Orientador: Carlos Alexandre Costa Crusciol / Banca: Gabriela Ferraz de Siqueira / Banca: Rafaela Rossetto / Banca: Gustavo Pavan Mateus / Banca: Sergio Gustavo Quassi de Castro / Resumo: Inúmeros são os patógenos que reduzem a produção da cana-de-açúcar, em particular os fitonematoides apresentam grande importância. Em campo, os sintomas de ataque de nematoides são reboleiras de plantas raquíticas e cloróticas, murchas nas horas mais quentes do dia e menos produtivas. Na tentativa de diminuir as populações de nematoides abaixo do nível de dano econômico, vários métodos de controle, em integração, têm sido pesquisados, dentre eles está o controle químico, biológico e adição de matéria orgânica ao solo. Objetivou-se, por meio do presente estudo, avaliar os efeitos estimulantes de crescimento de plantas de cana-de-açúcar pela aplicação de Trichoderma asperellum, Bacillus subtilis e Bacillus methylotrophicus no controle biológico de nematoides, em área sem e com aplicação de torta de filtro. O experimento foi conduzido em cana planta, soca e soca de 4o corte, nos anos agrícola de 2015/16 e 2016/17, em áreas pertencente à Usina da Barra, Grupo Raízen Energia S/A. O delineamento experimental utilizado foi em blocos casualizados, com quatro repetições: sendo em esquema fatorial 2x5, para cana planta e soca, constituído do fator torta de filtro (1 - sem e 2 - com) combinado com o fator nematicida (1 - controle, 2 - carbofurano, 3 - Quality + Rizos, 4 - Quality + Onix, 5 - Quality + Onix + Rizos), e apenas do fator nematicida para cana soca de 4o corte. As avaliações realizadas foram: teores foliares de macronutrientes, avaliações biométricas (altura, diâmetro, número... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: There are many pathogens that reduce the sugarcane production, especially the phytonematodes are of great importance. In the field, the nematode attack symptoms are the formation of reeds and stunted and chlorotic plants, wilted in the hottest hours of the day and less productive. In order to reduce nematode populations to a minimum level of economic damage, several integration methods have been researched, including chemical, biological control and addition of organic matter to the soil. The objective of this study was to evaluate the growth stimulating effects of sugarcane plants by the application of Trichoderma asperellum, Bacillus subtilis and Bacilus methylotrophicus in the biological control of nematodes, in an area without and with application of filter cake. The experiment was carried out in sugarcane plant, sugarcane ratoon and 4th ratoon, in the agricultural years of 2015/16 and 2016/17, in areas belonging to the Usina da Barra, Grupo Raízen Energia S/A. The experimental design was a randomized complete block design, with four replications: a 2x5 factorial scheme for sugarcane plant and ratoon, consisting of filter cake factor (1 - without and 2 - with) combined with nematicidal factor (1 - control, 2 - carbofuran, 3 - Quality + Rizos, 4 - Quality + Onix, 5 - Quality + Onix + Rizos), and only the nematicidal factor for sugarcane 4th ratoon. The evaluations were: foliar macronutrient contents, biometric evaluations (height, diameter, number of stalks per meter, mean internode length, number of internodes and shoot yield) and technological (pol, purity, fiber, reducing sugar, total sugar recoverable and sugar production). The incorporation of filter cake into the soil contributed to increases in crop productivity through the addition ... / Doutor

Orientador: Alvimar Jose da Costa / Banca: Gilson Pereira de Oliveira / Banca: Welber Daniel Zanetti Lopes / Resumo: Os objetivos específicos do presente trabalho foram diagnosticar espécies de nematódeos resistentes à ivermectina de alta concentração (3,15% e 3,5%) por meio de necropsias parasitológicas realizadas em bovinos procedentes de oitos municípios dos estados de Minas Gerais, São Paulo e Rio Grande do Sul; avaliar comparativamente as eficácias terapêuticas de duas formulações contendo altas concentrações de ivermectina 3,15% (630 mcg/Kg) e 3,5% (700 mcg/Kg), contra nematódeos parasitos de bovinos naturalmente infectados; fornecer subsídios para um melhor conhecimento sobre a distribuição da resistência de nematódeos à ivermectina, em diferentes regiões do Brasil. Para isto foram utilizados 108 animais, naturalmente infectados, os quais foram selecionados pelas médias de três pré-contagens (-3,-2 e -1) de ovos por grama de fezes (OPG), divididos em grupos experimentais, constituídos por seis repetições. Quatorze dias pós-tratamento (DPT), os animais foram eutanasiados e necropsiados. Resistência à ivermectina 3,15% (630 mcg/Kg) e 3,5% (700 mcg/Kg) foi diagnosticada obtendo eficácia terapêutica insuficientemente efetiva (<90%) pelas médias aritméticas ou 1000 espécimes sobreviveram nos grupos tratados, dentre as quais: Haemonchus placei (0,0-89,27%), Cooperia punctata (0,0-26,46%), Cooperia pectinata (0,0-90,16%), Cooperia spatulata (21,82-71,50%), Trichostrongylus axei (82,23%), Oesophagostomum radiatum (54,93-83,33%) e Trichuris discolor (64,71-82,58%); Considerando apenas os resultados analisados estatisticamente referentes aos oito experimentos realizados (116 necropsias) pode-se inferir que a resistência à ivermectina, em altas concentrações (630 e 700 μg/Kg) está amplamente disseminada, sobretudo nas espécies Haemonchus placei, Cooperia punctat... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The specific objectives of the present study were to diagnose high concentration ivermectin (3,15% and 3,5%) resistance in nematodes through parasitological necropsies conducted in bovines originating from eight cities from the states of Minas Gerais, São Paulo and Rio Grande do Sul; evaluate, by comparison, the therapeutic efficacies of two formulations containing high concentrations of ivermectin, respectively 3,15% (630 mcg/Kg) and 3,5% (700 mcg/Kg), against nematodes parasitizing naturally infected bovines; provide aid for a better knowledge about the distribution of ivermectin resistance in nematodes throughout different regions of Brazil. To achieve these goals 108 naturally infected animals were used, all selected using the average of three EPG counts (-3, -2 and -1), divided in experimental groups consisting of six repetitions. Fourteen days after treatment all animals were euthanized and necropsied. Resistance to 3,15% ivermectin (630 mcg/Kg) and 3,5% ivermectin (700 mcg/Kg) was diagnosed by obtaining insufficient therapeutical efficacy (< 90%) when analyzing arithmetic means or by finding at least 1000 specimens that survived in the treated groups, amongst which, Haemonchus placei (0,0-89,27%), Cooperia punctata (0,0-26,46%), Cooperia pectinata (0,0-90,16%), Cooperia spatulata (21,82-71,50%), Trichostrongylus axei (82,23%), Oesophagostomum radiatum (54,93-83,33%) e Trichuris discolor (64,71-82,58%). Considering only the statistically analyzed results related to the eight experiments conducted (116 necropsies) it can be inferred that resistance to high concentration ivermectin (630 and 700 μg/Kg) is highly disseminated, specially in Haemonchus placei, Cooperia punctata, Cooperia pectinata, Cooperia spatulata, Trichostrongylus axei, Oesophagostomum radiatum and... (Complete abstract click electronic access below) / Mestre

Orientador: Estevam Guilherme Lux Hoppe / Banca: Adjair Antônio do Nascimento / Banca: Paulo Aléscio Canola / Banca: Daniel Cortes Beretta / Banca: José Ribamar Privado Filho / Resumo: O monepantel, anti-helmíntico de uma nova classe, foi desenvolvido e colocado no mercado recentemente, como solução para criações com helmintos de ovinos multirresistentes. Após pouco tempo de utilização, foram relatados casos de resistência contra o produto. Objetivou-se avaliar o perfil de resistência ao monepantel em criações de ovinos em uma microrregião em torno de Jaboticabal-SP e caracterizar experimentalmente a resistência ao monepantel em isolado de Haemonchus contortus. Foi realizado o teste de redução de ovos em dez propriedades ovinocultoras próximas a Jaboticabal/SP, identificando a eficácia do monepantel e levantando o histórico da utilização do produto para determinação de fatores de risco associados à resistência. Obteve-se isolado de H. contortus resistente para teste crítico. Após coleta de fezes dos animais oriundos de propriedade onde o vermífugo foi ineficaz, obteve-se larvas infectantes que foram inoculadas em dois doadores para obtenção de fêmeas de H. contortus. Larvas infectantes deste isolado foram inoculadas em dez animais. Após confirmada a patência, cinco animais receberam monepantel, grupo tratamento, e cinco não receberam, grupo controle. Exames de OPG foram realizados nos dias 0 (dia do tratamento), 2, 4, 6, 8, 10, 12 e 14 e os animais eutanasiados no dia 14, avaliando o número de H. contortus no abomaso dos animais dos dois grupos. Revelou-se que em duas propriedades o monepantel foi ineficaz, em duas foi eficaz e nas outras seis foi altamente eficaz. Os principais fatores predisponentes detectados foram: intervalos curtos entre vermifugações, não alternância com outras bases químicas, tratamento massivo, raça e intensificação da criação. Observou-se H. contortus resistente após realização do teste crítico com eficácia de 24,65%. Conclui-se que helmintos de ovinos já estão resistentes ao monepantel em criações... / Abstract: Monepantel, anthelmintic of a new class, was recently developed and released to the international market, it was presented as a solution for properties with multirresistant helminthes. However, with little time utilization, were related cases of resistance against this product. The objectives of this work were evaluate the resistance profile of monepantel on sheep creations at a microregion around Jaboticabal-SP and characterize experimentally the monepantel resistance on Haemonchus contortus isolate. Initially, was accomplished egg count reduction test at ten ovine properties located around Jaboticabal-SP, identifying the monepantel efficacy and investigating the history of product use for determination risk factors resistance associated. Posteriorly, a H. contortus resistant isolate was obtained for the critic test. After animals collect stoll of a resistance suspected proprierty, there were obtained infective larvae that were inoculated on two donors for females of H. contortus obtainment and after, infective larvae production for inoculation in ten animals. After patency confirmed, five animals received monepantel, treatment group, and five didn't receive, control group. The treatment day was day 0. Fecal egg counts were performed on days 0, 2, 4, 6, 8, 10, 12 and 14 and the animals were euthanized on day 14, evaluating the H. contortus number in the sheep abomaso of both groups. The results of the fisrt part study revealed the monepantel was ineffective in two properties, was effective in two properties and highly effective on six properties. The risk factors detected were: short interval between anthelmintic treatments, no alternation with other chemical bases, massive treatment, breed, intensification of creation. In a second step, was observed H. contortus resistant after performed the critc test, with an efficacy of 24,65%. We concluded that sheep helminths are already monepantel ... / Doutor

Thesis (M.Sc.) - University of the North, 1977 / Refer to document

This thesis describes a series of three experiments designed to estimate the nutritional cost of the immune response to the gastrointestinal nematodes Trichostrongylus colubriformis and Teladorsagia circumcincta in sheep. For each experiment, animals were allocated hierarchically by liveweight into one of four groups that were either infected (group IF), similarly infected and concurrently immuno-suppressed with weekly intramuscular injections of 1.3mg kg liveweight (LW)-1 of methylprednisolone acetate (group ISIF), immuno-suppressed only (group IS) or remained as controls (group C). Body composition of all animals was estimated using x-ray computer tomography prior to infection and at the conclusion of each study with bodyweight and faecal nematode egg counts (FEC; eggs gram-1 of fresh faeces (epg)) measured along with blood samples taken for the determination of levels of serum proteins, phosphate and antibodies. In the first trial (Chapter 3), the nutritional cost of both the acquisition and maintenance of immunity to gastro-intestinal nematodes was investigated using immunologically naive 5-month-old lambs and immunologically competent 17-month-old ewes during infection with 2,000 and 4,000 L3 infective T. colubriformis larvae d-1, respectively (80 L3 T. colubriformis larvae kgLW-1 d-1). Profiles of FEC and comparative worm burdens at slaughter indicated an effective immune response was maintained in IF ewes and developed in IF lambs while successfully suppressed in both ISIF lambs and ISIF ewes and was confirmed by serum antibody titres. The typical reduction in voluntary feed intake as a consequence of infection was observed in IF lambs (0.30, p&lt0.001) but not in IF ewes, ISIF lambs or ISIF ewes, and appeared to be associated with L3 IgA. Gross efficiency of use of metabolizable energy (ME) for net energy (NE) deposition was reduced by 0.20 in lambs during acquisition of immunity and by 0.16 in ewes maintaining an established immunity. Infection in immuno-suppressed animals reduced efficiency by 0.05 and 0.15 for lambs and ewes. These findings allowed the hypothesis that the reduction in feed intake and nutrient utilization in young parasitized sheep is caused by physiological signalling associated with the acquisition phase of the host immune response to infection, rather than simply the damage caused by the parasite per se. The second trial (Chapter 4) investigated the influence of metabolizable protein (MP) supply on the metabolic disturbances associated with the acquisition phase of the immune response during infection with 2,000 L3 T. colubriformis d-1. Groups of lambs were offered either a low protein (L; 62g MP kgDM-1) or high protein diet (H; 95g MP kgDM-1). Patterns of total daily egg excretion indicated that an effective immune response was developed in HIF, but not LIF, HISIF nor LISF and was confirmed by comparative worm burdens. The proportionate reduction in feed intake in immunologically normal animals was reduced through the provision of additional protein, being 0.12 in HIF and 0.23 in LIF. Regardless of diet, infection did not cause a reduction in feed intake in immuno-suppressed animals (p&gt0.05). Infection proportionately reduced the gross efficiency of ME utilization in immunologically normal animals by 0.23 in HIF (p=0.09) and by 0.51 in LIF (p=0.01), but not in immuno-suppressed animals. Immuno-suppression did not suppress serum L3 IgA levels in seven of the eight HISIF and four of the eight LISIF animals. Furthermore, only four out of the eight immunologically normal animals from both the HIF and LIF groups displayed an L3 IgA response. Consequently, regardless of immuno-suppression treatment, animals were termed as IgA responders (HR or LR) or non-responders (HN or LN). Feed intake was proportionately reduced from day 22 by 0.15 in HR (p=0.03) and by 0.32 in LR (p=0.01), but was not significantly reduced in HN or LN. Gross efficiency of ME utilization was significantly reduced for LN animals only, being proportionately 0.59 (p&lt0.01). These findings allowed the conclusion that additional MP reduced the consequence of immunological signalling that was displayed in reduced feed intake and in nutrient utilization, both of which appeared to be associated with an IgA response. It is hypothesized that the lessening of nutritional disturbance observed in high protein and immuno-suppressed animals could be a consequence of altered physiological signalling during the immunological cascade. The third trial (Chapter 5) utilized lambs infected with the abomasal parasite T. circumcincta to explore the possibility that the reduction in feed intake and nutrient utilization is a universal phenomenon of the acquisition phase of the immune response to nematode parasites inhabiting different organs along the gastrointestinal tract. In addition, immunological changes at the site of parasite infestation in the abomasal mucosa were measured from serial biopsy tissue samples taken from a further twelve animals that were surgically fitted with an abomasal cannula and either infected (CIF) or concurrently infected and immuno-suppressed as described previously (CISIF). The development of immunity in IF animals was accompanied by a 0.17 proportional decrease in feed intake between days 15 to 28 of infection (p&lt0.05) and a 0.20 proportional reduction in nutrient utilization (p=0.07), none of which were observed in ISIF animals. While FEC and worm burdens indicated successful immuno-suppression in ISIF animals, both serum IgA and total antibody production were not reduced. The development of immunity in CIF was reflected in an increase in both mast cells and globule leukocytes in serial abomasal tissue biopsies, both of which were reduced in CISIF (p&lt0.01 for both). In serial biopsy tissue, immuno-suppression did prevent a rise in tissue IgA that was apparent in CIF animals (p&lt0.01) although these changes were not reflected in serum IgA levels. It appears that the alleviation of the reduction in feed intake and nutrient utilization in young lambs through the use of corticosteroid induced immuno-suppression may be a universal phenomenon for both intestinal and abomasal parasites, but the association with and/or role of IgA during infection with T. circumcincta is unclear. In summary, the reduction in feed intake and nutrient utilization in sheep during infection with both the abomasal nematode T. circumcincta and the small intestine nematode T. colubriformis appears to be associated with a component(s) of the acquisition phase of the host immune response, rather than, as conventionally assumed, the direct mechanical damage of the parasite per se. It is hypothesised that the nutritional disturbance as a consequence of infection in young lambs may be the result of pro-inflammatory cytokines involved in immunological signalling that may also be associated with the production of IgA, the effects of which can be reduced through the provision of adequate MP. These studies provide evidence that the immune response to gastrointestinal parasites is nutritionally costly to the animal and have implications for application of manipulations that are intended to promote the development of a strong immune reaction in high producing animals.

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

Bibliography: leaves 113-128 / xii, 128, [63] leaves, [6] leaves of plates : ill ; 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 Microbiology and Immunology, 1984

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

Bibliography: leaves 111-137.

The Colorado potato beetle (CPB), Leptinotarsa decemlineata (Say), is the most devastating foliage-feeding pest of potatoes in the United States. Potential biological control agents include the nematodes Heterorhabditis marelatus Liu & Berry and Steinernema riobrave Cabanillas, Poinar & Raulston, which provided nearly 100% CPB control in previous laboratory trials. In the present study, laboratory assays tested survival and infection by the two species under the soil temperatures CPB are exposed to, from 4-37°C. H. marelatus survived from 4-31°C, and S. riobrave from 4-37°C. Both species infected and developed in waxworm hosts from 13-31°C, but H. marelatus rarely infected hosts above 25°C, and S. riobrave rarely infected hosts below 19°C. H. marelatus infected an average of 5.8% of hosts from 13- 31°C, whereas S. riobrave infected 1.4%. Although H. marelatus could not survive at temperatures as high as S. riobrave. H. marelatus infected more hosts so is preferable for use in CPB control. Heterorhabditis marelatus rarely reproduced in CPB. Preliminary laboratory trials suggested the addition of nitrogen to CPB host plants improved nematode reproduction. Field studies testing nitrogen fertilizer effects on nematode reproduction in CPB indicated that increasing nitrogen from 226 kg/ha to 678 kg/ha produced 25% higher foliar levels of the alkaloids solanine and chaconine. However, the increased alkaloids did not affect nematode infection of, nor reproduction in, CPB prepupae. Nematodes applied to field plot soil at 50 infective juveniles/cm² reduced adult CPB by 50%, and increased numbers of dead prepupae in soil samples up to five times more than in non-nematode plots. Laboratory studies of H. marelatus and its symbiotic bacteria in CPB hemolymph indicated that immune responses did not limit nematode reproduction. A 58kD CPB hemolymph protein apparently caused the symbiotic bacteria to switch to the secondary form, which does not produce antibiotics and enzymes necessary for nematode growth and reproduction. Despite heat denaturation of the protein, the nematodes did not reproduce unless lipids were added to the hemolymph. Therefore, while H. marelatus may provide high levels of CPB control, nutritional constraints on the nematode and its bacteria inhibit reproduction in CPB and limit long-term multi-generation control. / Graduation date: 2003