Nematicidal potential of Artemisia annua and its main metabolites

Nematotoxic effect of an aqueous extract of Artemisia annua and its components caffeic acid, chlorogenic acid (5-caffeoylquinic acid, 5-CQA), artemisinin and the related semi-synthetic artesunate, was investigated on the root-knot nematode Meloidogyne incognita and the potato cyst nematode Globodera rostochiensis and on the virus-vector dagger nematode Xiphinema index. Juveniles of M. incognita and G. rostochiensis and females of X. index were exposed to 500, 250 and 125 μl ml^?1 solutions of the A. annua aqueous extract, caffeic acid, chlorogenic acid and artesunate and to 50 μl ml^?1 solution of artemisinin for 2, 4, 8 and 24 h. Egg masses of M. incognita and cysts of G. rostochiensis were exposed for 24, 48, 96 h and 1 or 2 weeks only to the extract solutions. Aqueous extract was highly effective on G. rostochiensis juveniles, whereas M. incognita juveniles were affected only at long exposure times. Adversely, egg hatch inhibition was strong on M. incognita and poor or minimal on G. rostochiensis. Females of X. index were sensitive only to long exposures to the highest extract concentration. Both caffeic and chlorogenic acid did not affect juveniles of M. incognita but were highly active on G. rostochiensis juveniles and X. index females even at the lowest concentration. Artesunate toxicity was almost zero on M. incognita and low on X. index females, but high on G. rostochiensis juveniles. Artemisinin solution was lethal to more than 50 % of G. rostochiensis juveniles within 24 h, but did not affect M. incognita juveniles and X. index females. Results suggest different roles of the tested compounds in the biocidal activity on each target nematode species. The extract of A. annua and its main phytochemicals seem to have a potential to be developed into new nematicidal formulates, though their activity should be validated in the soil.     

Verbascoside, isoverbascoside, and their derivatives recovered from olive mill wastewater as possible food antioxidants

Olive oil processing industries generate substantial quantities of phenolic-rich byproducts, which could be valuable natural sources of antioxidants. This work is focused on the recovery and structural characterization of antioxidant compounds from olive mill wastewater (OMWW), a polluting byproduct of the olive oil production process. Phenolics were extracted from the waste material using a membrane technology coupled to low-pressure gel filtration chromatography on a Sephadex LH-20. The LH-20 fraction was, in turn, characterized for its phenolic composition by HPLC-DAD-MS/MS analyses. Verbascoside, isoverbascoside, β-hydroxyverbascoside, β-hydroxyisoverbascoside, and various oxidized phenolics were identified. Uptake of verbascoside, purified from the LH-20 fraction, by HT-29 cells, an established model system for studying drug transport properties, was also assayed. Finally, the antioxidant activities of the LH-20 fraction and verbascoside were characterized by two different techniques. Individual verbascoside was more active as a scavenger of reactive oxygen species and as a chemopreventive agent protecting low-density lipoproteins from oxidative damage than the LH-20 fraction.     

Hydroponic cultivation improves the nutritional quality of soybean and its products

Hydroponic cultivation allows the control of environmental conditions, saves irrigation water, increases productivity, and prevents plant infections. The use of this technique for large commodities such as soybean is not a relevant issue on fertile soils, but hydroponic soybean cultivation could provide proteins and oil in adverse environmental conditions. In this paper, the compositions of four cultivars of soybean seeds and their derivates, soy milk and okara, grown hydroponically were compared to that of the same cultivar obtained from soil cultivation in an open field. Besides proximal composition, the concentrations of phytic acid and isoflavones were monitored in the seeds, soy milk, and okara. Results demonstrated that, independent from the cultivar, hydroponic compared to soil cultivation promoted the accumulation of fats (from 17.37 to 21.94 g/100 g dry matter) and total dietary fiber (from 21.67 to 28.46 g/100 g dry matter) and reduced isoflavones concentration (from 17.04 to 7.66 mg/kg dry matter), whereas protein concentration was unaffected. The differences found in seed composition were confirmed in the respective okara products, but the effect of cultivation system was not significant looking at the soy milk composition. Data showed that hydroponic cultivation improved the nutritional quality of soybean seeds with regard to fats and dietary fiber. They also suggest that specific cultivars should be selected to obtain the desired nutritional features of the soybean raw material depending on its final destination.     

Experimental cultivation of the Mediterranean calanoid copepods Temora stylifera and Centropages typicus in a pilot re‐circulating system

A pilot re‐circulating system was used for the cultivation of two Mediterranean calanoid copepods: Temora stylifera and Centropages typicus. The system automatically concentrated the naupliar and copepodite stages. Temora stylifera was fed the flagellate Rhodomonas baltica or Prorocentrum minimum, whereas C. typicus was fed with a mixture of R. baltica or P. minimum and Tetraselmis suecica. Both copepods also received Isochrysis galbana. After 21 days, the T. stylifera population increased 26‐fold, reaching a density of 38 000 individuals, mostly represented by nauplii (88%). The maximum density recorded was 380 ind. L^?1, with a production of 370 nauplii L^?1. On average, the egg hatching success for this copepod during the rearing period was 54%, with the highest viability in April and May (>75%). The C. typicus population increased more than 10‐fold after 7 weeks of rearing, reaching a density of 123 000 individuals, mainly represented by nauplii (>90%). The highest naupliar production was 100 ind. L^?1, with a mean egg hatching success of 68%. This system may be useful to produce nauplii and copepodite stages to be used as live, alternative or complementary food for fish larvae or to provide a ready source of organisms for physiological and bioassay studies.     

Comparison of two methods for field grow of puncture vine (Tribulus terrestris L.) in Slovakia

One of the most claimed aphrodisiac plant is puncture vine (Tribulus terrestris L.) known as ‘plant Viagra’. This herb, growing especially in South and East Europe, is used for pharmaceutical purposes, due to its content of furostanol saponins which have stimulant effects on natural testosterone levels. Puncture vine grows as a weed on sandy soils areas. A study aimed to the introduction of puncture vine into large cultivation scale was carried out. Two different methods of cultivation were compared and production of biomass and evaluation of the content of furostanol saponins were done. Transplantation of seedlings raised in the greenhouse into the open field was more effective for the production of high amount of biomass and active components than using plants that are directly sown. The transplantation of seedlings into open field is suitable for the puncture vine cultivation as it increases biomass production.     

Antiproliferative terpenoids from almond hulls (Prunus dulcis): identification and structure-activity relationships

Bioassay-guided fractionation of the EtOAc crude extract from Sicilian almond hulls, a waste material from Prunus dulcis crop, allowed identification of 10 constituents, isolated as pure compounds (1-5, 7, and 10) or unseparable mixtures (5 + 6 and 8 + 9). All compounds were subjected to spectroscopic analysis and 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide bioassay on MCF-7 human breast cancer cells. In addition to the main components oleanolic (1), ursolic (2), and betulinic (3) acids, the 2-hydroxy analogues alphitolic (4), corosolic (5), and maslinic (6) acids, as well as the related aldehydes, namely, betulinic (7), oleanolic (8), and ursolic (9), were identified. From a more polar fraction, the beta-sitosterol 3-O-glucoside (10) was also identified. A sample of commercially available betulin (11) was also included in bioassays as further support to a structure-activity relationship study. Betulinic acid showed antiproliferative activity toward MCF-7 cells (GI50 = 0.27 microM), higher than the anticancer drug 5-fluorouracil.     

Survey of mealybug (Hemiptera: Pseudococcidae) vectors of <Emphasis Type="Italic">Ampelovirus</Emphasis> and <Emphasis Type="Italic">Vitivirus</Emphasis> in vineyards of northwestern Italy

Mealybugs (Hemiptera: Pseudococcidae) are sap-sucking insects which infest a broad range of crops worldwide. They represent an important threat to viticulture as they are vectors of viruses associated with leafroll and rugose wood complex diseases. In this study, we surveyed the presence of mealybugs and their associated viruses in vineyards of the Piemonte and Liguria regions, northwestern Italy. In order to determine the collected specimens correctly, we added a species-specific marker for Heliococcus bohemicus to an existing molecular identification key. The only species collected in Piemonte was H. bohemicus, whereas in Liguria, H. bohemicus, Planococcus ficus and Pseudococcus longispinus were found; Ps. longispinus has never before been reported in Italian vineyards. Several specimens of all three species were infected by the ampeloviruses GLRaV-1 and GLRaV-3 and the vitivirus GVA. Both nymphs and adult females tested positive for the viruses and mixed infections were commonly found within the same insect. Population levels and virus incidence were higher in Liguria than in Piemonte, suggesting a greater risk of disease spread. We conclude that the mild, Mediterranean climate of Liguria favors the development of a diverse mealybug fauna while only H. bohemicus, known to be tolerant to the severe continental winter temperatures, colonize grapevines in colder viticultural areas.     

Detection of Streptococcus dysgalactiae subsp. equisimilis in equine nasopharyngeal swabs by PCR

Streptococcus (S.) dysgalactiae subsp. equisimilis is responsible for severe diseases in humans, including primary bacteraemia, pneumonia, endocarditis, and toxic shock syndrome. Infection in some animal species can also occur, although a few studies have looked into cross-species infectivity. In horses, S. equisimilis is generally considered infrequent or opportunistic, but has recently been isolated from cases of strangles-like disease. Rapid and sensitive diagnostic techniques could enable epidemiological studies and effective investigation of outbreaks involving these bacteria. In this study, PCR protocols previously described in cattle and in humans to detect the species S. dysgalactiae and the subspecies equisimilis were evaluated to detect specific sequences in equine samples. For this purpose, 99 monolateral nasal swabs were collected from horses from stud farms with a history of S. equisimilis infection and were tested blindly by bacteriological isolation and by single and duplex PCR. DNA for PCR was extracted both from the colonies grown on agar media and from enrichment broth aliquots after incubation with nasal swab samples. S. equisimilis was identified by bacteriological isolation in 23 out of 99 swab samples, and PCR assays on these colonies were fully concordant with bacteriological identification (kappa statistic = 1.00). In addition, PCR of the enrichment broth aliquots confirmed the bacteriological results and detected S. equisimilis in 6 samples more than the bacteriological examination (kappa statistic = 0.84). The PCR protocols appeared to be reliable for the rapid identification of S. equisimilis in equine nasal swab samples, and could be useful for microbiological diagnosis.