RESUMEN
The Strep Easy Kit, a bio-enrichment dual ICG-strip test, is a diagnostic tool designed for the detection of Streptococcus agalactiae, an important pathogenic bacterium in tilapia farming. The kit can simultaneously identify two different serotypes of S. agalactiae, serotype Ia and serotype III. This capability is crucial for the collection of valuable epidemiological data and facilitates strategic planning for effective vaccine development and deployment. The Strep Easy Kit consists of two main steps: pathogen enrichment and pathogen detection. The enrichment step increases the concentration of bacteria so that the bacterial load is raised to the level reliably detectable by the subsequent ICG strip test. This is achieved by incubating the fish samples in a suitable liquid medium under specified temperature and time conditions. The second step involves the use of the dual-ICG strip test. This strip test consists of two monoclonal antibodies and one polyclonal antibody that are specific to S. agalactiae and can distinguish between S. agalactiae serotype Ia and S. agalactiae serotype III. This dual capability enables the ICG strip test to simultaneously detect both serotypes of S. agalactiae in a single test kit. The detection limit of the test kit, which consists of a dual ICG-Strip test combined with an enrichment step, is 100 CFU/mL. The kit can be used to detect S. agalactiae in both live and dead fish samples, making it versatile for various testing scenarios. The test results obtained using the Strep Easy Kit have shown a 94.4% correlation with the standard method (Thai Agricultural Standard; TAS 10453-2010), with 90.2% sensitivity and 100% specificity. Significant advantages of the Strep Easy Kit lie in its simplicity and portability, allowing farmers to perform the test by themselves and on-site. This makes it a practical and accessible tool for the tilapia farming industry.
RESUMEN
Acidovorax citrulli, a seedborne bacterium and quarantine pest, causes the devastating bacterial fruit blotch disease in cucurbit plants. Immunological assays such as ELISA are widely used in routine field inspections for this bacterium. However, to the best of our knowledge, none of the currently available monoclonal antibodies (MAbs) can detect all common A. citrulli strains. We therefore aimed to produce a panel of MAbs and to develop an ELISA-based method capable of detecting all A. citrulli strains. We used a high-throughput bead array technique to screen and characterize A. citrulli-specific MAbs produced from hybridoma clones. The hybridoma library was simultaneously screened against five A. citrulli strains (PSA, KK9, SQA, SQB and P) and the closely related bacterium, Delftia acidovorans. Three MAbs exhibiting different binding patterns to A. citrulli were used to develop an ELISA-based method called "double antibody pairs sandwich ELISA" (DAPS-ELISA). DAPS-ELISA employing mixtures of MAbs was able to specifically detect all 16 A. citrulli strains tested without cross-reactivity with other bacteria. By contrast, our previously developed MAb capture-sandwich ELISA (MC-sELISA) and a commercial test kit detected only 15 and 14 of 16 strains, respectively. The sensitivity of the DAPS-ELISA ranged from 5×105 to 1×106 CFU/mL, while those of the MC-sELISA and the commercial test kit ranged from 5×104 to 1×107 CFU/mL and 5×104 to 5×105 CFU/mL, respectively. DAPS-ELISA thus represents an alternative method enabling rapid, accurate, and inexpensive detection of all A. citrulli strains. The method can be applied to seed testing prior to planting as well as to routine field inspections.
Asunto(s)
Anticuerpos Antibacterianos/inmunología , Anticuerpos Monoclonales/inmunología , Comamonadaceae/inmunología , Ensayo de Inmunoadsorción Enzimática/métodos , Serogrupo , Hibridomas , Límite de DetecciónRESUMEN
Nonomuraea rhodomycinica NR4-ASC07T is a rare actinomycete isolated from soil in Sirindhorn peat swamp forest. The crude extract of its culture broth exhibited antimicrobial and anticancer against diverse human pathogens and cancer cells. The chemical investigation of the crude extract led to the isolation of two new metabolites named nonomuric acid (1) and 3-hydroxy deoxydaunorubicinol aglycone (2), along with two known bioactive compounds [ε-rhodomycinone (3) and 7-deoxy-13-dihydrocarminomycinone (4)]. Compounds 1 and 3 showed antimalarial activity with the half maximal inhibitory concentration (IC50) of 8.00 and 8.88 µg mL-1, respectively. Compound 4 inhibited growth of Mycobacterium tuberculosis and Bacillus cereus at the minimum inhibitory concentrations of 50.0 and 12.50 µg mL-1, respectively. Every compound exhibited cytotoxicity against cancer cells tested at IC50 ≥ 6.34 µg mL-1. These finding are the first report of bioactive metabolites produced by strain NR4-ASC07T, suggesting that rare actinomycetes are yet promising sources for novel drug discovery.
Asunto(s)
Actinobacteria/metabolismo , Antiinfecciosos/aislamiento & purificación , Microbiología del Suelo , Antiinfecciosos/química , Antiinfecciosos/farmacología , Humanos , Pruebas de Sensibilidad MicrobianaRESUMEN
In a continuation of our research into antitubercular lanostane triterpenoids from submerged cultures of Ganoderma species, three strains, Ganoderma orbiforme BCC 22325, Ganoderma sp. BCC 60695, and Ganoderma australe BCC 22314, have been investigated. Fourteen new lanostane triterpenoids, together with 35 known compounds, were isolated. Antitubercular activities of these mycelium-associated Ganoderma lanostanoids against Mycobacterium tuberculosis H37Ra were evaluated. Taken together with the assay data of previously isolated compounds, structure-activity relationships of the antitubercular activity are proposed. Most importantly, 3ß- and 15α-acetoxy groups were shown to be critical for antimycobacterial activity. The most potent compound was (24E)-3ß,15α-diacetoxylanosta-7,9(11),24-trien-26-oic acid (35).
Asunto(s)
Antituberculosos/aislamiento & purificación , Antituberculosos/farmacología , Cuerpos Fructíferos de los Hongos/química , Ganoderma/aislamiento & purificación , Lanosterol/análogos & derivados , Micelio/química , Mycobacterium tuberculosis/química , Antituberculosos/química , Ganoderma/química , Lanosterol/química , Lanosterol/aislamiento & purificación , Lanosterol/farmacología , Estructura Molecular , Relación Estructura-ActividadRESUMEN
The terrestrial actinomycete strain BCC71188 was identified as Streptomyces by its morphology (having spiral chain spore on the aerial mycelium), chemotaxonomy (containing LL-diaminopimelic acid in the cell wall), and 16S rRNA gene sequence analysis [showing high similarity values compared with Streptomyces samsunensis M1463T (99.85 %) and Streptomyces malaysiensis NBRC 16446T (99.40 %)]. The crude extract exhibited antimalarial against Plasmodium falciparum (IC50 0.19 µg/ml), anti-TB against Mycobacterial tuberculosis (MIC 6.25 µg/ml), and antibacterial against Bacillus cereus (MIC 1.56 µg/ml) activities. Therefore, chemical investigation was conducted by employing bioassay-guided method and led to the isolation of 19 compounds including two cyclic peptides (1-2), five macrolides (3-7), new naphthoquinone (8), nahuoic acid C (9), geldanamycin derivatives (10-13), cyclooctatin (14), germicidins A (15) and C (16), actinoramide A (17), abierixin, and 29-O-methylabierixin. These isolated compounds were evaluated for antimicrobial activity, such as antimalarial, anti-TB, and antibacterial activities, and for cytotoxicity against both cancerous (MCF-7, KB, NCI-H187) and non-cancerous (Vero) cells. Compounds 1-7, 10-14 exhibited antimalarial (IC50 0.22-7.14 µg/ml), and elaiophylin analogs (4-6) displayed anti-TB (MIC 0.78-12.00 µg/ml) and B. cereus (MIC 0.78-3.13 µg/ml) activities. Compounds 1, 2, 14, and abierixin displayed weak cytotoxicity, indicating a potential for antimicrobial agents.
Asunto(s)
Antiinfecciosos/metabolismo , Productos Biológicos/metabolismo , Microbiología del Suelo , Streptomyces/metabolismo , Animales , Antiinfecciosos/química , Antiinfecciosos/aislamiento & purificación , Bacillus cereus/efectos de los fármacos , Productos Biológicos/química , Productos Biológicos/aislamiento & purificación , Línea Celular , Supervivencia Celular/efectos de los fármacos , Pared Celular/química , Citosol/química , ADN Bacteriano/química , ADN Bacteriano/genética , ADN Ribosómico/química , ADN Ribosómico/genética , Ácido Diaminopimélico/análisis , Humanos , Concentración 50 Inhibidora , Pruebas de Sensibilidad Microbiana , Microscopía , Mycobacterium tuberculosis/efectos de los fármacos , Plasmodium falciparum/efectos de los fármacos , Quinonas/análisis , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Streptomyces/clasificación , Streptomyces/genética , Streptomyces/aislamiento & purificaciónRESUMEN
Sixteen new lanostane triterpenoids (1-16), together with 26 known compounds (17-42), were isolated from cultures of the basidiomycete Ganoderma sp. BCC 16642. Antitubercular activities of these Ganoderma lanostanoids against Mycobacterium tuberculosis H37Ra were evaluated, and structure-activity relationships are proposed.
Asunto(s)
Antituberculosos/aislamiento & purificación , Antituberculosos/farmacología , Ganoderma/química , Lanosterol/análogos & derivados , Triterpenos/aislamiento & purificación , Triterpenos/farmacología , Antituberculosos/química , Cuerpos Fructíferos de los Hongos/química , Lanosterol/química , Lanosterol/aislamiento & purificación , Lanosterol/farmacología , Estructura Molecular , Mycobacterium tuberculosis/efectos de los fármacos , Resonancia Magnética Nuclear Biomolecular , Relación Estructura-Actividad , Tailandia , Triterpenos/químicaRESUMEN
The prostanoid pathway converts polyunsaturated fatty acids (PUFAs) into bioactive lipid mediators, including prostaglandins, thromboxanes and prostacyclins, all of which play vital roles in the immune and reproductive systems in most animal phyla. In crustaceans, PUFAs and prostaglandins have been detected and often associated with female reproductive maturation. However, the presence of prostanoid biosynthesis genes remained in question in these species. In this study, we outlined the prostanoid pathway in the black tiger shrimp Penaeus monodon based on the amplification of nine prostanoid biosynthesis genes: cytosolic phospholipase A2, hematopoietic prostaglandin D synthase, glutathione-dependent prostaglandin D synthase, prostaglandin E synthase 1, prostaglandin E synthase 2, prostaglandin E synthase 3, prostaglandin F synthase, thromboxane A synthase and cyclooxygenase. TBLASTX analysis confirmed the identities of these genes with 51-99% sequence identities to their closest homologs. In addition, prostaglandin F2α (PGF2α), which is a product of the prostaglandin F synthase enzyme, was detected for the first time in P. monodon ovaries along with the previously identified PUFAs and prostaglandin E2 (PGE2) using RP-HPLC and mass-spectrometry. The prostaglandin synthase activity was also observed in shrimp ovary homogenates using in vitro activity assay. When prostaglandin biosynthesis was examined in different stages of shrimp ovaries, we found that the amounts of prostaglandin F synthase gene transcripts and PGF2α decreased as the ovaries matured. These findings not only indicate the presence of a functional prostanoid pathway in penaeid shrimp, but also suggest a possible role of the PGF2α biosynthesis in shrimp ovarian development.
Asunto(s)
Proteínas de Artrópodos/genética , Vías Biosintéticas/genética , Enzimas/genética , Ovario/metabolismo , Penaeidae/metabolismo , Prostaglandinas/biosíntesis , Secuencia de Aminoácidos , Animales , Proteínas de Artrópodos/metabolismo , Cromatografía Líquida de Alta Presión , Cromatografía de Fase Inversa , Dinoprost/metabolismo , Dinoprostona/metabolismo , Enzimas/clasificación , Enzimas/metabolismo , Femenino , Regulación del Desarrollo de la Expresión Génica , Regulación Enzimológica de la Expresión Génica , Oxidorreductasas Intramoleculares/clasificación , Oxidorreductasas Intramoleculares/genética , Oxidorreductasas Intramoleculares/metabolismo , Espectrometría de Masas , Datos de Secuencia Molecular , Ovario/crecimiento & desarrollo , Penaeidae/crecimiento & desarrollo , Filogenia , Prostaglandina-E Sintasas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Homología de Secuencia de AminoácidoRESUMEN
Investigation of the chemical constituents of the root bark of Artocarpus rigidus BLUME subsp. rigidus has led to the isolation of six, structurally diverse phenolic compounds. These included two new compounds with modified skeletons, the flavonoid 7-demethylartonol E (1) and the chromone artorigidusin (2), together with four known phenolic compounds, the xanthone artonol B (3), the flavonoid artonin F (4), the flavonoid cycloartobiloxanthone (5), and the xanthone artoindonesianin C (6). Compounds 1, 4, and 5 exhibited antiplasmodial activity against Plasmodium falciparum. All compounds showed antimycobacterial activity against Mycobacterium tuberculosis, with 4 being the most active compound (MIC 6.25 microg/ml). Compounds 5 and 6 were active against KB cells, whereas 2, 5, and 6 showed varying toxicity to BC cells. Compounds 1-3, 5, and 6 were active in the NCI-H187 cytotoxicity assay, with 3 being the most active compound (IC(50) 1.26 microg/ml).
Asunto(s)
Artocarpus/química , Cromonas/química , Flavonoides/química , Fenoles/química , Corteza de la Planta/química , Raíces de Plantas/química , Xantonas/química , Animales , Línea Celular Tumoral , Proliferación Celular/efectos de los fármacos , Cromonas/aislamiento & purificación , Cromonas/farmacología , Ensayos de Selección de Medicamentos Antitumorales , Flavonoides/aislamiento & purificación , Flavonoides/farmacología , Humanos , Espectroscopía de Resonancia Magnética/métodos , Espectroscopía de Resonancia Magnética/normas , Pruebas de Sensibilidad Microbiana , Estructura Molecular , Mycobacterium tuberculosis/efectos de los fármacos , Fenoles/aislamiento & purificación , Fenoles/farmacología , Plasmodium falciparum/efectos de los fármacos , Estándares de Referencia , Sensibilidad y Especificidad , Estereoisomerismo , Relación Estructura-Actividad , Xantonas/aislamiento & purificación , Xantonas/farmacologíaRESUMEN
Chemical investigation of biologically active compounds of Getonia floribunda led to the isolation of two new macrocyclic lactones, combretastatins D-3 (1) and D-4 (2). The structures of these compounds were confirmed by spectroscopic analyses. Combretastatin D-3 (1) exhibited cytotoxicity towards the small-cell lung cancer cell line (NCI-H187, IC50=13.0 +/- 0.2 microg/mL) but was inactive against KB, BC-1, and Vero cell lines. Combretastatin D-3 (1) showed weak antitubercular activity with a minimum inhibitory concentration (MIC) of 100.0 microg/mL, and was inactive towards the malarial parasite. Combretastatin D-4 (2) was inactive in all antitubercular, antiplasmodial, and cytotoxic assays.