Differential effects of plant-beneficial fungi on the attraction of the egg parasitoid Trissolcus basalis in response to Nezara viridula egg deposition

There is increasing evidence that plant-associated microorganisms play important roles in defending plants against insect herbivores through both direct and indirect mechanisms. While previous research has shown that these microbes can modify the behaviour and performance of insect herbivores and their natural enemies, little is known about their effect on egg parasitoids which utilize oviposition-induced plant volatiles to locate their hosts. In this study, we investigated how root inoculation of sweet pepper (Capsicum annuum) with the plant-beneficial fungi Beauveria bassiana ARSEF 3097 or Trichoderma harzianum T22 influences the olfactory behaviour of the egg parasitoid Trissolcus basalis following egg deposition by its host Nezara viridula. Olfactometer assays showed that inoculation by T. harzianum significantly enhanced the attraction of the egg parasitoid, while B. bassiana had the opposite effect. However, no variation was observed in the chemical composition of plant volatiles. Additionally, fitness-related traits of the parasitoids (wasp body size) were not altered by any of the two fungi, suggesting that fungal inoculation did not indirectly affect host quality. Altogether, our results indicate that plant inoculation with T. harzianum T22 can be used to enhance attraction of egg parasitoids, which could be a promising strategy in manipulating early plant responses against pest species and improving sustainable crop protection. From a more fundamental point of view, our findings highlight the importance of taking into account the role of microorganisms when studying the intricate interactions between plants, herbivores and their associated egg parasitoids

Limited effects of plant-beneficial fungi on plant volatile composition and host-choice behavior of Nesidiocoris tenuis

Biological control using plant-beneficial fungi has gained considerable interest as a sustainable method for pest management, by priming the plant for enhanced defense against pathogens and insect herbivores. However, despite promising outcomes, little is known about how different fungal strains mediate these beneficial effects. In this study, we evaluated whether inoculation of tomato seeds with the plant-beneficial fungi Beauveria bassiana ARSEF 3097, Metarhizium brunneum ARSEF 1095 and Trichoderma harzianum T22 affected the plant’s volatile organic compound (VOC) profile and the host-choice behavior of Nesidiocoris tenuis, an emerging pest species in NW-European tomato cultivation, and the related zoophytophagous biocontrol agent Macrolophus pygmaeus. Results indicated that fungal inoculation did not significantly alter the VOC composition of tomato plants. However, in a two-choice cage assay where female insects were given the option to select between control plants and fungus-inoculated plants, N. tenuis preferred control plants over M. brunneum-inoculated plants. Nearly 72% of all N. tenuis individuals tested chose the control treatment. In all other combinations tested, no significant differences were found for none of the insects. We conclude that inoculation of tomato with plant-beneficial fungi had limited effects on plant volatile composition and host-choice behavior of insects. However, the observation that N. tenuis was deterred from the crop when inoculated with M. brunneum and attracted to non-inoculated plants may provide new opportunities for future biocontrol based on a push-pull strategy

Herbivore-Mediated Effects of Glucosinolates on Different Natural Enemies of a Specialist Aphid

The cabbage aphid Brevicoryne brassicae is a specialist herbivore that sequesters glucosinolates from its host plant as a defense against its predators. It is unknown to what extent parasitoids are affected by this sequestration. We investigated herbivore-mediated effects of glucosinolates on the parasitoid wasp Diaeretiella rapae and the predator Episyrphus balteatus. We reared B. brassicae on three ecotypes of Arabidopsis thaliana that differ in glucosinolate content and on one genetically transformed line with modified concentrations of aliphatic glucosinolates. We tested aphid performance and the performance and behavior of both natural enemies. We correlated this with phloem and aphid glucosinolate concentrations and emission of volatiles. Brevicoryne brassicae performance correlated positively with concentrations of both aliphatic and indole glucosinolates in the phloem. Aphids selectively sequestered glucosinolates. Glucosinolate concentration in B. brassicae correlated negatively with performance of the predator, but positively with performance of the parasitoid, possibly because the aphids with the highest glucosinolate concentrations had a higher body weight. Both natural enemies showed a positive performance-preference correlation. The predator preferred the ecotype with the lowest emission of volatile glucosinolate breakdown products in each test combination, whereas the parasitoid wasp preferred the A. thaliana ecotype with the highest emission of these volatiles. The study shows that there are differential herbivore-mediated effects of glucosinolates on a predator and a parasitoid of a specialist aphid that selectively sequesters glucosinolates from its host plant

Analysis of organochloro-pesticides in Eritrean water and sediment samples

Thesis (MSc)--Stellenbosch University, 2004.ENGLISH ABSTRACT: The choice of the sample preparation technique used in combination with a chromatographic technique in environmental analysis strongly influences the performance of the method as a whole. The main aim of this work was to evaluate methods like: micro liquid-liquid extraction (цLLE)and solid phase micro extraction (SPME) in combination with gas chromatography-electron capture detector (GCECD) and a recently developed technique named stir bar sorptive extraction (SBSE) combined with thermal desorption system-programmable temperature vaporizationgas chromatography-mass spectrometer (TDS-PTV-GC-MS) for the analysis of organochloro-pesticides (OCPs) in natural water and to make a comparison between them. The most suitable method was then selected for the analysis of water samples collected from Eritrea. An additional goal was to develop a sensitive method for the analysis ofOCPs in soil and sediment samples. цLLE-GC-ECD and SPME-GC-ECD methods were evaluated for the analysis of OCPs in water. The former allowed achieving a sensitivity of less than 100 ppt and the latter was able to detect down to the 1 ppt level. Both methods were found to be repeatable with %RSD of3% to 19% and 5% to 12% as well as linear with a range of 1:103 and 1:5x103 respectivey. SBSE-TDS-GC-MS was also investigated for the analysis of OCPs in water samples. With this technique a good sensitivity down to 1 ppt was also obtained. The less sensitive nature of the MS compared to ECD was hereby compensated for by the large amount of polydimethylsiloxane (PDMS) phase on the stir bar. The method was repeatable and linear with a range of 1:5000, which is similar to the one obtained for the SPME-GC-ECD method. All three methods were also tested with real contaminated water samples and a comparison of the three techniques in terms of sensitivity, linearity, repeatability, availability and cost effectiveness was done. As a method of choice in terms of the above criteria, SPMEGC- ECD was applied to the water samples collected from Eritrea. The presence of some oePs such as a-BBC (benzenehexachloride), heptachlor, heptachlor epoxide, endosulfan I, p,p'-DDE (dichlorodiphenylchloroethylene), endosulfan II, p,p'-DDD (dichlorodiphenyldichloroethane), endosu!fan sulfate and p,p'-DDT (dichlorodiphenyltrichloroethane) in some of the Eritrean water samples was demonstrated The SPME-GC-ECD technique for the analysis OCPs in soil and sediment samples was investigated. It was shown to be able to detect down to 1 pg/g (lppt) with good linearity and repeatability. The method was also evaluated for authentic soil samples. Due to a lack of time the method could not be applied for the analysis of the soil and sediment samples collected from Eritrea.AFRIKAANSE OPSOMMING: Die keuse van 'n gepaste monster-voorbereidingsmetode wat gebruik word tesame met 'n chromatografiese skeidingtegniek in omgewings-analise het 'n beduidende effek op die algehele metode. Die doel van hierdie werk was om metodes soos mikrovloeistof- vloeistof ekstraksie (цLLE) en soliede fase mikro ekstraksie (SPME) in kombinasie met gaschromatografie met elektronvangs deteksie (GC-ECD) te ontwikkel vir die analise van organochloor-pestisiede (OCPs) in natuurlike water. Vervolgens is hierdie metodes vergelyk met die onlangs-ontwikkelde roerstaaf sorptiewe ekstraksie tegniek (SBSE) in kombinasie met termiese desorbsie-GCmassaspektrometriese deteksie (TDS-GC-MSD). Die mees geskikte metode is vervolgens gebruik vir die analise van Eritreaanse watermonsters. Daarbenewens was die doelook om 'n sensitiewe metode te ontwikkel vir die analise van OCPs in gronden sedimentmonsters. Beide цLLE en SPME-GC-ECD metodes is gekarakteriseer deur goeie herhaalbaarheid en lineariteit. Die sensitiwiteit van bogenoemde metodes was minder as 100 dele per triljoen (ppt) en 1 ppt, respektiewelik. Ook met SBSE-TDS-GC-MSD kon 'n deteksielimiet van 1 ppt bereik word. In hierdie geval is vir die verlies van sensitiwiteit van die MSD in vergelyking met die ECD, gekompenseer deur die groter hoeveelheid PDMS op die roerstaaf in vergelyking met 'n SPME apparaat. Die herhaalbaarheid en lineariteit van die metode is vergelykbaar met dié van die SPMEGC- ECD metode. Al drie metodes is vergelyk vir die analise van gekontamineerde watermonsters in terme van sensitiwiteit, liniêre bereik, herhaalbaarheid, toeganklikheid en kosteeffektiwiteit. SPME-GC-ECD is vervolgens gekies as metode van voorkeur vir die analise van Eritreaanse watermonsters. Die teenwoordigheid van sekere OCPs soos a-BHC, heptachloor, heptachloorepoksied, endosulfan I, p,p'-DDE, endosulfan II, p,p' -DDD, endosulfan sulfaat en p,p' -DDT in die monsters is gedemonstreer. Die toepasbaarheid van die SPME-GC-ECD metode vir die analise van OCPs in grond-en sedimentmonsters is ook geëvalueer, en dit was moontlik om so min as 1 pg/g waar te neem. Weens tydbeperkings was dit nie moontlik om Eritreaanse grondmonsters met die ontwikkelde metode te analiseer nie

Application of modern chromatographic technologies for the analysis of volatile compounds in South African wines

Thesis (PhD (Chemistry and Polymer Science))--University of Stellenbosch, 2009.ENGLISH ABSTRACT: The present study was initiated by the wine industry of South Africa to overcome the lack of available information on the flavor and aroma of South African wines. The aim was to develop new analytical methods and improve existing ones for the analysis of volatile compounds in the South African wines. Initially a new analytical method based on stir bar sorptive extraction (SBSE) in the headspace mode for the analysis of 37 pre-selected volatile compounds was developed and validated. Consequently, the method was improved by making important modifications and increasing the number of compounds analyzed to 39. This method was successfully applied to a large number of Pinotage wines of vintages 2005 and 2006. The quantitative data of these wines were subjected to chemometric analysis in order to investigate possible co- /variances. A clear distinction was observed between the two vintages, where the 2005 wines were more characterized by wood-related compounds and the 2006 wines by the fermentation compounds. The developed method was further applied to other cultivars of vintage 2005, including two white (Sauvignon Blanc and Chardonnay) and three red (Shiraz, Cabernet Sauvignon and Merlot) cultivars. In a similar fashion, the quantitative data of the six cultivars of vintage of 2005 were analysed by chemometric methods. Significant differences were observed between the two white cultivars and among the four red cultivars. It was shown that among these cultivars, the major role-players were the wood and fermentation related volatiles. A striking observation was the confirmation of the unique character of the Pinotage wines compared to the other red cultivars, mainly influenced by the high level of isoamyl acetate and low level of isoamyl alcohol, the former being categorized as a varietal compound for Pinotage expressed by a fruity (banana) odor. In addition, advanced chromatographic technology in the form of comprehensive twodimensional gas chromatography (GC × GC) coupled to time-of-flight mass spectrometry (TOFMS) was investigated for the detailed analysis of volatile compounds in young South African wines. This work focused primarily on Pinotage wines. In the first instance, solid phase micro extraction (SPME) in the headspace mode in combination with GC × GC-TOFMS was used. Due to the high resolution and large peak capacity of GC × GC, more than 200 compounds previously reported as wine components were identified. These compounds were dominated by the highly volatile and less polar compounds, mainly due to the characteristics of SPME. In an attempt to further extend these results, another selective extraction method, solid phase extraction (SPE) was used in combination with GC × GC-TOFMS analysis. Using this technique, more than 275 compounds, most of them unidentified using the previous method, were detected. These groups of compounds include volatile phenols, lactones as well as mostly aromatic esters and norisoprenoids, which can potentially influence the aroma and flavor of wine. The techniques developed as part of this study have extended our knowledge of the volatile composition of South African wines.AFRIKAANSE OPSOMMING: Hierdie studie is geïnisieer deur die wyn industrie van Suid-Afrika om die tekort aan beskikbare inligting aangaande wyn aroma van Suid-Afrikaanse wyne te oorkom. Die doel was om nuwe analitiese metodes te ontwikkel en die huidige metodes te verbeter vir die analise van vlugtige verbindings in Suid-Afrikaanse wyne. Oorspronklik is ʼn nuwe analitiese metode ontwikkel en gevalideer gebaseer is op ‘stir bar sorptive extraction’ (SBSE) in die gas fase vir die analise van 37 vooraf geselekteerde vlugtige verbindings. Die metode is verbeter deur belangrike modifikasies aan te bring en die hoeveelheid verbindings wat analiseer word te vermeerder na 39. Hierdie metode is suksesvol aangewend op ʼn groot hoeveelheid Pinotage wyne van oesjare 2005 en 2006. Die kwantitatiewe data van hierdie wyne is onderwerp aan verskillende chemometriese analises om moontlike ko-/variasies te ondersoek. ʼn Duidelike onderskeid is opgemerk tussen die twee oesjare, waar die 2005 wyne gekarakteriseer is deur hout-verwante verbindings en die 2006 wyne weer meer deur fermentasie verbindings. Die verbeterde metode is verder aangewend vir analiese van ander kultivars van oesjare 2005, wat twee wit (Sauvignon Blanc en Chardonnay) en drie rooies (Shiraz, Cabernet Sauvignon en Merlot) ingesluit het. Die kwantitatiewe data van die ses kultivars van oesjaar 2005 is op ʼn soortgelyke wyse geanaliseer deur verskillende chemometriese metodes te gebruik. Beduidende verskille is opgemerk tussen die twee wit kultivars en tussen die vier rooi kultivars. Die hoof rolspelers tussen die ses kultivars was weereens die verbindings wat ʼn hout en fermentasie aard het. Die unieke karakter van die Pinotage wyne in vergelyking met die ander rooi kultivars was opvallend. Hierdie wyn word gekarakteriseer deur hoë vlakke van isoamiel asetaat en lae vlakke van isoamiel alkohol, waar eersgenoemde gekatogiseer word as ʼn verbinding wat ʼn vrugte (piesang) geur in Pinotage uitdruk. Verder is gevorderde chromatografiese tegnologie in die vorm van ‘comprehensive two-dimentional gas chromatography’ (GC x GC) gekoppel met ‘time-of-flight mass spectroscopy’ (TOFMS) ondersoek vir die analiese van vlugtige verbindings in jong Suid-Afrikaanse wyne. Hierdie werk het hoofsaaklik op Pinotage wyne gefokus. Eerstens is ‘solid phase micro extraction’ (SPME) in die gas fase gekombineer met GC x GC-TOFMS. As gevolg van die hoë resolusie en groot piek kapasiteit van GC x GC is meer as 200 verbindings wat voorheen gerapporteer is as wyn komponente geïdentifiseer. Hierdie verbindings is gedomineer deur hoë vlugtige polêre verbindings, hoofsaaklik as gevolg van die karaktersitieke van SPME..In ʼn poging om die metode verder te verbeter is ʼn selektiewe ekstraksie metode naamlik ‘solid phase extraction’ (SPE) in kombinasie met GC x GC-TOFMS gebruik. Met hierdie tegniek is meer as 275 verbindings geïdentifiseer, waarvan die meeste nie met die vorige metode waargeneem is nie. Hierdie verbindings sluit vlugtige fenole, laktone en meestal aromatiese esters en norisoprenoïdes in, wat moontlik die reuk en smaak van wyn kan beïnvloed. Die metodes ontwikkel gedurende die studie het nuwe informasie verskaf aangaande die vlugtige komponente teenwoordig in Suid Afrikaanse wyne

PERFORMANCE OF FULL-WAVER SYSTEM IN DEEP ELECTRICAL RESISTIVITY TOMOGRAPHY FOR SUBSURFACE CHARACTERIZATION IN THE COSTERMANO AREA, LAKE GARDA.

This thesis discovers the performance of the deep electrical resistivity tomography by applying a full-waver system and integrating it with the seismic reflection method applied in the same area of investigation. Data acquisition involves thorough survey design using the Full-waver system for deep investigation, data collection is conducted by injecting electrical currents into the ground and measuring voltage responses, and Synchronization via GPS technology in challenging terrains. and data processing using Geostudi Astier s.r.l's ERTlab studio software. The comparative analysis with seismic reflection surveys confirms DERT's reliability in delineating subsurface structures and tectonic discontinuities, such as the Valpolicella Fault - Sant'Ambrogio and Sirmione faults. This research targets how performed the DERT method to characterize the subsurface of the investigation area and asses it’s contribution to environmental geohazards and geothermal potential, applying the Full-Waver DERT method in the area was lacking. The Valpolicella – Sant'Ambrogio Fault indicated by the geophysical survey is segmented along the route, with portions coinciding with what is reported in other findings and, conversely, parts displaced by 100-200 meters in NE direction and in the south direction of the investigation area it is observed that the presence of the Sirmione fault and limestone reservoir’s favoring potential research activities. Therefore, this area of investigation is in need of further and deep geophysical investigation