Monitoring of Bt-Maize pollen exposure in the vicinity of the nature reserve Ruhlsdorfer Bruch in northeast Germany 2007 to 2008

Background, aim, and scope Basically, technological innovations are associated with benefits and risks. This is also true for the introduction of genetically modified organisms (GMO) into agriculture. In Germany, precautionary regulations currently demand isolation distances (i.?e. buffer zones) for the cultivation of genetically modified maize (Bt-maize) in the vicinity of conventional (150?m) and organic maize fields (300?m). The Bt-toxin may harm non-target organisms (NTO) such as Lepidoptera. Despite this, corresponding regulations for the protection of nature reserves are lacking to date. Conventional and Bt-maize have been grown in the vicinity of the Flora-Fauna-Habitat (FFH) Ruhlsdorfer Bruch in Brandenburg, Germany. The aim of this study was to investigate whether exposure of maize pollen from surrounding Bt-maize fields to NTOs in the nature protection area could be excluded or not. Two types of exposure were investigated: Firstly, whether maize pollen was dispersed by wind into the nature reserve area exposing resident NTOs. Secondly, foraging NTOs from the nature reserve are exposed by roaming the surrounding fields and collecting maize pollen. In order to fulfil the precautionary principle defined by law, the study should help to determine appropriate isolation distances for the cultivation of Bt-maize with regard to sustainable protection of NTOs in the FFH Ruhlsdorfer Bruch. In 2007, the local authorities issued an isolation distance between Bt-maize fields and nature reserves of 100?m and in 2008, this became 250?m in the northern and 500?m in the westerly direction, respectively. Materials and methods Standardised methods for biological and technical pollen sampling issued by the Association of German Engineers (VDI 4330 Part 3, 2007 and VDI 4330 Part 4, 2006) were applied providing a quality controlled and methodologically harmonised database which does not only serve the needs to be fulfilled by the present case-specific monitoring study but can also be used as a reference database for further investigations. Maize pollen exposure was measured within the FFH Ruhlsdorfer Bruch and its immediate vicinity in July and August 2007 and 2008. In 2007, the sampling was performed at three sites using 12 technical samplers (Sigma-2/PMF) placed at five measuring points at distances from 5?m to 120?m from the maize field edges. Additionally, for biological pollen sampling six bee colonies were situated at these three sites (two colonies at each site). The technical sampler Sigma-2/PMF enables point sampling which is primarily influenced by wind and topography providing information on effective maize pollen input (flow and deposition) at the measuring sites. Honey-bees roaming in the surrounding area with typical foraging distances of several kilometres may act as planar collectors. They may serve as indicators for the exposure to pollen-collecting NTOs. Furthermore, biological sampling is more selective due to the organism’s preferences, whereas the technical sampling is neutral. Hence, both the technical and the biological sampling complement each other in their scope of application. The pollen samples of both matrices were analysed microscopically and the maize pollen loads were quantified. Pollen-DNA was analysed by means of the quantitative PCR-method (qPCR) identifying conventional and Bt-maize pollen by two independent laboratories. In 2008, the monitoring was repeated with additional sites. Eighteen technical samplers were exposed at five sites with eight measuring points at distances from 5?m to 250?m from the maize field edges. Two honey-bee colonies for biological sampling were placed at one site for control purposes. PCR-analyses were performed to measure the amounts of Bt-maize pollen in the samples. Results The results of pollen monitoring at the Ruhlsdorfer Bruch revealed maize pollen exposure for all monitoring sites in both surveys. In 2007, up to 1.75 million maize pollen/m² were deposited at sites closest to the maize field. At 120?m from field edge in the middle of the FFH-reserve, 99,000 maize pollen/m² were detected. In 2008, similar results were found, at distances up to 250?m from the field edges deposition of 164,000 maize pollen/m² was detected. Data on maize pollen deposition show a clear distance relationship and are in accordance with results of further comprehensive surveys based on the same methodology (Hofmann 2007). The results of the microscopic analysis of the pollen pellets demonstrated that bees collected maize pollen at all sites, in 2007 and 2008. Although maize pollen is not the main food source (2007:0.1–0.3?%; 2008:2–3?%) the collection efficiency of the bee colonies resulted in high amounts of sampled maize pollen with 4 to 11 million per site in 2007 and up to 467 million in 2008. Molecular-biological analysis of maize pollen DNA by qPCR demonstrated that transgenic Bt-MON810 DNA was present in all technical and biological samples, corroborated by two independent laboratories. In 2007, the GMO-content in the samples ranged up to 44?% in the bioaerosols and 49?% in the pollen pellets. In 2008, GMO-proportions of up to 18?% were detected. Discussion The results of this study provide evidence that NTOs in the Ruhlsdorfer Bruch were exposed to Bt-maize pollen under the cultivation conditions in 2007 with a buffer zone of 100?m. The GMO-content reached up to 48?%. The results of the monitoring in 2008 confirmed these findings. Even though the exposure could be reduced by increasing the isolation distances to 250?m and 500?m respectively, the results still show percentages of up to 18?% Bt-MON810 in the pollen samples. The results on maize pollen deposition at the Ruhlsdorfer Bruch in 2007 and 2008 correspond to the results of an investigation which was conducted over several years applying the same standardised method, but covering a wider range of distances. The correlation between maize pollen deposition (n/m²) and distance to the source field (m) fitted best to a power function of the type y = 1.2086 · 10⁶ · x ^–0.548. Despite the same trend, the pollen deposition in the Ruhlsdorfer Bruch revealed above-average findings. Also the analysis of the pollen pellets collected by the bees showed an exposure in 2007 with values for the GMO-content of up to 49?%. For both methods, the exposure decreased in 2008 due to the greater buffer zones up to 500?m. Whereas the GMO-content for the biological sampling were reduced to values below 10?%, the values for the technical sampling were still higher indicating that greater buffer zones would be necessary for safety reasons under the precautionary principle. Conclusions The results of this investigation proved that maize pollen were dispersed by wind to distances farther than 250?m from field edge leading to maize pollen exposure in the centre of the nature reserve. The results also demonstrated that foraging NTOs living in the nature reserve were exposed to maize pollen from surrounding fields. Considering the cultivation of Bt-maize MON810, the assumption of the Environmental Risk Assessment (ERA) that there will be no relevant exposure beyond the Bt-maize fields, cannot be confirmed. Considering the results of this and related studies and with respect to the precautionary principle, one can state that buffer zones between Bt-maize fields and protected areas are an effective measure to minimise the exposure of Bt-maize pollen to NTOs and, thus, to prevent from adverse effects. Recommendations and perspectives Because of still insufficient ecotoxicological data for the risk assessment of Bt-maize MON810 considering butterflies and other protected NTOs, protection standards assuring the precautionary principle have to be implemented to avoid Bt-maize pollen exposure to NTOs. This applies for the case Ruhlsdorfer Bruch and for nature reserve areas in general. In order to exclude risks to protected NTOs occurring in nature reserves, sufficient buffer zones for Bt-maize cultivation should be considered. The statistical analysis revealed that distances of more than 500?m are necessary to decisively reduce exposure to foraging insects. In fact, distances of more than 1,000?m are necessary to prevent maize pollen deposition from values above 100,000 pollen/m² with a certainty of 90?%. An adequate risk assessment can only be attained if based on field measurements accounting for the high variation of pollen deposition due to local environmental site conditions and field management. The monitoring should be based on standardised methods. It should include locations with the highest expected deposition rates, the boundaries of the protected areas and sites of interest within those boundaries, e.?g., specific habitats of sensitive species.     

Understanding the Lessons and Limitations of Conservation and Development

Abstract: The lack of concrete instances in which conservation and development have been successfully merged has strengthened arguments for strict exclusionist conservation policies. Research has focused more on social cooperation and conflict of different management regimes and less on how these factors actually affect the natural environments they seek to conserve. Consequently, it is still unknown which strategies yield better conservation outcomes? We conducted a meta‐analysis of 116 published case studies on common resource management regimes from Africa, south and central America, and southern and Southeast Asia. Using ranked sociodemographic, political, and ecological data, we analyzed the effect of land tenure, population size, social heterogeneity, as well as internally devised resource‐management rules and regulations (institutions) on conservation outcome. Although land tenure, population size, and social heterogeneity did not significantly affect conservation outcome, institutions were positively associated with better conservation outcomes. There was also a significant interaction effect between population size and institutions, which implies complex relationships between population size and conservation outcome. Our results suggest that communities managing a common resource can play a significant role in conservation and that institutions lead to management regimes with lower environmental impacts.     

Assessing Sustainability at Multiple Scales in a Rotational Bushmeat Hunting System

Abstract: Results of many studies show unsustainable levels of bushmeat hunting across West/Central Africa. Nevertheless, these results are usually derived from snapshot sustainability indices in which critical parameters are often taken from the literature. Simple, more informative tools for assessing sustainability are needed. We evaluated the impact of bushmeat hunting across a range of temporal, spatial, and taxonomic scales in a comparison of different measures of sustainability. Over 15 months in 2002–2004 in and around a village close to Equatorial Guinea's Monte Alén National Park, we collected data via a village offtake survey, hunter‐camp bushmeat‐consumption diaries, hunter interviews, and following hunters during hunts. We compared 2003 data with a previous offtake survey (1998–1999) and interview reports back to 1990. In the past 14 years, average distance from the village at which hunters operated remained constant, with hunters switching back and forth between long‐established camps, although trapping effort increased. In the past 5 years, overall offtake and number of active hunters did not change substantially, although catch per unit effort (CPUE) decreased slightly. Although the proportion of the two most commonly trapped species (Cephalophus monticola and Atherurus africanus) and gun‐hunted primates increased in the offtake, species presumably less robust to trapping decreased slightly. Apparent sustainability in economic terms may be masking gradual local extirpation of more vulnerable species before and during this study. Our results suggest that changes in prey profiles and CPUE may be the most accurate indicators of actual sustainability; these indices can be monitored with simple village‐based offtake surveys and hunter interviews to improve community management of bushmeat hunting.     

Habitat Split as a Cause of Local Population Declines of Amphibians with Aquatic Larvae

Abstract:  Most amphibian species have biphasic life histories and undergo an ontogenetic shift from aquatic to terrestrial habitats. In deforested landscapes, streams and forest fragments are frequently disjunct, jeopardizing the life cycle of forest-associated amphibians with aquatic larvae. We tested the impact of habitat split—defined as human-induced disconnection between habitats used by different life-history stages of a species—on four forest-associated amphibian species in a severely fragmented landscape of the Brazilian Atlantic Forest. We surveyed amphibians in forest fragments with and without streams (referred to as wet and dry fragments, respectively), including the adjacent grass-field matrix. Our comparison of capture rates in dry fragments and nearby streams in the matrix allowed us to evaluate the number of individuals that engaged in high-risk migrations through nonforested habitats. Adult amphibians moved from dry fragments to matrix streams at the beginning of the rainy season, reproduced, and returned at the end of the breeding period. Juveniles of the year moved to dry fragments along with adults. These risky reproductive migrations through nonforested habitats that expose individuals to dehydration, predation, and other hazards may cause population declines in dry fragments. Indeed, capture rates were significantly lower in dry fragments compared with wet fragments. Declining amphibians would strongly benefit from investments in the conservation and restoration of riparian vegetation and corridors linking breeding and nonbreeding areas.     

Molecular prey identification in wild Octopus vulgaris paralarvae

The trophic ecology of Octopus vulgaris paralarvae collected in 2008 off the Ría de Vigo, NW Spain (42° 12.80′ N–9° 00.00′ W), was approached by both morphological and molecular methods. External digestion of prey and posterior suction of the liquefied contents by wild O. vulgaris paralarvae made the morphological identification of gut contents impossible. Thus, a PCR-based method using group-specific primers was selected to identify prey consumed by O. vulgaris paralarvae in the pelagic realm. The mitochondrial ribosomal 16S gene region was chosen for designing group-specific primers, which targeted a broad range of crustaceans and fishes but avoided the amplification of predator DNA. These primers successfully amplified DNA of prey by using a semi-nested PCR-based approach and posterior cloning. Homology search and phylogenetic analysis were then conducted with the 20 different operational taxonomic units obtained to identify the putative organisms ingested. The phylogenetic analysis clustered ingested prey into 12 families of crustaceans (11 belonging to the order Decapoda and 1 to the order Euphausiacea) and two families of fishes (Gobiidae and Carangidae). According to the Czekanowski’s Index (CI), the trophic niche breadth of O. vulgaris paralarvae is low (CI = 0.13), which means that these paralarvae are specialist predators at least during the first weeks of their life cycle. It is the first time that natural prey has been identified in O. vulgaris paralarvae collected from the wild, and such knowledge may be critical to increasing the survival of O. vulgaris hatchlings in captivity, a goal that has been actively pursued since the 1960s by aquaculture researchers.     

Development of a new standardised method for sustainable monitoring of the vulnerable pink sea fan Eunicella verrucosa

The aim of this survey was to test a standardised method to follow the demographic evolution of a dense aggregative ‘forest’ population of the temperate gorgonian Eunicella verrucosa (Octocorallia, Gorgoniidae) using in situ photographic recordings. Distribution, density, growth and demographic evolution of the colonies was compared along two parallel transects. Computer treatment allowed the estimation of the total branch fan surface area, and the individual growth of tagged colonies was determined by measuring the increase in this surface area, using consecutive photographs taken at two-year intervals. To integrate the potential bias of branch overlapping, we proposed a correction factor between the in situ photographic surface area and the surface area of the gorgonian calculated from ex situ photographic surface area with all branches deployed. The surface-frequency distribution of colonies was converted to estimated-age-frequency distribution using an estimated growth curve based on the net growth rate. The technique used revealed significant differences in population structure and the dynamics of gorgonian colonies, as the two transects appeared to be influenced by different environmental conditions. The recruitment also seemed to vary according to year and transect, resulting in different densities. Our work showed clear results in characterising the variations of gorgonian demographic evolution at a small spatial scale; thus, it is assumed that this method could be a sustainable tool for coastal environmental managers.     

Arsenic bioaccessibility in a gold mining area: a health risk assessment for children

High concentrations of total arsenic (As) have been measured in soils of gold mining areas of Brazil. However, bioaccessibility tests have not yet been conducted on those materials, which is essential for better health risk estimates. This study aimed at?evaluating As bioaccessibility in samples from a gold mining area located in Brazil and assessing children's exposure to As-contaminated materials. Samples were collected from different materials (a control and four As-contaminated soils/sediments) found in a gold mine area located in Paracatu (MG), Brazil. Total and bioaccessible As concentrations were determined for all samples. The control soil presented the lowest As concentrations, while all other materials contained high total As concentrations (up to 2,666?mg?kg(-1)) and low bioaccessible As percentage (<4.2%), indicating a low risk from exposure of resident children next to this area. The calculated dose of exposure indicated that, except for the pond tailings, in all other areas, the exposure route considering soil ingestion contributed at most to 9.7% of the maximum As allowed ingestion per day (0.3?μg?kg(-1) BW day(-1)).     

A universal approach to estimate biomass and carbon stock in tropical forests using generic allometric models

Allometric equations allow aboveground tree biomass and carbon stock to be estimated from tree size. The allometric scaling theory suggests the existence of a universal power-law relationship between tree biomass and tree diameter with a fixed scaling exponent close to 8/3. In addition, generic empirical models, like Chave's or Brown's models, have been proposed for tropical forests in America and Asia. These generic models have been used to estimate forest biomass and carbon worldwide. However, tree allometry depends on environmental and genetic factors that vary from region to region. Consequently, theoretical models that include too few ecological explicative variables or empirical generic models that have been calibrated at particular sites are unlikely to yield accurate tree biomass estimates at other sites. In this study, we based our analysis on a destructive sample of 481 trees in Madagascar spiny dry and moist forests characterized by a high rate of endemism (> 95%). We show that, among the available generic allometric models, Chave's model including diameter, height, and wood specific gravity as explicative variables for a particular forest type (dry, moist, or wet tropical forest) was the only one that gave accurate tree biomass estimates for Madagascar (R2 > 83%, bias < 6%), with estimates comparable to those obtained with regional allometric models. When biomass allometric models are not available for a given forest site, this result shows that a simple height-diameter allometry is needed to accurately estimate biomass and carbon stock from plot inventories.     

Landscape context and scale differentially impact coffee leaf rust, coffee berry borer, and coffee root-knot nematodes

Crop pest and disease incidences at plot scale vary as a result of landscape effects. Two main effects can be distinguished. First, landscape context provides habitats of variable quality for pests, pathogens, and beneficial and vector organisms. Second, the movements of these organisms are dependent on the connectivity status of the landscape. Most of the studies focus on indirect effects of landscape context on pest abundance through their predators and parasitoids, and only a few on direct effects on pests and pathogens. Here we studied three coffee pests and pathogens, with limited or no pressure from host-specific natural enemies, and with widely varying life histories, to test their relationships with landscape context: a fungus, Hemileia vastatrix, causal agent of coffee leaf rust; an insect, the coffee berry borer, Hypothenemus hampei (Coleoptera: Curculionidae); and root-knot nematodes, Meloidogyne spp. Their incidence was assessed in 29 coffee plots from Turrialba, Costa Rica. In addition, we characterized the landscape context around these coffee plots in 12 nested circular sectors ranging from 50 to 1500 m in radius. We then performed correlation analysis between proportions of different land uses at different scales and coffee pest and disease incidences. We obtained significant positive correlations, peaking at the 150 m radius, between coffee berry borer abundance and proportion of coffee in the landscape. We also found significant positive correlations between coffee leaf rust incidence and proportion of pasture, peaking at the 200 m radius. Even after accounting for plot level predictors of coffee leaf rust and coffee berry borer through covariance analysis, the significance of landscape structure was maintained. We hypothesized that connected coffee plots favored coffee berry borer movements and improved its survival. We also hypothesized that wind turbulence, produced by low-wind-resistance land uses such as pasture, favored removal of coffee leaf rust spore clusters from host surfaces, resulting in increased epidemics. In contrast, root-knot nematode population density was not correlated to landscape context, possibly because nematodes are almost immobile in the soil. We propose fragmenting coffee plots with forest corridors to control coffee berry borer movements between coffee plots without favoring coffee leaf rust dispersal.     

Catastrophe and the life span of coral reefs

A strong earthquake in the western Caribbean in 2009 had a catastrophic impact on uncemented, unconsolidated coral reefs in the central sector of the shelf lagoon of the Belizean barrier reef. In a set of 21 reef sites that had been observed prior to the earthquake, the benthic assemblages of 10 were eradicated, and one was partially damaged, by avalanching of their slopes. Ecological dynamics that had played out over the previous 23 years, including the mass mortalities of two sequentially dominant coral species and a large increase in the cover of an encrusting sponge, were instantaneously rendered moot in the areas of catastrophic reef-slope failure. Because these prior dynamics also determined the benthic composition and resilience of adjacent sections of reef that remained intact, the history of disturbance prior to the earthquake will strongly influence decadal-scale recovery in the failed areas. Geological analysis of the reef framework yielded a minimum return time of 2000-4000 years for this type of high-amplitude event. Anthropogenic degradation of ecosystems must be viewed against the backdrop of long-period, natural catastrophes, such as the impact of strong earthquakes on uncemented, lagoonal reefs.