Use of plant extracts in the control of common bean anthracnose

Three hundred (300) methanol extracts of barks, leaves, flowers and stems of 200 plant species from Alto Rio Grande and Vale do São Francisco regions, State of Minas Gerais, Brazil, were prepared and submitted to an in vitro growth test with the fungus Colletotrichum lindemuthianum, causal agent of anthracnose in common bean. Extracts from 13 plant species (Astronium fraxinifolium, Inga marginata, Malva sylvestris, Matayba elaeagnoides, Miconia argyrophylla, Myrcia fallax, Ocimum gratissimum, Origanum vulgare, Rollinia emarginata, Siparuna arianeae, Styrax pohlii, Tabebuia serratifolia and Trichilia pallida) presented antifungal activity and were used in other in vitro assays. Extracts from M. argyrophylla, M. fallax, O. vulgare, S. arianeae and S. pohlii were the most promising for the inhibition of the mycelial growth while the extracts of M. argyrophylla, M. elaeagnoides and O. gratissimum presented the best results for the inhibition of conidial germination. Under greenhouse conditions, the extracts of M. argyrophylla and O. vulgare caused the greatest reductions (41.82% and 37.65%, respectively) in disease severity when a local effect assay was carried out. In the systemic effect assay, also in a greenhouse, the most promising extracts were those from I. marginata, M. argyrophylla, M. fallax, M. sylvestris, O. gratissimum, O. vulgare and S. arianeae, which reduced the severity of the anthracnose to values below 35% of the observed for the control. Therefore, future studies with these plant species should be carried out to develop new products to control the common bean anthracnose.     

Comparative assessment of antioxidant and cholinesterase inhibitory properties of the marigold extracts from Calendula arvensis L. and Calendula officinalis L.

Calendula sp. is an important medicinal and industrial plant with various bioactivities. In this study, we examined enzyme inhibitory effects of the n-hexane, dichloromethane, acetone, ethyl acetate, methanol, and water extracts of the leaf and flowers of Calendula arvensis L. and C. officinalis L. against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The extracts were screened for their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion-chelating capacity and ferric-reducing antioxidant power (FRAP) assays at 250, 500, and 1000 μg mL⁻¹. Total phenol and flavonoid quantification of the extracts was achieved using Folin-Ciocalteau and AlCl₃ reagents, respectively. The ethyl acetate extract of C. arvensis flowers was the most active in AChE inhibition assay (31.24 ± 1.29%), while the n-hexane extract of C. officinalis leaves exerted the highest ferric ion-chelating capacity (74.27 ± 2.25%). Thin layer chromatographic analysis indicated presence of flavonoid and triterpene derivatives mainly in the extracts.     

Insecticidal activities of crude extracts and phospholipids from Chenopodium ficifolium against melon and cotton aphid, Aphis gossypii

Several organic solvent extracts of Chenopodium ficifolium were tested for their insecticidal activity against melon and cotton aphid, Aphis gossypii, on cucumber plants. Both methanol and ethanol extracts, at 5000 μg ml⁻¹, were highly active giving over 80% control. The other crude extracts displayed moderate or weak insecticidal activity giving control in the range of 16–69%. Two phospholipids were isolated as insecticidal active substances from C. ficifolium. Their chemical structures were identified as 1-palmitoyl-2-(3-trans)-hexadecenoyl-sn-glycero-3-phosphoglycerol and 1-palmitoyl-2-linoleoyl-3-glycerophosphocholine by GC–MS, EDS, mass and NMR spectral analyses. Both compounds displayed a dose-dependent mortality of A. gossypii. Furthermore, the liquid formulation that was obtained by partitioning with n-hexane from the methanol extract of C. ficifolium controlled melon and cotton aphid on cucumber plants effectively. These results indicate that extracts of C. ficifolium have potential for development as botanical insecticides for controlling A. gossypii infesting cucumber plants.     

Potassium phosphite for control of downy mildew of soybean

Downy mildew of soybean, caused by Peronospora manshurica, is widely spread throughout Brazil. The objective of this study was to evaluate the use of potassium phosphite to control this disease. Field experiments were conducted during the growing season of 2006/2007 and 2007/2008 in the state of Parana in southern Brazil. The experimental design consisted of completely randomized blocks in a factorial arrangement (4 × 2) with four replications. Four rates of potassium phosphite (0, 375, 750 and 1500 g P₂O₅ + K₂O ha⁻¹) were applied at two growth stages, V6 (fifth trifoliolate leaf) and R2 (full flowering), followed by one or two applications of pyraclostrobin and epoxiconazole (66.5 + 25 g a.i. ha⁻¹) at R3 (pod development) or R2 and R5.1 (10% of pod filling), mainly for the control of Asian soybean rust (Phakopsora pachyrhizi) and powdery mildew (Microsphaera diffusa). Field experiments were conducted to quantify the severity of downy mildew on leaves, nutrient content in leaf tissue (N, P and K), leaf area index (LAI), yield and seed weight. The maximum severity of downy mildew was observed at a growth stage of R5.3 (50% of pods were ripe), with 14% and 46% of the leaf area affected in 2006/2007 and 2007/2008, respectively. Also it was detected some effect of phosphite on Asian rust control but it was mostly in the trial of 2007/08 when the epidemic was very low (9.7-21.8% of severity). There was a linear reduction in the severity of downy mildew and a significant improvement in the LAI with an increase in the rate of phosphite applied. During the 2006/2007 growing season, a significant yield improvement was observed due to the application of the highest rate of phosphite. Two fungicide applications following phosphite application significantly improved the control of Asian soybean rust and powdery mildew, yield and seed weight when compared to a single fungicide application.     

Efficacy of medicinal plant extracts against Formosan subterranean termite, Coptotermes formosanus

The Formosan subterranean termite, Coptotermes formosanus Shiraki, is among the most devastating termite pests. Natural products derived from plant extracts were tested in a discovery programme for effective, environment friendly termite control agents. Screening for anti-termitic activity of plant extracts with some known medicinal attributes could lead to the discovery of new agents for termite control. The aim of this study was to investigate the anti-termitic activity of crude leaf hexane, ethyl acetate, acetone and methanol extracts of Andrographis lineata Wallich ex Nees. (Acanthaceae), Andrographis paniculata (Burm.f.) Wall. ex Nees. (Acanthaceae), Argemone mexicana L. (Papaveraceae), Aristolochia bracteolata Lam. (Aristolochiaceae), Datura metel L. (Solanaceae), Eclipta prostrata L. (Asteraceae), Sesbania grandiflora (L.) Pers. (Fibaceae) and Tagetes erecta L. (Compositae) against C. formosanus. An impregnated filter paper no-choice bioassay method was followed. All the crude extracts showed anti-termitic activity in a dose-dependent manner and exhibited a significant activity after 24 h and 48 h of exposure; the highest termite mortality was found in leaf hexane extract of A. bracteolata, ethyl acetate extract of A. paniculata, D. metel, E. prostrata, methanol extract of A. lineata and D. metel after 24 h (LD₅₀ = 363, 371, 298, 292, 358 and 317 ppm; LD₉₀ = 1433, 1659, 1308, 1538, 1703 and 1469 ppm), respectively. The hexane extract of T. erecta, acetone extract of A. mexicana, methanol extract of S. grandiflora and T. erecta showed activity after 48 h (LD₅₀ = 245, 253, 289, 409 ppm; LD₉₀ = 1378, 1511, 1508 and 2425 ppm), respectively. Among the natural products tested, may provide a renewable source of safe natural wood preservatives. These findings corroborate traditional insecticidal application of selected plants and the results can be extended for the control of termites. The primary objective of the present study was to identify novel, natural chemotypes from biologically active crude plant extracts that may be useful as part of termite treatment regimens in their natural form or as synthons for structure-activity studies in the future. The results reported here open the possibility of further investigations of efficacy on their anti-termitic properties of natural product extracts.     

Variation in essential oil and fatty acid composition in different organs of cultivated and growing wild Ruta chalepensis L.

The essential oil and fatty acid composition of two provenances of Ruta chalepensis from four organs (leaves, flowers, stems and fruits) was determined. The effect of the plant part on total fatty acid contents, essential oil yields, fatty acid and volatile constituents was significant.Fatty acid profiles varied significantly among the studied provenances and organs. Linolenic acid had the highest amount in leaves of the two provenances. From R. chalepensis, in all organs, the main fatty acids were palmitic (13.10-25.31%), followed by palmitoleic (0-15.72%), stearic (1.03-6.85%), oleic (1.90-24.04%), arachidic (0.11-4.03%), eicosatetraenoic (0.10-5.60%) and behenic (0.47-6.09%) acids. Saturated fatty acids had the highest amounts in growing wild R. chalepensis flowers, and cultivated R. chalepensis stems were characterized by the predominance of polyunsaturated fatty acids. Oil composition of all studied organs has a healthy and nutritionally value. Essential oil yields varied from 0.39% to 2.46% and showed a remarkable variation with plant organs. Thirty-six volatile compounds were identified in different analyzed essential oils; 2-undecanone, 2-nonanol and 2-dodecanone had the highest percentages.     

Variability in essential oil content and composition of Origanum hirtum L., Origanum onites L., Coridothymus capitatus (L.) and Satureja thymbra L. populations from the Greek island Ikaria

Plants belonging to four “oregano” plant species (Origanum hirtum L., Origanum onites L., Coridothymus capitatus L., and Satureja thymbra L.) were collected during flowering from 33 sites located in the eastern part of the Greek island of Ikaria in the Eastern Aegean during April, May and July 2008. C. capitatus and O. hirtum were mostly observed in higher altitudes, whereas O. onites and S. thymbra in lower ones. The spatial distribution of all species was depicted on a GIS map. All four species exhibited essential oil concentrations higher than those reported in earlier literature, namely O. onites 3-4.3%, S. thymbra 4-6.5%, C. capitatus 3.7-5.6% and O. hirtum 5.5-10.0% (v/w). Carvacrol was the main constituent of the essential oils of all species, followed by γ-terpinene, p-cymene and caryophyllene, while thymol was not detected. All constituents varied remarkably among the four species, with carvacrol exhibiting the lowest variation. Carvacrol content varied between 72.3 and 89.2% in O. onites; 46.5 and 58.0% in S. thymbra; 82.9 and 90.9% in C. capitatus; and 84.4 and 93.8% in O. hirtum. By applying hierarchical cluster analysis on the basis of the essential oil constituents two main groups, divided into four subgroups of the taxa were evident. The first group consisted of O. onites and S. thymbra, while the second one of C. capitatus and O. hirtum. The results are discussed in terms of topography and climatic variation.     

Identification of an insecticidal polyacetylene derivative from Chrysanthemum macrotum leaves

Compounds responsible for insecticidal properties of Chrysanthemum macrotum (D.R.) Ball. leaves against Spodoptera littoralis Boiduval caterpillars have been investigated. The screening of the insecticidal activity was performed by incorporating methanol, buthanol or ethyl acetate extracts, or some chromatographic fractions to the caterpillars’ artificial diet. It was noted that extracts and fractions ameliorated or disturbed nutritional indexes, being not always toxic for caterpillars. Among the tested fractions, one pure compound with a high insecticidal activity (percentage of mortality 66.7%) was purified. The nuclear magnetic resonance study allowed its identification as a polyacetylene derivative, in particular a spiroketal enol ether one.     

Improvement of essential oil yield of oil-bearing (Rosa damascena Mill.) due to surfactant and maceration

The essential oil content of the oil-bearing rose (Rosa damascene Mill.) is relatively low, around 0.3-0.4 mL kg⁻¹ in fresh flowers. There is a need to increase essential oil yield of oil-bearing rose. The objective was to examine the effect of Tween 20 (polyoxyethylene sorbitan monolaurate) applied with, or without, maceration of flowers on oil content and composition of oil-bearing rose harvested at beginning of flowering, full bloom, and end of flowering. Addition of Tween 20 at 1000 mL L⁻¹ and 2500 mL L⁻¹ increased essential oil yield by 26% (to 0.44 mL kg⁻¹) and 54% (to 0.54 mL kg⁻¹) respectively relative to the untreated control that gave 0.35 mL kg⁻¹ yield. Maceration, in combination with the addition of Tween 20 at 1000 mL L⁻¹ and Tween 20 at 2500 mL L⁻¹, increased oil yield by 69% (to 0.59 mL kg⁻¹) and 94% (to 0.68 mL kg⁻¹) respectively. Among the three phenological phases of harvest, harvesting at the beginning of flowering gave the highest yield followed by the full bloom and then by the end of flowering phases. Since the interaction effect was not significant, the differences obtained among the treatments were regardless of the phase, and vice versa. Treatments did not significantly alter composition of the essential oil. Postharvest pre-extraction application of Tween 20 in combination with maceration could be used in the rose industry for increasing the essential oil yield.     

Determinants of fenhexamid effectiveness against grey mould on grapevine: Respective role of spray timing,fungicide resistance and plant defences

Botrytis bunch rot of grapes is mainly controlled by applying fungicides at three crop growth stages: the end of flowering (BBCH 68), bunch closure (BBCH 77) and the beginning of veraison (BBCH 81). The hydroxyanilide derivative fenhexamid is among the most effective fungicides registered to control Botrytis cinerea. Its effectiveness was examined in relation to spray timing, fungicide resistance and defense responses of grapevine. Overall, the earlier fenhexamid was applied, the more effective it was at controlling B. cinerea. Frequencies of B. cinerea strains which were resistant to fungicides were evaluated at harvest. The frequencies of resistant phenotypes were similar among treatments and years with the exception of a class of multidrug resistant strain (MDR 2) whose frequency appeared to increase after fenhexamid applications. If current spray programs including fenhexamid appear to control bunch rot at the current MDR frequency, a propagation of MDR 2 strains might lead to a decline in disease control. Finally, defense responses were studied in grapevine flowers/berries following fenhexamid application. None of the defense processes tested was induced in flowers/berries at stages 68 and 77. Only an increase in chitinase activity was observed in treated-berries at stage 81, suggesting that fenhexamid effectiveness was not related to a stimulation of defense responses.     

Potential of extracts of the tropical plant Balanites aegyptiaca (L) Del. (Balanitaceae) to control the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae)

The effects of extracts of different parts of the perennial tropical plant Balanites aegyptiaca (L) Del., including various solvent extracts of roots, methanol extracts from leaves, fruits, flowers and roots, partially purified saponins obtained from its roots and a standard saponin were studied on the life cycle (adult longevity, number of eggs, crawlers, adults, weight of adults and % wax content) of a laboratory-reared parthenogenic line of the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae). Extracts derived from various parts of B. aegyptiaca (leaves, fruits, flowers, and roots in methanol) affected the life cycle of M. hirsutus with a methanol root extract being the most effective at a concentration of 500 μg ml⁻¹. Partially purified saponin of B. aegyptiaca and the commercial bark saponin extract (Sigma) from Quillaja saponaria at a concentration of 500 μg ml⁻¹ were effective in reducing the longevity of M. hirsutus. Significant reductions in oviposition by M. hirsutus were found for all the extracts at a concentration of 500 μg ml⁻¹. Extracts also affected the number of emerging crawlers, number of adults as well as the weight and wax content of emerging adults. These studies suggest that B. aegyptiaca plant extracts and saponins can be useful botanical insecticides for the protection of crops from mealy bugs.     

Genetic characterization of kernel polyphenol oxidases in wheat and related species

Polyphenol oxidase (PPO) activity causes undesirable darkening of raw Asian noodles and other wheat products. In this study we investigate the genetic origins and diversity of wheat kernel PPO. PPO was characterized via activity assays, antigenic staining, and Southern blots in Triticum aestivum, Triticum dicoccoides, Triticum durum, Triticum dicoccum, Triticum monococcum, Triticum urartu, Aegilops speltoides, and Aegilops tauschii. Among these species, PPO activity was well-correlated with antigenic staining intensity toward a wheat kernel-type PPO antibody. High PPO activity was observed in all three T. monococcum accessions (A^m genome), one Ae. speltoides accession, one T. durum accession, and two hexaploid wheat cultivars. Southern blots suggested the presence of two or more kernel-type PPO genes in diploid progenitors of the hexaploid A, B, and D genomes. Whole-kernel PPO activity was evaluated in disomic substitution lines derived from three T. dicoccoides accessions in the background of T. durum ‘Langdon’. PPO activity was primarily associated with chromosome 2A and to a much lower degree with chromosome 2B. DNA sequence comparisons showed that the intron associated with the high PPO allele on chromosome 2AL of hexaploid wheat had 94% nucleotide identity with the homeologous intron found in T. monococcum, a species with high kernel PPO activity. This implies that the ancestral PPO allele on the A genome is one of the high activity, and the low PPO allele found in hexaploid wheat represents a relatively recent genetic alteration. Results confirm the presence of multiple kernel-type PPO genes in the diploid and tetraploid progenitors and relatives of hexaploid wheat. However, it is likely that relatively few of the many kernel-type PPO genes present in wheat contribute substantially to kernel PPO activity. A single genetic locus on homeologous group 2 chromosomes may be the primary cause of high PPO activity in wheat kernels.     

Time-mortality relationships of larvae and adults of grain beetles exposed to extreme cold

Mortality effects of low temperature exposure to −16 °C were investigated on larvae and adults of the grain beetles Tribolium confusum Jacquelin du Val (Coleoptera: Tenebrionidae), Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae) and Trogoderma granarium (Everts) (Coleoptera: Dermestidae), in the laboratory. The main effects and interactions of exposure time (0.2, 0.5, 1, 2, 4, 8, 12, 24 and 48 h), developmental stage (larva and adult) and individual’s age (young and old) were examined. After 4 h of exposure 100% mortality has been achieved in most of the treatments. The only exceptions were that of old larvae of T. granarium and O. surinamensis, and adults of T. granarium which needed 48, 8, and 24 h, respectively. T. granarium was the most cold-hardy species irrespective of developmental stage, and age, followed in decreasing order by O. surinamensis and T. confusum. Younger adults were generally more susceptible to cold in the cases of O. surinamensis and T. confusum. On the contrary, older adults of T. granarium suffered higher mortality than younger ones. Larvae of T. granarium were generally more cold-tolerant than adults but the opposite pattern was observed in O. surinamensis and T. confusum. Main effects of exposure time, developmental stage and individual’s age on mortality proved to be significant for all species, with the exception of T. granarium where the effect of developmental stage proved to be insignificant. Lethal time values were estimated via probit analysis. Values of LT₅₀ ranged between 0.7 and 1.0 h for T. confusum, 0.7-1.9 h for O. surinamensis and 1.2-3.4 h for T. granarium. Our results are discussed with findings from relevant studies and the possibility of including extreme cooling to IPM programs against stored product pests is investigated.     

Antifungal activity of leaf extracts from South African trees against plant pathogens

The antifungal activity of acetone, methanol, hexane and dichloromethane leaf extracts of six plant species (Bucida buceras, Breonadia salicina, Harpephyllum caffrum, Olinia ventosa, Vangueria infausta and Xylotheca kraussiana) were evaluated for antifungal activity against seven plant pathogenic fungal species (Aspergillus niger, Aspergillus parasiticus, Colletotricum gloeosporioides, Penicillium janthinellum, Penicillium expansum, Trichoderma harzianum and Fusarium oxysporum). These plant species were selected from 600 evaluated inter alia, against two animal fungal pathogens. All plant extracts were active against the selected plant pathogenic fungi. Of the six plant species, B. buceras had the best antifungal activity against four of the fungi, with minimum inhibitory concentration (MIC) values as low as 0.02 mg/ml and 0.08 mg/ml against P. expansum, P. janthinellum, T. harzianum and F. oxysporum. Some of the plant extracts had moderate to low activity against other fungi, indicating that the activity is not based on a general metabolic toxicity. P. janthinellum, T. harzianum and F. oxysporum were the most sensitive fungal species, with a mean MIC of 0.28 mg/ml, while the remaining four fungi were more resistant to the extracts tested, with mean MICs above 1 mg/ml. The number of active compounds in the plant extracts was determined using bioautography with the listed plant pathogens. No active compounds were observed in some plant extracts with good antifungal activity as a mixture against the fungal plant pathogens, indicating possible synergism between the separated metabolites, B. salicina and O. ventosa were the most promising plant species, with at least three antifungal compounds. Leaf extracts of different plant species using different methods (acetone, hexane, DCM and methanol) had antifungal compounds with the same R_f values. The same compounds may be responsible for activity in extracts of different plant species. Based on the antifungal activity, crude plant extracts may be a cost effective way of protecting crops against fungal pathogens. Because plant extracts contain several antifungal compounds, the development of resistant pathogens may be delayed.     

Assessment of the Antimicrobial Activity of Algae Extracts on Bacteria Responsible of External Otitis

External otitis is a diffuse inflammation around the external auditory canal and auricle, which is often occurred by microbial infection. This disease is generally treated using antibiotics, but the frequent occurrence of antibiotic resistance requires the development of new antibiotic agents. In this context, unexplored bioactive natural candidates could be a chance for the production of targeted drugs provided with antimicrobial activity. In this paper, microbial pathogens were isolated from patients with external otitis using ear swabs for over one year, and the antimicrobial activity of the two methanol extracts from selected marine (Dunaliella salina) and freshwater (Pseudokirchneriella subcapitata) microalgae was tested on the isolated pathogens. Totally, 114 bacterial and 11 fungal strains were isolated, of which Staphylococcus spp. (28.8%) and Pseudomonas aeruginosa (P. aeruginosa) (24.8%) were the major pathogens. Only three Staphylococcus aureus (S. aureus) strains and 11 coagulase-negative Staphylococci showed resistance to methicillin. The two algal extracts showed interesting antimicrobial properties, which mostly inhibited the growth of isolated S. aureus, P. aeruginosa, Escherichia coli, and Klebsiella spp. with MICs range of 1.4 × 10⁹ to 2.2 × 10¹⁰ cells/mL. These results suggest that the two algae have potential as resources for the development of antimicrobial agents.     

Hybanthus parviflorus (Violaceae): Insecticidal activity of a South American plant

Hybanthus parviflorus is a perennial shrub, widely distributed in the tropical and subtropical regions of America, and is known as ‘violetilla’ in Argentina. Previous phytochemical studies of this species led us to the isolation and determination of the primary structure of a novel macrocyclic polypeptide, the cyclotide hypa A. Here the insecticidal activities of extracts of H. parviflorus were determined against Ceratitis capitata Wied., the Mediterranean fruit fly or ‘Medfly’. Mortality in the different life stages of Medfly, total mortality and modifications of the insect’s physiology caused by 50% EtOH and CH₂Cl₂ extracts of H. parviflorus were evaluated. In addition, we determined the occurrence of ursolic acid, β-sitosterol and the polyphenols quercetin, quercetin-3-methyl ether, apigenin, luteolin, kaempferol, rutin, caffeic acid and chlorogenic acid. The promising insecticidal activity of 50% EtOH extracts of H. parviflorus and its purified fractions was related to the presence of cyclotides. The insecticidal activity of CH₂Cl₂ extracts could be related to the presence of polyphenols, ursolic acid and β-sitosterol.     

Antioxidant and Antimicrobial Potential of the Bifurcaria bifurcata Epiphytic Bacteria

Surface-associated marine bacteria are an interesting source of new secondary metabolites. The aim of this study was the isolation and identification of epiphytic bacteria from the marine brown alga, Bifurcaria bifurcata, and the evaluation of the antioxidant and antimicrobial activity of bacteria extracts. The identification of epiphytic bacteria was determined by 16S rRNA gene sequencing. Bacteria extracts were obtained with methanol and dichloromethane (1:1) extraction. The antioxidant activity of extracts was performed by quantification of total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and oxygen radical absorbance capacity (ORAC). Antimicrobial activities were evaluated against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae and Candida albicans. A total of 39 Bifurcaria bifurcata-associated bacteria were isolated and 33 were identified as Vibrio sp. (48.72%), Alteromonas sp. (12.82%), Shewanella sp. (12.26%), Serratia sp. (2.56%), Citricoccus sp. (2.56%), Cellulophaga sp. (2.56%), Ruegeria sp. (2.56%) and Staphylococcus sp. (2.56%). Six (15.38%) of the 39 bacteria Bifurcaria bifurcata-associated bacteria presented less than a 90% Basic Local Alignment Search Tool (BLAST) match, and some of those could be new. The highest antioxidant activity and antimicrobial activity (against B. subtilis) was exhibited by strain 16 (Shewanella sp.). Several strains also presented high antimicrobial activity against S. aureus, mainly belonging to Alteromonas sp. and Vibrio sp. There were no positive results against fungi and Gram-negative bacteria. Bifurcaria bifurcata epiphytic bacteria were revealed to be excellent sources of natural antioxidant and antimicrobial compounds.     

Toxicity of quinones against two-spotted spider mite and three species of aphids in laboratory and greenhouse conditions

Toxicities of the eight quinones were evaluated through leaf dip bioassays conducted against Tetranychus urticae, Myzus persicae, Myzocallis walshii, and Illinoia liriodendri. Based on LC₅₀ values, plumbagin (LC₅₀ = 0.001%) was the most active compound against T. urticae and ubiquinone Q₀ (LC₅₀ = 0.005%), plumbagin (LC₅₀ = 0.010%), and dibromothymoquinone (LC₅₀ = 0.012%) were the most active compounds against M. persicae. The most active compounds against M. walshii were juglone (LC₅₀ = 0.011%) and ubiquinone Q₀ (LC₅₀ = 0.019%), whereas dibromothymoquinone (LC₅₀ = 0.030%), plumbagin (LC₅₀ = 0.033%) and ubiquinone Q₀ (LC₅₀ = 0.058%) were the most toxic to I. liriodendri. Ecotrol (positive control) was the least toxic compound (LC₅₀ = 0.39%) against T. urticae and M. persicae (LC₅₀ = 0.447%). Although the majority of the compounds tested were toxic to all four test species in residual bioassays, there was little overlap among the test species in terms of susceptibility to the compounds and interspecific differences were observed. Regarding structure-activity relationships for quinones, the addition of a hydroxyl group resulted in a significant increase in the toxicity of the 1,4-naphthoquinones, and those possessing a methyl group exhibited the highest levels of activity in T. urticae. The bromine atom at the 2- and 5-positions of the benzoquinone ring is crucial to the toxicity of the compounds against I. liriodendri. Toxicity was greatly affected not only by the number of hydroxyl groups, but also by their positions in the ring in the case of M. walshii. Juglone and plumbagin as residual toxins in the laboratory also reduced the population of two-spotted spider mites compared to EcoTrol™ (positive control) and the negative control in the greenhouse experiment. Some quinones tested may have potential as commercial insecticides and miticides, or alternatively, could serve as lead compounds for the development of more potent crop protection agents.     

Isolation of entomopathogenic Bacillus from a biodynamic olive farm and their pathogenicity to lepidopteran and coleopteran insect pests

The occurrence of Bacillus entomopathogenic bacteria on a Tunisian biodynamic farm was determined by examining 75 samples from olive tree (Olea europaea L.) habitats. A total of 40 Bacillus isolates were characterized according to their phenotypic, physiological and biochemical parameters. Isolates of the species Bacillus subtilis, Bacillus mycoides, Brevibacillus brevis, Paenibacillus polymyxa, Bacillus licheniformis, Bacillus sp. (1), Bacillus sp. (2) and a standard strain Btk HD-1 were used separately in feeding bioassays on fresh artificial diet against larvae of lepidopterans Prays oleae (Bernard) and Palpita unionalis (Hübner) and coleopterans Hylesinus oleiperda (F.) and Phloeotribus scarabaeoides (Bernard), which are olive tree pests. Larvae were successfully reared on an artificial diet with 25 g powdered olive tree leaves. Compared to the control data, only Btk and the isolates of B. licheniformis, P. polymyxa and B. brevis were entomopathogenic. Larval mortality assessed 7 days post-treatment showed high mortality rates with Btk to lepidopteran larvae (86.6% for P. oleae and 80.9% for P. unionalis) and low mortality against coleopteran pests. B. brevis isolates showed high mortality rates against P. oleae (up to 67.9%). B. licheniformis isolates caused up to 59.2% larval mortality for P. oleae and 43.6% for P. unionalis. Highest coleopteran mortality was achieved by P. polymyxa isolates (up to 55%). According to the 16S rDNA results, isolates of each of the three entomopathogenic strains were similar. Proteins in the strain supernatants were toxic to P. oleae larvae with LC50 values of 10.0 (B. brevis), 12.5 (B. licheniformis) and 37.6 μg/ml (P. polymyxa). Also, P. polymyxa showed an LC50 of 12.4 mg/l against P. scarabaeoides. Our results suggest that entomopathogenic Bacillus present locally in the biodynamic farm could be used in biological control programmes of olive tree pests.     

Laminarin from Irish Brown Seaweeds Ascophyllum nodosum and Laminaria hyperborea: Ultrasound Assisted Extraction,Characterization and Bioactivity

Ultrasound assisted extraction (UAE), purification, characterization and antioxidant activity of laminarin from Irish brown seaweeds Ascophyllum nodosum and Laminarina hyperborea were investigated. UAE was carried out using 60% ultrasonic power amplitude and 0.1 M hydrochloric acid for 15 min. Separately, solid-liquid extraction was carried in an orbital shaker using 0.1 M hydrochloric acid at 70 °C for 2.5 h. UAE with hydrochloric acid resulted in the highest concentration of laminarin, 5.82% and 6.24% on dry weight basis from A. nodosum and L. hyperborea, respectively. Purification of all extracts was carried out using molecular weight cut off dialysis at 10 kDa. Characterization of the laminarin fraction was carried out using matrix assisted laser desorption/ionization time-of-flight mass spectrometry. Antioxidant activity of A. nodosum and L. hyperborea extracts had 2,2-diphenyl-1-picrylhydrazyl (DPPH) inhibition levels of 93.23% and 87.57%, respectively. Moreover, these extracts have shown inihibition of bacterial growth of Staphylcoccus aureus, Listeria monocytogenes, Escherichia coli and Salmonella typhimurium.