso-C15 : 0 and iso-C17 : 0 3-OH, while those of strain C2-7T were iso-C15 : 0 and C16 : 1 ω7c/C16 : 1 ω6c. MK-6 was the only respiratory quinone that was compatible with the genus of strain M39T. The predominant menaquinone of strain C2-7T was MK-7. The major polar lipids of strain M39T were phosphatidylethanolamine and glycolipids, and those of strain C2-7T were phosphatidylethanolamine, one unidentified aminolipid and four unidentified lipids. The DNA G+C contents of strains M39T and C2-7T were 46.9 and 40.8 mol%, respectively. Based upon the results presented in this study, strains M39T and C2-7T represent novel species of the genera Robiginitalea and Algoriphagus, respectively, for which the names Robiginitalea aurantiaca sp. nov. and Algoriphagus sediminis sp. nov. are proposed with the type strains M39T (=MCCC 1H00498T=KCTC 92014T) and C2-7T (=MCCC 1H00414T=KCTC 92027T).

Streptomyces changanensis sp. nov. Isolated from Soil in China.

An aerobic, Gram-positive, and non-motile actinomycete, designated HL-66T, was isolated from a soil sample collected in the Meridian Valley, Shaanxi Province, China. Morphological, chemotaxonomic, and phylogenetic characteristics showed a high similarity to the genus Streptomyces. Based on 16S rRNA gene sequence analysis, the closest phylogenetic neighbour of HL-66T were Streptomyces lavendofoliae NBRC 12882T (99.17%), Streptomyces gobitricini NBRC 15419T (99.03%) and Streptomyces roseolilacinus NBRC 12815T (98.96%). Genome relatedness indexes revealed that the average nucleotide identity and digital DNA-DNA hybridization values between HL-66T and its closest phylogenomic relative (S. roseolilacinus JCM 4335T) were 88.61% and 32.10%, respectively. The cell-wall peptidoglycan contains LL-diaminopimelic acid. Predominant menaquinones are MK-9 (H6), MK-9(H4) and MK-9(H8). The major cellular fatty acids were iso-C16:0, anteiso-C15:0, iso-C16:1 H, and C16:1 ω7c. The polar lipid pattern consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylinositol, phosphatidylinositol mannosides, and an unknown phospholipid. Based on phylogenetic analyses, genome-genome distance calculation, and average nucleotide identity, strain HL-66T represents a novel species of the genus Streptomyces. Therefore, a new species Streptomyces changanensis sp. nov. is proposed with strain HL-66T (= CGMCC 22674 = JCM 35800) as the type strain.

Candidatus Kirkpatrickella diaphorinae gen. nov., sp. nov., an uncultured endosymbiont identified in a population of Diaphorina citri from Hawaii

Diaphorina citri is the hemipteran pest and vector of a devastating bacterial pathogen of citrus worldwide. In addition to the two core bacterial endosymbionts of D. citri, Candidatus Carsonella ruddii and Candidatus Profftella armatura, the genome of a novel endosymbiont and as of yet undescribed microbe was discovered in a Hawaiian D. citri population through deep sequencing of multiple D. citri populations. Found to be closely related to the genus Asaia in the family Acetobacteraceae by 16S rRNA gene sequence analysis, it forms a sister clade along with other insect-associated 16S rRNA gene sequences from uncultured bacterium found associated with Aedes koreicus and Sogatella furcifera. Multilocus sequence analysis confirmed the phylogenetic placement sister to the Asaia clade. Despite the culturable Asaia clade being the closest phylogenetic neighbour, attempts to culture this newly identified bacterial endosymbiont were unsuccessful. On the basis of these distinct genetic differences, the novel endosymbiont is proposed to be classified into a candidate genus and species ‘Candidatus Kirkpatrickella diaphorinae’. The full genome was deposited in GenBank (accession number CP107052; prokaryotic 16S rRNA OP600170).

Isolation and identification of pathogens of Morchella sextelata bacterial disease.

Morel mushroom (Morchella spp.) is a rare edible and medicinal fungus distributed worldwide. It is highly desired by the majority of consumers. Bacterial diseases have been commonly observed during artificial cultivation of Morchella sextelata. Bacterial pathogens spread rapidly and cause a wide range of infections, severely affecting the yield and quality of M. sextelata. In this study, two strains of bacterial pathogens, named M-B and M-5, were isolated, cultured, and purified from the tissues of the infected M. sextelata. Koch's postulates were used to determine the pathogenicity of bacteria affecting M. sextelata, and the pathogens were identified through morphological observation, physiological and biochemical analyses, and 16S rRNA gene sequence analysis. Subsequently, the effect of temperature on the growth of pathogenic bacteria, the inhibitory effect of the bacteria on M. sextelata on plates, and the changes in mycelial morphology of M. sextelata mycelium were analyzed when M. sextelata mycelium was double-cultured with pathogenic bacteria on plates. The results revealed that M-B was Pseudomonas chlororaphis subsp. aureofaciens and M-5 was Bacillus subtilis. Strain M-B started to multiply at 10-15°C, and strain M-5 started at 15-20°C. On the plates, the pathogenic bacteria also produced significant inhibition of M. sextelata mycelium, and the observation of mycelial morphology under the scanning electron microscopy revealed that the inhibited mycelium underwent obvious drying and crumpling, and the healthy mycelium were more plump. Thus, this study clarified the pathogens, optimal growth environment, and characteristics of M. sextelata bacterial diseases, thereby providing valuable basic data for the disease prevention and control of Morchella production.

Comparative analysis of the intestinal microbiome in Rattus norvegicus from different geographies.

Rat species Rattus norvegicus, also known as the brown street rat, is the most abundant mammal after humans in urban areas, where they co-exist with humans and domestic animals. The reservoir role of R. norvegicus of zoonotic pathogens in cities among rodent-borne diseases that could endanger the lives of humans and other mammals. Therefore, understanding the normal microbiome of R. norvegicus is crucial for understanding and preventing zoonotic pathogen transmission to humans and animals. We investigated the intestinal microbiome of free-living R. norvegicus collected from the Ruili, Nujiang, and Lianhe regions of Yunnan, China, using 16S rRNA gene sequence analysis. Proteobacteria, followed by Firmicutes, and Bacteroidetes were abundant in the intestines of R. norvegicus; however, bacterial compositions varied significantly between samples from different locations. Following a similar trend, Gammaproteobacteria, Bacilli, and Clostridia were among the top bacterial classes in most intestinal samples. The situation differed slightly for the Lianhe and Nujiang samples, although Phyla Bacteroidota and Spirochaetota were most prevalent. The Alpha diversity, Chao1, and Simpson indexes revealed microbial richness among the R. norvegicus samples. A slight variation was observed among the samples collected from Ruili, Nujiang, and Lianhe. At species levels, several opportunistic and zoonotic bacterial pathogens, including Lactococcus garvieae, Uruburuella suis, Bartonella australis, Clostridium perfringens, Streptococcus azizii, Vibrio vulnificus, etc., were revealed in the R. norvegicus intestines, implying the need for a regular survey to monitor and control rodent populations. In conclusion, we explored diverse microbial communities in R. norvegicus intestines captured from different regions. Further, we identified several opportunistic and potential bacterial pathogens, which still need to be tested for their underlying pathogenesis. The findings of our current study should be considered a warning to the health authorities to implement rat control and surveillance strategies globally.

Alteriqipengyuania flavescens sp. nov., isolated from Pearl River Estuary sediment.

Two Gram-stain-negative, aerobic, rod-shaped, non-motile and golden yellow pigmented bacteria, designated as SCSIO 75105T and SCSIO 75732, were isolated from sediment in the Pearl River Estuary, Guangdong Province, PR China. Cells were positive for catalase and oxidase. Growth occurred at 10-37 °C (optimum, 28 °C), pH 6.0-10.0 (optimum, pH 7.0) and 0-5.0 % (w/v) NaCl (optimum, 2.0-3.0 %). The 16S rRNA gene analysis indicated that these two isolates shared a similarity of 100 % each other. The 16S rRNA gene sequence analysis indicated that these two isolates showed highest similarity to Altererythrobacter ishigakiensis CGMCC 1.14979T (97.3 %). However, a phylogenetic tree based on 288 orthologous clusters indicated that these two isolates were closely related to Alteriqipengyuania halimionae CPA5T. The average nucleotide identity, average amino acid identity, digital DNA-DNA hybridization and evolutionary distance values between the two isolates and Alteriqipengyuania halimionae CPA5T were 73.7-74.0 %, 65.2 %, 19.5 % and 0.24, respectively. The genomic DNA G+C content of both isolates was 65.2 mol%. The major cellular fatty acids were C18 : 1 ω7c, C17 : 1 ω6c and summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), and Q-10 was the respiratory quinone. The polar lipid profile contained diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylcholine, sphingoglycolipid and one unidentified glycolipid. On the basis of the results of phenotypic, physiological, chemotaxonomic and genotypic characterization, strains SCSIO 75105T and SCSIO 75732 are considered to represent a novel species in the genus Alteriqipengyuania, for which the name Alteriqipengyuania flavescens sp. nov. is proposed. The type strain is SCSIO 75105T (=KCTC 92502T=MCCC 1K07993T).

Pontibacter harenae sp. nov., isolated from the soil of a Populus euphratica forest.

A Gram-negative, rod-shaped, aerobic and light pink-pigmented bacterium, designated XAAS-A31T, was isolated from the soil of a Populus euphratica forest located near Hotan City, Xinjiang, PR China. Polyphasic, taxonomic and phylogenomic analyses were used to determine the taxonomy position of the strain. Phylogenetic analysis based on 16S rRNA gene sequence analysis indicated that XAAS-A31T belongs to the genus Pontibacter, family Hymenobacteraceae, and shows highest sequence similarity to Pontibacter silvestris XAAS-R86T (96.2 %). The digital DNA-DNA hybridization (22.0 %-19.2 %) and orthologous average nucleotide identity (74.1 %-72.7 %) values relative to closest validly published Pontibacter species were lower than the recommended thresholds of 70 and 96 %, respectively. The cells grew at 4-40 °C (optimum, 28-30 °C), at pH 6.5-8.5 (pH 7.0-7.5) and with 0-8% NaCl (0.5-2.0% NaCl). The main respiratory quinone of XAAS-A31T is MK-7, and the principal cellular fatty acids are iso-C15 : 0, iso-C17 : 0 3OH and summed feature 4 (iso-C17 : 1 I and/or anteiso-C17 : 1 B). The major polar lipids are phosphatidylethanolamine, three unidentified amino-phospholipids, one unidentified glycolipid and eight unidentified lipids. The genome length of strain XAAS-A31T is 5.48 Mbp with a DNA G+C content of 44.2 mol% and 4013 protein-coding genes. Phenotypic and genotypic data suggested that XAAS-A31T represents a novel Pontibacter species, for which we propose the name Pontibacter harenae sp. nov. and type strain XAAS-A31T (=CCTCC AB 2017162T=KCTC 62049T).