Study of the effects of ß-myrcene on rat fertility and general reproductive performance

ß-Myrcene (MYR) is a monoterpene found in the oils of a variety of aromatic plants including lemongrass, verbena, hop, bay, and others. MYR and essential oils containing this terpenoid compound are used in cosmetics, household products, and as flavoring food additives. This study was undertaken to investigate the effects of MYR on fertility and general reproductive performance in the rat. MYR (0, 100, 300 and 500 mg/kg) in peanut oil was given by gavage to male Wistar rats (15 per dose group) for 91 days prior to mating and during the mating period, as well as to females (45 per dose group) continuously for 21 days before mating, during mating and pregnancy, and throughout the period of lactation up to postnatal day 21. On day 21 of pregnancy one-third of the females of each group were submitted to cesarean section. Resorption, implantation, as well as dead and live fetuses were counted. All fetuses were examined for external malformations, weighed, and cleared and stained with Alizarin Red S for skeleton evaluation. The remaining dams were allowed to give birth to their offspring. The progeny was examined at birth and subsequently up to postnatal day 21. Mortality, weight gain and physical signs of postnatal development were evaluated. Except for an increase in liver and kidney weights, no other sign of toxicity was noted in male and female rats exposed to MYR. MYR did not affect the mating index (proportion of females impregnated by males) or the pregnancy index (ratio of pregnant to sperm-positive females). No sign of maternal toxicity and no increase in externally visible malformations were observed at any dose level. Only at the highest dose tested (500 mg/kg) did MYR induce an increase in the resorption rate and a higher frequency of fetal skeleton anomalies. No adverse effect of MYR on postnatal weight gain was noted but days of appearance of primary coat, incisor eruption and eye opening were slightly delayed in the exposed offspring. On the basis of the data presented in this paper the no-observed-adverse-effect level (NOAEL) for toxic effects on fertility and general reproductive performance can be set at 300 mg of ß-myrcene/kg body weight by the oral route.

Effects of terpineol on the compound action potential of the rat sciatic nerve

Terpineol, a volatile terpenoid alcohol of low toxicity, is widely used in the perfumery industry. It is an important chemical constituent of the essential oil of many plants with widespread applications in folk medicine and in aromatherapy. The effects of terpineol on the compound action potential (CAP) of rat sciatic nerve were studied. Terpineol induced a dose-dependent blockade of the CAP. At 100 µM, terpineol had no demonstrable effect. At 300 µM terpineol, peak-to-peak amplitude and conduction velocity of CAP were significantly reduced at the end of 180-min exposure of the nerve to the drug, from 3.28 ± 0.22 mV and 33.5 ± 7.05 m/s, respectively, to 1.91 ± 0.51 mV and 26.2 ± 4.55 m/s. At 600 µM, terpineol significantly reduced peak-to-peak amplitude and conduction velocity from 2.97 ± 0.55 mV and 32.8 ± 3.91 m/s to 0.24 ± 0.23 mV and 2.72 ± 2.72 m/s, respectively (N = 5). All these effects developed slowly and were reversible upon 180-min washout.

Bumblebees in a Changing Climate: Evaluating the Effects of Temperature on Queen Performance

Bumblebees are a very essential group of pollinating insects, but their populations have declined drastically during the past decades. We need to understand why their numbers are decreasing and what can be done to reverse this trend. Climate change-related phenomena, such as changes in the overwintering temperatures and spring conditions, are among the most prominent threats to bumblebees. Queens have a special role in the lifecycle of bumblebees because they overwinter and start new colonies the next year. Their successful performance: survival, overwintering ability, longevity, immune competence, and nest establishing capability in spring, is highly important for bumblebee populations. However, the effects of climate change on bumblebee queen performance remain unknown. The main objective of this thesis was to assess how temperature affects the performance of bumblebee queens during and after overwintering. The effects of warm temperature predicted by climate change scenarios on queen survival and stress-tolerance were studied by a four-month artificial diapause of bumblebee queens at two temperatures (9°C and 1.8°C). Bumblebee colonies were also reared in a laboratory and factors affecting colony characteristics were examined. In addition, queen performance during spring was studied in a starvation experiment using two temperatures (15°C as normal; 24°C as warmer than average) and queens collected from nature right after their emergence. My research revealed how temperature affects queen performance, and queen size was found to be an important factor determining the direction of some of these effects. We found a 0.4g weight threshold for bumblebee queens to be able to survive overwintering. In addition, during mild winters, larger queens have a higher chance than smaller ones to survive through winter and also to cope with immunological stresses after overwintering. During cold conditions, which are normal in the current climatic situation, this advantage disappears. In the spring starvation experiment, the starved queens survived approximately eight days longer in 15°C than in 24°C, which means that starvation risk rises significantly with increasing spring temperature, in a situation where food is scarce due to for example frost damage or asynchrony between bumblebees and their important food plants. These results could mean that in the future climate, larger queens are better able to survive the winter, initiate their nests and start rearing their offspring. This may be problematic, because I also detected two alternative strategies of colony development that differ between large and small queens; larger queens start to lay eggs earlier at nest initiation, their colonies mature later, they produce more workers, and they have a more strongly male biased sex allocation compared with smaller queens. If larger queens have a greater change of producing offspring after a mild winter, this could lead to a significant decline in the total production of new queens at a population level. Thus, it seems that queen size could act as one mechanism regulating the population level outcomes in different temperatures. The new information presented in my thesis reinforces that basic research, monitoring, and local species conservation of bumblebees both in Finland and globally must be increased to ensure that this highly important pollinator group survives in the face of climate change.

Prebiotics modulate the effects of antibiotics on gut microbial diversity and functioning in vitro

Intestinal bacteria carry out many fundamental roles, such as the fermentation of non-digestible dietary carbohydrates to produce short chain fatty acids (SCFAs), which can affect host energy levels and gut hormone regulation. Understanding how to manage this ecosystem to improve human health is an important but challenging goal. Antibiotics are the front line of defence against pathogens, but in turn they have adverse effects on indigenous microbial diversity and function. Here, we have investigated whether dietary supplementation—another method used to modulate gut composition and function—could be used to ameliorate the side effects of antibiotics. We perturbed gut bacterial communities with gentamicin and ampicillin in anaerobic batch cultures in vitro. Cultures were supplemented with either pectin (a non-fermentable fibre), inulin (a commonly used prebiotic that promotes the growth of beneficial bacteria) or neither. Although antibiotics often negated the beneficial effects of dietary supplementation, in some treatment combinations, notably ampicillin and inulin, dietary supplementation ameliorated the effects of antibiotics. There is therefore potential for using supplements to lessen the adverse effects of antibiotics. Further knowledge of such mechanisms could lead to better therapeutic manipulation of the human gut microbiota.

Effects of morin on snake venom phospholipase A(2) (PLA(2))

Flavonoids are potent anti-inflammatory compounds isolated from several plant extracts, and have been used experimentally against inflammatory processes. In this work, a PLA(2) isolated from the Crotalus durissus cascavella venom and rat paw oedema were used as a model to. study the effect of flavonoids on PLA(2). We observed that a treatment of PLA(2) with morin induces several modifications in the aromatic amino acids, with accompanying changes in its amino acid composition. In addition, results from circular dichroism spectroscopy and UV scanning revealed important structural modifications. Concomitantly, a considerable decrease in the enzymatic and antibacterial activities was observed, even though anti-inflammatory and neurotoxic activities were not affected. These apparent controversial results may be an indication that PLA(2) possess a second pharmacological site which does not affect or depend on the enzymatic activity. (c) 2005 Elsevier Ltd. All rights reserved.

Effects of caffeine on time to exhaustion in exercise performed below and above the anaerobic threshold

Controversy still exists concerning the potential ergogenic benefit of caffeine (CAF) for exercise performance. The purpose of this study was to compare the effects of CAF ingestion on endurance performance during exercise on a bicycle ergometer at two different intensities, i.e., approximately 10% below and 10% above the anaerobic threshold (AT). Eight untrained males, non-regular consumers of CAF, participated in this study. AT, defined as the intensity (watts) corresponding to a lactate concentration of 4 mM, was determined during an incremental exercise test from rest to exhaustion on an electrically braked cycle ergometer. on the basis of these measurements, the subjects were asked to cycle until exhaustion at two different intensities, i.e., approximately 10% below and 10% above AT. Each intensity was performed twice in a double-blind randomized order by ingesting either CAF (5 mg/kg) or a placebo (PLA) 60 min prior to the test. Venous blood was analyzed for free fatty acid, glucose, and lactate, before, during, and immediately after exercise. Rating of perceived exertion and time to exhaustion were also measured during each trial. There were no differences in free fatty acids or lactate levels between CAF and PLA during and immediately after exercise for either intensity. Immediately after exercise glucose increased in the CAF trial at both intensities. Rating of perceived exertion was significantly lower (CAF = 14.1 ± 2.5 vs PLA = 16.6 ± 2.4) and time to exhaustion was significantly higher (CAF = 46.54 ± 8.05 min vs PLA = 32.42 ± 14.81 min) during exercise below AT with CAF. However, there was no effect of CAF treatment on rating of perceived exertion (CAF = 18.0 ± 2.7 vs PLA = 17.6 ± 2.3) and time to exhaustion (CAF = 18.45 ± 7.28 min vs PLA = 19.17 ± 4.37 min) during exercise above AT. We conclude that in untrained subjects caffeine can improve endurance performance during prolonged exercise performed below AT and that the decrease of perceived exertion can be involved in this process

Effects of microhabitat on palm seed predation in two forest fragments in southeast Brazil

The establishment of plants depends crucially on where seeds are deposited in the environment. Some authors suggest that in forest understory seed predation is lower than in gaps, and higher than at the forest edge. However, most studies have been carried out in large forest patches and very little is known about the effects of microhabitat conditions on seed predation in forest fragments. We evaluated the effects of three microhabitats (gaps, forest edge, and understory) on seed predation of two palm species (Euterpe edulis and Syagrus romanzoffiana) in two semi-deciduous forest fragments (230 and 2100 hat in southeast Brazil. Our objective was to test two hypotheses: (1) Low rodent abundance in small fragments as a result of meso-predator action levels leads to lower seed predation in small fragments. (2) Most mammal species in small fragments are generalists with respect to diet and habitat, so that seed predation is similar in different microhabitats (gaps, forest edge and understory) in the small fragment, but not in the larger one. The study community of small fragments is usually composed of generalist species (in diet and habitat aspects), so we expected the same rate of seed predation among microhabitats (gaps, forest edge and understory) in the tested smaller fragment. The experiment was carried out in the dry season (for E. edulis) and in the wet season (for S. romanzoffiana) in 1999. We conclude that post-dispersal seed predation in forest fragments can be directly connected with mammal communities, reflecting their historical and ecological aspects. (C) 2004 Elsevier SAS. All rights reserved.

Effects of monocrotaline on energy metabolism in the rat liver

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

Toxicological and histopathological effects of hydramethylnon on Atta sexdens rubropilosa (Hymenoptera: Formicidae) workers

The leaf-cut ants are important agricultural pest, because they can cause intense defoliation in plants and destroy large areas cultivated. Although there are several works for the control of these insects by examining the toxicity of natural chemical compounds on various species of ants, few are focused on analyses of morphological changes caused in the affected organs. The aim of this study was to evaluate the effects of hydramethylnon on Atta sexdens rubropilosa workers through toxicological bioassays and morphological analysis of the post-pharyngeal glands, midgut, and Malpighian tubules of these ants. Hydramethylnon dissolved either in acetone (HA) or in a mixture of acetone and soy oil (HAO) was added to the artificial diet at a concentration of 200 μg/mL. The workers fed daily with the diet containing hydramethylnon showed higher mortality than the controls, especially when HAO was used. Moreover, light and electron microscopy revealed morphological alterations in the midgut and Malpighian tubules of workers treated with HA, whereas alterations of the post-pharyngeal glands were observed in the HAO-treated group. These results indicated that the presence of soy oil provided an alternate route for the ingestion of the formicide's active ingredient and corroborated previous studies that suggested a role for the post-pharyngeal glands in lipid metabolism. Our findings suggest that the oil may carry hydramethylnon to the gland lumen, resulting in lower quantity of the active ingredient in the intestinal lumen and Malpighian tubules that explains the lower degree of morphological alterations in these structures in the workers treated with HAO. These results may provide insight into the toxicological effects of hydramethylnon on leaf-cutting ants and the use of vegetable oil as an adjuvant in baits to control ants. © 2012 Elsevier Ltd.

Morphological effects of flooding on Styrax pohlii and the dynamics of physiological responses during flooding and post-flooding conditions

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Lethal effects of abamectin on the aquatic organisms Daphnia similis, Chironomus xanthus and Danio rerio

Abamectin is used as an acaricide and insecticide for fruits, vegetables and ornamental plants, as well as a parasiticide for animals. One of the major problems of applying pesticides to crops is the likelihood of contaminating aquatic ecosystems by drift or runoff. Therefore, toxicity tests in the laboratory are important tools to predict the effects of chemical substances in aquatic ecosystems. The aim of this study was to assess the potential hazards of abamectin to the freshwater biota and consequently the possible losses of ecological services in contaminated water bodies. For this purpose, we identified the toxicity of abamectin on daphnids, insects and fish. Abamectin was highly toxic, with an EC50 48 h for Daphnia similis of 5.1 ng L-1, LC50 96 h for Chironomus xanthus of 2.67 mu g L-1 and LC50 48 h for Danio rerio of 33 mu g L-1. (C) 2011 Elsevier Ltd. All rights reserved.

Effects of flavomycin on ruminal fermentation, In situ degradability and In vivo digestibility in bovine fed sugarcane diets

Problem statement: The aim of the present study was to characterize and differentiate the effects of addition of flavomycin or monensin on ruminal fermentation and degradability as well as on total digestibility in bovine. Approach: Twelve non-pregnant and non-lactating cows (736 kg of BW) were randomly assigned to three treatments: control, flavomycin (20 mg animal-1 day-1) and monensin (300 mg animal-1 day-1). The trial lasted 21 days. The last 10 days were used for external marker administration (15 g of chromic oxide animal-1 day-1). The last 5 days of the trial were used for feces collection and evaluation of corn grain, soybean meal or sugarcane ruminal degradability and the 21st day was used for ruminal fluid sampling. Results: Monensin increased 27.2%, on average, propionate molar proportion at 0, 4, 6, 8, 10 and 12 h after feeding, compared to control and flavomycin groups. When compared to control, flavomycin reduced the degradation rate of soybean meal CP in 31.0%, decreasing the effective degradability when passage rates of 5 and 8% h-1 were used. Dry matter intake, pH, total Short Chain Fatty Acids (tSCFA) or ammoniacal Nitrogen (NH3-N) concentration were not influenced by the addition of either antibiotics. Effective degradability of sugarcane NDF was not influenced by the use of either antibiotic; neither were the TDN nor the digestibility of DM, CP, EE, NFE, ADF, NDF, GE or starch of the diet. Conclusion/Recommendations: In the present study, it was possible to show the beneficial effects of monensin but not of flavomycin, on rumen fermentation

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.

Effects of Harvesting on Herbaceous Layer Diversity of a Central Appalachian Hardwood Forest

Clearcutting is a common harvesting practice in many eastern hardwood forests. Among the vegetation strata of these forests, the herbaceous layer is potentially the most sensitive in its response to harvest-mediated disturbances and has the highest species diversity. Thus, it is important to understand the response of herbaceous layer diversity to forest harvesting. Previous work on clearcut and mature stands at the Fernow Experimental Forest (FEF), West Virginia, has shown that, although, harvesting did not alter appreciably herbaceous layer cover, it influenced the relationship of cover to biotic and abiotic factors, such as tree density and soil nutrients, respectively. The purpose of this study was to examine the response of species diversity of the herbaceous layer to harvesting at FEF. Fifteen circular, 0.04 ha sample plots were established in each of four watersheds (60 plots in total) representing two stand age categories: two watersheds with 20 years even-age stands following clearcutting and two watersheds with mature second growth stands. All woody stems ≥2.5 cm diameter at breast height were identified, tallied, and measured for diameter. The herbaceous layer was sampled by identifying all vascular plants ≤1 m in height and estimating cover for each species in each of 10 (1 m2) circular sub-plots per sample plot (600 sub-plots total). Species diversity for each plot was calculated from herbaceous layer data using the ln-based Shannon Index (H′) equation. Ten stand and soil variables also were measured on each plot. Mean herbaceous layer cover for clearcut versus mature stands was 27.2±14.3% versus 20.2±8.1% (P>0.05), respectively and mean H′ was 1.67±0.42 versus 1.55±0.48 (P>0.05), respectively. Herbaceous layer diversity was negatively correlated with cation exchange capacity and extractable Ca and Mg in the mineral soil in clearcut stands. In contrast, herbaceous layer diversity was positively correlated with soil organic matter and clay content. Although, 20 years of recovery after clearcutting did not have significant effects on the species diversity of the herbaceous layer when examining stand age means alone, harvesting did appear to influence the spatial relationships between herbaceous layer diversity and biotic factors (e.g. tree density) and abiotic factors (e.g. soil nutrients).

Effects of management on native and exotic plant communities in Pictured Rocks National Lakeshore in the Upper Peninsula of Michigan

Ecological disturbances may be caused by a range of biotic and abiotic factors. Among these are disturbances that result from human activities such as the introduction of exotic plants and land management activities. This dissertation addresses both of these types of disturbance in ecosystems in the Upper Peninsula of Michigan. Invasive plants are a significant cause of disturbance at Pictured Rocks Natural Lakeshore. Management of invasive plants is dependent on understanding what areas are at risk of being invaded, what the consequences of an invasion are on native plant communities and how effective different tools are for managing the invasive species. A series of risk models are described that predict three stages of invasion (introduction, establishment and spread) for eight invasive plant species at Pictured Rocks National Lakeshore. These models are specific to this location and include species for which models have not previously been produced. The models were tested by collecting point data throughout the park to demonstrate their effectiveness for future detection of invasive plants in the park. Work to describe the impacts and management of invasive plants focused on spotted knapweed in the sensitive Grand Sable Dunes area of Pictured Rocks National Lakeshore. Impacts of spotted knapweed were assessed by comparing vegetation communities in areas with varying amounts of spotted knapweed. This work showed significant increases in species diversity in areas invaded by knapweed, apparently as a result of the presence of a number of non-dune species that have become established in spotted knapweed invaded areas. An experiment was carried out to compare annual spot application of two herbicides, Milestone® and Transline® to target spotted knapweed. This included an assessment of impacts of this type of treatment on non-target species. There was no difference in the effectiveness of the two herbicides, and both significantly reduced the density of spotted knapweed during the course of the study. Areas treated with herbicide developed a higher percent cover of grasses during the study, and suffered limited negative impacts on some sensitive dune species such as beach pea and dune stitchwort, and on some other non-dune species such as hawkweed. The use of these herbicides to reduce the density of spotted knapweed appears to be feasible over large scales.