The Effects of a Nematode Lungworm (Rhabdias hylae) on its Natural and Invasive Anuran Hosts.

Biological invasions can bring both the invader and native taxa into contact with novel parasites. As cane toads ( Rhinella marina ) have spread through Australia, they have encountered lungworms (Rhabdias hylae) that occur in native frogs. Field surveys suggest that these lungworms have not host-switched to toads. In our laboratory studies, R. hylae infected cane toads as readily as it infected native frogs, but failed to reach the lungs of the novel host (i.e., were killed by the toads' immune response). Plausibly, then, R. hylae might reduce the viability both of their native hosts (frogs, that can exhibit high parasite burdens) and cane toads (that must deal with infective larvae traveling through the host body). Our laboratory trials suggest, however, that the impacts of the parasite on infected anuran hosts (both frogs and toads) were minimal, with no significant decrements to host survival, activity, growth, or locomotor performance. Ironically, the lack of impact of the parasite on its native hosts appears to be an outcome of co-evolution (frogs tolerate the lungworm), whereas the lack of impact on the novel host is due to a lack of co-evolution (toads can recognize and eliminate the lungworm).

Defensive Metabolites from Antarctic Invertebrates: Does Energetic Content Interfere with Feeding Repellence?

Many bioactive products from benthic invertebrates mediating ecological interactions have proved to reduce predation, but their mechanisms of action, and their molecular identities, are usually unknown. It was suggested, yet scarcely investigated, that nutritional quality interferes with defensive metabolites. This means that antifeedants would be less effective when combined with energetically rich prey, and that higher amounts of defensive compounds would be needed for predator avoidance. We evaluated the effects of five types of repellents obtained from Antarctic invertebrates, in combination with diets of different energetic values. The compounds came from soft corals, ascidians and hexactinellid sponges; they included wax esters, alkaloids, a meroterpenoid, a steroid, and the recently described organic acid, glassponsine. Feeding repellency was tested through preference assays by preparing diets (alginate pearls) combining different energetic content and inorganic material. Experimental diets contained various concentrations of each repellent product, and were offered along with control compound-free pearls, to the Antarctic omnivore amphipod Cheirimedon femoratus. Meridianin alkaloids were the most active repellents, and wax esters were the least active when combined with foods of distinct energetic content. Our data show that levels of repellency vary for each compound, and that they perform differently when mixed with distinct assay foods. The natural products that interacted the most with energetic content were those occurring in nature at higher concentrations. The bioactivity of the remaining metabolites tested was found to depend on a threshold concentration, enough to elicit feeding repellence, independently from nutritional quality.

Natural born weight gainers: The mechanisms of obesity in transgenic mice overexpressing neuropeptide Y

Neuropeptide Y (NPY) is a neurotransmitter promoting energy storage by activating Y-receptors and thus affecting food intake, thermogenesis and adipose tissue metabolism. NPY is expressed both in the central and sympathetic nervous system. Hypothalamic NPY is known to stimulate feeding, but the effects of noradrenergic neuron NPY are more ambiguous. Chronic stress stimulates fat accumulation via NPY release from noradrenergic neurons. Furthermore, polymorphism in the human Npy gene has been associated with metabolic disturbances and increased NPY secretion after sympathetic stimulation. The main objective of this study was to clarify the mechanisms of noradrenergic neuron NPY in the development of obesity. The metabolic phenotype of a homozygous mouse overexpressing NPY in the brain noradrenergic neurons and sympathetic nervous system (OE-NPYDβH mouse) was characterized. OE-NPYDβH mice had an increased fat mass and body weight, which caused impairments of glucose metabolism and hyperinsulinaemia with age. There were no differences in energy intake or expenditure, but the sympathetic tone was down-regulated and the endocannabinoid system activated. Furthermore, peripheral Y2-receptors in energy-rich conditions played an important role in mediating the fat-accumulating effect of NPY. These results indicate that noradrenergic neuron NPY promotes obesity via direct effects in the periphery and by modulating the sympatho-adrenal and endocannabinoid systems. Additionally, NPY in the central noradrenergic neurons is believed to possess many important roles. The phenotype of the OE-NPYDβH mouse resembles the situations of chronic stress and Npy gene polymorphism and thus these mice may be exploited in testing novel drug candidates for the treatment of obesity.

Potential contribution of natural enemies to patterns of local adaptation in plants

Genetic differentiation among plant populations and adaptation to local environmental conditions are well documented. However, few studies have examined the potential contribution of plant antagonists, such as insect herbivores and pathogens, to the pattern of local adaptation. Here, a reciprocal transplant experiment was set up at three sites across Europe using two common plant species, Holcus lanatus and Plantago lanceolata. The amount of damage by the main above-ground plant antagonists was measured: a rust fungus infecting Holcus and a specialist beetle feeding on Plantago, both in low-density monoculture plots and in competition with interspecific neighbours. Strong genetic differentiation among provenances in the amount of damage by antagonists in both species was found. Local provenances of Holcus had significantly higher amounts of rust infection than foreign provenances, whereas local provenances of Plantago were significantly less damaged by the specialist beetle than the foreign provenances. The presence of surrounding vegetation affected the amount of damage but had little influence on the ranking of plant provenances. The opposite pattern of population differentiation in resistance to local antagonists in the two species suggests that it will be difficult to predict the consequences of plant translocations for interactions with organisms of higher trophic levels.

Ecological specificity of arbuscular mycorrhizae: evidence from foliar- and seed-feeding insects

Arbuscular mycorrhizal (AM) fungi have a variety of effects on foliar-feeding insects, with the majority of these being positive, although reports of negative and null effects also exist. Virtually all previous experiments have used mobile insects confined in cages and have studied the effects of one, or at most two, species of mycorrhizae on one species of insect. The purpose of this study was to introduce a greater level of realism into insect-mycorrhizal experiments, by studying the responses of different insect feeding guilds to a variety of AM fungi. We conducted two experiments involving three species of relatively immobile insects (a leaf-mining and two seed-feeding flies) reared in natural conditions on a host (Leucanthemum vulgare). In a field study, natural levels of AM colonization were reduced, while in a phytometer trial, we experimentally colonized host plants with all possible combinations of three known mycorrhizal associates of L. vulgare. In general, AM fungi increased the stature (height and leaf number) and nitrogen content of plants. However, these effects changed through the season and were,dependent on the identity of the fungi in the root system. AM fungi increased host acceptance of all three insects and larval performance of the leaf miner, but these effects were also season- and AM species-dependent. We suggest that the mycorrhizal effect on the performance of the leaf miner is due to fungal-induced changes in host-plant nitrogen content, detected by the adult fly. However, variability in the effect was apparent, because not all AM species increased plant N content. Meanwhile, positive effects of mycorrhizae were found on flower number and flower size, and these appeared to result in enhanced infestation levels by the seed-feeding insects. The results show that AM fungi exhibit ecological specificity, in that different. species have different effects on host-plant growth and chemistry and the performance of foliar-feeding insects. Future studies need to conduct experiments that use ecologically realistic combinations of plants and fungi and allow insects to be reared in natural conditions.

13C-2H correlation NMR spectroscopy studies of the in vivo transformations of natural products from Artemisia annua

13C-2H correlation NMR spectroscopy (13C-2H COSY) permits the identification of 13C and 2H nuclei which are connected to one another by a single chemical bond via the sizeable 1JCD coupling constant. The practical development of this technique is described using a 13C-2H COSY pulse sequence which is derived from the classical 13C-1H correlation experiment. An example is given of the application of 13C-2H COSY to the study of the biogenesis of natural products from the anti-malarial plant Artemisia annua, using a doubly-labelled precursor molecule. Although the biogenesis of artemisinin, the anti-malarial principle from this species, has been extensively studied over the past twenty years there is still no consensus as to the true biosynthetic route to this important natural product – indeed, some published experimental results are directly contradictory. One possible reason for this confusion may be the ease with which some of the metabolites from A. annua undergo spontaneous autoxidation, as exemplified by our recent in vitro studies of the spontaneous autoxidation of dihydroartemisinic acid, and the application of 13C-2H COSY to this biosynthetic problem has been important in helping to mitigate against such processes. In this in vivo application of 13C-2H COSY, [15-13C2H3]-dihydroartemisinic acid (the doubly-labelled analogue of the natural product from this species which was obtained through synthesis) was fed to A. annua plants and was shown to be converted into several natural products which have been described previously, including artemisinin. It is proposed that all of these transformations occurred via a tertiary hydroperoxide intermediate, which is derived from dihyroartemisinic acid. This intermediate was observed directly in this feeding experiment by the 13C-2H COSY technique; its observation by more traditional procedures (e.g., chromatographic separation, followed by spectroscopic analysis of the purified product) would have been difficult owing to the instability of the hydroperoxide group (as had been established previously by our in vitro studies of the spontaneous autoxidation of dihydroartemisinic acid). This same hydroperoxide has been reported as the initial product of the spontaneous autoxidation of dihydroartemisinic acid in our previous in vitro studies. Its observation in this feeding experiment by the 13C-2H COSY technique, a procedure which requires the minimum of sample manipulation in order to achieve a reliable identification of metabolites (based on both 13C and 2H chemical shifts at the 15-position), provides the best possible evidence for its status as a genuine biosynthetic intermediate, rather than merely as an artifact of the experimental procedure.

Association between feeding rate and parasitoid resistance in Drosophila melanogaster

Replicate lines of Drosophila melanogaster have been selected for increased resistance against one of two species of parasitoid wasp, Asobara tabida and Leptopilina boulardi. In both cases, it has been shown that an improved ability to mount an immunological defense against the parasitoid's egg is associated with reduced survival when the larvae are reared under conditions of low resource availability and thus high competition. We show here that in both sets of selected lines, lower competitive ability is associated with reduced rates of larval feeding, as measured by the frequency of retractions of the cephalopharyngeal skeleton. This suggests that the same or similar physiological processes are involved in the trade-off between competition and resistance against either parasitoid and shows how the interaction between adaptations for competition and natural enemy resistance may be mediated.

Use of pelleted sericea lespedeza (Lespedeza cuneata) for natural control of coccidia and gastrointestinal nematodes in weaned goats

Infection with Eimeria spp. (coccidia) can be devastating in goats, particularly for young, recently-weaned kids, resulting in diarrhea, dehydration, and even death. Feeding dried sericea lespedeza [SL; Lespedeza cuneata (Dum.-Cours.) G. Don.] to young goats has been reported to reduce the effects of internal parasites, including gastrointestinal nematodes (GIN) but there have been no reports of the effects of feeding this forage on Eimeria spp. in goats. Two confinement feeding experiments were completed on recently-weaned intact bucks (24 Kiko-cross, Exp. 1; 20 Spanish, Exp. 2) to determine effects of SL pellets on an established infection of GIN and coccidia. The bucks were assigned to 1 of 2 (Exp. 1) or 3 (Exp. 2) treatment groups based upon the number of Eimeria spp. oocysts per gram (OPG) of feces. In Exp. 1, the kids were fed 1 of 2 pelleted rations ad libitum; 90% SL leaf meal + 10% of a liquid molasses/lignin binder mix and a commercial pellet with 12% crude protein (CP) and 24% acid detergent fiber (n = 12/treatment group, 2 animals/pen). For Exp. 2, treatment groups were fed 1) 90% SL leaf meal pellets from leaves stored 3 years (n = 7), 2) 90% SL pellets from leaf meal stored less than 6 months, (n = 7), and the commercial pellets (n = 6) ad libitum. For both trials, fecal and blood samples were taken from individual animals every 7 days for 28 days to determine OPG and GIN eggs per gram (EPG) and packed cell volume (PCV), respectively. In Exp. 2, feces were scored for consistency (1 = solid pellets, 5 = slurry) as an indicator of coccidiosis. In Exp. 1, EPG (P < 0.001) and OPG (P < 0.01) were reduced by 78.7 and 96.9%, respectively, 7 days after initiation of feeding in goats on the SL pellet diet compared with animals fed the control pellets. The OPG and EPG remained lower in treatment than control animals until the end of the trial. In Exp. 2, goats fed new and old SL leaf meal pellets had 66.2 and 79.2% lower (P < 0.05) EPG and 92.2 and 91.2% lower (P < 0.05) OPG, respectively, than control animals within 7 days, and these differences were maintained or increased throughout the trial. After 4 weeks of pellet feeding in Exp. 2, fecal scores were lower (P < 0.01) in both SL-fed groups compared with control animals, indicating fewer signs of coccidiosis. There was no effect of diet on PCV values throughout either experiment. Dried, pelleted SL has excellent potential as a natural anti-coccidial feed for weaned goats.

A non-invasive method for silencing gene transcription in honeybees maintained under natural conditions

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Viabilidade do uso de secadores solar de convecção natural e forçada para a secagem do coco Licuri

Licuri is a palm tree from the semiarid regions of Bahia State, Brazil. It is an important source of food and feed in that region, since their nuts are commonly eaten by humans and used as maize substitute for poultry feeding. The aim of this dissertation is to study the feasibility for use of natural convection solar dryers and forced being compared with the traditional drying outdoors for drying coconut licuri Syagrus coronate. The study led to the construction of two prototype solar dryer for carrying out experiments proving: model Solar Drying System Direct Exposure to Natural Convection built with wood, has a drying chamber with direct cover transparent glass laminates 4 mm, using techniques for proper isolation of the drying chamber. The two prototypes were comparatively analyzed for performance and drying efficiency with traditional extractive use by the community. Were evaluated the variables: time and drying rates and quality of the final samples of coconut licuri. The fruits were harvested and brought the town of Ouricuri, in the city of Caldeirão Grande, BA for the experiments comparing the three methods of drying was used a standard load of 4.0 kg The quantitative analysis for the result of the drying rate was found in 74% yield and 44% for natural and forced convection respectively compared with the traditional drying. These drying rates represent variation 3-5 times lower. Drying using forced convection licuri showed better quality, was found in a reddish pulp, representing the quantities that were kept of the nutrient beta carotene, and not notice the flavor change from the previous system, the final cost of construction of this system were higher . The prototypes built competitive advantage and had testified fully to resolve the technical difficulties previously encountered in the production of products made of coconut licuri. Allowing add value and increase their potential use for the fruit extractive communities of semi-arid region of Bahia

Growth curve of Nellore calves reared on natural pasture in the Pantanal

Weight-age and hip height-age relations of Nellore calves, from birth to 10 months old were fitted using a logistic model including sex and year of birth as fixed effects. Calves and their dams were reared on natural pasture using continuous grazing system. The crude protein content and total digestible nutrients were analyzed for pasture selected by the animals. The weights of the calves were adjusted to 205 days and 365 days. There were no significant effects of sex and birth year on the growth curve parameters, but there were significant effects of sex on hip height. The average weight (a parameter) at 10 months of age was 170 kg and the inflection point was observed at 93.5 days old. When weight-age and hip height-age curves were combined in the same graph, the intersection occurred at 142 days. The number of days to gain 160 kg from birth to 205 days of age (adjusted) and number of days to gain 240 kg from 205 days to slaughter was different between the birth years, which were probably due to the quality of the natural pastures. It is necessary to implement nutritional management strategies such as high quality pasture and/or feeding supplementation for calves once they reach three months of age.

Influence of feeding regime and finishing system on lamb muscle fiber and meat quality

Avaliou-se a influência dos níveis nutricionais da ovelha antes do parto, da idade à desmama e do sistema de terminação sobre as fibras musculares e a qualidade da carne dos cordeiros. Utilizaram-se matrizes mestiças Ile de France × Bergamacia criadas a pasto. As ovelhas foram divididas em dois grupos: um com suplementação alimentar 30 dias antes do parto e outro sem suplementação. Os animais (ovelha e cordeiros) foram subdivididos de acordo com a idade à desmama (45 ou 60 dias de idade) e os cordeiros desmamados foram submetidos a três sistemas de terminação: confinamento com dieta completa; confinamento somente com feno; e confinamento somente no pasto. O abate dos cordeiros foi realizado aos 30 kg de peso vivo ou aos 150 dias de idade. As características de carcaça analisadas foram morfofisiologia da fibra do músculo longissimus dorsi, fragmentação miofibrilar pós-morte (maturados nos dias 0, 3 e 7 pos mortem) e redução do pH e da temperatura da carne pós-abate. Não houve efeito da suplementação às ovelhas ou da idade à desmama sobre nenhuma das características avaliadas. A composição em fibras de contração lenta e oxidativa, contração rápida e oxidativa ou contração rápida e glicolítica não foi alterada pelos fatores em estudo, mas a área transversal das fibras foi menor nos cordeiros terminados com feno, assim como os valores de pH, temperatura e as medidas dos fragmentos de miofibrila. Com o aumento do tempo de maturação de 0 para 3 e 7 dias post mortem, houve redução do comprimento dos fragmentos de miofibrilas. O sistema de terminação de cordeiros em confinamento somente com feno interfere mais intensamente na qualidade da carne que o sistema de terminação a pasto ou em confinamento com dieta completa.

Stable carbon (delta(13)C) and nitrogen (delta(15)N) isotopes as natural indicators of live and dry food in Piaractus mesopotamicus (Holmberg, 1887) larval tissue

This study proposed the use of the stable isotope technique to track the type of food utilized by pacu Piaractus mesopotamicus larvae during their development, and to identify the moment when the larvae start using nutrients from the dry diet by retaining its carbon and nitrogen atoms in their body tissues. Five-day-old pacu larvae at the onset of exogenous feeding were fed Artemia nauplii or formulated diet exclusively; nauplii+formulated diet during the entire period; or were weaned from nauplii to a dry diet after 3, 6 or 12 days after the first feeding. delta(13)C and delta(15)N values for Artemia nauplii were -15.1 parts per thousand and 4.7 parts per thousand, respectively, and -25.0 parts per thousand and 7.4 parts per thousand for the dry diet. The initial isotopic composition of the larval tissue was -20.2 parts per thousand and 9.5 parts per thousand for delta(13)C and delta(15)N respectively. Later, at the end of a 42-day feeding period, larvae fed Artemia nauplii alone reached values of -12.7 parts per thousand and 7.0 parts per thousand for delta(13)C and delta(15)N respectively. Larvae that received the formulated diet alone showed values of -22.7 parts per thousand for delta(13)C and 9.6 parts per thousand for delta(15)N. The stable isotope technique was precise, and the time at which the larvae utilized Artemia nauplii, and later dry diet as a food source could be clearly defined.

Does gestation or feeding affect the body temperature of the golden lancehead, Bothrops insularis (Squamata: Viperidae) under field conditions?

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The opportunistic feeding and reproduction strategies of the annual fish Cynopoecilus melanotaenia (Cyprinodontiformes: Rivulidae) inhabiting ephemeral habitats on southern Brazil

A maioria dos peixes da família Rivulidae são popularmente conhecidos como anuais por completarem todo seu ciclo biológico em pequenos corpos de água temporários que secam obrigatoriamente em determinados períodos do ano causando a morte dos indivíduos adultos. Possuem características biológicas peculiares como pequeno porte, maturação sexual precoce, reprodução contínua, um elaborado padrão de corte e uma grande capacidade reprodutiva entre os peixes. Os rivulídeos se encontram amplamente distribuídos nas Américas do Norte, Central e Sul. Este trabalho analisou a dieta e a biologia reprodutiva de Cynopoecilus melanotaenia. Foram coletados 263 exemplares e a análise de 233 conteúdos gastrointestinais revelou uma dieta invertívora composta principalmente por microcrustáceos (Cladocera, Amphipoda e Ostracoda) e insetos imaturos (Chaoboridae, Culicidae, Syrphidae, mas principalmente larvas de Chironomidae). Foi registrada a lepidofagia na dieta dos machos. A fecundidade foi estimada pela análise de 59 pares de ovários maduros e variou entre 2 a 157 ovócitos (média, 19 ± 26[DP]). A espécie apresentou desova parcelada, uma estratégia para aumentar a chance de sobrevivência a depleções prolongadas. Este trabalho apresenta a primeira investigação sobre a biologia reprodutiva de C. melanotaenia. Os resultados encontrados sugerem a estratégia reprodutiva oportunista de C. melanotaenia e fornecem novas informações biológicas que podem contribuir para pesquisas de conservação e manutenção da espécie em seu ambiente natural.