The phytochemical investigation of the crude methanolic extracts roots and stem bark of Anthonotha cladantha (Harms) J.Léonard led to the isolation and identification of twelve secondary metabolites: 2,3-dihydroxypropyl hexacosanoate (1), hederagenine (2), cycloeucalenol (3), 2α-hydroxylupeol (4), betulinic acid (5), lupeol (6), heptacosan-2-one (7), triacontanoic acid (8), stigmast-4-en-3-one (9), β-sitosterol (10), stigmasterol (11), and stigmasterol-3-O-β-D-glucopyranoside (12). Their structures were elucidated with the help of their spectroscopic and physical data and by comparison with those reported in the literature. To the best of our knowledge, from all those compounds, 2,3-dihydroxypropyl hexacosanoate (1), hederagenine (2), cycloeucalenol (3), 2α-hydroxylupeol (4), and betulinic acid (5) are being reported for the first time from this genus. In addition, the acetylation of compound 1 afforded a new derivative 3-(hexacosanoyloxy)propane-1,2-diyl diacetate (1a). Compound 1 possessed a moderate α-glucosidase inhibitory activity with an IC50 value of 39.2 ± 0.22 μM; it neither showed antioxidant activity nor inhibition against the enzyme urease. Compound 1a exhibited weak antioxidant activity in the DPPH assay with an IC50 value of 80.3 ± 0.83 μM but was inactive against α-glucosidase and urease. Furthermore, both compounds 1 and 1a were inactive against seven pathogenic bacterial strains.
Hala Khyami-Horani;Amal Al-Aboudi;Musa Abu Zarga;Monther Sadder;Halima Othman
Microbiology and Biotechnology Letters
/
v.51
no.4
/
pp.474-483
/
2023
Members of the genus Streptomyces produce more than 70% of antibiotics. The rise in antibiotic resistance globally enhanced the search for novel species with the ability to produce new bioactive compounds. This study was initiated to investigate different regions in Jordan for previously uncultured and rare Streptomyces species capable of producing novel antimicrobial compounds especially active against bacteria resistant to antibiotics. A total of 191 Streptomyces strains were isolated from 26 soil samples collected from different geographic regions in Jordan. Isolates were characterized based on colony and cellular morphology as well as using 16S rRNA gene sequencing. These isolates were screened for their ability to produce antibiotics by the perpendicular-cross streak method, and then tested by well diffusion method against tested pathogens. Fifty-four isolates showed potential to produce antimicrobial products especially active against resistant bacteria, 20.1% of the isolates showed inhibitory effect against Staphylococcus aureus, 16.9% against clinical MSSA strains, and 18.0% against MRSA: whereas only 4.2% against Esherichia coli, 3.2% against Klebsiella pneumonia, 2.7% against Pseudomonas aeruginosa, and 10.0% against clinical Candida albicans. Three isolates were selected for further identification due to their antibacterial activity against S. aureus, MRSA, and MSSA. These isolates were identified as follows; Streptomyces aburaviensis DSa3, Streptomyces alboniger SAb7 and Streptomyces misionensis ZAb2, based on cultural, biochemical characteristics and molecular analysis of the 16S rRNA.
It has intended to identify the members of the Genus Lespedeza in Korea by a chemical colour reaction, and the following five species of the Genus Lespedeza grown in the garden have been used in this experiment. 1. Lespedeza bicolor Turcz 2. Lespedeza bicolor var. melanantha (Nak.) T. Lee 3. Lespedeza cyrtobotrya Miq. 4. Lespedeza japonica var. intermedia Nakai 5. Lespedeza maritima Nakai 6. Lespedeza maximowiczii Schneider 7. Lespedeza maximowiczii var. tomentella Nakai A few drops of each solution of $K_2Cr_2O_7$. $FeSO_4{\cdot}7H_2O$, $FeCl_3$, $KH_2PO_4$, $KMnO_4$, $NH_4OH$, and HCl was added to the methanol extracts of wood dust to get the specific colour reaction. HCl-infused wood was also used for the identification of L. bicolor var. melanantha and L. bicolor. The results can be summarized as the following key; 1. Chrome lemon by $K_2Cr_2O_7$${\cdots}{\cdots}$2 1. Sun flower yellow by $K_2Cr_2O_7$${\cdots}{\cdots}$Lespedeza maximowiczii var. tomentella Nakai 2. $KH_2PO_4$ Oystem white by $KH_2PO_4$; golden yellow by $FeCl_3$${\cdots}{\cdots}$=3 2. Cream colour by $KH_2PO_4$=6 3. Oyster white by $NH_4OH$; corn colour by $FeSO_4{\cdot}7H_2O$${\cdots}{\cdots}$4 3. Cream colcur by $NH_4OH$${\cdots}{\cdots}$5 4. Van dyke brown by $KMnO_4$${\cdots}{\cdots}$; sea shell pink by HCl injection under heating ${\cdots}{\cdots}$Lespedeza japonica var. intermedia Nakai 4. Sepia colour by $KMnO_4$; honey colour by HCl injection under heating ${\cdots}{\cdots}$Lespedeza maritima Nakai 5. Golden red by $FeSO_4{\cdot}7H_2O$; andover green by HCl-infused wood dust ${\cdots}{\cdots}$Lespedeza bicolor var. melanantha (Nak.) T. Lee 5. Yellow ochre by $FeSO_4{\cdot}7H_2O$; sand warm gray by HCl-infused wood dust ${\cdots}{\cdots}$Lespedeza bicolor Turcz 6. Amber green by $FeCl_3$${\cdots}{\cdots}$Lespedeza cyrtobotrya Miq. 6. Leather brown by $FeCl_3$${\cdots}{\cdots}$Lespedeza maximowiczii Schneider.
Green mold disease caused by Trichoderma species has recently caused considerable damage to oyster mushroom industries in Korea. This disease Trichoderma, Penicillium, Aspergillus, such as in (genus) to be included in a disease caused by a species that collectively the largest incidence and damage is caused by the pathogen Trichoderma genus. T. longibrachiatum, Trichoderma koningii, Trichoderma virens, T. hazianum, T. atroviride, and T. pseudokoningii were detected on oyster mushroom beds and, of them, T. virens, T. hazianum, T. longibrachiatum was the most frequently detected. The knowledge concerning physiological and ecological properties of Trichoderma spp. was essential for their effective control. T. longibrachiatum hyphal growth is very fast, spore formation, and, particularly well-chlamydospore formation characteristics, and reviews are dark green discoloration. T. koningii, fast mycelial growth, aerial hyphae and spores in aerial hyphae formation is concentrated. T. virens, especially if the color change caused by spore-forming, slow, late in infection, the more severe the damage is discovered. T. hazianum fast mycelial growth, white aerial hyphae and late turns dark green. After spore formation hyphae glob of white pustules or tufts on the top of the formation. T. atroviride. aerial hyphae usually the mycelial growth and spore formation in the unlikely event of the formation and smells similar to the smell of coconut is that. Fast T. pseudokoningii mycelial growth, spore formation is formed around the inoculation site, discoloration of the medium color and well formed chlamydospores.
The southern coast of Korea is important for the ark shell ($Scapharca$$broughtonii$) aquaculture, but the productivity was rapidly reduced during the previous decade by mass mortality. To overcome this economic loss, investigations only focused on environmental factors, and microbiological researches were performed insufficiently. In this study, two sites (Gangjin and Jinhae bay) were selected for their high and low rate of mortality, respectively, and the existence of microflora from underwater sediments in the bodies of $S.$$broughtonii$ was analyzed. We screened the whole body of each sample and chose unique colonies, which exhibit alpha- and beta-hemolytic activity, for identification. The microflora in $S.$$broughtonii$ was less variable than sediments, and restricted species were isolated. We identified 17 genera of 88 species and 16 genera of 64 species from the two bays, respectively. A major proportion was comprised of $Bacillus$ species, with the $Bacillus$$cereus$ group being the most common species among the $Bacillus$ strains, while $Paenibacillus$, $Lynsilbacillus$, and $Vibrio$ species were the second most abundant species. At the genus level, there were no significant microbial differences between the two coastal regions. 64 species were isolated from rare site (Jinhae bay), but more species (88) with greater variety were isolated from the frequent site (Gangjin bay). Therefore, it was assumed that the cause of mass mortality lay in the difference in specie-level diversity, and conducting investigations on the diagnosis of pathogenic species by challenging tests using isolated unique species.
Jang, Mun Hee;Kim, Keun-Yong;Lee, Yu Hee;Oh, Yun Kyung;Lee, Jeong-Ho;Song, Jun-Young
Journal of fish pathology
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v.33
no.2
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pp.111-118
/
2020
The genus Edwardsiella belonging to the family Enterobacteriaceae is a member of Gram-negative rod-shaped bacteria that cause disease in diverse aquatic organisms such as fish, amphibians and reptiles as well as avians and mammals including human throughout the world. This genus had been composed of three species, E. hoshinae, E. ictaluri and E. tarda, but recent researches erected two novel species, E. anguillarum and E. piscicida that were conventionally identified as E. tarda. In this study, we isolated seven strains belonging to the genus Edwardsiella from freshwater fishes that had been reared at inland fish farms in South Korea and investigated their biochemical characteristics and molecular phylogenetic relationships. The seven strains showed typical characteristics of four Edwardsiella species, E. anguillarum, E. ictaluri, E. piscicida and E. tarda, by biochemical analyses of Gram staining, indole and hydrogen sulfide (H2S) production, and API (Analytic Profile Index) 20E test. Molecular phylogenetic analyses inferred from DNA sequence data of both 16S ribosomal RNA (rRNA) and DNA gyrase subunit B (gyrB) genes were congruent with the biochemical characteristics. As a result, both biochemical and molecular phylogenetic analyses identified four strains isolated from three Anguilla species as E. anguillarum, E. piscicida and E. tarda, two strains from Pelteobagrus fulvidraco and Silurus asotus as E. ictaluri, and one strain from Moroco oxycephalus as E. piscicida. In this study, we isolated and successfully identified recently newly erected species, E. anguillarum and E. piscicida in addition to historically notorious pathogenic species, E. ictaluri and E. tarda. In the future study, systematic and comprehensive monitoring of the four Edwardsiella species are required for studying differences in pathogenicity among freshwater fishes.
Chinese cabbage ('Baechu') Kimchi was fermented at the three different temperatures right after it was prepared. Samples were taken everyday for measuring bacterial populations, pH, and titratable acidity through the whole periods of fermentation up to 50 days. pH values and developed acidity were significantly affected by the fermenting temperatures of 4, 10, and $20^{\circ}C$, suggesting that different bacterial flora has been established by the temperatures exposed. The modified MRS agar containing vancomycin (300 $\mu$g/mL) was used for isolating the vancomycin-resistant LAB strains and 127 isolates were finally obtained. Of the LAB isolates, 13 isolates were subjected to the identification experiments based on the biochemical characteristics and the molecular-typing approach, an ITS-PCR, whether they belong to the genus Leuconostoc or not. The data obtained from API 50 CHL kit resulted that six isolates were identified as the members of Leuconostoc and six as Lactobacillus brevis strains except for a single isolate YKI 30-0401, which was not able to be identified because its biochemical traits were not matched to the database of API 50 CHL kit. It was noted that some isolates were distinct in a couple of some biochemical characteristics compared with those of the reference Leuconostoc species. To overcome the limitations experienced in the commercial identification products above, an ITS-PCR experiment was also conducted for the isolates, resulting that eight isolates belong to Leu. mesenteroides ssp. mesenteroides or dextranicum with a single band of 564 bp, and four to L. brevis strains. The ITS-PCR profiles clearly differentiated the closely-related LAB isolates for which same results were obtained by the biochemical method. This molecular approach, however, failed to produce the amplicons for the YKI 20-1003, leaving the strain unidentified. Judging from the identification data obtained in the Kimchi fermented at $4^{\circ}C$ or $10^{\circ}C$, Leuconostoc spp. including Leu. mesenteroides/dextranicum were likely predominant species in the earlier stage and L. brevis occurred at the high level through the whole period. By contrast, L. brevis, as one of the major flora, possibly lead the fermentation from the beginning in the Kimchi fermented at $20^{\circ}C$.}C$.
DNA barcoding without assessing reliability and validity causes taxonomic errors of species identification, which is responsible for disruptions of their conservation and aquaculture industry. Although DNA barcoding facilitates molecular identification and phylogenetic analysis of species, its availability in clariid catfish lineage remains uncertain. In this study, DNA barcoding was developed and validated for clariid catfish. 2,970 barcode sequences from mitochondrial cytochrome c oxidase I (COI) and cytochrome b (Cytb) genes and D-loop sequences were analyzed for 37 clariid catfish species. The highest intraspecific nearest neighbor distances were 85.47%, 98.03%, and 89.10% for COI, Cytb, and D-loop sequences, respectively. This suggests that the Cytb gene is the most appropriate for identifying clariid catfish and can serve as a standard region for DNA barcoding. A positive barcoding gap between interspecific and intraspecific sequence divergence was observed in the Cytb dataset but not in the COI and D-loop datasets. Intraspecific variation was typically less than 4.4%, whereas interspecific variation was generally more than 66.9%. However, a species complex was detected in walking catfish and significant intraspecific sequence divergence was observed in North African catfish. These findings suggest the need to focus on developing a DNA barcoding system for classifying clariid catfish properly and to validate its efficacy for a wider range of clariid catfish. With an enriched database of multiple sequences from a target species and its genus, species identification can be more accurate and biodiversity assessment of the species can be facilitated.
To examine the leaf epidermal microstructure of three genera (Scopolia s.s., Anisodus, AtroPanthe, including Przewalskia as an outgroup) in the genera Scopolia Jacq. s.l., leaves of 10 species (37 specimens) were investigated by the light microscopy (LM) and scanning electron microscopy (SEM). The stomata of studied taxa were 'amphistomatic type' and the size (guard cell) range was $18-64{\times}11-48{\mu}m$. The size of stomata is slightly differed from between the taxa; the smallest size of stomata were found in the monotypic genus, Przewalskia ($24-27{\times}16-17{\mu}m$), on the other hand the largest one was found in Anisodus carniolicoides ($62-64{\times}43-48{\mu}m$). The stomatal complex was mostly anomocytic (in Scopolia s.s., Anisodus taxa : A. luridus, A. carniolicoides, A. acutangulus) and sometimes anisocytic (in Anisodus tanguticus, Przewalskia, Atropanthe). The stomata is mostly crescent in shape, but rarely circular, especially in Przewalskia tangutica. The shapes of epidermal cells are similar in both adaxial and abaxial sides, and mostly undulate/sinuate polygonal anticlinal wall, but rarely arched in Przewalskia tangutica. The epicuticular wax was not well developed in most studied taxa, except Anisodus tanguticus which is well developed cuticular striae around the stomatal complex. The elongate-headed glandular trichomes were found in Scopolia s.s. and Przewalskia. While the taxa of Anisodus and Atropanthe have not any trichomes (i. e., glabrous), except Anisodus luridus, which has simple or sometimes branched (dendritic- type) non-glandular trichome. Finally, the systematic and ecological significance of the leaf micromorphological features (stomata complex, trichome, etc.) in identification and elucidation of Scopolia s.l. including Przewalskia, especially between or within the genera including among the species is also discussed.
Taxonomic and genetic analysis of Phytophthora species belonging to six different morphological groups (GI, GII, GIII, GIV, GV, GVI) was conducted using RAPD method. Amplified fragments ranged $0.3{\sim}3.2$ kb in their molecular weights. Among total of 145 bands, there were 109 polymorphic bands. Seven isolates of P. infestans showed high similarities of $0.92{\sim}0.99$, and P. infestans isolate 3 from potato showed similarities of $0.93{\sim}0.95$ compared with other P. infestans. Among isolates of P. capsici, similarities of $0.77{\sim}0.86$ were observed and they were grouped in 80% level. P. cinnamomi and P. cryptogea isolates which belonging to group GVI showed very similar RAPD fingerprinting pattern. Primers OPA-04, OPA-17, OPA-18, OPA-19, and OPB-12 showed high level of differences among the tested isolates in major bands and molecular weights. The similarity between the isolates was 0.67. P. megasperma and P. sojae in group GV showed similarity of 0.65. These two isolates showed big differences in single major band in reactions with primers OPA-08, OPA-17, and OPA-19. Phytophthora-specific and P. infestans-specific molecular markers were also selected with one of the random primers tested. In reaction with primer OPA-20, all the genus Phytophthora showed common band at 600 bp, and all the P. infestans isolates showed specific band at 680 bp. These markers can be useful for identification of Phytophthora speices or P. infestans. As a result, P. infestans isolated from tomato and/or potato can easily be differentiated from other Phytophthora species with this primer.
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