A trial was carried out in a plastic house on tomato to assess the efficacy of the chitinolytic fungus Aphanocladium album isolate MX-95 (AA MX-95) against the soil borne pathogen Pyrenochaeta lycopersici and the root-knot nematode Meloidogyne incognita. Treatments were: a) AA MX-95 applied in sub irrigation at 2,5 L./plot (1,2 x 107 CFU/mL., conidial suspension) in pre transplant (20 days), transplant and in post transplant (2 times every 20 days) + root dipping (5 min - conc.1,2 x 107 CFU/mL) at transplant); b) as before indicated without treatment in pre transplant; c) root dipping and d) dazomet (chemical control) applied 30 days before transplant at 600 Kg./ha.Untreated plots served as control. As significant yield increase and a reduction of severity of corky root and nematode attacks were observed in AA MX-95 and dazomet treated plots in comparison to control. High positive correlations were found between the symptoms caused on tomato root by M. incognita and P. lycopersici.

The efficacies of a new promising control agent Aphanocladium album isolate MX-95 (patent N° 0001374382 of the University "A. Moro" of Bari) and quebracho tannins were tested in an open field trial on egglaplant (cv. Black Beauty) in the control of combined attacks of Verticillium dahliae and Meloidogyne incognita. A soil at Valenzano (province of Bari, Apulia region) naturally infested by the soil borne plant pathogen and the root-knot nematode was deeply ploughed, rotavated and subdivided in 3 x 4 m plots spaced 0.5 m each other and distributed in a randomized block design with three replicates/treatment. Treatments were: a) A. album isolate MX-95 (8 l/plot; 1.5 x 107 CFU/ml); b) quebracho tannins (400 Kg/ha); c) Azoxystrobin (20 l/ha, fungicide) + Fosthiazate (30 Kg/ha, nematicide) and d) an untreated control. The isolate MX-95 of A. album was applied in pre and post-transplant (14 days and 10 days, respectively). Tannins and chemicals were applied at transplant. Treatments a) and b) were applied by injector in localised form to the base of each plant. During the growing season the crop received the cultural practices that are common for the area. Fruits were harvested (six times) during crop cycle and main yield parameters such as fruit growth in order to verify ripening kinetics, fruit weight, fruit number/plant and marketable yield were assessed. At the end of the experimental trial, plants from the central row in each plot were uprooted to estimate root gall index caused by the nematode attack according to a 0-5 scale (0 = health root system and 5 = root system completely deformed by numerous and large galls). Severity of Verticillium wilt was assessed according to a 0-5 scale (0 = health plant; 1 = yellowing of the basal leaves; 2 = widespread yellowing; 3 = wilting; 4 = widespread wilting and 5 = death plant). Stem of each plant was transversely cut 3 cm above the soil level and the severity of vascular discoloration (% area affect) (VD) was recorded according a 0-4 scale (0= no vascular discoloration; 1 = 1-10% VD; 2 = 11-50% VD; 3 = 51-75% VD and 4 = 76-100% VD). Nematodes of each plot were extracted from soil samples processing 500 mL soil sub-sample with the Coolen's method. Data were statistically analysed and means compared by Duncan's Multiple Range Test. All treatments had a positive influence on the yield parameters and significantly increased marketable yield in comparison to the untreated control. No statistical difference was evident among the three considered treatments (Table 1). The severity of wilting and symptoms of vascular discoloration were also significantly reduced on plants of treated plots in comparison to the untreated control. A. album isolate MX-95 and quebracho tannins were effective to reduce root gall index and final nematode population density in comparison to the untreated control (P = 0.05) (Table 1).Results demonstrate that the use of the chitynolitic fungus A. album isolate MX-95 and quebracho t

I nematodi galligeni del genere Meloidogyne sono estremamente distruttivi e difficili da controllare, a causa della loro polifagia e della larga diffusione a livello mondiale. La crescente esigenza di una maggiore sostenibilità ambientale delle produzioni agricole impone la sostituzione dei nematocidi di sintesi tradizionalmente impiegati per il controllo di tali parassiti con prodotti a minore impatto sull'ambiente. I formulati basati su metaboliti secondari ad attività nematocida presenti in numerose specie vegetali offrono una concreta alternativa all'uso dei suddetti pesticidi. In particolare, gli olii essenziali ed i loro costituenti presentano un elevato potenziale applicativo, sia se impiegati direttamente in formulazioni nematocide che per lo sviluppo di analoghi derivati di maggiore efficacia. Vengono qui presentati i risultati di due esperimenti su pomodoro, diretti a verificare comparativamente l'attività nematocida nei confronti del nematode galligeno Meloidogyne incognita di trattamenti al terreno con soluzioni di differenti olii essenziali. Nel primo esperimento sono stati messi a confronto gli olii essenziali di Schinus molle, Cinnamomum camphora, Eugenia caryophillata, Cinnamomum zeylanicum e Citrus aurantium, mentre nella seconda prova sono stati utilizzati gli olii essenziali di due specie di Eucalyptus, E. citriodora e E. globulosus. In entrambi gli esperimenti gli olii sono stati distribuiti in acqua alle concentrazioni di 0,05, 0,1 e 0,2 mg kg-1 di terreno quattro settimane prima del trapianto di pomodoro della cv. Rutgers. Le piante sono state mantenute in serra a 25 °C per due mesi, al termine del quale periodo sono stati rilevati i parametri di crescita delle piante, l'indice di formazione di galle sulle radici e la popolazione del nematode sulle radici e nel terreno. Nel primo esperimento tutti i trattamenti con i cinque olii essenziali, tranne quelli con C. zeylanicum alle dosi di 0,05 e 0,1 mg kg-1 di terreno, hanno significativamente ridotto il numero di uova e larve di M. incognita sulle radici di pomodoro rispetto al testimone non trattato. L'effetto dei trattamenti è risultato meno evidente sulla formazione di galle sulle radici. Gli olii di S. molle e E. caryophillata sono complessivamente risultati più efficaci rispetto agli altri tre olii in prova. Nella seconda prova, tutti i trattamenti con gli olii essenziali di entrambe le specie hanno significativamente ridotto il numero di uova e larve e la formazione di galle sulle radici di pomodoro ed il tasso di riproduzione del nematode rispetto al testimone non trattato. L'olio essenziale di E. citriodora ha complessivamente mostrato un' attività nematocida superiore rispetto a quello di E. globulosus. Gran parte dei trattamenti con i due olii essenziali hanno inoltre indotto uno sviluppo dell'apparato radicale delle piante significativamente maggiore rispetto al testimone non trattato, mentre non hanno avuto alcun significativo effetto sulla parte aerea. I risultati

Strains of Streptomycetes isolated from soils of R. Moldova (National Collection of Non-pathogenic Microoganisms IMB ASM) were tested on germination of tomato seeds (Fakeland Marmande varieties), antibacterial and antifungal activities, and suppression of plant parasitic nematodes. For variety Fakel (R. Moldova) strains Streptomyces sp. 11, Streptomyces sp. 47, Streptomyces sp. 49, Streptomyces sp. 154 and Streptomyces sp. 33 significantly increased germination of tomato seeds at 0.5% and 1% dilutions and stimulated formation of root system. However, germination of seeds of variety Marmande (Belgium) were inhibited by strains exametabolites (EM) at 100%, 75%, 50%, 25%, 10% and 5% dilutionsand there was decreasing incline in suppression from 100% to 0.5% dilutions for all the strains. EM of Streptomyces strains showed antagonistic effect on phytopathogenic bacteria and fungi. All the strains demonstrated capacity to decrease the growth of bacteria (Xanthomonas campestris, Agrobacterium tumefaciens, Clavibacter michiganensis) and fungi (Aspergillus niger, Fusarium solani, F. graminearum and Botrytis cinerea). The delayed zone of growth of phytopathogenic bacteria was 15-23 mm and for fungi 10-15 mm. Among strains of Streptomyces from the National Collection of Non-pathogenic Microoganisms, ASM, isolated from soils of R. Moldova the selected strains (Streptomyces sp. 11, Streptomyces sp. 47, Streptomyces sp. 49, Streptomyces sp. 154 and Streptomyces sp. 33) showed nematicidal activities against the root-knot nematodes (Meloidogyne spp.)

The effectiveness of different rates of Dimethyl disulfide (DMDS) applied tothe soil in drip irrigation was tested against the root-knot nematode Meloidogyneincognita in two plastic-house trials on melon and tomato in southern Italy. Thetrials were carried out in naturally infested sandy soils according to a randomizedblock design with 5 replicates per treatment. In both trials the DMDS ECformulation was applied at rates of 400 and 300 L/ha by drip irrigation with PVCdrip lines covered with virtually impermeable film (VIF). DMDS treatments werecompared with 1,3-dichloropropene (140 L/ha) and untreated controls. Plastic VIFwere removed 2 weeks after fumigations. Melons and tomatoes were harvested 5 and7 times, respectively, and the yields recorded. At the end of the crop cycles, thenematode attack on the roots was evaluated according to a 0-5 scale. Soil sampleswere collected in each plot to evaluate soil nematode population. All nematicidaltreatments significantly increased marketable yields of melon and tomato incomparison to the untreated controls (0.48 and 0.16 kg/m2). In the melon trial bothrates of DMDS (400 and 300 L/ha) did not significantly differ from each other (0.71and 0.70 kg/m2, respectively) and from 1,3-D (0.70 kg/m2). The same pattern wasobserved in the tomato trial, in which 2.78 and 2.16 kg/m2 of tomatoes were recordedwith the highest and lowest DMDS rates, respectively, in comparison to 2.78 kg/m2in 1,3-D treated plots. In the melon trial, final nematode population densities inDMDS and 1,3-D treated plots (0.2 eggs and juveniles/ml soil) were significantlylower than that observed in the untreated control (1.7). In the tomato trial, the soilnematode population density observed in the untreated control (18.4 eggs andjuveniles/ml soil) was significantly higher than those recorded in all DMDS and1,3-D fumigated plots (1 eggs and juveniles/ml soil). Also, in both trials and in alltreatments, root gall indices were significantly lower than those observed in theuntreated controls with no significant differences among the fumigant treatments.

Dimethyl disulfide (DMDS), found in nature in several plants belonging to Alliaceae and Brassicaceae families, is able to control soilborne pathogens, weeds and phytoparasitic nematodes. To verify its efficacy three field trials were carried out on carrot in Abruzzo, Apulia and Lazio regions (Italy) (trials A, B and C, respectively) against the carrot cyst nematode Heterodera carotae. The soil was subdivided in plots 50 x 4.8 m for shank application with machines. Each plot was subdivided in 5 sub-plots to provide replications for each treatment. In trials A and B treatments were: i, ii and iii, DMDS as shank formulation (Accolade 99.1%) at the rates of 180, 280 and 380 L/ha. In the trial C treatments were DMDS at rates of 280 and 470 L/ha. In all trials 1,3 D (Telone 97) at 140 L/ha was used as standard with untreated as controls. DMDS was applied three weeks before sowing in a wet soil (70% of the water holding capacity, soil T > 20 °C). Fumigated plots were covered with plastic films virtually impermeable (VIF) for 2 weeks. One week later soil aeration, carrot seeds were sown in each plot. Soil samples were collected before and after treatments and at the carrot harvest. Cysts were extracted by the Fenwick can and their number/100 g dried soil and number of eggs and juveniles inside cysts determined. In the trial B, after fumigations, the extracted cysts from soil of all plots were subjected to an hatching test to verify the vitality of the eggs inside the treated or untreated cysts. At the harvest the weights of marketable and unmarketable carrots were recorded. After treatments and at the harvest no significant differences were evident among treatments including the untreated controls in the number of cysts/100 g dried soil. However, from the hatching tests resulted that the number of viable eggs in the untreated cysts was significantly higher (35%) than those in the different treatments (0.01 - 0.36%). Marketable carrot yield in all treatments and trials was significantly higher than those in the untreated controls. In the trials A and C no significant difference was evident among the DMDS tested rates also compared with 1.3 D, whereas in the trial B the significant highest marketable carrot yield was observed in the highest DMDS rate (380 L/ha). Finally, from our data, DMDS can be considered a very effective fumigant to control carrot cyst nematode.

Two field trials were carried out in Apulia region (Southern Italy) in the years 2012 and 2013 to verify the efficacy of dimethyl disulfide (DMDS) in shank application against the carrot cyst nematode Heterodera carotae and the root-knot nematode Meloidogyne javanica, respectively. DMDS is an innovative soil fumigant for a modern and sustainable soil management. In both trials soil was subdivided in 4 x 50 m plots for shank application of the fumigant. Each plot was subdivided in 5 sub-plots to provide replications for each treatment. DMDS was applied at rates of 180, 280 and 370 L/ha and then covering the soil with VIF plastic film. Untreated soils were used as controls. As chemical controls were considered 1,3 dichloropropene (140 L/ha) in the year 2012 and oxamyl (50 Kg/ha) in the year 2013. In the second trial regarding the control of M. javanica, for the lowest dose of DMDS (180 L/ha) the soil was covered after shank fumigation not only with VIF film but also with TIF film. Films were removed in both trials 15 days later their applications and the soil aired for further 15 days. After soil aeration carrot (cv Bolero in 2012 and cv Exelso in 2013) was sown. In the first year before sowing, soil samples were collected in each plot to extract cysts by the Fenwick can. Cysts, collected from treated and untreated soils, were subjected to a hatching test to verify vitality of their eggs. Hatch percentage in untreated control was significantly higher (35.2%) than those observed in all other treatments (< 0.36%) that were not significantly different each other (P=0.01). At the end of the experiment no significant differences were observed in the number of cysts among the different treatment included the untreated control (P=0.01). The higher and significant marketable carrot yield was recorded in plots treated with DMDS at 370L/ha (575 q/ha) (P=0.01). Other treatments were significantly lower than DMDS at 370 L/ha. In the untreated control no marketable yield was harvested. In the second trial on M. javanica, at the end of the trial, soil samples were collected to extract eggs and juveniles by the Coolen's method. The lowest soil nematode population was observed in soils of plots treated with DMDS at rate of 370 L/ha and it was significant lower than those observed in oxamyl and untreated plots (P=0.01). However no statistical differences were observed among the different rates of DMDS and between the two plastic films VIF and TIF (P=0.01). The highest carrot marketable yield was observed in all three applied doses of DMDS covered with VIF (P=0.01). In untreated control marketable yield was significantly lower than those observed in all other treatments (P=0.01). Results providing evidence that DMDS represents a sustainable technical solution for controlling root-knot and cyst carrot nematodes. Further trials to control nematodes, combining DMDS with not chemical solutions, are ongoing for a sustainable soil management.

The potato cyst nematode (PCN) Globodera pallida causes considerable yield losses to the main Italian and Dutch potato growing areas. Dimethyl disulfide (DMDS), on the base of recent studies can control phytoparasitic nematodes, soilborne pathogens and weeds. Two field trials were carried out to verify its efficacy on potato in Italy (Conversano, Bari province) and in The Netherlands (Nieuwe Pekela, Groningen province) (trial IT and NL, respectively) against G. pallida. The soil was subdivided in plots of 200 m2 for shank application with machines. Each plot was subdivided in 5 (IT) or 4 (NL) sub-plots to provide replications for each treatment. In both trials DMDS treatments were: i, ii and iii, Accolade in shank formulation (DMDS, 99.1%) at the rates of 180, 280 and 380 L/ha. Mocap (ethoprofos 10%) was used at 100 Kg/ha and Monam (metam sodium 510 g/L) at 300 L/ha, as standard controls in Italy and The Netherlands, respectively. The lowest dose of DMDS in the trial NL was also applied mixing the soil by a spading machine and then pressing it with a roller. In both trials untreated soils were used as controls. DMDS was applied three (IT) and five (NL) weeks before sowing. Fumigated treated plots were covered for two (IT) and four (NL) weeks with plastic films virtually impermeable (VIF) with the exception of the metam-Na. One week later soil aeration, potato were sown in each plot. Soil samples were collected before and after treatments and at the potato harvest. Cysts were extracted by the Fenwick can and their number/100 g dried soil and number of eggs and juveniles inside cysts determined. At the harvest the weights of potato tubers were recorded. No significant differences were evident among treatments including the untreated controls in the number of cysts/100 g dried soil in the I trial. In NL trial all treatments significantly reduced the number of cysts in comparison to the untreated control. Marketable potato yields (MPY) in all DMDS and Ethoprofos (IT) and metam-Na (NL) treatments were significantly higher than in the untreated control. Among DMDS treatments at different rates in both trials no significant differences were recorded. MPY in Ethoprofos treatment was significantly lower than those recorded in all DMDS treatments. Concluding, DMDS treatments could be positively and favourably considered in PCN sustainable control.

Abamectin is a mixture of macrocyclic lactones produced by the actinomycete Streptomyces avermitilis especially known for its insecticidal and acaricidal activities. The effectiveness of an abamectin formulation (Vertimec® EC) for the control of the widespread and polyphagous root-knot nematode Meloidogyne incognita was investigated in a pot experiment on tomato (cv. Rutgers) in glasshouse. Different concentrations (0, 0.125, 0.25, 0.5, 0.75 and 1 mL/L) of an aqueous solution of the abamectin formulation were applied as a single treatment at transplant or at transplant plus 15 days later to a soil uniformly infested by M. incognita (7 eggs and juveniles/mL soil). Fenamiphos (62.5 c.p. L/Ha) and untreated soil were used as controls. Pots were arranged on benches in a glasshouse at 25 + 2°C, according to a randomized block design with five replications for each treatment. After two months tomato plants were uprooted and fresh and dry top weights were recorded. Severity of the nematode attack (root gall index - RGI) was assessed on each tomato root system according to a 0 - 10 scale. Final nematode population density was calculated summing nematodes extracted from tomato roots by sodium hypochlorite's method and those extracted from 500 mL soil with the Coolen's method. Reproduction rate (r=Pf/Pi) was calculated as ratio between final and initial population density. Data from the experiment were subjected to analysis of variance and means compared by the Least Significance Difference's test (LSD) using the PlotIT program. All abamectin solutions significantly increased fresh and dry top weight in comparison to the untreated control. Root gall index, eggs and juveniles/g root, soil nematode population density and reproduction rate were significantly reduced by all the abamectin treatments in comparison to the untreated control (P=0.05). Nematode population density and reproduction rate in the soil treated with the three highest concentrations of abamectin solution were not significantly different from those calculated in the fenamiphos treated soil.

Among the natural products extracted from plants, tannins have been reported to possess antihelmintic properties especially for gastrointestinal nematodes in ruminants. Also, they are toxic to a wide range of fungi, bacteria and yeasts. Therefore, an in vitro and a glasshouse pot experiments were undertaken to evaluate the effect of chestnut tannins on the control of the root-knot nematode Meloidogyne javanica. In the in vitro experiment, different tannin concentrations in a geometric scale (from 0.32 to 20.48 g/l), were tested for their effect on hatching of the nematode, whereas in the pot experiment, 100, 250 and 450 g/m2 of tannins in aqueous solutions, were used in pots at transplant or at transplant and two weeks later for their effect on nematode control. In both experiments treatments were compared to untreated and fenamiphos-treated controls. In vitro a nematostatic effect of tannins was observed, whereas in the pot experiment a significant reduction of eggs and juveniles/g root, total population density and reproduction rates of the nematode were recorded. The anatomical changes induced by M. javanica in tomato roots treated with tannins did not differ from those produced by this and other Meloidogyne species on various hosts reported earlier.

Essential oils (EOs) have been largely investigated for their biocidal activity on root-knot nematodes, Meloidogyne species, the most economically relevant group of phytoparasitic nematodes. Adversely, experimental activity on the effect of EOs on cyst-forming nematode species is very poor, though sustainable management of these nematode species should need new control tools alternative to chemicals, such as EOs-based formulates. EOs of Eugenia caryophyllata (L.) Merr. & L.M. Perry and Schinus molle L. are reported for a wide spectre of biological activities, among which also a biological activity against plant insect pests and fungal pathogens. The major component of the EO of E. caryophyllata is usually considered to be eugenol, with ?-caryophyllene and lower amounts of other components such as benzyl alcohol. The major constituents in S. molle EO were ?-phellandrene and ?-phellandrene, with variable amounts of ?-pinene, p-cymene and ?-pinene.An experimental activity was carried out to assess the in vitro effect of treatments with EOs on egg hatch of different cyst nematodes. Results of a hatching test with cysts of the potato and carrot cyst nematodes, Globodera rostochiensis and Heterodera carotae, respectively, treated with EOs of E. caryophyllata and S. molle are reported in this work. Batches of 100 cysts of each nematode species were exposed to 125, 250 and 500 ?l L-1 solutions of each EO for 12, 24 and 48 hours. There were 4 replications for each concentration x exposure time. Non-treated cysts were used as control. After treatments, hatching test of G. rostochiensis and H. carotae continued over 7 weeks in sodium metavanadate (0.6 mM) or zinc chloride (10 mM) solutions, respectively, counting emerged juveniles at weekly intervals. At the end of the experiment, unhatched eggs were determined and final cumulative percentage hatch was calculated. The EO of E. caryophyllata significantly reduced hatch of eggs of G. rostochiensis only after a 24 or 48 hour exposure to the highest concentration, whereas cyst treatment with the 125 ?l L-1 solution resulted in a significant increase of hatched eggs compared to the non-treated control. Adversely, all treatments with the same EO significantly reduced the percentage hatch of H. carotae eggs, with the exception of the 12 hour exposure to the 125 ?l L-1 solution. Compared to non-treated cysts, all treatments with the EO of S. molle did not significantly affect the hatch of G. rostochiensis, whereas significantly reduced the percentage hatch of H. carotae.

This paper reports the effects of several amendments on the root-knot nematode Meloidogyne incognitapopulation, the melon production and some soil features. Treatments were: (i) virgin olive pomace (VOP)at rate of 22 t ha-1, (ii) olive pomace based compost (COP) at rate of 9 t ha-1, (iii) chicken manure (CM)at dose of 3 t ha-1, (iv) CM plus the nematophagous fungus Paecilomyces lilacinus, (CMB) at 3 t ha-1and4 kg ha-1, respectively and (v) the naturally infested and untreated control (CON). The doses of the amend-ments were calculated to provide the same amount of total nitrogen (ca. 120 kg ha-1). The experimentwas carried out following a randomized block design with four replications for each treatment.The melon production significantly increased with CM and CMB in comparison to all other treatments(P <= 0.05). VOP and COP enlarged the soil organic C, the total and organic N, and the exchangeable Kcontent, while CM and CMB increased the inorganic N and available P content. The root gall index, the soilnematode population density and the reproduction rate were significantly reduced in all amended plotsin comparison to CON. The use of P. lilacinus did not influence significantly the M. incognita population,suggesting that it is possible to manage the soil fertility and the root-knot nematode population by usingorganic amendments alone.

The influence of ten initial population levels (0, 0.1, 0.2, 0.4, 0.8, 1.6, 3.2, 6.4, 12.8 and 25.6 second stage juveniles/cm3 soil) of Meloidogyne incognita and M. javanica on olive cultivar Zard growth was studied in two pot trials. Ten month old self-rooted olive cuttings were individually transplanted into 2 000 cm(3) pot and inoculated with the defined initial populations, of both nematode species. Plants were grown in glasshouse for 13 months, then they were uprooted and plant growth (percent growth increase of main shoot length, number of nodes on main shoot, top and root fresh and dry weights and root length) and nematode parameters (root gall index, J(2)/g root, final nematode population density and reproduction rate Pf/Pi) were recorded. Results showed that cv. Zard was more susceptible to M. javanica than to M. incognita. A significant reduction of main shoot length growth 37.6 % and 10.7 % was observed at 0.1 and 12.8 juveniles/cm3 soil of M. javanica and M. incognita, respectively, in comparison to uninfested plants. Root systems of olive plants grown in M. incognita or M. javanica infested soils were galled within the gall index range 1.4 - 6. No significant differences were observed in the number of nodes on main shoot, top and root fresh weights and root dry weight at high levels of M. incognita Pi. A tolerance limit (T) of 0.4 juveniles/cm3 soil was estimated for olive plants cv. Zard to M. javania. The use of resistant olive rootstock or selected cultivars is recommended to minimize or to limit damage of nematode infections in nurseries and to prevent secondary attacks of soil borne plant pathogens especially Verticillium dahliae.

Dimethyl disulfide (DMDS), in shank application, was used in two field trialson carrot in Emilia-Romagna and Apulia regions (Italy) against the root-knotnematode Meloidogyne incognita and the cyst nematode Heterodera carotae. The soilwas subdivided in plots 30 or 50 × 4.8 m for shank application with machines. Eachplot was subdivided in 4 or 5 sub-plots to provide replications for each treatment. Inthe trial with M. incognita treatments were: i) DMDS at the rate of 250 L/ha, ii)granular oxamyl (60 kg/ha) followed by the commercial product Micosat F WP NE(6 kg/ha) and iii) granular oxamyl (60 kg/ha) followed by Bioact WG includingPaecilomyces lilacinus (4 kg/ha). In the trial with H. carotae treatments were: i, iiand iii, DMDS at the rates of 370, 280 and 180 L/ha and iv) 1,3 dichloropropene(140 L/ha). In both trials untreated soil was used as control. DMDS was applied3 weeks before sowing in a wet soil (70% of the water holding capacity) with atemperature higher than 20°C. The DMDS and 1,3-D treated plots were thencovered for 2 weeks with a very impermeable plastic film (VIF). After one week ofsoil aeration carrot seeds were sown in each plot. At the end of the crop cycle in theexperiment with M. incognita, DMDS treatment significantly increased marketablecarrot yield (9.7 kg/m2) in comparison to oxamyl followed by either Bioact WG(7.2 kg/m2) or Micosat F WP NE (1.8 kg/m2) and the untreated control (3.9 kg/m2).Also M. incognita soil population density was significantly decreased by DMDStreatment (Pf/Pi=0.2) in comparison to the untreated control (Pf/Pi=14.4) giving thelowest reproduction factor. M. incognita infestation on the roots was assessedaccording to a scale of 6 classes from 0 to 5. In the experiment in Apulia thesignificantly highest marketable carrot yield (5.8 kg/m2) was observed in plotstreated with 400 L/ha of DMDS in comparison to all other treatments and to theuntreated control (0.0 kg/m2). H. carotae soil population density in the untreated control (10.8 eggs and juveniles/g soil) was significantly higher than those observedin the highest DMDS dose (4.5 eggs and juveniles/g soil).

An in vitro study was conducted to investigate the effect of soil exposure for 2 - 2,048 hours to a 20 - 45 °C temperature range on the hatching behaviour of eggs of the cyst nematodes Heterodera carotae and Heterodera schachtii. A complete mortality of H. carotae eggs occurred for treatments from 512 h at 40 °C to 64 h at 45 °C, whereas almost 99% of H. schachtii eggs were killed by 256, 128 and 16 h exposures at 40, 42.5 and 45 °C, respectively.

Plant parasitic nematodes attack a wide range of crop species. The survival of one quarantine plant parasitic nematode species attacking potatoes, Globodera rostochiensis patotype Ro1 was evaluated in in vitro and pot experiments by tannin aqueous solutions at different concentrations. In the in vitro experiment a different tannin concentration were tested for effect on the egg hatch of the nematode. All the tannin concentrations tested exerted a highly significant reduction of egg viability from 56 to 87% in comparison to the untreated control. In the pot experiment, tannins, as aqueous solutions at rates of 100. 250 and 450 g.m"2, were applied to soil at two different application times (at sowing and at sowing and two weeks later). Ali tested doses were effective to reduce the number of cyst.100 g"' soil, eggs and juveniles.g"1 soil and reproduction rate in comparison to untreated control. The number of eggs and juveniles/cyst was not influenced by the different applied rates of tannins.

Winter wheat is the largest cultivated crop in the world with over 220 million hectares. In the Republic of Moldova about 25% of the arable land, corresponding to 40% of the surface occupied by cereal crops, is cultivated to winter wheat. Considering the importance of winter crops in all agro-ecological areas of R. Moldova it is important to evaluate the severity of pathogens and plant parasitic attacks (number of species, frequency and host ranges) in order to adopt useful control measures. Recently different researches on the use of exometabolites (EMs) of genus Streptomyces, with nematostatic/nematicidal, antifungal and phyto-stimulating plant growth properties, have been carried out against soil borne plant pathogens and phytoparasitic nematodes. Combined attacks of pathogens and parasites on winter wheat are particularly significant because of they produce yellowing, rots, necrosis of roots and stems and severe yield losses in many Western and Eastern Europe areas including all ecological zones of R. Moldova. Winter cereals are affected by more than 100 species of pests, including phytopathogenic agents (Fusarium graminearun, F. avenaceum, F. nivale, F. culmorum, F. oxysporum, etc.) alone or in combination with complexes of phyto-nematodes of the genus Pratylenchus, Tylenchus, Aglenchus, Rotylechus, Helicotylenchus, Paratylenchus, Tylenchorhynchus, Ditylenchus, Merlinius, Criconemoides and micofages, ectoparasitic species which level range between 5 and 30 individuals/g soil with incidence from 3 to 35% of the total cultivated area according to the period, zone, sector and grain growth phase. Fusariosis with symptoms of root and stem rots (regarding 10-40% of the cereal cultivated surface) are particularly dangerous in the early stages of wheat growth in the late autumn - early spring. Streptomycetes are widely distributed in nature, especially in many soils, and they have been considered in the recent years not only as antibiotic producers, but also for the production of other biologically active substances as enzymes, amino acids, vitamins, lipids and phyto-hormones which influence seed germination, plant growth and crop yield.Considering previous studies on streptomycetes EMs on tomatoes, cucumbers, tobacco and corn seeds, new experiments were carried out on winter wheat seeds using EMs produced by streptomycetes isolated from soils of R. Moldova. EMs of the Streptomyces strains 9, 10, 12, 66 and 205 had positive effect, increasing number of roots (14.5-19.8%) and root length (20.3-26.1%) in comparison to the untreated control. Also an antifungal activity was highlighted against Alternaria alternata, Fusarium solani and F. graminearum. Nematicidal activity against root-knot nematodes was observed using Streptomyces sp. 9 and 205 in comparison to untreated control.On the base of our results the use of EMs from Streptomyces strains isolated from soils of the R. Moldova could be favorably considered in cereal crops protection.

Acetophenones are a class of aromatic compounds frequently produced by plants as a response to a stress or as a protection reaction against parasites or herbivors. In the present research, we investigated the role of the carbonyl moiety in a series of commercially available aromatic ketones tested for their nematicidal activity against the root-knot nematode Meloidogyne incognita (Kofoid et White) Chitwood. Interestingly, we found that the presence of electron withdrawing groups onto the aromatic ring were highly efficient in inducing nematodeparalysis and death (EC 50/24h between 2.5 and 54.8 mg/L and EC50/72h between 2.3 and 65.6 mg/L, respectively).Moreover, a chlorine atom in ?-position to the carbonyl functionality emerged as a key residue in improving acetophenones activity. In this paper, the nematicidal ability of a series of differently substituted acetophenones is reported and the preliminary structure-activity relationship studies are also discussed. We also performed an in vitro GC-MS metabolomics analysis on the potato cyst nematode Globodera pallida, after treatment with 2,4?-dichloroacetophenone (6) at 100 mg/L for 24 h. The 1-dodecanol and talose were evidencedas the main upregulated metabolites, suggesting a possible V-ATPase dysfunction.

Annually significant tomato yield losses are caused by climate changes and pests and diseases attacks. Root-knot nematodes (Meloidogyne spp.) are the most damaging pests on the base of their wide distribution in tropic and sub-tropic climates and their wide host range. They modify host root tissue using effector proteins to create feeding sites as their source of nutrition. To develop alternative control strategies to nematode infections, we have considered aqueous ozone treatments (AOT) as a possible tool. Although gaseous ozone is phytotoxic, previous studies demonstrated that ozone in aqueous phase is effective to contain pests. The purposes of this study were to confirm the impact of AOT on tomato-M. incognita interaction and to analyse its role in plant defence response. Ozonated water was produced in situ by an ozone generator at 10 ppm and directly applied to tomato roots. Treatments were performed daily (10 ml/pot) for 4 days as soil drench on 14 days old tomato plants maintained in a growth cabinet (25 ± 2 °C). Nematode infection control by ozone treatments was set up by infecting untreated and treated plants with J2s of M. incognita. A number of plants were used 2, 4 and 7 days after nematode infection for biochemical and molecular analysis. Other plants were transferred in a glasshouse at 25 ± 2 °C and after 60 days they were uprooted and dry shoot and root fresh weights recorded. Root gall index, eggs and J2s/g root, total nematode population density and reproduction rate were evaluated. AOT significantly decreased severity of root gall index and soil nematode population in comparison to untreated plants. Tomato sensitivity to AOT was verified as reactive oxygen species production (ROS) in both treated and untreated root galls and compared with uninfected treated and untreated roots. A higher significant ROS production was observed in ozone treated uninfected roots in comparison to treated and untreated infected roots. As antioxidant mechanisms play an important role in the response of plant to the combination of abiotic and biotic stresses, the effect of AOT on the expression of different genes involved in ROS scavenging, such as CAT, SOD and APX, was also evaluated. AOT by altering gene expression, ROS production and inducing a non specific defense response can be considered a useful tool to contain nematode infection.Aknowledgement: This work was supported with funds provided by the National Research Council for the project "Innovazione e Sviluppo del Mezzogiorno - Conoscenze Integrate per Sostenibilità ed Innovazione del Made in Italy Agroalimentare".

Melia azedarach fruit extracts have recently raised a substantial interest for their use in crop protection against phytoparasitic nematodes. The effect of M. azedarach on the root-knot nematode Meloidogyne incognita on cucumber, as well as the effect on the plant deference mechanism, is reported herein. Crushed fruits of M. azedarach, tested in the soil at the rates of 30 and 60 g kg(-1), exhibited nematicidal activity similar to the one of fenamiphos (0.02 g a.i. kg(-1)) in terms of nematode population in roots and soil as well as reproduction rate. M. azedarach water extracts, rich in aldehydes, alcohols and carboxylic acids, showed nematicidal activity against M. incognita. Moreover, all M. azedarach treatments decreased the activities of catalase (CAT) and peroxidase (POX) involved in host H2O2 detoxification. Soil application of M. azedarach fruits could be favourably considered in the control of M. incognita on cucumber in a sustainable agriculture, since they act directly as nematicidals. Furthermore, M. azedarach elicits plant defence and helps the host to fight the nematodes infestation in an indirect waY

Phytoparasitic nematodes can cause severe yield losses to numerous vegetables therefore it is necessary their control. To evaluate the efficacy of two essential oils (EOs) isolated from Thymus vulgaris and Cinnamomum camphora in the control of the potato cyst nematode Globodera rostochiensis an in vitro experiment (hatching test) was carried out.Batches of 50 G. rostochiensis cysts, of similar size, extracted by the Fenwick can from infested soil, were placed in Eppendorf tubes (2mL) in 1 mL of distilled water before treatments with EOs. The two EOs solutions (10% ethanol, v/v) in appropriate amount were diluted in distilled water containing 0.3% polysorbate surfactant 20 (Tween-20) (v/v) to obtain 4, 8 and 16 ?L mL-1 solutions. The EOs were then used to treat G. rostochiensis cysts adding to each Eppendorf tube 1 mL of each solution. Then, tubes were closed and cysts exposed to the EOs treatment for 48 hrs. Untreated cysts were used as control. Four replications for each treatment were considered. After treatments cysts were removed from the Eos solutions, rinsed in distilled water and put in 2 cm diam sieves (215 ?m aperture) placed in 3.5 cm diam Petri dishes. In the hatching test potato root leachate was used as natural hatching agent and the test was carried out in a growth cabinet at 21 °C. Emerged juveniles were removed and counted every week over a 6 week period. The hatching agent was renewed at each observation time. Numbers of second stage juveniles emerging every week were expressed as cumulative percentages of the total egg content of the cysts.Percentage hatch of G. rostochiensis eggs was significantly reduced (P < 0.05) by the different concentrations of both Eos, although significantly higher eggs mortality was observed in cysts immersed in thyme oil. Our results suggest that essential oils of Thymus and Cinnamon could be favourably considered in a sustainable control of the potato cyst nematode, although further studies are needed.Acknowledgement: This study was supported by the VEGA scientific grant agency, grant No. 2/0079/13 and by the Agreement between CNR (Italy) and ASM (R. Moldova) for the years 2015-16 for the project "Agro technical and biological methods for a sustainable control of quarantine cyst nematodes in Italy and in R. Moldova

The fertilizing and nematicidal effects of three organic amendments were evaluated in a pot experiment on melon plants infested by the root-knot nematode Meloidogyne incognita. A soil artificially infested with 4 eggs and juveniles/ ml soil of the nematode was amended with: a) virgin olive pomace (VOP); b) composted olive pomace (COP); c) chicken manure based fertilizer (CM) and d) chicken manure based fertilizer combined with the biological control agent Paecilomyces lilacinus strain 251, brand name BioAct WG (CMB). VOP was applied at doses of 11 (VOP-A), 22 (VOP-B) and 44 t/ha (VOP-C); COP at 4.5 (COP-A), 9 (COP-B) and 18 t/ha (COP-C); CM at 3 t/ha and CMB at 3 t/ha combined with 4 kg/ha of BioAct WG. Untreated soil was used as control. The treatments CM, CMB, VOP-B and COP-B were established on the basis of N requirement of melon plants (120 kg/ha) taking into account soil and amendments N availability. Two weeks later amendment application and nematode inoculation, the soil was poured in 4.8 l clay pots which were arranged in a greenhouse according to a randomized block design with ten replications for each treatment. A one-month old melon seedling (cv. Galia) was transplanted in each pot and organic farming management practices were used during the growing period. At the end of the experiment, 60 days after transplant, plants were uprooted and height, fresh and dry shoot and root weights were recorded. Root gall index, on the roots, caused by the nematode attack, was estimated according to a 0-5 scale. Final nematode population density and reproduction rate were also calculated for each pot. All data were subjected to statistical analysis of variance (ANOVA) and means compared according to Least Significant Difference's Test. Nematode population and root infestation were significantly suppressed by the addition of all amendments, compared to untreated control. However, CM and CMB resulted in a total more suppressive effect and in a significantly higher plant growth in comparison to all the other treatments. A significant correlation was found between root gall index and eggs and juveniles/g root and final nematode population density. No signifycant correlations were found between nematological parameters or plant growth parameters and amendment doses.

New pesticides based on plant extracts have recently gained interest in the development of nontoxic crop protection chemicals. Numerous research studies are focused on the isolation and identification of new active compounds derived from plants. In this manuscript we report about the use of the Mediterranean species Capparis spinosa as a potent natural nematicidal agent against the root knot nematodes Meloidogyne incognita. Leaves, stems, and caper buds of Cap paris spinosa were used to obtain their methanol extracts (LME, SME, BME) that were successively in vitro tested against second stage nematode juveniles (J2). In terms of paralysis induction, the methanol extract of the stem part (SME) was found more effective against M. incognita and then the caper methanol buds and leaves extracts. The chemical composition analysis of the extracts carried out by GC/MS and LC/MS techniques showed that methylisothiocyanate was the main compound of SME. The EC50 for SME after 3 days of immersion was 215 +/- 36 mg/L. The constituent components of SME such as 2-thiophenecarboxaldehyde and methylisothiocyanate were successively in vitro tested for their nematicidal activity against J2. Both compounds induced paralysis on root knot nematodes ranking first (EC50 = 7.9 +/- 1.6, and 14.1 +/- 1.9 mg/L respectively) for M. incognita. Moreover, 2-thiophenecarboxaldehyde showed a strong fumigant activity.

Recently, tannins have been reported for their nematicidal activity against the root-knot nematode Meloidogyne javanica both in vitro and in pot experiments in addition to a biocidal effect on a wide range of fungi, bacteria and yeasts. However, no information is available on the effect of these polyphenols on plant parasitic cyst nematodes. Therefore, an in vitro and a pot experiments on potato were undertaken to investigate the nematicidal activity of tannin aqueous solutions at different concentrations on the potato cyst nematode Globodera rostochiensis. In the in vitro experiment different tannin concentrations in a geometric scale (from 0.32 to 20.48 g/l) were tested for their effect on the egg hatch of the nematode. All tested tannin concentrations were effective to reduce egg viability from 56 to 87%, in comparison to the untreated control. In the pot experiment, tannins, as aqueous solutions at rates of 100, 250 and 450 g/m(2), were applied to soil at two different application times (at sowing and at sowing and two weeks later). All tested doses were effective to reduce the number of cyst/100 g soil, eggs and juveniles/g soil and reproduction rate in comparison to untreated control. The number of eggs and juveniles/cyst was not influenced by the different applied rates of tannins.

Numerous studies on the interaction between soil borne plant pathogens and phytoparasitic nematodes both on vegetables and fruit trees have been conducted. In the last two decades, in several areas of the R. Moldova, devoted to tomato cultivation in plastic houses, severe disease symptoms, caused by soil borne pathogens, have been found in association with root-knot nematode attacks (Meloidogyne spp.). In many countries to limit damages caused by these plant pathogens and parasites, plant protection is based on chemical treatments (dazomet, oxamyl, fosthiazate and more recently DMDS). In every case new control integrated strategies are required and they must be environmentally safe and economically convenient at the same time. Among these alternatives, the use of new biological active substances, with phyto-stimulating, antimicrobial and nematicidal properties, synthesized by soil microorganisms of the genera Streptomyces, Azotobacter, Pseudomonas and Bacillus can reduce the population level of harmful organisms (plant-parasitic nematodes, phytopathogenous insects, phytopathogenic bacteria and virus) below the threshold of damage. The intensive and repeated use of plastic-houses dramatically increases Meloidogyne spp. attacks with severe yield losses and economic damages. Implementation and application of biological control measures and new cultivation agro-technologies are justified by the expensive chemical treatments which cause soil pollution and a scarce qualitative and security food production.Estimation of phyto-sanitary situation on tomato cultivated in plastic-houses (surface 20 ha) in R. Moldova was carried out during winter-spring period. Some dangerous sedentary endoparasites as Meloidogyne incognita (Kofoid et White) Chitw. and Pratylenchus pratensis (de Man) were extracted from soil samples collected during the survey. In some plastic-houses the root-knot nematode populations exceeding the tolerance limit of tomato plants to the nematode (3.3 eggs and juveniles/cm3 soil). Density of nematode communities including plant parasitic and free-living nematodes varied from 6.75 to 10.25 individuals/cm3 soil. Virus infections (10-15% infected plants) were also observed. Symptoms were mainly due to: a) viral infections by Lycopersicum virus 3 Smith and b) Marmor tabaci var. lycopersicum.Moreover, tomato plants were also affected by Phytophtora infestant (Mont.) de Bary (15-25 %) showing yellow spots and necrosis on leaves and stems (late blight of tomato) and by Fusarium oxysporum Schl. f. sp. lycopersi (5-10 %) with plants showing wilting symptoms.Effectuated monitoring by us served basic element in providing protection integrates methods by decrease phytoparasitic and phytopathologic affections on tomatoes in greenhouse.The researches were made by bilateral project Italy/R. of Moldova Potential of biological active substances to reduce Meloidogyne harmfulness on horticultural crops in greenhouses". Registry code No 13.8

Few studies have been carried out on the effect of ozonated water (O3wat) on the oxidative stress of root systems and, in particular,in combination with biotic stress. The aim of this study was to determine whether aqueous ozone is effective in the control of root-knot nematode (RKN) infection and to investigate the concomitant changes in the basal defence system. A tomato cultivar susceptible to Meloidogyne incognita was treated with O3wat as a soil drench. No negative effects were seen following ozone application in comparison with the control under the exposure conditions used. The treatment reduced significantly the nematode infection rate and induced changes in the morphology of nematode feeding sites, some of which were characterized by visible symptoms of senescence. The antioxidant response, as well as parameters of oxidative damage, were examined in untreated and Owat-treated galls at 2, 4 and 7 days after inoculation and compared with uninfected roots. High levels of reactive oxygen species (ROS), H32O and malondialdehyde were generated in galls in response to combined abiotic and biotic stresses. Throughout the experimental period, the activities and relative transcript levels of the antioxidant enzymes catalase, superoxide dismutase and ascorbate peroxidase produced different responses when exposed to ozone treatment and/or infection. The results demonstrate how O32wat protects tomato against the RKN M. incognita through the modulation of basal defence mechanisms.

According to the necessity to find new control strategies at low environmental impact against plants parasitic nematodes, a pot experiment on tomato (cv. Super Marmande) was carried out to verify the efficacy of soil ozonated water treatments against the root-knot nematode Meloidogyne incognita. Treatments were applied at transplant and for 15, 30 and 60 days and compared with: a) untreated control and b) fenamiphos treatment (synthetic nematicide) at 62.5 L/Ha used as controls. Pots were arranged, according to a randomised block design, in a glasshouse (25 + 2°C) on benches with ten replications for each treatment. Ozonated water for watering was produced in situ by an ozone generator dissolving ozone gas into the water until an ORP of 800 mV. At the end of the experiment, 60 days after transplant, plants were uprooted from each pot to estimate root gall index (RGI), caused by nematode attack, according to a 0-10 scale. Nematodes extracted from roots by the sodium hypochlorite's method and those extracted from 500 mL soil with the Coolen's method were used to calculate for each pot the final nematode population density and the reproduction rate (r=Pf/Pi). All data from the experiment were subjected to analysis of the variance (ANOVA) and means compared by the Least Significant Difference's Test using the software PlotIT. Root gall index was significantly reduced in all treatments with ozonated water in comparison to the untreated control, although it was significantly higher than those observed in the chemical control. Final nematode population density and reproduction rate were significantly lower in soil ozonated water treatments for 30 and 60 days compared with the untreated control. No significant difference was observed between these two last treatments and fenamiphos. Histological observations on M. incognita galled tomato roots, treated with ozonated water for different application times, were made to compare the anatomical changes induced by the nematode among the different treatments. In the roots of the untreated control and in those treated only for a limited time (15 days) were observed well developed polinucleate giant cells (feeding sites) associated with fully developed females able to deposit a large number of eggs. Undersized giant cell feeding sites associated with immature females, with reduced ability to produce large number of eggs, were noted in tomato roots treated with ozonated water for a longer exposure time (30 and 60 days). In the chemical control was observed well developed giant cells and mature female with limited egg production.No statistical difference was noted in the percentage of hatch from egg-masses produced in the different treatments, including the controls, subjected to an hatching test.

Severe infections of parsley plants and soil infestations with Meloidogyne javanica during an autumn surveys for the pathogenic root-knot nematode infestations were found in Monopoli at Bari province in Southern Italy. This unusual severe infection of parsley, considered a winter crop, was possibly instigated by a very warm autumn from the previous year. Nematodes were extracted from soil samples according to the Coolen's method. Morphological analysis (based on stylet length, tail length and shape, adult females perineal pattern, excretory pore position and Ep/stylet ratio) and molecular studies were used for the nematode characterization and identification. In the soil of infested area a severely deformed root systems were observed, showing a galling rate = 2.5 - 4 (scale 0-5) and a soil nematode population densities ranging from 350 to 2,730 eggs and J2 per 5 g of fresh root. M. javanica attack on parsley roots is a limiting factor for plant growth. Considering that curly-leaf parsley varieties resistant to the nematodes are not yet available control strategies must be focused on reduction of soil infestation level below tolerance limit of the target nematode species. Due to the higher cost and reduced availability of fumigant and non-fumigant nematicides, soil solarization, organic amendments or biological control approaches should be preferably used as alternatives.

Stunted cabbage ('Lupini') associated with severe soil infestations by a cyst-forming nematode were observed in large patches of open fields in Castellaneta, province of Taranto, southern Italy. Morphological traits based on mature cysts, males, and second-stage juveniles (J2s) and molecular analysis of ribosomal DNA (D2 to D3 expansion segments of 28S and internal transcribed spacer [ITS]1 region) were used to identify the species. ITS1 sequence information supported the identity of Heterodera cruciferae, also showing a high degree of similarity to other species of the Heterodera Goettingiana group, including H. goettingiana, H. carotae, and H. urticae. Nematodes successfully established permanent feeding sites in cabbage roots which caused cellular alterations in the root cortex, endodermis, pericycle, and vascular cylinder by inducing typical multinucleate syncytia. Syncytial cytoplasm was granular and dense, with variously sized vacuoles and hypertrophied nuclei with nucleoli. Cabbage plant growth was also reduced in pathogenicity tests. The relationship between the initial nematode population density in soil and shoot plant weight was well described by the Seinhorst's equation. Tolerance limits with respect to shoot plant weight of cabbage to H. cruciferae was estimated as 1.50 units of eggs plus J2s/cm3 of soil. The minimum relative value (m) for plant height was 0.71 at an initial nematode population density of (Pi) >= 64 units of eggs plus J2s/cm3 of soil. The maximum nematode reproduction rate (Pf/Pi) was 4.6 times that of the initial population density of 8 units of eggs plus J2s/cm3 of soil.

Different maleimide derivatives were synthesized and assayed for their in vitro activity on the soil inhabiting, plant-parasitic nematode Meloidogyne incognita, also known as root-knot nematode. The compounds maleimide, N-ethylmaleimide, N-isopropylmaleimide, and N-isobutylmaleimide showed the strongest nematicidal activity on the second stage juveniles of the root-knot nematode with EC50/72h values of 2.6 +/- 1.3, 5.1 +/- 3.4, 16.2 +/- 5.4, and 19.0 +/- 9.0 mg/L, respectively. We also determined the nematicidal activity of copper sulfate, finding an EC50 value of 48.6 +/- 29.8 mg/L. When maleimide at 1 mg/L was tested in combination with copper sulfate at SO mg/L, we observed 100% mortality of the nematodes. We performed a GC-MS metabolomics analysis after treating nematodes with maleimide at 8 mg/L for 24 h. This analysis revealed altered fatty acids and diglyceride metabolites such as oleic acid, palmitic acid, and 1-monopalmitin. Our results suggest that maleimide may be used as a new interesting building block for developing new nematicides in combination with copper salts.

The nematicidal activity of selected aromatic aldehydes was tested against the root knot nematode Meloidogyne incognita. The most active aldehyde was phthalaldehyde (1) with an EC50 value of 11 +/- 6 mg/L followed by salicylaldehyde (2) and cinnamic aldehyde (3) with EC50 values of 11 +/- 1 and 12 +/- 5 mg/L, respectively. On the other hand, structurally related aldehydes such as 2-methoxybenzaldehyde (21), 3,4-dimethoxybenzaldehyde, and vanillin (23) were not active at the concentration of 1000 mg/L. By liquid chromatography-mass spectrometry the reactivity of tested aldehydes against a synthetic peptide resembling the nematode cuticle was characterized. At the test concentration of 1 mM, the main adduct formation was observed for 3,4-dihydroxybenzaldehyde (22), 2-methoxybenzaldehyde (21), and 3,4-dimethoxybenzaldehyde. Considering that 2-methoxybenzaldehyde (21) and 3,4-dimethoxybenzaldehyde were not active against M. incognita in in vitro experiments led us to hypothesize a different mechanism of action rather than an effect on the external cuticle modification of nematodes. When the toxicity of the V-ATPase inhibitor pyocyanin (10) was tested against M. incognita J2 nematodes, an EC50 at 24 h of 72 +/- 25 mg/L was found. The redox-active compounds such as phthalaldehyde (1) and salicylaldehyde (2) may share a common mode of action inhibiting nematode V-ATPase enzyme. The results of this investigation reveal that aromatic redox-active aldehydes can be considered as potent nematicides, and further investigation is needed to completely clarify their mode of action.

Root-knot nematodes, Meloidogyne species, are strongly destructive and extremely difficult to manage due to their poliphagy and worldwide spread. Control of these parasites has been effectively based on synthetic fumigants, but environmental concerns are imposing the adoption of safer management strategies. Plant essential oils may have a high potential of application as nematicidal soil biofumigants, due to their content of biocidal volatile compounds. Soil fumigation treatments with pure essential oils from five different species, Eucalyptus citriodora, Eucalyptus globulus, Mentha pulegium, Pelargonium asperum and Ruta graveolens were comparatively tested at the rates of 50, 100 and 200 ?l kg-1 soil against the root-knot nematode Meloidogyne incognita on tomato in pot. Plant growth parameters, gall infestation and nematode multiplication on tomato roots were evaluated after a two-month permanence of plants in a greenhouse at 25 °C. Soil fumigation treatments with essential oils of E. citriodora, E. globulus and P. asperum significantly reduced the number of eggs and juveniles of M. incognita on tomato roots compared to the non-treated control at rates higher than 50 ?l kg-1 soil, whereas oils of M. pulegium and R. graveolens were suppressive only at the highest rate. All treatments, but 50 ?l kg-1 soil of E. citriodora, P. asperum and M. pulegium, significantly reduced also the gall formation on tomato roots. Tomato top and root biomass was significantly increased by all treatments with oils of P. asperum and E. globulus, whereas the highest rate of the oil of R. graveolens and all rates of the oill of M. plulegium resulted in some phytotoxicity. Soil biofumigation with the five essential oils may be an effective tool for a sustainable management of root-knot nematodes, though the essential oils of P. asperum and E. globulus may be preferable, in consideration of their overall higher suppressiveness and positive effects on plant growth.

Bacteria appear to exert the nematicide effects in respect to root-knot nematodes limiting their population. One have proposed the simple and viable procedure for assessing of nematicide activity of bacterial strains

Aqueous ozone treatments were applied to tomato plants (cv. Roma VF) before and after Meloidogyne incognita infection in controlled growth conditions. Ozonated water was produced in situ by an ozone generator at 8 ppm. Treatments were applied for 3 and 4 days as soil drench at the rate of 10 ml/pot (50 ml). Untreated plants were used as control. The sensitivity to aqueous ozone treatments was verified by the production of reactive oxygen species (ROS) in roots. A significant ROS production was observed after two days of ozone treatments in comparison to the untreated plants, without adversely influencing the growth of the treated plants. Visual examination of the root systems revealed no symptomatic evidence of root browning or other damages indicative of oxidative stress. Tomato plants infected with M. incognita before and after ozone treatments were grown in a glasshouse at 25 ± 2°C. After 60 days the plants were uprooted, and dry shoot and root fresh weights were recorded. Root gall index, eggs and juveniles/g root, total nematode population density and reproduction rates were evaluated. Root gall index was assessed according to a 0-10 scale, whereas nematodes were extracted from roots and soil by centrifugation and the Coolen's method, respectively. Reproduction rate was calculated by the ratio between final and initial nematode population density (r =Pf/Pi). Data from the experiments were subjected to ANOVA and means compared by Least Significant Difference's test. Root gall index, eggs and juveniles/g root, total nematode population density and the reproduction rate were significantly reduced in both ozone treatments (3 and 4 days of treatments), either before or after nematode inoculation, in comparison to the untreated control.

Five composts (C1: fresh olive pomace, straw, chicken manure, urea; C2: fresh olive pomace, lettuce residues, cow manure, straw, sawdust; C3: sewage sludge, municipal green residues; C4: grass, leaves, tree branches, soil; C5: by-product from penicillin production (mycelium), straw and sawdust) were tested in a pot experiment to investigate their short-term effect on the nematode community of a grassland soil. Composts were mixed with soil at the rates of 10, 25, 50 and 100 g (kg soil)-1 and barley was sown in each potting mixture after a 2-month decomposition period. Nematodes were extracted from each pot 5 months after barleysowing, identified at genus level and grouped into bacterial, fungal and root fungal feeders, predators, omnivores and plant parasites. Bacterial feeders, predators and omnivores were favoured by amendments with composted penicillin substrate and partly by compost from olive pomace, whereas composts from green wastes and sewage sludge suppressed these trophic groups except for predators. All compost treatments suppressed fungal feeders as well as significantly reducing the density of plant-parasitic nematodes compared with non-amended soil. The highest suppressiveness on plant-parasitic nematodes was found for the composts from urban green residues, penicillin substrate and olive pomace plus cow manure. Suppressive effect was hypothesised to be related to ammoniacal nitrogen content of the composts. Composts from fungal mycelium or olive pomace seem to be the most suitable for application in nematode management strategies due to their low impact on beneficial trophic groups.

Suppressiveness of soil amendments with different rates of composted biowaste materials, olive pomace, municipal green wastes, sewage sludge and spent mushroom substrate, was evaluated against the root-knot nematode Meloidogyne incognita on tomato in potting mixtures. Soil amendments were applied at 0, 10, 25, 50 and 100 g kg-1 soil, according to a randomized block design with five replications for each treatment. Sixty days after tomato transplanting, nematode population density on plant roots and in soil and root gall infestation were assessed on each root system, and plant top and root weight were also recorded. Soil pH, dry and organic matter content, total and ammoniacal nitrogen were analyzed at the same time. Olive pomace-based composts resulted in the highest nematode suppression (73 – 97 %, according to the rate) and significantly reduced gall formation on tomato roots. Olive-waste compost affected positively tomato growth only in combination with sheep wool wastes, but it caused phytotoxicity when mixed with chicken manure and urea. Soil amendments with composted mushroom substrate also provided a consistent nematode suppression and a significant increase of plant growth, whereas composted municipal green wastes were more suppressive and positively affected tomato growth when combined with sewage sludge. Soil chemical parameters were scarcely affected by compost amendments, as organic matter was significantly increased only by the olive pomacederived composts and nitrogen content only at the highest rate of the five composts. Data from the experiment confirmed the potential of compost amendments for sustainable management of root-knot nematodes both in field and greenhouse container media, though their technical effectiveness and economic convenience are strictly dependent on a correct proportion and local availability of raw materials used in the composting process

Incorporation into the soil of olive mill wastes after an appropriate composting process (composted - OMW) can represent a possible solution to the problem of their disposal and in addition can improve plant resistance to nematode attack by stimulating root development and plant growth because of their large content of nutritive elements. Therefore, a field experiment was undertaken in southern Italy to investigate the suppressive effect of incorporation into the soil of olive composted pomace, both fresh and exhausted, on the root-knot nematode Meloidogyne incognita. Composted olive pomace obtained by mixing fresh solid cake with farmyard manure and another compost obtained by mixing exhausted solid cake with poultry manure and wheat straw at doses of 10, 20 and 40 t ha-1, were compared with two controls: i) untreated control; ii) treatment with fenamiphos (traditional nematicide) at 0.3 t ha-1 . All treatments were applied before transplanting tomato plants on a sandy soil infested by the nematode. Tomato crop yield, soil nematode population and root gall index were recorded in all plots. The results obtained showed that crop yield in amended plots was enhanced and that both composted - OMW were suppressive on M. incognita.

Root-knot nematodes, Meloidogyne species, are strongly destructive and extremely difficult to manage due to their poliphagy and worldwide spread (1). Control of these parasites has been prevalently based on chemical treatments, but environmental concerns are imposing the adoption of safer management strategies. Plant-derived compounds can represent a concrete alternative to chemicals, as nematicidal secundary metabolites are present in many plant families (2). Plant essential oils and their constituents have a great potential for nematode control since they can be developed for the use as nematicides themselves or can be used as model compounds for the development of more active derivatives (3, 4). Soil drench treatments with 50, 100 and 200 ml kg-1 soil solutions of the essential oils from five different species, i.e. Schinus molle, Cinnamomum camphora, Eugenia caryophillata, Cinnamomum zeylanicum and Citrus aurantium were comparatively tested against the root-knot nematode Meloidogyne incognita on tomato in pot. Plant growth parameters, gall infestation and nematode multiplication on tomato roots were evaluated after a two-month permanence of plants in a greenhouse at 25 °C. All the treatments with the five essential oils but the 50 and 100 and 200 ml kg-1 soil rates of C. zeylanicum significantly suppressed the number of eggs and juveniles of M. incognita on tomato roots compared to the non-treated control. Effect of treatments was less evident on gall formation on tomato roots. Essential oils of S. molle and E. caryophillata were overall more suppressive than the other three oils and generally as nematotoxic as fenamiphos chemical control. Tomato top and root biomass was not affected by all the tested treatments. Soil treatments with essential oils formulations were confirmed to be an effective tool for a sustainable management of root-knot nematode infestations.

A pot experiment on potato was carried out to verify the nematicidal effect of four composts of different origin (C1:70 % horse manure + 15 % sugar beet pomace + 5 % poultry manure + 10% grape pomace; C2: 100 % pig manure decomposed by juveniles of Musca domestica; C3:100 % vermicompost from medical plants wastes; C4:100% vermicompost from cattle manure) on the potato cyst nematodes G. rostochiensis (Ro1) and G. pallida (Pa2 and Pa3). Composts at different rates (1.0, 2.5 and 5.0 % w/w) were mixed with the nematode infested soils. Pots with unamended soils were used as control. Pots (4 l) were arranged in a glasshouse according to a randomized block design with four replications per each treatment.A significant reduction in number of cysts, eggs and juveniles/cyst and eggs and juveniles/g soil was observed in each compost in comparison to unamended soil. The suppressive nematode effect increased according to the compost NH4 + content and compost rate.

Carbonyl groups are known to form covalent adducts with endogenous proteins, but so far, their nematicidal mechanism of action of has been overlooked. The nematicidal activity of ten lactones was tested in vitro against the root knot nematodes Meloidogyne incognita and Meloidogyne arenaria. In particular, the saturated lactones alpha-methylene-gamma-butyrolactone or tulipaline A (1) and gamma-butyrolactone (3) were active against M. incognita with an EC50/48h of 19.3 +/- 10.0 and 40.0 +/- 16.2 mg/L respectively. Moreover the alpha, beta-unsaturated lactone 5,6-dihydro-2H-pyran-2-one (2) exhibited the strongest nematicidal activity against the two species with EC50/48h 14.5 +/- 5.3 and 21.2 +/- 9.7 mg/L respectively. Here we propose that the toxic effects of lactones and aldehydes on M. incognita and M. arenaria might be a consequence of their vacuolar-type H+-ATPase (V-ATPase) inhibition activity; in fact alpha-methylene-gamma-butyrolactone (1) and salicylaldehyde (12) produced an increased pH in lysosomal-like organelles on HeLa human cell line and this alteration was most likely related to a V-ATPase impairment

After 2 months from the infestation of tomato plants with the root-knot nematode (RKN) Meloidogyne incognita,we performed a gas chromatography-mass spectrometry untargeted fingerprint analysis for the identification of characteristicmetabolites and biomarkers. Principal component analysis, and orthogonal projections to latent structures discriminant analysissuggested dramatic local changes of the plant metabolome. In the case of tomato leaves, ?-alanine, phenylalanine, and melibiosewere induced in response to RKN stimuli, while ribose, glycerol, myristic acid, and palmitic acid were reduced. For tomato stems,upregulated metabolites were ribose, sucrose, fructose, and glucose, while fumaric acid and glycine were downregulated. Thevariation in molecular strategies to the infestation of RKNs may play an important role in how Solanum lycopersicum and otherplants adapt to nematode parasitic stress

The present invention relates to a portable apparatus for detecting falls and immobility of a subject. The apparatus comprises at least one sensor generating a signal indicative of the accelerations of at least one portion of the subject's body, a control unit suitable to process this signal and communication means for sending a call for help to a remote control station. Advantageously, the control unit automatically activates the communication means only when a fall of the subject and his/her subsequent immobility are detected based on said processing.