Antibacterial, mosquito larvicidal, and cytotoxicity potential of AgNPs synthesized using Pittosporum undulatum under in vitro conditions.

In this study, the phytochemical profile and silver nanoparticle (AgNPs)-synthesizing ability of Pittosporum undulatum methanol extract were investigated. Furthermore, biological applications of the AgNPs, such as antibacterial effect (against Klebsiella pneumoniae, Staphylococcus aureus, Bacillus subtilis, and Escherichia coli), mosquito larvicidal effect (against Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti), and cytotoxicity (against fibroblast cell line L929) were evaluated using in vitro experiments. The phytochemical analysis revealed that the methanol extract contained cardiac glycosides, terpenoids, saponins, alkaloids, flavonoids, glycosides, coumarins, phenolics, and tannins. Furthermore, standard characterization techniques such as UV-Vis spectrometry, SEM, TEM, FTIR, and XRD confirmed that the methanol extract of P. undulatum effectively synthesized the AgNPs. The synthesized AgNPs had a spherical shape and size of 20-200 nm. The bactericidal analysis revealed that the AgNPs have dose-dependent antibacterial activity. The MTT assay showed that the AgNPs were bio-compatible up to a dosage of 250 µg mL-1 in the normal fibroblast cell line L929. Furthermore, the LC50 values for AgNPs against larvae of An. stephensi, Cx. quinquefasciatus, and Ae. aegypti were 0.4, 4.7, and 1.2 ppm, respectively. Field trials demonstrated that the larvicidal effect was enhanced within 24-72 h, and the rate of reduction increased over time. Thus, our findings provide an ideal sustainable AgNP bio-pesticide to combat filarial, dengue, and malaria vectors.

Authentication of incense (Pittosporum undulatum Vent.) honey from the Azores (Mel dos Açores) by a novel real-time PCR approach.

'Mel dos Açores' is a unique nectar honey produced from the exceptional and diverse flora of the Azores archipelago, categorised as incense honey ('mel de incenso') or multifloral honey ('mel multiflora'). Incense honey should contain over 30 % of pollen grains of Pittosporum undulatum Vent. In this work, a real-time PCR method targeting the ITS region was proposed for the first time to detect P. undulatum in the honey from the Azores. The approach exhibited high analytical performance, achieving a quantification limit of 0.01 pg of incense DNA. The method was successfully applied to 22 honey samples, from which incense was detected in all 9 monofloral incense honeys and in 5 out of 10 multifloral samples from the Azores. Generally, the quantitative results for incense DNA were in good agreement with the melissopalynological data. Therefore, a simple, cost-effective and reliable tool was herein proposed to authenticate and valorise the Azores honey.

Nematicidal activity of essential oils and volatiles derived from Portuguese aromatic flora against the pinewood nematode, Bursaphelenchus xylophilus.

Twenty seven essential oils, isolated from plants representing 11 families of Portuguese flora, were screened for their nematicidal activity against the pinewood nematode (PWN), Bursaphelenchus xylophilus. The essential oils were isolated by hydrodistillation and the volatiles by distillation-extraction, and both were analysed by GC and GC-MS. High nematicidal activity was achieved with essential oils from Chamaespartium tridentatum, Origanum vulgare, Satureja montana, Thymbra capitata, and Thymus caespititius. All of these essential oils had an estimated minimum inhibitory concentration ranging between 0.097 and 0.374 mg/ml and a lethal concentration necessary to kill 100% of the population (LC(100)) between 0.858 and 1.984 mg/ml. Good nematicidal activity was also obtained with the essential oil from Cymbopogon citratus. The dominant components of the effective oils were 1-octen-3-ol (9%), n-nonanal, and linalool (both 7%) in C. tridentatum, geranial (43%), neral (29%), and ß-myrcene (25%) in C. citratus, carvacrol (36% and 39%), γ-terpinene (24% and 40%), and p-cymene (14% and 7%) in O. vulgare and S. montana, respectively, and carvacrol (75% and 65%, respectively) in T. capitata and T. caespititius. The other essential oils obtained from Portuguese flora yielded weak or no activity. Five essential oils with nematicidal activity against PWN are reported for the first time.

Anti-inflammatory guaiane-type sesquiterpenes from the fruits of Pittosporum undulatum.

Two unprecedented guaiane-type sesquiterpene glycosides (undulatumosides A and B) were isolated by bioassay-guided fractionation from the MeOH extract of Pittosporum undulatum fruits, along with six known compounds, including the guaiane isomers 5-guaien-11-ol and 4-guaien-11-ol. The structures of the compounds were established as 4-guaiene-11-O-ß-d-(3'-angeloxy-6'-deoxy)-glucopyranoside and 1(5)-guaiene-11-O-ß-d-(3'-angeloxy-6'-deoxy)-glucopyranoside by spectroscopic methods, including 1D and 2D homo- and heteronuclear NMR experiments (COSY, HSQC, HMBC and NOESY), and HR-mass spectrometry. P. undulatum is a highly invasive weed that often outcompetes other plants, yet its fruits have become a traditional anti-inflammatory medicine in Azores. Therefore, aiming to investigate the claimed properties, the in vitro anti-inflammatory activity of guaiane-type sesquiterpenes was evaluated by analyzing their inhibitory effects on chemical mediators released by the LPS activated RAW 264.7 murine macrophages cell line. In addition, the cytotoxicity of these compounds was also evaluated in this cell line. Undulatumoside A, 5-guaien-11-ol and 4-guaien-11-ol displayed anti-inflammatory activity with IC50 values of 16.4, 8.1 and 7.2µM, respectively, comparable to that of the positive control, indomethacin (IC50=18.2 µM), with no cytotoxic effects (IC50 ≥ 198 µM). Furthermore, the same set of compounds was also assessed for anti-proliferative activity in lung large cell carcinoma COR-L23 and amelanotic melanoma C32 cells.