Submitted by Daniella Santos (daniella.santos@ufma.br) on 2017-08-07T12:56:26Z No. of bitstreams: 1 Sandra Alves.pdf: 2374477 bytes, checksum: bf34f6a346f4775e29afabb54e505859 (MD5) / Made available in DSpace on 2017-08-07T12:56:26Z (GMT). No. of bitstreams: 1 Sandra Alves.pdf: 2374477 bytes, checksum: bf34f6a346f4775e29afabb54e505859 (MD5) Previous issue date: 2017-04-04 / CAPES / FAPEMA / Haemonchus contortus (RUDOLPHI, 1803) is a species of gastrointestinal nematode of great importance due to the damages caused in livestock. Natural products have been studied as an alternative to the use of commercial anthelmintics, responsible for the development of resistance in parasites. This study has as main objective to verify the anthelmintic activity of protein extracts of Leucaena leucocephala (Linn.) and Spigelia anthelmia (Linn.) against the nematode H. contortus. The present work was divided in two chapters. In chapter 1, L. leucocephala seeds were ground and the resulting flour, homogenized in 100 mM sodium phosphate buffer, pH 7. The suspension obtained was centrifuged (15,000 x g at 4 ° C for 30 min). After centrifugation, the obtained supernatant was denominated: Total extract (TE), cotyledon extract (CE) and shell extract (SE). The protein content, proteolytic, protease inhibitory, chitinolytic and anthelmintic activity of L. leucocephala extracts on H. contortus were verified. For each treatment, the effective concentration (EC) of the samples on the nematode was calculated. The extracts TE and CE presented a high protein content, besides having proteolytic, chitinolytic and protease inhibitory activity and inhibiting egg hatching (EC50 0.33 and 0.48 mg mL -1, respectively). However, L. leucocephala samples did not inhibit the larvae of H. contortus. It is concluded that proteins are correlated with the action of L. leucocephala on H. contortus. In Chapter 2, the parts of S. anthelmia were separated into leaves, roots and stem and, washed with distilled water, lyophilized and subsequently crushed. Subsequently the proteins were solubilized in 100 mM sodium phosphate buffer, pH 7. The suspension was centrifuged (15,000 x g at 4 ° C for 30 min) and the supernatant collected for analysis. From the ammonium sulfate precipitation (0-90%) of S. anthelmia extracts and membrane dialysis with 14 kDa molecular exclusion, protein fractions were obtained and denominated: Leaf protein fraction (LPF), stem protein fraction (SPF), root protein fraction (RPF). The samples were submitted to analysis by high performance liquid chromatography for the detection of compounds of secondary metabolism and mass spectrometry to identify possible bioactive proteins present. The protein content, proteolytic, protease inhibitory, chitinolytic, haemagglutinating and anthelmintic action of S. anthelmia protein fractions on H. contortus were verified. For each treatment, the effective concentration (EC) of the samples on the nematode was calculated. Fractions of S. anthelmia had an inhibitory effect on egg hatching, and this effect was more pronounced by LPF (EC50 0.17 mg mL -1). In addition, greater inhibition of larvae by LPF and RPF (EC50 0.27 and 0.25 mg mL -1, respectively) was observed. The protein fractions of root, stem and leaf of S. anthelmia were effective on the inhibition of larval migration (EC50 0.11, 0.14 and 0.21 mg mL -1, respectively). No secondary metabolite compounds were detected in the S. anthelmia fractions, while several proteins with anthelmintic potential were identified by mass spectrometry. It is concluded that proteins are correlated with the action of S. anthelmia on H. contortus, having potential for the development of anthelmintic products. Thus, the bioactive proteins present in L. leucocephala and S. anthelmia have promising pharmacological properties for the control of the H. contortus nematode. / Haemonchus contortus (RUDOLPHI, 1803) é uma espécie de nematoide gastrintestinal de grande importância devido aos prejuízos que causa na pecuária. Produtos naturais vêm sendo estudados como alternativa ao uso de anti-helmínticos comerciais, responsáveis pelo desenvolvimento de resistência nos parasitos. Este estudo tem como principal objetivo verificar a atividade anti-helmíntica de extratos proteicos de Leucaena leucocephala (Linn.) e Spigelia anthelmia (Linn.) contra o nematoide H. contortus. O presente trabalho foi dividido em dois capítulos. No capítulo 1, sementes de L. leucocephala foram trituradas e a farinha resultante, homogeneizada em tampão fosfato de sódio 100 mM, pH 7. A suspenção obtida foi centrifugada (15.000 x g a 4 ºC por 30 min). Após centrifugação, o sobrenadante obtido foi denominado: Extrato total (TE), extrato de cotilédone (CE) e extrato de casca (SE). O teor de proteínas, atividade proteolítica, inibitória de protease, quitinolítica e ação anti-helmíntica dos extratos de L. leucocephala sobre H. contortus foram verificados. Para cada tratamento foi calculada a concentração efetiva (EC) das amostras sobre o nematoide. Os extratos TE e CE apresentaram elevado teor proteico, além de possuírem atividade proteolítica, quitinolítica e inibitória de protease e inibirem a eclosão dos ovos (EC50 0,33 e 0,48 mg mL-1, respectivamente). No entanto, amostras de L. leucocephala não inibiram o desembainhamento larvar de H. contortus. Conclui-se que proteínas estão correlacionadas com a ação de L. leucocephala sobre H. contortus. No capítulo 2, as partes de S. anthelmia foram separadas em folhas, raízes e caule e, lavadas com água destilada, liofilizadas e subsequentemente trituradas. Posteriormente as proteínas foram solubilizadas em tampão fosfato de sódio 100 mM, pH 7. A suspensão foi centrifugada (15.000 x g a 4 ºC por 30 min) e o sobrenadante recolhido para análises. A partir da precipitação com sulfato de amônio (0-90%) dos extratos de S. anthelmia e diálise em membrana com exclusão molecular de 14 kDa, frações proteicas foram obtidas e denominadas: Fração proteica de folha (LPF), fração proteica de caule (SPF), fração proteica de raiz (RPF). As amostras foram submetidas a análise por cromatografia líquida de alta eficiência para detecção de compostos do metabolismo secundário e, espectrometria de massas para identificação de possíveis proteínas bioativas presentes. O teor de proteínas, atividade proteolítica, inibitória de protease, quitinolítica, hemaglutinante e ação anti-helmíntica das frações proteicas de S. anthelmia sobre H. contortus foram verificados. Para cada tratamento foi calculada a concentração efetiva (EC) das amostras sobre o nematoide. As frações de S. anthelmia apresentaram efeito inibitório sobre a eclosão dos ovos, sendo este efeito mais pronunciado pela LPF (EC50 0,17 mg mL-1). Além disso, foi verificada maior inibição do desembainhamento larvar por LPF e RPF (EC50 0,27 e 0,25 mg mL-1, respectivamente). As frações proteicas de raiz, caule e folha de S. anthelmia foram efetivas sobre a inibição da migração larvar (EC50 0,11; 0,14 e 0,21 mg mL-1, respectivamente). Não foram detectados compostos do metabolismo secundário nas frações de S. anthelmia, enquanto diversas proteínas com potencial anti-helmíntico foram identificadas por espectrometria de massas. Conclui-se que proteínas estão correlacionadas com a ação de S. anthelmia sobre H. contortus, tendo potenciais para o desenvolvimento de produtos anti-helmínticos. Assim, as proteínas bioativas presentes em L. leucocephala e S. anthelmia possuem propriedades farmacológicas promissoras para o controle do nematoide H. contortus.

Made available in DSpace on 2016-04-28T20:15:32Z (GMT). No. of bitstreams: 1 2009 - Carla Alves Soleiro.pdf: 3960368 bytes, checksum: 0dd24a3adc28cf6201d540a47b33d074 (MD5) Previous issue date: 2009-02-11 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / Ostriches are susceptible to many diseases and until recently, many of them are unknown and can affect these animals when it comes to commercially breeding. In Brazil, the commercial establishment has been initiated 20 years ago, but already there are reports about the diseases that can affect these birds. The objectives of this study were identify morphologically gastrointestinal protozoans and nematodes of ostriches and see the variations: the parasitic infection that occurs in accordance to the dry and rainy seasons, and the elimination of eggs of nematodes and oocysts / cysts of protozoa by age, in a commercial establishment located in Itabora? City, Microregion of Rio de Janeiro, State of Rio de Janeiro. During the period from June 2004 to May 2006 ostriches were separated into three age groups: up to 90 days, 91 to 365 days and over 365 days (adults). The collection was held monthly and fecal samples were individually obtained, placed in plastic bags, identified and kept under refrigeration until the moment to be examined. There were employed the techniques of centrifuge-flotation in saturated sucrose solution, centrifuge-sedimentation in formalin-ether, culture of feces and stained by Safranin-Metilen Blue, and Gomori trichrome of iron Hematoxylin. Meteorological data were also obtained to determine the dry and rainy seasons. There were identified the nematode Codiostomum struthionis and three genera of intestinal protozoa: Blastocystis, Entamoeba and Cryptosporidium. All birds younger than 365 days showed a higher frequency of the genus Cryptosporidium. Among the mixed infections the most common was the association of C. struthionis with Cryptosporidium. (17.74%) in adults. There was a greater shedding of Strongylida eggs (p <0.05) and also larger number of animals that shed oocysts / cysts of intestinal protozoans in feces during the rainy season (p <0.05). A larger number of animals less than 365 days eliminated cysts / oocysts of protozoa, which is significantly correlated to the genus Entamoeba in animals up to 90 days compared with 91 to 365 (p=0.036) and to the genus Blastocystis, in this case when compared with the over 365 days (p=0.09). / Avestruzes s?o suscet?veis a diversas enfermidades e at? h? pouco tempo se desconheciam aquelas que podem acometer esses animais quando criados comercialmente. No Brasil a cria??o comercial iniciou-se h? cerca de 20 anos, mas j? existem relatos sobre as doen?as que podem acometer essas aves. Os objetivos do presente trabalho foram identificar morfologicamente protozo?rios e nemat?ides gastrintestinais de avestruzes e verificar as varia??es: da infec??o parasit?ria de acordo com as ?pocas seca e chuvosa, e da elimina??o de ovos de nemat?ides e de oocisto/cisto de protozo?rios por faixa et?ria, em uma cria??o comercial localizado no Munic?pio de Itabora?, Microrregi?o do Rio de Janeiro, Estado do Rio de Janeiro. Durante o per?odo de junho de 2004 a maio de 2006 foram coletadas fezes de avestruzes de tr?s faixas et?rias: at? 90 dias, de 91 a 365 dias e acima de 365 dias (adultos). A coleta foi realizada mensalmente e as amostras fecais eram obtidas individualmente, colocadas em sacos pl?sticos, identificadas e mantidas sob refrigera??o at? o memento de serem examinadas. Foram empregadas as t?cnicas de centrifugo-flutua??o em solu??o saturada de a??car, centrifugo-sedimenta??o em formol-?ter, coprocultura e colora??es por safranina-azul de metileno, tricr?mio de gomori e hematoxilina f?rrica. Tamb?m foram obtidos dados meteorol?gicos para determinar as ?pocas seca e chuvosa. Foram identificados o nemat?ide Codiostomum struthionis e tr?s g?neros de protozo?rios intestinais: Blastocystis, Cryptosporidium e Entamoeba. Todas as aves com menos de 365 dias apresentaram maior freq??ncia do g?nero Cryptosporidium. Dentre as infec??es mistas a mais comum foi a associa??o de C. struthionis com Cryptosporidium sp. (17,74%) nos adultos. Houve maior elimina??o de ovos da ordem Strongylida (p<0,05) e tamb?m um maior n?mero de animais que eliminaram oocistos/cistos de protozo?rios intestinais nas fezes durante a ?poca chuvosa (p<0,05). Um maior n?mero de animais com menos de 365 dias eliminaram cistos/oocistos de protozo?rios, sendo essa correla??o significativa para o g?nero Entamoeba nos animais com at? 90 dias quando comparados com os de 91 a 365 e para o g?nero Blastocystis, nesse caso quando comparados com os acima de 365 dias.

Made available in DSpace on 2017-07-10T17:36:55Z (GMT). No. of bitstreams: 1 2014_Diss_Rafael_Augusto_Swarowsky.pdf: 1265471 bytes, checksum: b7b6c650055a2ba1f2346656fda7a03d (MD5) Previous issue date: 2014-02-24 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Considering the importance of the tomato crop and the high costs of the control of Meloidogyne incognita with resistant cultivar or nematicides, grows the search of new alternatives to control the root-knot disease. The homeopathy may be an alternative way of control, by inducing resistance. This study aimed to evaluate the influence of the homeopathic drug Cina in the number of root galls, eggs and J2 present in the roots of tomato and soil, as well as the growth of tomato plants. The dynamizations used were 12, 24, 50, 100, 200, and 400CH (centesimal hahnemanian dilutions). It was also used the chemical carbofuran, 70% ethanol and water as control treatments. The assay was conducted in pots in greenhouse for the in vivo assay and in laboratory for the in vitro ones. In both assays, homeopathic treatments were diluted with distilled water to 0.1% before application. On the in vivo assay, the homeopathic drug was applied weekly by foliar spray. The first treatments were done three days before pathogen inoculation with approximately 5000 eggs and 500 J2 per pot. The evaluations were made 50 days after transplanting of the tomato seedlings and 44 days after inoculation. The Cina demonstrated effectiveness in stimulating root development, once the results of root volume were higher in homeopathic treatments than in control. Cina 100CH also favored the growth of the stalk diameter of the plants. However, the homeopathic product showed no nematostatic nor nematicide effect / Considerando a importância da cultura do tomateiro e os custos com variedades resistentes e com nematicidas para controle de Meloidogyne incognita, são necessárias alternativas para manejo da doença das galhas radiculares. A homeopatia pode ser uma alternativa de controle, através da indução de resistência de plantas. Este trabalho objetivou avaliar a influência do medicamento homeopático Cina quanto ao número de galhas radiculares, ovos e J2 presentes nas raízes do tomateiro e no solo, bem como variáveis de crescimento das plantas de tomate. As dinamizações usadas foram 12, 24, 50, 100, 200 e 400CH (centesimal hahnemanniana). Também foram utilizados como tratamentos para comparação o nematicida carbofurano, etanol 70% e água. O experimento foi conduzido em vasos dentro de estufa climatizada para ensaio in vivo e em laboratório para o ensaio in vitro. Em ambos os ensaios, os tratamentos homeopáticos foram diluídos em água destilada a 0,1% antes da aplicação. No ensaio in vivo o medicamento homeopático foi aplicado semanalmente por aspersão foliar. A primeira aplicação dos tratamentos foi realizada três dias antes da inoculação, com aproximadamente 5000 ovos e 500 J2 por vaso. As avaliações foram feitas 50 dias após o transplante das mudas de tomate e 44 dias após a inoculação. A Cina demonstrou eficiência em estimular o desenvolvimento radicular, pois os resultados de volume de raiz foram maiores nos tratamentos homeopáticos do que nas testemunhas. Cina a 100CH também favoreceu o crescimento do diâmetro de caule das plantas. Contudo, o medicamento homeopático não demonstrou nenhum efeito nematostático e nem nematicida

There has been an increase in the incorporation of silver nanomaterials into consumer products due to their antimicrobial properties. Therefore there is potential for silver nanoparticles (Ag-NPs) to leach out into the environment during different life-cycle stages of these nanomaterial-containing products. Concern about the toxicity of Ag-NPs has led to investigations into their toxic effects on a variety of organisms mainly using acute and sub-chronic, single-generation exposures. The focus of this project was to understand the effects of long-term continuous multigenerational exposure to AgNO3 and Ag-NPs in both pristine and environmentally transformed forms, on the model organism, Caenorhabditis elegans, a soil nematode. A previous multigenerational C. elegans study, showed increased sensitivity in terms of reproductive toxicity, in response to AgNO3 and Ag-NPs, but not sulfidized Ag-NPs (sAg-NPs), with increasing generations of exposure. The reproductive toxicity persisted in subsequently unexposed generations even after rescue from the exposure. We hypothesized that genomic mutations and/or epigenetic changes were possible mechanisms by which the reproductive toxicity was inherited. We investigated the potential for induction of germline mutations in C. elegans after exposures for ten generations to AgNO3, Ag-NPs, and sAg-NPs using whole genome DNA sequencing. Epigenetic changes at histone methylation markers, (H3K4me2 and H3K9me3), and DNA methylation at adenosine (N6-methyl-2’-deoxyadenosine) were investigated after multigenerational exposure as well as after rescue from the exposure using enzyme-linked immunosorbent assays (ELISA) and liquid chromatography with tandem mass spectrometry (LC-MS/MS), respectively. Expression levels of the genes of methyltransferases and demethylases, associated with the histone methylation markers and DNA methylation, were also examined. Our results for germline mutations reveal no significant differences between the nematodes exposed to AgNO3 or pristine Ag-NPs when compared to controls. The significant increase in the number of transversion was observed only for sAg-NPs. However, a trend toward an increase in the total number of mutations was observed in all Ag treatments with some of those mutations having a predicted moderate or high impact. This potentially contributed towards reproductive as well as growth toxicity shown previously after ten generations of exposure in every treatment.. These results did not entirely support the multigenerational reproductive toxicity observed previously. Epigenetic responses at histone methylation markers revealed opposite patterns between pristine and transformed Ag-NPs with Ag-NPs causing a significant increase while exposure to sAg-NPs resulted in significant decrease in methylation at H3K4me2 mark. The increase in H3K4me2 levels was also inherited by subsequent unexposed generations rescued from Ag-NP exposure. Only sAg-NPs caused a significant decrease in methylation at H3K9me3 mark. Changes in mRNA levels for histone methyltransferases and demethylase corresponded with the histone methylation levels affected by Ag-NPs and sAg-NPs. For DNA methylation, a significant increase was observed only for AgNO3, which was not inherited after the rescue. In conclusion, while germline mutations with a high or moderate impact may affect reproduction, our results do not support this as a mechanism for the heritable increase in C. elegans sensitivity to reproductive toxicity from AgNO3 and pristine Ag-NPs. The epigenetic changes, however, do show partial correlation with the observed reproductive toxicity. The reproductive multigenerational effects of AgNO3 can be attributed to changes in DNA methylation whereas that of Ag-NPs can be attributed to changes in histone methylation. Further studies, focused on the investigation of changes in histone and DNA methylation levels at specific loci using chromatin immunoprecipitation sequencing (ChIP-Seq) and methylated DNA immunoprecipitation sequencing (MeDIP-Seq), respectively, are warranted for a better understanding of the impact of such changes.

Thesis (M.Agric. (Horiticulture)) --Unversity of Limpopo, 2013 / The ground leaching technology (GLT) system, using crude extracts of wild cucumber (Cucumis myriocarpus) and wild watermelon (Cucumis africanus) fruits, had been widely researched and developed in management of the root-knot (Meloidogyne species) nematodes in tomato (Solanum lycopersicon) production. In the GLT system, experiments were harvested at 56 days after inoculation with nematodes, which was approximately three generations of Meloidogyne species. Also, studies in GLT systems demonstrated that effective micro-organisms (EM) were not essential in the release of chemicals from crude extracts for nematode suppression, with suggestions that the system exclusively relied upon irrigation or rainwater for leaching out chemicals. However, the system had hardly been tested on other nematode species with longer life cycles and crops. The objective of this study was to investigate the influence of crude extracts of C. myriocarpus (cucurbitacin A-containing phytonematicide) and C. africanus (cucurbitacin B-containing phytonematicide) with and without EM on suppression of population densities of the citrus nematode (Tylenchulus semipenetrans) on rough lemon (Citrus jambhiri) over three generations of the nematode. Two studies, one on C. myriocarpus and the other on C. africanus, with and without EM, were conducted separately using the GLT system under greenhouse conditions with three generations of T. semipenetrans as the standard of application interval of the materials. Citrus seedlings were transplanted in 7-L plastic pots contain 6.5-L pasteurised river sand and Hygromix (3:1 v/v) and inoculated with approximately 25 000 second-stage juveniles (J2s) in 2 × 2 × 2 factorial experiment, where treatments were arranged in a randomised complete block design (RCBD), with six replications. At 150 days after treatment, nematode and plant variables were collected and subjected to factorial analysis of variance. Under C. myriocarpus (Cm), EM × Cm interaction was not significant for nematodes (juveniles + eggs) in roots and juveniles in soil, while under C. africanus (Ca), EM × Ca interaction was highly significant for nematodes, but not for juveniles. Crude extracts of C. myriocarpus and C. africanus fruits contributed 21-36% and 38-59% to total treatment variation in nematodes, respectively. Relative to untreated control, crude extracts of C. myriocarpus fruit reduced nematodes by 22% in roots, but increased juveniles in soil by 93%. Similarly, C. africanus fruit reduced nematodes in roots by 80%, but increased juveniles in soil by 178%. The increase of juveniles in the soil was explained on the basis of opposing forces on nematode population densities under crude extracts of Cucumis and untreated control, along with the inherent nature of cyclic population growth in plant-parasitic nematodes. In plant variables, certain significant (P ≤ 0.05) interactions consistently occurred under both Cucumis species. However, effects of the interactions were not consistent under the two Cucumis species. In most of the variables, the non-significant effects of EM × Cm interactions supported the view that the GLT systems were independent of microbial activities, while significant (P ≤ 0.05) EM × Ca interactions suggested that the systems under C. africanus fruit could be viewed as being dependent upon microbial degradation activities. Growth of rough lemon rootstock was, to a certain extent, suppressed by application of crude extracts from Cucumis fruits, suggesting that the material were phytotoxic to this citrus rootstock. Under low nematode population densities, T. semipenetrans infection supported the view that nematode numbers below the damage threshold levels have stimulatory effects on growth of plants as observed in plant height under conditions of this study. In conclusion, the approximately three nematode-generation-application interval of 150 days for crude extracts of fruits in Cucumis species was rather too long for the efficacies of the materials on suppression of the population densities of T. semipenetrans in rough lemon seedlings. Consequently, shorter application intervals, as demonstrated for Meloidogyne species would be appropriate, although caution has to be taken to ensure that phytotoxicity to the rootstock was avoided.

The interaction of herbicides and Northern root-knot nematodes (Meloidogyne hapla Chitwood) on onions and beans was studied in the greenhouse. Seeds of onions (Allium cepa L.) were planted in soil treated with defferent levels of dimethyl 2,3,5,6-tetrachloroterephthalate (DCPA). Seeds of beans (Phaseolus vulgaris L.) were planted in soil treated with different levels of DCPA or α, α, α-trifluoro-2,6-dinitro-N,N-dipropyl-p-toluidine (trifluralin). Seedlings were inoculated with root-knot nematode larvae after 2 week's growth. DCPA reduced root and shoot growth on onions but did not significantly affect bean growth. Trifluralin significantly reduced root and shoot growth of beans. Growth reduction was directly related to the concentration of DCPA or trifluralin. Anatomical studies on onion roots indicated that the epidermal cells were seriously affected by DCPA treatment. They were greatly misshapen, overlapped, and in various stages of apparent dissolution or collapse. Symptoms were more severe under higher concentrations. Macroscopic examination of roots treated with DCPA or trifluralin showed soil particles strongly adhered to root tissue. DCPA-treated and non-treated nematode larvae were equally pathogenic on onions. Both DCPA and trifluralin significantly reduced the number of nematode galls formed on beans and onions grown in treated soil.

Includes bibliographical references (leaves 241-25). Aims to evaluate sampling procedures; assess the extent and magnitude of yield loss caused by Pratylenchus neglectus; assess the population dynamics of Pratylenchus neglectus in cereals; determine whether resistance occurs in field crops; and, assess whether variation occurs between geographically isolated species of Pratylenchus neglectus

Attempts were made to grow Fergusobia nematodes in a dual aseptic culture with Eucalyptus camaldulensis. Callus tissue was grown from E. camaldulensis stem pieces in aseptic conditions. Calli were prone to deterioration after 14 days unless transferred to fresh growth medium. Lower levels of solutes (25% Murashige and Skoog salts, 25% plant vitamins and 0.5% sucrose) were more successful than published concentrations. Fergusobia J2 nematodes were surface sterilised with either Hibitane or washed with water to prepare them for inoculation of callus (Hay, 1994). Fergusobia subsequently recovered from plates of callus were all dead, which raised questions of how the nematodes are suited to Murashige and Skoog salt solutions. The survival of Fergusobia in aqueous solutions was then observed. It has been assumed that Fergusobia live about 2 days after dissection. Amphimictic nematodes from E. camaldulensis axial bud galls were used for a survival study. Nematodes lived for as long as 12 days in fresh tap water and 11 days in 1% M.S. solution. They were more active in tap water than in 1% M.S. solution. Nematodes in a dish together with dissected gall material died within 2 days. Nematodes in a separate dish with fresh tap water and clean gall fibres were observed to gather around the fibres. Observations of Fergusobia could be made within fresh solutions providing deteriorating gall tissues were removed from the dish. Gall production was attempted on E. camaldulensis grown in the glasshouse. These tree saplings were pruned to encourage new growth and periodically treated for infestations of scale insects leading to growth of sooty mould. Two forms of cage construction were used: (1) 1 m ³ screened cages and (2) acetate sleeves as used by Goolsby et al., 2000. Within the 1 m ³ cages containing flies, the growing points on saplings were blackened, possibly due to over-exploitation by ovipositing flies. One growing point caged in an acetate sleeve showed oviposition scars but did not produce a mature gall. Production of galls in the glasshouse was hindered by a lack in coincidence of flies emerging from mature galls and the flush of new growth following pruning. The production of galls within the glasshouse was not achieved. The phenology of E. camaldulensis, a host for the Fergusonina/Fergusobia mutualism, and gall ecology were observed in a two-year, non-destructive, field study in the Urrbrae Wetlands, Adelaide, South Australia. Tree growth and gall development was observed in the lower regions (0 – 2 m) of young trees. Three bud forms, terminal leaf bud, axial leaf bud and flower bud galls were monitored on the trees. The densities of galls were highly seasonal. Greatest density of growing points, axial leaf bud and flower bud galls occurred mostly during mid-winter to spring, whereas that of terminal leaf bud galls occurred during mid-spring to summer. Galling of flower buds did not appear to influence flowering and more flower buds and flowers occurred in the second year of the study as the trees matured. Trees mostly had medium (30-70%) levels of leaf damage, but there were seasonal trends in damage levels. Low scores for leaf damage were associated with increases in flower bud and flower production. Leaf damage, including sooty mould, appeared to increase during the cooler winter months. There were no significant seasonal relationships between levels of leaf damage and either growing point density or the occurrence of galls. When trees were compared with each other, those with lower leaf damage were more likely to have more growing points. The appearance of the canopy and the likelihood of a tree to have galls varied greatly between the trees. One tree was particularly susceptible to leaf attack, rarely had new leaves and produced no mature galls during the study. The colouring of leaves varied between trees, which indicates possible genetic variations causing some trees to be more likely hosts for Fergusonina/Fergusobia. Both new vegetative growth and terminal leaf bud galls were concentrated on the northern and eastern quadrants. Axial leaf bud and flower bud galls occurred more on the western or southern quadrants where they were possibly more protected from sun exposure on the northern or eastern quadrants. Axial galls on the northern side of one tree became reddened while those in the southern and western quadrants remained green. Reddening of axial galls may increase their likelihood of parasitism and predation by birds. Each of the three gall forms occurred within certain positions in the canopy. The tree host resource is partitioned effectively, with the three gall forms occurring on three different host structures. Additionally, the two vegetative forms terminal leaf bud and axial leaf bud galls occur on different shoot regions and in different seasons. The numbers of the galls is probably also affected by biotic and climatic influences. Parasitism, plant canopy shading, nutrient levels and host genetics are possible influences. Assuming an interval of 4 weeks between oviposition and first observation within the current study, terminal leaf bud galls had an average longevity of 11 weeks and axial leaf bud galls an average longevity of 14 weeks. Flower bud galls had longevities of 14 to 27 weeks from oviposition to senescence, assuming an interval of 6 weeks between oviposition and first observation within the current study. Flowers and flower buds occur irregularly within the eucalypts so it would be advantageous for flies and nematodes developing within flower bud galls to have extended or variable longevities to allow fly emergences to coincide with new flower buds. Not all of the galls recorded matured to produce adult flies. Nearly half of the terminal leaf bud galls initiated were aborted, recorded as absent, parasitised or eaten (45% of initiated galls). Of the three gall forms, they were the most prone to obvious parasitism and as many as 12 hymenopteran species have been reared from terminal leaf bud galls on E. camaldulensis (Taylor et al., 1996). These galls obviously provide a resource for many species within multiple trophic levels. Fourteen percent of axial leaf bud galls were absent or eaten and birds were seen breaking off and feeding on the galls. More than half (55%) of the initiated flower bud galls disappeared during the period of observation, possibly due to the foraging of birds. Destructive sampling and rearing out of parasitoids from both axial leaf bud and flower bud gall forms is needed to establish what species exist within them. Terminal leaf bud galls ranged from 7.5 to 30.1 mm in diameter and 10.0 to 43.6 mm in length. Flower bud gall size varied, with the largest being 15.0 mm by 22.3 mm. Axial leaf bud galls, ranged from 2.6 to 13.0 mm in diameter and length ranged from 2.3 to 10.5 mm. The larger axial leaf bud galls were nodular and appeared to have multiple locules. Destructive sampling and rearing out of flies is needed to establish the relationship between size and numbers of flies emerging. Terminal leaf bud galls increased in size, including many locules and exit holes per gall. Axial leaf bud galls were much smaller than the terminal leaf bud galls and 99% had only one to three exit holes. The rounded shape and presence of few locules within the axial leaf bud galls indicate that this form is limited to a shape and size producing few flies. The observation of greater size of terminal leaf bud galls suggests that these galls may have multiple foundresses. Twelve of the 13 flower bud galls with exit holes had either one or two holes. In flower bud galls on E. camaldulensis. the operculum remains sealed and the characteristic Fergusonina “window” appears at the side of the flower bud before fly emergence through a single hole. Destructive sampling is also necessary to determine parasitism of each of the gall forms. / http://proxy.library.adelaide.edu.au/login?url= http://library.adelaide.edu.au/cgi-bin/Pwebrecon.cgi?BBID=1331016 / Thesis (M.Sc.) -- University of Adelaide, School of Agriculture, Food and Wine, 2008

Includes bibliography / xxvii, 278, xxiv leaves, [43] leaves of plates : ill ; 31 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Plant Pathology, 1987

Copy of author's previously published article inserted. / Bibliography: leaves 60-75. / viii, 75, [40] leaves, [24] leaves of plates : ill. (some col.) ; 30 cm. / Title page, contents and abstract only. The complete thesis in print form is available from the University Library. / Thesis (Ph.D.)--University of Adelaide, Dept. of Crop Protection, 1995?