Prophylaxis against pneumocystis pneumonia in rheumatoid arthritis patients treated with b/tsDMARDs: insights from 3787 cases in the FIRST registry.

OBJECTIVES: The use of biologic and targeted synthetic (b/ts) DMARDs in the treatment of RA is increasing. Therefore, prevention of b/tsDMARDs-induced infection is important. Here we describe a prophylaxis protocol for preventing pneumocystis pneumonia (PCP) in RA patients treated with b/tsDMARDs. METHODS: The study subjects were 3787 RA patients from the FIRST registry. They were divided into cohort 1 (n = 807, requiring prophylaxis against PCP based on physicians' assessment at the point of new treatment with or switch to b/tsDMARDs) and cohort 2 (n = 2980, receiving strategic PCP prophylaxis). The incidence and risk factors for PCP were investigated. RESULTS: Twenty-six PCP cases were observed throughout the study. After the introduction of strategic PCP prophylaxis, PCP incidence diminished from 0.51/100 person-years (PYs) to 0.21/100 PYs (risk ratio = 0.42). Sulfamethoxazole and trimethoprim in combination (SMX-TMP) showed greater efficacy in the prevention of PCP than pentamidine inhalation (P <0.0001). The prophylaxis rate increased chronologically despite the falls in the average SMX-TMP dose and in the incidence of PCP. Subanalysis of the data for 929 patients from both groups who did not receive prophylaxis showed that old age, high BMI, coexisting lung diseases, low lymphocyte count, and low serum IgG levels increased the risk of PCP development. Development of PCP could be predicted (using an equation based on these variables) in patients not treated with glucocorticoids [area under the curve (AUC) = 0.910)], but less accurately in those on glucocorticoids (AUC = 0.746). CONCLUSIONS: Our study clarified the risk factors for PCP in RA patients on b/tsDMARDs treatment and highlighted and defined the criteria for effective prophylaxis against PCP.

Sphingobium aromaticivastans sp. nov., a novel aniline- and benzene-degrading, and antimicrobial compound producing bacterium.

A strictly aerobic, orange-pigmented strain was isolated and designated as UCM-25T. This strain is capable of degrading aniline and benzene, while is also producing antimicrobial compounds which inhibit the growth of some common pathogenic microbes. A near full-length 16S rRNA gene sequence revealed similarity to Sphingobium chlorophenolicum NBRC 16172T (98.6%). The level of DNA-DNA hybridization between the new isolate and the related species suggests UCM-25T to be a new species belonging to the genus Sphingobium. The bacterial cells contained phosphatidylglycerol, phosphatidylethanolamine, sphingoglycolipid, phosphatidylcholine, phosphatidylmonomethylethanolamine, phosphatidyldimethylethanolamine, three unidentified polar lipids, and an unidentified aminophospholipid. Ubiquinone Q-10 was the major quinone and spermidine was the major polyamine. The G+C content in the DNA of strain UCM-25T was 62.9 mol%. Cells contained summed feature 8 (C18:1ω7c and/or C18:1ω6c), summed feature 3 (C16:1ω7c and/or C16:1ω6c), C16:0, and C14:0 2-OH as major fatty acids. Based on the comparison of phenotypic, genotypic, and chemotaxonomic characteristics, strain UCM-25T represents a new member of the genus Sphingobium, for which the name S. aromaticivastans sp. nov. is proposed. The type strain is UCM-25T (=KACC 19288T =DSM 105181T).

Effective Soil Extraction Method for Cultivating Previously Uncultured Soil Bacteria.

Here, a new medium, named intensive soil extract medium (ISEM), based on new soil extract (NSE) using 80% methanol, was used to efficiently isolate previously uncultured bacteria and new taxonomic candidates, which accounted for 49% and 55% of the total isolates examined (n = 258), respectively. The new isolates were affiliated with seven phyla (Proteobacteria, Acidobacteria, Firmicutes, Actinobacteria, Verrucomicrobia, Planctomycetes, and Bacteroidetes). The result of chemical analysis showed that NSE included more diverse components of low-molecular-weight organic substances than two conventional soil extracts made using distilled water. Cultivation of previously uncultured bacteria is expected to extend knowledge through the discovery of new phenotypic, physiological, and functional properties and even roles of unknown genes.IMPORTANCE Both metagenomics and single-cell sequencing can detect unknown genes from uncultured microbial strains in environments, and either method may find the significant potential metabolites and roles of these strains. However, such gene/genome-based techniques do not allow detailed investigations that are possible with cultures. To solve this problem, various approaches for cultivation of uncultured bacteria have been developed, but there are still difficulties in maintaining pure cultures by subculture.

Rhodococcus pedocola sp. nov. and Rhodococcus humicola sp. nov., two antibiotic-producing actinomycetes isolated from soil.

Two novel actinobacterial strains, UC12T and UC33T, were isolated from forest topsoil in Suwon, Gyeonggi-Do, South Korea. Comparative analysis of nearly full-length 16S rRNA gene sequences of UC12T and UC33T revealed close pairwise similarity with species of the genus Rhodococcus, and the UC12T and UC33T sequences were most closely related to Rhodococcus canchipurensis MBRL 353T (98.91 % 16S rRNA gene sequence similarity) and Rhodococcus triatomae IMMIB RIV-085T (97.71 %), respectively. DNA-DNA hybridization showed 33.05-35.60 % genomic similarity between strains UC12T and UC33T, while strain UC12T shared DNA-DNA relatedness values of 32.71-41.29 % with the closest species of the genus Rhodococcus and strain UC33T shared 29.12-37.91 % genomic relatedness with the closest species of the genus Rhodococcus. Both strains showed similar chemotaxonomic characteristics. The major fatty acids were C16 : 0, summed feature 3, C18 : 1ω9c and C18 : 0 10-methyl. The polar lipids were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol and phosphatidylinositol mannoside. The major quinone derived was MK-8(H2). The cell-wall peptidoglycan contained meso-diaminopimelic acid, and galactose, glucose, arabinose and ribose were detected in whole cells. Mycolic acids were detected. The DNA G+C content of strains UC12T and UC33T was 72.7 mol% and 68.8 mol%, respectively. Both strains produced antibiotic(s) that inhibited bacterial pathogens but not fungi. Based on the physiological, biochemical and genotypic features and the DNA-DNA hybridization between the isolates and type strains of closely related species, we propose that these bacteria be classified as novel species of the genus Rhodococcus with the names Rhodococcus pedocola sp. nov. (type strain UC12T=KACC 18499T=NBRC 111580T) and Rhodococcus humicola sp. nov. (type strain UC33T=KACC 18500T=NBRC 111581T).

Description of Variovorax humicola sp. nov., isolated from a forest topsoil.

Employing a modified cultivation method, we studied two bacterial strains, UC10 and UC38T, found on the Kyonggi University campus, Suwon in Gyeonggi-Do province, South Korea. These strains were non-spore-forming, Gram-stain-negative, motile and rod-shaped. Growth occurred in the presence of 0-2 % (w/v) NaCl, at pH 4-9 and a temperature range of 4-35 °C. On an R2A agar plate incubated for 5 days at 28 °C, irregular, raised and pale-yellowish colonies were observed. Comparative analysis of nearly full-length 16S rRNA gene sequences indicated that these strains were closely related to Variovorax guangxiensis GXGD002T, with 98.6 % similarity. Strains UC10 and UC38T were 98.0 % similar to V.ariovorax soli GH9-3T; 97.8 % to V.ariovorax dokdonensis DS-43T; 97.3-97.7 % to V.ariovorax ginsengisoli Gsoil 3165T; 97.7-98.0 % to V.ariovorax paradoxus IAM 12373T; 97.4-97.6 % to V.ariovorax defluvii 2C1-bT; and 97.3-97.4 % to V.ariovorax boronicumulans BAM-48T. The predominant ubiquinone was Q-8. The primary polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. The major fatty acids were C16 : 0, summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c), summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) and C17 : 0 cyclo. DNA-DNA hybridization assays indicated 89.2-91.4 % genomic DNA similarity between strains UC10 and UC38T. Moreover, genomic DNA similarity between these novel strains and reference strains of the genus Variovoraxwas less than the 70 %. Based on these results, strain UC38T was designated a representative of a novel species of the genus Variovorax, with the proposed name Variovorax humicola sp. nov. The type strain is UC38T (=KACC 18501T=NBRC 111520T).

Flavobacterium fulvum sp. nov., Flavobacterium pedocola sp. nov. and Flavobacterium humicola sp. nov., three new members of the family Flavobacteriaceae, isolated from soil.

Four Gram-stain-negative, non-endospore-forming, non-motile strains were found in soil, South Korea. Based on their 16S rRNA gene sequences, strains UCM-R15T and UCM-R21 are most closely related to Flavobacterium enshiense DK69T (97.4-97.5 %, pairwise similarity) while strains UCM-R36T and UCM-46T are most closely related to Flavobacterium suncheonense GH29-5T (97.5 % and 98.3 %, respectively), with all four strains sharing less than 97 % pairwise similarity to the type strain of any other species of the genus Flavobacterium. None of the four strains can reduce/digest nitrate or urea. The only menaquinone detected was MK-6 and the major fatty acids were iso-C15 : 0, iso-C17 : 0 3-OH, iso-C15 : 1 G and summed feature 9 in all the type strains. Phosphatidylethanolamine was found in three strains as the major polar lipid, phosphatidylserine was found in both strains UCM-R15T and UCM-R36T, but not UCM-46T, and phosphatidylmonomethylethanolamine only occurred in strain UCM-R15T. The genomic DNA G+C content values of strains UCM-R15T, UCM-R21, UCM-R36T and UCM-46T were 35.3-39.0 mol%. Taking into account their physiological and biochemical characteristics, we suggest that three of the strains are novel members of the genus Flavobacterium. We propose the names Flavobacterium fulvum sp. nov. for type strain UCM-R15T (=KACC 18666T=NBRC 111764T), and strain UCM-R21 as an additional strain Flavobacterium pedocola sp. nov. for type strain UCM-R36T (=KACC 18668T=NBRC 111765T), and Flavobacterium humicola sp. nov. for type strain UCM-46T (=KACC 18575T=NBRC 111657T).

Description of Novosphingobiumflavum sp. nov., isolated from soil.

A novel yellow bacterial strain, designated UCM-28T, was isolated from forest soil in Gyeonggi-Do, South Korea. The isolated strain was Gram-stain-negative, aerobic, non-spore-forming, non-motile and rod-shaped, and grew at 10-37 °C, pH 5.5-9 and with 0-1 % NaCl. It could reduce nitrate to nitrite and hydrolyse aesculin. We determined the taxonomic position of strain UCM-28T; based on the 16S rRNA gene sequence, the strain belongs to the genus Novosphingobium. The bacterium showed the highest similarity to Novosphingobiumpiscinae SLH-16T (98.9 %), Novosphingobium rhizosphaerae JM-1T (97.7 %), Novosphingobium taihuense T3-B9T (97.2 %), Novosphingobium subterraneum DSM 12447T (97.1 %), Novosphingobium aromaticivorans DSM 12444T (97.1 %) and Novosphingobium capsulatum GIFU 11526T (96.7 %). Phylogenic trees also confirmed that strain UCM-28T is most closely related to Novosphingobiumpiscinae SLH-16T and others, and is positioned within the genus Novosphingobium. The DNA relatedness of strain UCM-28T with its references was in the range of 20.9-35.2 %. The polar lipid profile revealed diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, sphingoglycolipid, phosphatidylcholine, phosphatidylmonomethylethanolamine, six unidentified polar lipids and two unknown glycolipids. The major quinone was ubiquinone Q-10, and the major polyamine was spermidine. The DNA G+C content was 63.5 mol%. The major fatty acids included (>10 %) summed feature 8 (C18 : 1ω7c and/or C18 : 1ω6c) (46.3 %), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) (24.9 %) and C14 : 0 2-OH (11.8 %). Based on the phylogenetic and phenotypic data, strain UCM-28T should be classified within the genus Novosphingobium as a representative of a novel species, named Novosphingobium flavum sp. nov. The type strain is UCM-28T (=KACC 18571T=NBRC 111647T).

Description of Streptomyces fabae sp. nov., a producer of antibiotics against microbial pathogens, isolated from soybean (Glycine max) rhizosphere soil.

An actinomycete, designated strain T66T and isolated from soybean rhizosphere soil at Gyeonggi Siheung Sorae in the Republic of Korea, has antibiotic activity against a broad range of microbial pathogens. The strain was determined to be closely related to several known species in the genus Streptomyces on the basis of 16S rRNA gene sequence data (97.73-98.07 % similarity). The strain exhibited cell-wall chemotype I and phospholipid type II. The menaquinones present were MK-9 (H6), MK-9 (H8) and MK-10 (H2). Major fatty acids were anteiso-C15 : 0, iso-C16 : 0, iso-C15 : 0, and anteiso-C17 : 0. The level of DNA-DNA relatedness between strain T66T and closely related type strains was determined to be below 40 %. Strain T66T had spiral spore chains and a rugose spore surface that is different from its closest relatives. Comparison of the genotypic and phenotypic features confirmed that strain T66T ( = KEMB 9005-219T = KACC 18226T = NBRC 110902T) should be considered as the type strain of a novel species in the genus Streptomyces, for which the name Streptomyces fabae sp. nov. is proposed.

Streptomyces olivicoloratus sp. nov., an antibiotic-producing bacterium isolated from soil.

Strain T13T, isolated from forest soil in Jeollabuk-do, South Korea, exhibited antibiotic production on yeast extract-malt extract-glucose (YMG) medium containing magnesium chloride as a trace mineral, and inhibited the growth of Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis, Paenibacillus larvae, Escherichia coli, Candida albicans and Aspergillus niger. Growth occurred at 15-45 °C, pH 4-11 and in the presence of up to 2 % (w/v) NaCl. Biochemical analyses indicated that the predominant menaquinones produced by this strain were MK-9(H6) and MK-9(H8); small amounts of MK-10(H2) and MK-10(H4) were also detected. The polar lipid profile comprised diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylcholine, and the cell-wall peptidoglycan contained ll-diaminopimelic acid, glutamic acid, alanine and glycine. Whole-cell hydrolysates contained glucose, galactose, ribose and rhamnose. The fatty-acid profile of strain T13T was made up predominantly of iso- and anteiso-branched fatty acids. Genetic analyses demonstrated that strain T13T is closely related to Streptomyces gramineus JR-43T (98.29 % 16S rRNA gene sequence similarity), S. graminisoli JR-19T (97.99 %), S. rhizophilus JR-41T (97.86 %), S. longwoodensis LMG 20096T (97.84 %), S. graminifolii JL-22T (97.79 %) and S. yaanensis Z4T (97.56 %), and DNA-DNA hybridization yielded relatedness values of 35.27-43.42 % when T13T was compared to related strains. The results of morphological, chemotaxonomic, phylogenetic and phenotypic analyses confirm that this strain represents a novel species of the genus Streptomyces, for which the name Streptomyces olivicoloratus sp. nov. is proposed. The type strain is T13T ( = KEMB 9005-210T = KACC 18227T = NBRC 110901T).

Streptomyces gilvifuscus sp. nov., an actinomycete that produces antibacterial compounds isolated from soil.

This study describes a novel actinomycete, designated T113T, which was isolated from forest soil in Pyeongchang-gun, Republic of Korea, and is an aerobic, Gram-stain-positive actinobacterium that forms flexibilis chains of smooth, elliptical or short rod-shaped spores. The results of 16S rRNA sequence analysis indicated that strain T113T exhibited high levels of similarity to previously characterized species of the genus Streptomyces (98.19­98.89 %, respectively). However, the results of phylogenetic and DNA­DNA hybridization analyses confirmed that the organism represented a novel member of the genus Streptomyces. Furthermore, using chemotaxonomic and phenotypic analyses it was demonstrated that the strain exhibited characteristics similar to those of other members of the genus Streptomyces. The primary cellular fatty acids expressed by this strain included anteiso-C15 : 0, anteiso-C17 : 0, iso-C15 : 0 and iso-C16 : 0. While diphosphatidylglycerol and phosphatidylethanolamine were the predominant lipids expressed by strain T113T, moderate amounts of phosphatidylinositol and phosphatidylinositol mannoside were also detected. Whole-cell hydrolysates contained glucose and ribose, and the predominant menaquinone detected was MK-9 (H6); however, moderate amounts of MK-9 (H8) and trace amounts of MK-10 (H2) and MK-10 (H4) were also detected. We therefore propose that strain T113T be considered as representing a novel species of the genus Streptomyces and propose the name Streptomyces gilvifuscus sp. nov. for this species, with strain T113T ( = KEMB 9005-213T = KACC 18248T = NBRC 110904T) being the type strain.

Antifungal and antibacterial activities of Streptomyces polymachus sp. nov. isolated from soil.

Strain T258T was isolated from forest soil at Bongnae Falls, South Korea. The strain exhibited antimicrobial and antifungal activity against the following strains: Bacillus subtilis, Staphylococcus aureus, Pseudomonas aeruginosa, Paenibacillus larvae, Escherichia coli, Candida albicans and Aspergillus niger. Growth occurred on all ISP media tested (2, 3, 4, 5, 6 and 7), Czapek-Dox agar, potato dextrose agar, trypticase soy agar, Bennett's modified agar and nutrient agar at 28 °C. Aerial spores were produced solely on ISP Medium 4; the colour of the aerial mycelium was white and the substrate mycelium was ivory. Melanin production was negative on peptone-yeast extract iron agar (ISP Medium 6). The cell-wall peptidoglycan contained ll-diaminopimelic acid, glutamic acid, alanine and glycine. Whole-cell hydrolysates contained glucose, ribose and galactose. The predominant menaquinones were MK-9(H6) and MK-9(H8) while the minor menaquinone was MK-10(H2). The polar lipids included diphosphatidylglycerol, phosphatidylethanolamine and phosphatidylinositol. The major fatty acids (>10%) were C16 : 0 (29.8%), summed feature 3 (C16 : 1ω7c and/or C16 : 1ω6c) (15.1%), anteiso-C15 : 0 (13.5%) and iso-C15 : 0 (10.3%). DNA-DNA similarity with other strains ranged between 37.84 ± 1.15% and 50.25 ± 1.91 %. On the basis of these data, we suggest that strain T258T represents a novel species that belong to the genus Streptomyces, for which we propose a name Streptomyces polymachus sp. nov. The type strain is T258T ( = KACC 18247T = KEMB 9005-212T = NBRC 110905T).

Bacillus polymachus sp. nov., with a broad range of antibacterial activity, isolated from forest topsoil samples by using a modified culture method.

A new, modified culture method that utilizes a transwell plate with a 0.4 µm pore-size microporous membrane was developed. This system allows only trace nutrients from the soil into the liquid culture through the microporous membrane. The method is a more powerful tool for the discovery of novel species from soils than are traditional methods. Such newly identified species could potentially produce useful metabolites. A bacterial strain, T515(T), was isolated using this modified culture method. Growth of strain T515(T) occurred at pH 4-9 in a temperature range between 20 °C and 40 °C and in the presence of 0-2 % (w/v) NaCl on R2A agar. Colonies on the agar plates were tiny, white, and convex after 5 days incubation at 28 °C. Comparative analysis of the nearly full-length 16S rRNA gene sequence of strain T515(T) revealed close pairwise similarity with species of the genus Bacillus, and strain T515(T) was most closely related to Bacillus panaciterrae Gsoil 1517(T) (96.7 %) and Bacillus funiculus NAF001(T) (96.0 %). The major quinone of strain T515(T) was menaquinone-7 (MK-7) and the major fatty acids were iso-C15 : 0 (45.5 %), anteiso-C15 : 0 (23.2 %) and C16 : 0 (10.9 %). The predominant polar lipids were diphosphatidylglycerol, phosphatidylglycerol and phosphatidylethanolamine. Strain T515(T) was sensitive to streptomycin and tetracycline, but resistant to rifampicin (0.125 µg ml(-1)), ampicillin (0.5 µg ml(-1)) and chloramphenicol (1 µg ml(-1)). The strain showed antimicrobial activities against the six strains tested: Bacillus subtilis KEMB 51201-001, Staphylococcus aureus KEMB 4659, Pseudomonas aeruginosa KACC 10185, Staphylococcus epidermidis KACC 13234, Paenibacillus larvae KACC 14031 and Escherichia coli KEMB 212-234. Based on these results, strain T515(T) represents a novel species of the genus Bacillus with the proposed name, Bacillus polymachus sp. nov. The type strain is T515(T) ( = KEMB 9005-168(T) = KACC 18242(T) = NBRC 110614(T)).

Mesorhizobium soli sp. nov., a novel species isolated from the rhizosphere of Robinia pseudoacacia L. in South Korea by using a modified culture method.

Strain NHI-8(T) was isolated from a forest soil sample, collected in South Korea, by using a modified culture method. Comparative analysis of its nearly full-length 16S rRNA gene sequence showed that strain NHI-8(T) belongs to the genus Mesorhizobium and to be closely related to Mesorhizobium chacoense PR5(T) (97.32 %). The levels of DNA-DNA relatedness between strain NHI-8(T) and reference type strains of the genus Mesorhizobium were 32.28-53.65 %. SDS-PAGE of total soluble proteins and the sequences of the housekeeping genes recA, glnII, and atpD were also used to support the clade grouping in rhizobia. The new strain contained summed feature 8 (57.0 %), cyclo-C19:0ω8c (17.3 %), and C18:0 (11.0 %) as the major fatty acids, as in genus Mesorhizobium. The strain contained cardiolipin, phosphatidylglycerol, ornithine-containing lipid, phosphatidylethanolamine, phosphatidyl-N-dimethylethanolamine, and phosphatidylcholine. Morphological and physiological analyses were performed to compare the characteristics of our strain with those of the reference type strains. Based on the results, strain NHI-8(T) was determined to represent a novel member of the genus Mesorhizobium, and the name Mesorhizobium soli is proposed. The type strain is NHI-8(T) (=KEMB 9005-153(T) = KACC 17916(T) = JCM 19897(T)).

Streptomyces bambusae sp. nov., Showing Antifungal and Antibacterial Activities, Isolated from Bamboo (Bambuseae) Rhizosphere Soil Using a Modified Culture Method.

Strain T110(T) was isolated from a bamboo rhizosphere soil sample in the Republic of Korea and was found to produce antibiotics and secondary metabolites against a broad range of bacterial and fungal pathogens. It is a gram-positive actinobacterium with a straight and smooth, spore chain morphology. Morphological, physiological, and biochemical characterization suggest that T110(T) belongs to the genus Streptomyces. The predominant menaquinones of strain T110(T) were MK-9 (H6), MK-9 (H8), and MK-10 (H4). The cell wall peptidoglycan contained L L-diaminopimelic acid, glutamic acid, alanine, and glycine. Ribose and glucose were detected as whole-cell hydrolysates. The polar lipids consisted of diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol, and phosphatidylinositol. The main fatty acids were anteiso-C(15:0), anteiso-C(17:0), C(16:0), and iso-C(16:0). Sequence analysis of the 16S rRNA gene (GenBank accession no. KM229361) combined with multiple alignment tools revealed that T110(T) shared the highest degree of similarity with Streptomyces albosporeus subsp. labilomyceticus NBRC 15387(T) (97.9%). However, DNA-DNA hybridization and phylogenetic analysis indicate that strain T110(T) is distinct from its most closely related species. Therefore, we conclude that strain T110(T) is a novel species of the genus Streptomyces and propose naming it Streptomyces bambusae. The type strain is T110(T) (=KEMB 9005-214(T) = KACC 18225(T) = NBRC 110903(T)).

First study on capsular serotypes and virulence factors of Pasteurella multocida isolates from Phan Rang sheep in Vietnam.

Background and Aim: Pasteurella multocida is considered as a main factor mediating pneumonic pasteurellosis in ruminants, including sheep. It is also a current threat to Phan Rang sheep in Vietnam. This study aimed to characterize P. multocida isolated from Phan Rang sheep, their antibiotic resistance profile, and the prevalence of some virulence-associated genes of these strains. Materials and Methods: Bacteria were isolated on brain heart infusion, 10% sheep blood agar plates, and screened by biochemical tests. The polymerase chain reaction technique was used with specific primers to identify P. multocida, the presence of virulence-associated genes, and serotypes of isolates. Antimicrobial susceptibility and biofilm formation of isolates were examined using the disk diffusion method and crystal violet-based method, respectively. Results: A total of 41 P. multocida strains were isolated from 485 samples from clinically sick and healthy sheep. Of the isolates, 58.53% were serotype A, 9.75% were serotype B, and 31.71% were serotype D. Healthy animals were infected with serotype D only. All 15 virulence genes were identified in all strains isolated from clinically sick sheep, while strains isolated from healthy sheep carried 11/15 virulence genes tested. Among virulence-associated genes exbB, exbD, tonB, ompA, oma87, fimA, hgbA, and nanB were detected in over 90% of isolates, whereas hgbB, nanH, tbpA and pfhA were less frequent. Interestingly, pmHAS and tadD were highly prevalent in capsular type A strains, whereas the toxA gene was detected in capsular type D strains only. All of the isolated strains were fully susceptible to enrofloxacin, ciprofloxacin, neomycin, and ofloxacin. About 92.68% were susceptible to chloramphenicol and 90.24% to amikacin, but there was high resistance to erythromycin, tetracycline, and amoxicillin. Our results reveal that 53.65% of 41 isolates could produce biofilm, whereas 46.34% could not. Conclusion: Pasteurella multocida from Phan Rang sheep possess many virulence genes and resistance to several common antibiotics such as erythromycin, tetracycline, and amoxicillin. The results are an important warning regarding antibiotic resistance of P. multocida.

Antioxidant and Antimicrobial Properties of Fruiting Body and Submerged Mycelium of Medicinal Mushroom Phellinus robiniae (Agaricomycetes).

This study was conducted to evaluate extraction yield, antioxidant content, antioxidant capacity and antibacterial activity of extracts obtained from submerged mycelium (ME) and fruiting body (FBE) of Phellinus robiniae NTH-PR1. The results showed that yields of ME and FBE reached 14.84 ± 0.63 and 18.89 ± 0.86%, respectively. TPSC, TPC, and TFC were present in both mycelium and fruiting body, and the more contents of them were found in fruiting body. The concentrations of TPSC, TPC and TFC in ME and FBE were 17.61 ± 0.67 and 21.56 ± 0.89 mg GE g-1, 9.31 ± 0.45 and 12.14 ± 0.56 mg QAE g-1, and 8.91 ± 0.53 and 9.04 ± 0.74 mg QE g-1, respectively. EC50 values for DPPH radical scavenging revealed FBE (260.62 ± 3.33 µg mL-1) was more effective than ME (298.21 ± 3.61 µg mL-1). EC50 values for ferrous ion chelating in ME and FBE were 411.87 ± 7.27 and 432.39 ± 2.23 µg mL-1, respectively. Thus, both extracts were able to inhibit Gram-positive and Gram-negative pathogenic bacterial strains, at concentrations ranging in 25-100 mg mL-1 of ME and 18.75-75 mg mL-1 of FBE for Gram-positive bacteria; ranging in 75-100 mg mL-1 of ME and 50-75 of FBE for Gram-negative bacteria. Overall submerged mycelial biomass and fruiting bodies of Ph. robiniae NTH-PR1 can be considered as useful natural sources for development of functional food, pharmaceuticals and cosmetic products or cosmeceuticals.

Risk factors for cannula-associated arterial thrombosis following extracorporeal membrane oxygenation support: a retrospective study.

BACKGROUND: Hemostatic dysfunction during extracorporeal membrane oxygenation (ECMO) due to blood-circuit interaction and the consequences of shear stress imposed by flow rates lead to rapid coagulation cascade and thrombus formation in the ECMO system and blood vessels. We aimed to identify the incidence and risk factors for cannula-associated arterial thrombosis (CaAT) post-decannulation. METHODS: A retrospective study of patients undergoing arterial cannula removal following ECMO was performed. We evaluated the incidence of CaAT and compared the characteristics, ECMO machine parameters, cannula sizes, number of blood products transfused during ECMO, and daily hemostasis parameters in patients with and without CaAT. Multivariate analysis identified the risk factors for CaAT. RESULTS: Forty-seven patients requiring venoarterial ECMO (VA-ECMO) or hybrid methods were recruited for thrombosis screening. The median Sequential Organ Failure Assessment score was 11 (interquartile range, 8-13). CaAT occurred in 29 patients (61.7%), with thrombosis in the superficial femoral artery accounting for 51.7% of cases. The rate of limb ischemia complications in the CaAT group was 17.2%. Multivariate analysis determined that the ECMO flow rate-body surface area (BSA) ratio (100 ml/min/m2) was an independent factor for CaAT, with an odds ratio of 0.79 (95% confidence interval, 0.66-0.95; P=0.014). CONCLUSIONS: We found that the incidence of CaAT was 61.7% following successful decannulation from VA-ECMO or hybrid modes, and the ECMO flow rate-BSA ratio was an independent risk factor for CaAT. We suggest screening for arterial thrombosis following VA-ECMO, and further research is needed to determine the risks and benefits of such screening.

The effect of Non-Motor symptoms on Health-Related quality of life in patients with young onset Parkinson's Disease: A single center Vietnamese Cross-Sectional study.

BACKGROUND: Young onset Parkinson's disease (YOPD) is a distinct entity from typical late onset Parkinson's disease (LOPD). The influene of non-motor features on the health - related quality of life (HRQoL) in LOPD has been previously reported, but little is known about the impact of non-motor features in YOPD. OBJECTIVE: The aim of this study was to explore the relationship between non-motor burden and HRQoL in patients with YOPD. METHODS: This was an observational, cross-sectional study in patients with a PD, whose age at disease onset ranged from 21 to 40 years (YOPD). Participants were assessed with the MDS Unified Parkinson's Disease Rating Scale (MDS-UPDRS), Non-Motor Symptoms Scale (NMSS) and the 39-item Parkinson's Disease Questionnaire (PDQ-39; range 0-100). Spearman's rank test was used to identify correlations between NMSS domains and several dimension of HRQoL. Stepwise multiple linear regression analysis was performed to identify the independent predictors of HRQoL as measured by PDQ-39 summary index. RESULTS: 89 patients with YOPD mean (SD) age = 42.15 (5.84) participated. Patients reported 10.17 (4.74) non-motor symptoms, the most common (75%) and severe (median = 3) of which was was fatigue (IQR = 7). The most frequently reported and severely affected NMSS domain was sleep/fatigue (89.9%, median = 8; IQR = 13) followed by mood/cognition (83.1%, median = 6; IQR = 18) and attention/memory (82%, median = 5; IQR = 8). The mean (SD) summary index of PDQ-39 was 32.89 (16.8). The means (SD) of each PDQ-39 dimensions were: mobility 37.33 (21.96), ADL 42.93 (25.33), emotional well-being 39.77 (25.47), stigma 38.19 (28.44), social support 19.03 (22.89), cognition 29.59 (20.63), communication 26.96 (23.57), and bodily discomfort 29.96 (23.19). With the exception of gastrointestinal tract and sexual function, all other NMSS domain scores were correlated with the PDQ-39 summary index. The multivariate model revealed that three NMSS domains including sleep/fatigue, mood/cognition and attention/memory accompanied with UPDRS part III were independent predictors of HRQoL as measured by PDQ-39SI. CONCLUSIONS: Non-motor symptoms pertaining to sleep disturbances/fatigue, mood/cognition and attention/memory negatively impact HRQoL in patients with YOPD.

Monitoring Unfractionated Heparin in Adult Patients Undergoing Extracorporeal Membrane Oxygenation (ECMO): ACT, APTT, or ANTI-XA?

BACKGROUND: During ECMO, anticoagulants, in particular, unfractionated heparin (UFH), are commonly used and monitored by laboratory tests, including ACT, APTT, and anti-Xa level. METHOD: A single-center retrospective observational study was conducted on adult patients undergoing ECMO between January 2019 and January 2020 at a tertiary hospital. The correlations between ACT, APTT, anti-Xa, antithrombin, and UFH dose were assessed. RESULTS: 129 sets of measurements from 37 patients were obtained including ACT, APTT, anti-Xa, antithrombin, and UFH dose measured simultaneously. 102 out of 129 sets of values were interpreted as antithrombin deficiencies. The correlation coefficient between APTT and anti-Xa; ACT and anti-Xa are 0.72 and 0.33, respectively, p < 0.001. The patients with normal antithrombin levels exhibited a significant correlation between APTT and anti-Xa (r = 0.80, p < 0.001). ACT, on the other hand, was poorly correlated with UFH dose, whether there is AT deficiency or not. Anti-Xa and APTT are only moderately correlated with UFH dose in the group without antithrombin deficiency, with correlation coefficients of 0.62 and 0.57, respectively, p < 0.05. CONCLUSION: APTT value is strongly correlated with anti-Xa value, particularly in patients with normal antithrombin levels. However, the ACT value was poorly correlated with anti-Xa and not with the UFH dose. In groups without antithrombin deficiency, APTT and anti-Xa values only moderately correlated with UFH dose.

The genome insights of Streptomyces lannensis T1317-0309 reveals actinomycin D production.

The members of Streptomyces have been identified as a major source of antimicrobial agents with broad spectrum. This study is mainly focused on bioactivity-guided isolation and characterization of bioactive molecule from strain Streptomyces sp. T1317-0309 and its whole-genome sequence analysis for possible isolation of novel natural products. Strain Streptomyces sp. T1317-0309 showed 100% sequence similarity with strain Streptomyces lannensis TA4-8T consisting 10, 453,255 bp of genome with 5 scaffolds and 69.9 mol% G + C content. The genome analyses revealed a total of 17 putative biosynthetic gene clusters (BGCs) responsible for various secondary metabolites including actinomycin, bacteriocin, ectoine, melanin, terpene, siderophore, betalactone, NRPS, T2PKS, and T3PKS. The BGC and bioactivity-guided purification of ethyl acetate extract of strain T1317-0309 showed the great potency of antimicrobial activities against various gram-positive multi-drug resistant human pathogens including MRSA. The BGC-predicted bioactive secondary metabolite was identified by various NMR analyses and confirmed as actinomycin D. In addition, this study reveals the first genome study of Streptomyces lannensis as a novel source for actinomycin D.