Host plant recognition by the root feeding clover weevil, Sitona lepidus (Coleoptera : Curculionidae)

This study investigated the ability of neonatal larvae of the root-feeding weevil, Sitona lepidus Gyllenhal, to locate white clover Trifolium repens L. (Fabaceae) roots growing in soil and to distinguish them from the roots of other species of clover and a co-occurring grass species. Choice experiments used a combination of invasive techniques and the novel technique of high resolution X-ray microtomography to non-invasively track larval movement in the soil towards plant roots. Burrowing distances towards roots of different plant species were also examined. Newly hatched S. lepidus recognized T. repens roots and moved preferentially towards them when given a choice of roots of subterranean clover, Trifolium subterraneum L. (Fabaceae), strawberry clover Trifolium fragiferum L. (Fabaceae), or perennial ryegrass Lolium perenne L. (Poaceae). Larvae recognized T. repens roots, whether released in groups of five or singly, when released 25 mm (meso-scale recognition) or 60 mm (macro-scale recognition) away from plant roots. There was no statistically significant difference in movement rates of larvae.

Tracking larval insect movement within soil using high resolution X-ray microtomography

1. In contrast to above-ground insects, comparatively little is known about the behaviour of subterranean insects, due largely to the difficulty of studying them in situ. 2. The movement of newly hatched (neonate) clover root weevil (Sitona lepidus L. Coleoptera: Curculinidae) larvae was studied non-invasively using recently developed high resolution X-ray microtomography. 3. The movement and final position of S. lepidus larvae in the soil was reliably established using X-ray microtomography, when compared with larval positions that were determined by destructively sectioning the soil column. 4. Newly hatched S. lepidus larvae were seen to attack the root rhizobial nodules of their host plant, white clover (Trifolium repens L.). Sitona lepidus larvae travelled between 9 and 27 mm in 9 h at a mean speed of 1.8 mm h(-1). 5. Sitona lepidus larvae did not move through the soil in a linear manner, but changed trajectory in both the lateral and vertical planes.

Effects of carbon dioxide on the searching behaviour of the root-feeding clover weevil Sitona lepidus (Coleoptera : Curculionidae)

The respiratory emission of CO2 from roots is frequently proposed as an attractant that allows soil-dwelling insects to locate host plant roots, but this role has recently become less certain. CO2 is emitted from many sources other than roots, so does not necessarily indicate the presence of host plants, and because of the high density of roots in the upper soil layers, spatial gradients may not always be perceptible by soil-dwelling insects. The role of CO2 in host location was investigated using the clover root weevil Sitona lepidus Gyllenhall and its host plant white clover (Trifolium repens L.) as a model system. Rhizochamber experiments showed that CO2 concentrations were approximately 1000 ppm around the roots of white clover, but significantly decreased with increasing distance from roots. In behavioural experiments, no evidence was found for any attraction by S. lepidus larvae to point emissions of CO2, regardless of emission rates. Fewer than 15% of larvae were attracted to point emissions of CO2, compared with a control response of 17%. However, fractal analysis of movement paths in constant CO2 concentrations demonstrated that searching by S. lepidus larvae significantly intensified when they experienced CO2 concentrations similar to those found around the roots of white clover (i.e. 1000 ppm). It is suggested that respiratory emissions of CO2 may act as a 'search trigger' for S. lepidus, whereby it induces larvae to search a smaller area more intensively, in order to detect location cues that are more specific to their host plant.

Modelling the movement and survival of the root-feeding clover weevil, Sitona lepidus, in the root-zone of white clover

White clover (Trifolium repens) is an important pasture legume but is often difficult to sustain in a mixed sward because, among other things, of the damage to roots caused by the soil-dwelling larval stages of S. lepidus. Locating the root nodules on the white clover roots is crucial for the survival of the newly hatched larvae. This paper presents a numerical model to simulate the movement of newly hatched S. lepidus larvae towards the root nodules, guided by a chemical signal released by the nodules. The model is based on the diffusion-chemotaxis equation. Experimental observations showed that the average speed of the larvae remained approximately constant, so the diffusion-chernotaxis model was modified so that the larvae respond only to the gradient direction of the chemical signal but not its magnitude. An individual-based lattice Boltzmann method was used to simulate the movement of individual larvae, and the parameters required for the model were estimated from the measurement of larval movement towards nodules in soil scanned using X-ray microtomography. The model was used to investigate the effects of nodule density, the rate of release of chemical signal, the sensitivity of the larvae to the signal, and the random foraging of the larvae on the movement and subsequent survival of the larvae. The simulations showed that the most significant factors for larval survival were nodule density and the sensitivity of the larvae to the signal. The dependence of larval survival rate on nodule density was well fitted by the Michealis-Menten kinetics. (c) 2005 Elsevier B.V All rights reserved.

PGR forum: serving the crop wild relative user community

The Euro-Mediterranean region is an important centre for the diversity of crop wild relatives. Crops, such as oats (Avena sativa), sugar beet (Beta vulgaris), apple (Malus domestica), annual meadow grass (Festuca pratensis), white clover (Trifolium repens), arnica (Arnica montana), asparagus (Asparagus officinalis), lettuce (Lactuca sativa), and sage (Salvia officinalis) etc., all have wild relatives in the region. The European Community funded project, PGR Forum (www.pgrforum.org) is building an online information system to provide access to crop wild relative data to a broad user community; including plant breeders, protected area managers, policy-makers, conservationists, taxonomists and the wider public. The system will include data on uses, geographical distribution, biology, population and habitat information, threats (including IUCN Red List assessments) and conservation actions. This information is vital for the continued sustainable utilisation and conservation of crop wild relatives. Two major databases have been utilised as the backbone to a Euro-Mediterranean crop wild relative catalogue, which forms the core of the information system: Euro+Med PlantBase (www.euromed.org.uk) and Mansfeld’s World Database of Agricultural and Horticultural Crops (http://mansfeld.ipk-gatersleben.de). By matching the genera found within the two databases, a preliminary list of crop wild relatives has been produced. Around 20,000 of the 30,000+ species listed in Euro+Med PlantBase can be considered crop wild relatives, i.e. those species found within the same genus as a crop. The list is currently being refined by implementing a priority ranking system based on the degree of relatedness of taxa to the associated crop.

Efficacy of condensed tannins against larval Hymenolepis diminuta (Cestoda) in vitro and in the intermediate host Tenebrio molitor (Coleoptera) in vivo

Natural anti-parasitic compounds in plants such as condensed tannins (CT) have anthelmintic properties against a range of gastrointestinal nematodes, but for other helminths such effects are unexplored. The aim of this study was to assess the effects of CT from three different plant extracts in a model system employing the rat tapeworm, Hymenolepis diminuta, in its intermediate host, Tenebrio molitor. An in vitro study examined infectivity of H. diminuta cysticercoids (excystation success) isolated from infected beetles exposed to different concentrations of CT extracts from pine bark (PB) (Pinus sps), hazelnut pericarp (HN) (Corylus avellana) or white clover flowers (WC) (Trifolium repens), in comparison with the anthelmintic drug praziquantel (positive control). In the in vitro study, praziquantel and CT from all three plant extracts had dose-dependent inhibitory effects on cysticercoid excystation. The HN extract was most effective at inhibiting excystation, followed by PB and WC. An in vivo study was carried out on infected beetles (measured as cysticercoid establishment) fed different doses of PB, HN and praziquantel. There was a highly significant inhibitory effect of HN on cysticercoid development (p = 0.0002). Overall, CT showed a promising anti-cestodal effect against the metacestode stage of H. diminuta.

Potent anthelmintic activity of galloylated proanthocyanidins from shea meal (Vitellaria paradoxa)

Proanthocyanindins (PAs) from shea meal (SM), a by-product obtained after lipid extraction of the nuts, contained B-type linkages, had a high ratio of prodelphinidins (73%) and were galloylated (42%). The average polymer size was 8 flavan-3-ol subunits (≈2384 Daltons) and epigallocatechin gallate was the major subunit. Purified PA fractions from SM were tested in vitro for anthelmintic properties against gastrointestinal nematodes from ruminants (H. contortus and T. colubriformis) [1] by the larval exsheathment inhibition assay and from pigs (A. suum) by the larval migration inhibition assay. Results showed that PAs from SM have a potent anthelmintic activity against those parasites similar to white clover (Trifolium repens) flowers (WCF) [1, 2] (EC50 µg/mL; SM: 55.1, 16.5, 75.9; WCF: 37.4, 14.5, 110.1 for A. suum, H. contortus and T. colubriformis respectively). WCF PAs are constituted almost exclusively of prodelphinidin (PD) compared to SM (98% vs. 73%) but do not contained galloylated PAs. Studies [1, 2] have shown that anthelmintic activity of PAs was mainly associated with their PD ratio but our current results suggest that galloylation can be a major factor to anthelmintic activity and SM as a potential nutraceutical anthelmintic feed for controlling parasitic nematodes.

Desempenho de novilhas expostas à reprodução aos 14/15 meses de idade

Foram conduzidos dois experimentos para avaliar o comportamento reprodutivo de novilhas de corte pertencentes a quatro grupos genéticos (Hereford, ½ Nelore½ Hereford; ¼ Nelore ¾ Hereford; ½ Angus ½ Hereford) acasaladas aos 14/15 meses de idade. No experimento iniciado em 2001 foram utilizadas quatro alternativas de alimentação no outono/inverno: a) suplementação do campo nativo (CN) a 1,5% do peso vivo (PV) com ração comercial (RC) contendo 14% de proteína bruta (PB) e 68% de NDT (S68); b) suplementação do CN a 1,5% do PV com RC contendo 14% de PB e 75% de NDT (S75); c) suplementação em pastagem de azevém (Lolium multiflorum Lan), trevo branco (Trifolium repens) e cornichão (Lotus corniculatus) a 0,5% do PV de milho em grão (SPAS);d) confinamento a céu aberto com silagem de milho e 0,7% do PV de ração (CON). Após, em um só grupo, permaneceram em pastagem de azevém e aveia (Avena sativa) até o início do período reprodutivo. Durante o período de aplicação dos tratamentos alimentares o ganho médio diário (GMD) do tratamento SPAS e CON (0,755 vs 0,784 kg/dia) não apresentou diferença significativa (P>0,05), nem os S68 e S75 (0,511 vs 0,489 kg/dia). Durante o período conjunto em pastagem não foi determinada diferença significativa (P>0,05) no GMD entre S68, S75 e SPAS, nem entre S68, SPAS e CON. O peso ao início do período reprodutivo foi de 233,7 e 232,3 kg para S68 e S75 (P>0,05) e de 260,6 e 254,3 kg para SPAS e CON (P>0,05). As taxas de prenhez do SPAS e COM não apresentaram diferença entre si (62,16% e 53,86%), mas foram significativamente mais elevadas (P<0,05) em relação às S68 e S75 (20,51% e 25,64%) as quais também não diferiram entre si (P>0,05). As novilhas que conceberam foram mais pesadas, apresentaram maiores GMD de peso, condição corporal (CC) e escore trato reprodutivo (ETR) ao início do período reprodutivo em relação às que não conceberam. No experimento iniciado em 2002 foram utilizadas três alternativas de alimentação no outono/inverno: a) suplementação do CN com RC (SUR) a 1% do PV, contendo 14% de PB e 75% de NDT; b) suplementação do CN com farelo de arroz (SUFA) a 0,5% PV; c) pastejo contínuo em pastagem cultivada de azevém (PAST) e aveia. O tratamento PAST (0,478 kg/dia) apresentou GMD de peso mais elevado (P<0,05), seguido do tratamento SUR (0,326 kg/dia) superior (P<0,05) ao tratamento SUFA (0,100 kg/dia). Durante o período em conjunto em pastagem de azevém e aveia os tratamentos SUFA e PAST não apresentaram diferença significativa no GMD de peso (P>0,05) mas, foram superiores (P<0,05) ao tratamento SUR. O peso ao início do período reprodutivo foi de 265,4; 236,7 e 222,6 kg para os tratamentos PAST, SUR e SUFA, respectivamente (P<0,05). A taxa de prenhez do tratamento PAST (61,36%) foi superior (P<0,05) a dos tratamentos SUR e SUFA (20,0 e 22,73%; P>0,05). As novilhas que conceberam foram as mais pesadas e mais velhas, do início dos tratamentos alimentares até o final do período reprodutivo.

Identificação e avaliação de sistemas de produção de leite em pastagens cultivadas na região do Planalto do Rio Grande do Sul

A pecuária leiteira tem uma importante participação na economia do setor primário do Sul do Brasil, porém, custos de produção elevados, impacto ambiental e qualidade dos produtos tem limitado sua viabilidade econômica. Entretanto, na busca por sistemas sustentáveis de produção depara-se com uma grande escassez de informações. Este trabalho pretendeu contribuir com a geração de conhecimentos que possam ser facilmente adotados pelos produtores da Região do Planalto Riograndense, através da condução da pesquisa em duas etapas: inicialmente foi desenvolvido um estudo baseado em procedimentos de pesquisa diagnóstica aplicados em 46 propriedades que incluiu características infra-estruturais, ambientais, sociais e técnico-econômicas. Em um segundo momento, realizou-se a comparação de dois sistemas de condução dos animais em pastagens, propostos a partir de características apontadas como relevantes no diagnóstico realizado, definindo os seguintes tratamentos: T 1: onde os animais tiveram acesso à pastagem por um período de 3 h/dia, e, T 2: onde os animais tiveram livre acesso à pastagem, ambos, mantidos em pastejo sobre sorgo forrageiro (Sorghum vulgare) e trevo branco (Trifolium repens L.) em pastejo rotativo, recebendo suplementação protéica e energética Os resultados do diagnóstico indicaram uma elevada importância das características de infraestrutura e de questões técnicas relativas à exploração leiteira, onde destacam-se variáveis relacionadas à questões de manejo e alimentação do rebanho. Os sistemas de pastejo apresentaram diferença significativa (p<0,01), na produção de leite, produzindo 16,6 e 18,9 L/dia, para T1 e T2, respectivamente. Com a obtenção desses resultados, fica evidenciado que animais de bom potencial genético, proporcionam respostas positivas de produtividade a medida que lhes é permitido maior tempo de permanência em pastagens de qualidade, ofertadas em quantidade suficiente possibilitando a expressão do potencial do animal em selecionar a sua dieta.

Desempenho produtivo e comportamento de vacas holandesas com a utilização de duas fontes protéicas na alimentação

O trabalho experimental foi desenvolvido em uma propriedade de produção leiteira no município de Camargo, na região fisiográfica denominada de Planalto Médio do RS, com o objetivo de estudar e quantificar o efeito da substituição do componente protéico (farelo de soja) da ração, comumente fornecida aos animais, por pastejo controlado de trevo branco (Trifolium repens L.) em animais mantidos em pastagens de azevém (Lolium multiflorum L.), na performance produtiva, comportamento e qualidade do leite de vacas holandesas. Para tal, foram utilizados dois grupos de animais agrupados de forma homogênea após estratificação por produção, período de lactação e peso corporal, em um delineamento experimental completamente casualizado em blocos. Os dois grupos eram mantidos em pastagens de azevém, por um determinado período de tempo durante o dia (aproximadamente 5 horas). Um grupo recebia diariamente 3 kg de suplemento energético constituído de farelo de milho, farelo de trigo, casca de soja e sal mineral (subtraída de farelo de soja) e tinha acesso controlado à pastagem de trevo branco (aproximadamente 2,5 horas), denominado tratamento TB; outro recebia a mesma quantidade de suplemento, adicionada de quantidade de proteína equivalente ao consumo diário no tratamento anterior via trevo, na forma de farelo de soja, denominado tratamento FS. As medições eram realizadas em avaliações quinzenais, procedendo-se controles leiteiros individuais, medidas de perímetro torácico, coletas de amostras de sangue e leite dos animais e registro de atividades dos animais em pastejo. Os resultados demonstraram semelhanças significativas entre os dois tratamentos, para a maioria das variáveis estudadas, com exceção do teor de proteína do leite que foi maior para o tratamento TB, e os valores de teor de lactose, tempo de pastejo e freqüência de bocados maiores para o tratamento FS. Estes resultados indicam que a utilização do trevo branco, como fonte protéica para os animais, foi tecnicamente viável e eficiente.

Biological and molecular characterization of an isolate of Tobacco streak virus obtained from soybeans in Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Comportamento ingestivo de cordeiros em terminação a pasto

The objective of this study was to evaluate intake behavior of lambs in pasture systems: (1) lambs kept with their dams in ryegrass (Lolium multiflorum Lam.) without supplementation; (2) lambs kept with their dams in the same pasture but supplemented in creep feeding; (3) lambs kept with their dams in the same pasture but supplemented in creep grazing with white clover. Three evaluations of intake behavior were performed to check the activities carried out by animals (grazing, rumination, suckling and other activities). Grazing, rumination and other activities times were different (p < 0.05) for non-supplemented lambs. For these lambs, grazing and rumination times were longer than other activities time compared to other systems. Creep feeding reduced grazing time (p < 0.05), probably due to easy access and quality of concentrate. Lambs in creep grazing had reduced (p < 0.05) rumination time. Bite mass in creep grazing was higher (p < 0.05), probably due to easy harvesting of leaves by lambs. It was concluded that the production system affects feeding strategy of lambs and the preference for legumes suggests the use of creep grazing for finishing lambs.

Greenhouse gas balance in the conversion from extensive pasture to other agricultural systems in Andean region of Colombia

Pós-graduação em Agronomia (Ciência do Solo) - FCAV

Phytoremediation of mercury by terrestrial plants

Mercury (Hg) pollution is a global environmental problem. Numerous Hg-contaminated sites exist in the world and new techniques for remediation are urgently needed. Phytoremediation, use of plants to remove pollutants from the environment or to render them harmless, is considered as an environment-friendly method to remediate contaminated soil in-situ and has been applied for some other heavy metals. Whether this approach is suitable for remediation of Hg-contaminated soil is, however, an open question. The aim of this thesis was to study the fate of Hg in terrestrial plants (particularly the high biomass producing willow, Salix spp.) and thus to clarify the potential use of plants to remediate Hg-contaminated soils. Plants used for phytoremediation of Hg must tolerate Hg. A large variation (up to 30-fold difference) was detected among the six investigated clones of willow in their sensitivity to Hg as reflected in their empirical toxicity threshold (TT95b), the maximum unit toxicity (UTmax) and EC50 levels. This gives us a possibility to select Hg-tolerant willow clones to successfully grow in Hgcontaminated soils for phytoremediation. Release of Hg into air by plants is a concern when using phytoremediation in practice. No evidence was found in this study that Hg was released to the air via shoots of willow, garden pea (Pisum sativum L. cv Faenomen), spring wheat (Triticum aestivum L. cv Dragon), sugar beet (Beta vulgaris L. cv Monohill), oil-seed rape (Brassica napus L. cv Paroll) and white clover (Trifolium repens L.). Thus, we conclude that the Hg burden to the atmosphere via phytoremediation is not increased. Phytoremediation processes are based on the ability of plant roots to accumulate Hg and to translocate it to the shoots. Willow roots were shown to be able to efficiently accumulate Hg in hydroponics, however, no variation in the ability to accumulate was found among the eight willow clones using CVAAS to analyze Hg content in plants. The majority of the Hg accumulated remained in the roots and only 0.5-0.6% of the Hg accumulation was translocated to the shoots. Similar results were found for the five common cultivated plant species mentioned above. Moreover, the accumulation of Hg in willow was higher when being cultivated in methyl-Hg solution than in inorganic Hg solution, whereas the translocation of Hg to the shoots did not differ. The low bioavailability of Hg in contaminated soil is a restricting factor for the phytoextraction of Hg. A selected tolerant willow clone was used to study whether iodide addition could increase the plant-accumulation of Hg from contaminated soil. Both pot tests and field trials were carried out. Potassium iodide (KI) addition was found to mobilize Hg in contaminated soil and thus increase the bioavailability of Hg in soils. Addition of KI (0.2–1 mM) increased the Hg concentrations up to about 5, 3 and 8 times in the leaves, branches and roots, respectively. However, too high concentrations of KI were toxic to plants. As the majority of the Hg accumulated in the roots, it might be unrealistic to use willow for phytoextraction of Hg in practice, even though iodide could enhance the phytoextraction efficiency. In order to study the effect of willow on various soil fractions of Hg-contaminated soil, a 5-step sequential soil extraction method was used. Both the largest Hg-contaminated fractions, i.e. the Hg bound to residual organic matter (53%) and sulphides (43%), and the residual fraction (2.5%), were found to remain stable during cultivations of willow. The exchangeable Hg (0.1%) and the Hg bound to humic and fulvic acids (1.1%) decreased in the rhizospheric soil, whereas the plant accumulation of Hg increased with the cultivation time. The sum of the decrease of the two Hg fractions in soils was approximately equal to the amount of the Hg accumulated in plants. Consequently, plants may be suitable for phytostabilization of aged Hg-contaminated soil, in which root systems trap the bioavailable Hg and reduce the leakage of Hg from contaminated soils.

Effects of flooding on coastal vegetation and salt marshes

This thesis examines the effects of flooding on coastal and salt marsh vegetation. I conducted a field experiment in Bellocchio Lagoon to test the effects of different inundation periods (Level 1 = 0.468 or 11.23 hours; Level 2 = 0.351 or 8.42 hours; Level 3 = 0.263 or 6.312 hours; Level 4 = 0.155 or 3.72 hours; Level 5 = 0.082 or 1.963 hours; Level 6 = 0.04 or 0.96 hours) on the growth responses and survival of the salt marsh grass Spartina maritima in summer 2011 and 2012. S. maritima grew better at intermediate inundation times (0,351 hours; 0,263 hours, 0,115 hours; 0,082 hours), while growth and survival were reduced at greater inundation periods (0,468 hours). The differences between the 2011 and 2012 experiment were mainly related to differences in the initial number of shoots (1 and 5, respectively in 2011 and 2012). In the 2011 experiment a significant lower number of plants was present in the levels 1 and 6, the rhizomes reached the max pick in level 4, weights was major in level 4, spike length reached the pick in level 3 while leaf length in level 2. In the 2012 experiment the plants in level 6 all died, the rhizomes were more present in level 3, weights was major in level 3, spike length reached the pick in level 3, as well as leaf length. I also conducted a laboratory experiment which was designed to test the effects of 5 different inundation periods (0 control, 8, 24, 48, 96 hours) on the survival of three coastal vegetation species Agrostis stolonifera, Trifolium repens and Hippopae rhamnoides in summer 2012. The same laboratory experiment was repeated in the Netherlands. In Italy, H. rhamnoides showed a great survival in the controls, a variable performance in the other treatments and a clear decrease in treatment 4. Conversely T. repens and A. stolonifera only survive in the control. In the Netherlands experiment there was a greater variability responses for each species, still at the end of the experiment survival was significantly smaller in treatment 4 (96 h of seawater inundation) for all the three species. The results suggest that increased flooding can affect negatively the survival of both saltmarsh and coastal plants, limiting root system extension and leaf growth. Flooding effect could lead to further decline and fragmentation of the saltmarshes and coastal vegetation, thereby reducing recovery (and thus resilience) of these systems once disturbed. These effects could be amplified by interactions with other co-occurring human impacts in these systems, and it is therefore necessary to identify management options that increase the resilience of these systems.