Showing papers by "Wageningen University and Research Centre published in 2008"

Small CRISPR RNAs guide antiviral defense in prokaryotes

Abstract: Prokaryotes acquire virus resistance by integrating short fragments of viral nucleic acid into clusters of regularly interspaced short palindromic repeats (CRISPRs). Here we show how virus-derived sequences contained in CRISPRs are used by CRISPR-associated (Cas) proteins from the host to mediate an antiviral response that counteracts infection. After transcription of the CRISPR, a complex of Cas proteins termed Cascade cleaves a CRISPR RNA precursor in each repeat and retains the cleavage products containing the virus-derived sequence. Assisted by the helicase Cas3, these mature CRISPR RNAs then serve as small guide RNAs that enable Cascade to interfere with virus proliferation. Our results demonstrate that the formation of mature guide RNAs by the CRISPR RNA endonuclease subunit of Cascade is a mechanistic requirement for antiviral defense.

Strigolactone inhibition of shoot branching

Abstract: A carotenoid-derived hormonal signal that inhibits shoot branching in plants has long escaped identification. Strigolactones are compounds thought to be derived from carotenoids and are known to trigger the germination of parasitic plant seeds and stimulate symbiotic fungi. Here we present evidence that carotenoid cleavage dioxygenase 8 shoot branching mutants of pea are strigolactone deficient and that strigolactone application restores the wild-type branching phenotype to ccd8 mutants. Moreover, we show that other branching mutants previously characterized as lacking a response to the branching inhibition signal also lack strigolactone response, and are not deficient in strigolactones. These responses are conserved in Arabidopsis. In agreement with the expected properties of the hormonal signal, exogenous strigolactone can be transported in shoots and act at low concentrations. We suggest that endogenous strigolactones or related compounds inhibit shoot branching in plants. Furthermore, ccd8 mutants demonstrate the diverse effects of strigolactones in shoot branching, mycorrhizal symbiosis and parasitic weed interaction.

The LEDA Traitbase: A database of life-history traits of the Northwest European flora

Abstract: Summary 1. An international group of scientists has built an open internet data base of life-history traits of the Northwest European flora (the LEDA-Traitbase) that can be used as a data source for fundamental research on plant biodiversity and coexistence, macro-ecological patterns and plant functional responses. 2. The species-trait matrix comprises referenced information under the control of an editorial board, for ca. 3000 species of the Northwest European flora, combining existing information and additional measurements. The data base currently contains data on 26 plant traits that describe three key features of plant dynamics: persistence, regeneration and dispersal. The LEDA-Traitbase is freely available at www.leda-traitbase.org. 3. We present the structure of the data base and an overview of the trait information available. 4. Synthesis. The LEDA Traitbase is useful for large-scale analyses of functional responses of communities to environmental change, effects of community trait composition on ecosystem properties and patterns of rarity and invasiveness, as well as linkages between traits as expressions of fundamental trade-offs in plants.

Microbial Electrolysis Cells for High Yield Hydrogen Gas Production from Organic Matter

Abstract: The use of electrochemically active bacteria to break down organic matter, combined with the addition of a small voltage (>0.2 V in practice) in specially designed microbial electrolysis cells (MECs), can result in a high yield of hydrogen gas. While microbial electrolysis was invented only a few years ago, rapid developments have led to hydrogen yields approaching 100%, energy yields based on electrical energy input many times greater than that possible by water electrolysis, and increased gas production rates. MECs used to make hydrogen gas are similar in design to microbial fuel cells (MFCs) that produce electricity, but there are important differences in architecture and analytical methods used to evaluate performance. We review here the materials, architectures, performance, and energy efficiencies of these MEC systems that show promise as a method for renewable and sustainable energy production, and wastewater treatment.

Genome sequence of the metazoan plant-parasitic nematode Meloidogyne incognita

Abstract: Plant-parasitic nematodes are major agricultural pests worldwide and novel approaches to control them are sorely needed. We report the draft genome sequence of the root-knot nematode Meloidogyne incognita, a biotrophic parasite of many crops, including tomato, cotton and coffee. Most of the assembled sequence of this asexually reproducing nematode, totaling 86 Mb, exists in pairs of homologous but divergent segments. This suggests that ancient allelic regions in M. incognita are evolving toward effective haploidy, permitting new mechanisms of adaptation. The number and diversity of plant cell wall-degrading enzymes in M. incognita is unprecedented in any animal for which a genome sequence is available, and may derive from multiple horizontal gene transfers from bacterial sources. Our results provide insights into the adaptations required by metazoans to successfully parasitize immunocompetent plants, and open the way for discovering new antiparasitic strategies.

Peatlands and the carbon cycle: from local processes to global implications - a synthesis

Abstract: Although peatlands cover only 3% of the Earth's land surface, boreal and subarctic peatlands store about 15?30% of the world's soil carbon as peat. Despite their potential for large positive feedbacks to the climate system through sequestration and emission of greenhouse gases, peatlands are not explicitly included in global climate models and therefore in predictions of future climate change. In April 2007 a symposium was held in Wageningen, the Netherlands, to advance our understanding of peatland C cycling through integration across disciplines and research approaches and to develop a more synthetic picture of the present and future role of peatlands in the global C cycle and their interactions with the climate system. This paper aims to synthesize the main findings of the symposium, focusing on (i) small-scale processes, (ii) C fluxes at the landscape scale, and (iii) peatlands and climate. The paper concludes with a summary of the main drivers of the C balance of peatlands, and proposes directions for new research to reduce key uncertainties in our knowledge of C cycling in peatlands in order to facilitate the explicit inclusion of these ecosystems in a new generation of earth system models.

Proxy global assessment of land degradation

Abstract: Land degradation is always with us but its causes, extent and severity are contested. We define land degradation as a long-term decline in ecosystem function and productivity, which may be assessed using long-term, remotely sensed normalized difference vegetation index (NDVI) data. Deviation from the norm may serve as a proxy assessment of land degradation and improvement – if other factors that may be responsible are taken into account. These other factors include rainfall effects which may be assessed by rain-use efficiency, calculated from NDVI and rainfall. Results from the analysis of the 23-year Global Inventory Modeling and Mapping Studies (GIMMS) NDVI data indicate declining rain-use efficiency-adjusted NDVI on ca. 24% of the global land area with degrading areas mainly in Africa south of the equator, South-East Asia and south China, north-central Australia, the Pampas and swaths of the Siberian and north American taiga; 1.5 billion people live in these areas. The results are very different from previous assessments which compounded what is happening now with historical land degradation. Economic appraisal can be undertaken when land degradation is expressed in terms of net primary productivity and the resultant data allow statistical comparison with other variables to reveal possible drivers.

Slowing down as an early warning signal for abrupt climate change

Abstract: In the Earth's history, periods of relatively stable climate have often been interrupted by sharp transitions to a contrasting state. One explanation for such events of abrupt change is that they happened when the earth system reached a critical tipping point. However, this remains hard to prove for events in the remote past, and it is even more difficult to predict if and when we might reach a tipping point for abrupt climate change in the future. Here, we analyze eight ancient abrupt climate shifts and show that they were all preceded by a characteristic slowing down of the fluctuations starting well before the actual shift. Such slowing down, measured as increased autocorrelation, can be mathematically shown to be a hallmark of tipping points. Therefore, our results imply independent empirical evidence for the idea that past abrupt shifts were associated with the passing of critical thresholds. Because the mechanism causing slowing down is fundamentally inherent to tipping points, it follows that our way to detect slowing down might be used as a universal early warning signal for upcoming catastrophic change. Because tipping points in ecosystems and other complex systems are notoriously hard to predict in other ways, this is a promising perspective.

Heterosubtypic neutralizing monoclonal antibodies cross-protective against H5N1 and H1N1 recovered from human IgM+ memory B cells.

Abstract: Background The hemagglutinin (HA) glycoprotein is the principal target of protective humoral immune responses to influenza virus infections but such antibody responses only provide efficient protection against a narrow spectrum of HA antigenic variants within a given virus subtype. Avian influenza viruses such as H5N1 are currently panzootic and pose a pandemic threat. These viruses are antigenically diverse and protective strategies need to cross protect against diverse viral clades. Furthermore, there are 16 different HA subtypes and no certainty the next pandemic will be caused by an H5 subtype, thus it is important to develop prophylactic and therapeutic interventions that provide heterosubtypic protection. Methods and Findings Here we describe a panel of 13 monoclonal antibodies (mAbs) recovered from combinatorial display libraries that were constructed from human IgM+ memory B cells of recent (seasonal) influenza vaccinees. The mAbs have broad heterosubtypic neutralizing activity against antigenically diverse H1, H2, H5, H6, H8 and H9 influenza subtypes. Restriction to variable heavy chain gene IGHV1-69 in the high affinity mAb panel was associated with binding to a conserved hydrophobic pocket in the stem domain of HA. The most potent antibody (CR6261) was protective in mice when given before and after lethal H5N1 or H1N1 challenge. Conclusions The human monoclonal CR6261 described in this study could be developed for use as a broad spectrum agent for prophylaxis or treatment of human or avian influenza infections without prior strain characterization. Moreover, the CR6261 epitope could be applied in targeted vaccine strategies or in the design of novel antivirals. Finally our approach of screening the IgM+ memory repertoire could be applied to identify conserved and functionally relevant targets on other rapidly evolving pathogens.

Comparing the input, output, and validation maps for several models of land change

Abstract: This paper applies methods of multiple resolution map comparison to quantify characteristics for 13 applications of 9 different popular peer-reviewed land change models. Each modeling application simulates change of land categories in raster maps from an initial time to a subsequent time. For each modeling application, the statistical methods compare: (1) a reference map of the initial time, (2) a reference map of the subsequent time, and (3) a prediction map of the subsequent time. The three possible two-map comparisons for each application characterize: (1) the dynamics of the landscape, (2) the behavior of the model, and (3) the accuracy of the prediction. The three-map comparison for each application specifies the amount of the prediction’s accuracy that is attributable to land persistence versus land change. Results show that the amount of error is larger than the amount of correctly predicted change for 12 of the 13 applications at the resolution of the raw data. The applications are summarized and compared using two statistics: the null resolution and the figure of merit. According to the figure of merit, the more accurate applications are the ones where the amount of observed net change in the reference maps is larger. This paper facilitates communication among land change modelers, because it illustrates the range of results for a variety of models using scientifically rigorous, generally applicable, and intellectually accessible statistical techniques.

Treatment of input uncertainty in hydrologic modeling: doing hydrology backward with Markov chain Monte Carlo simulation.

Abstract: [1] There is increasing consensus in the hydrologic literature that an appropriate framework for streamflow forecasting and simulation should include explicit recognition of forcing and parameter and model structural error. This paper presents a novel Markov chain Monte Carlo (MCMC) sampler, entitled differential evolution adaptive Metropolis (DREAM), that is especially designed to efficiently estimate the posterior probability density function of hydrologic model parameters in complex, high-dimensional sampling problems. This MCMC scheme adaptively updates the scale and orientation of the proposal distribution during sampling and maintains detailed balance and ergodicity. It is then demonstrated how DREAM can be used to analyze forcing data error during watershed model calibration using a five-parameter rainfall-runoff model with streamflow data from two different catchments. Explicit treatment of precipitation error during hydrologic model calibration not only results in prediction uncertainty bounds that are more appropriate but also significantly alters the posterior distribution of the watershed model parameters. This has significant implications for regionalization studies. The approach also provides important new ways to estimate areal average watershed precipitation, information that is of utmost importance for testing hydrologic theory, diagnosing structural errors in models, and appropriately benchmarking rainfall measurement devices.

Accuracy of predicting the genetic risk of disease using a genome-wide approach.

Abstract: Background: The prediction of the genetic disease risk of an individual is a powerful public health tool. While predicting risk has been successful in diseases which follow simple Mendelian inheritance, it has proven challenging in complex diseases for which a large number of loci contribute to the genetic variance. The large numbers of single nucleotide polymorphisms now available provide new opportunities for predicting genetic risk of complex diseases with high accuracy. Methodology/Principal Findings: We have derived simple deterministic formulae to predict the accuracy of predicted genetic risk from population or case control studies using a genome-wide approach and assuming a dichotomous disease phenotype with an underlying continuous liability. We show that the prediction equations are special cases of the more general problem of predicting the accuracy of estimates of genetic values of a continuous phenotype. Our predictive equations are responsive to all parameters that affect accuracy and they are independent of allele frequency and effect distributions. Deterministic prediction errors when tested by simulation were generally small. The common link among the expressions for accuracy is that they are best summarized as the product of the ratio of number of phenotypic records per number of risk loci and the observed heritability. Conclusions/Significance: This study advances the understanding of the relative power of case control and population studies of disease. The predictions represent an upper bound of accuracy which may be achievable with improved effect estimation methods. The formulae derived will help researchers determine an appropriate sample size to attain a certain accuracy when predicting genetic risk.

High-throughput diversity and functionality analysis of the gastrointestinal tract microbiota

Abstract: The human gastrointestinal (GI) tract microbiota plays a pivotal role in our health. For more than a decade a major input for describing the diversity of the GI tract microbiota has been derived from the application of small subunit ribosomal RNA (SSU rRNA)-based technologies. These not only provided a phylogenetic framework of the GI tract microbiota, the majority of which has not yet been cultured, but also advanced insights into the impact of host and environmental factors on the microbiota community structure and dynamics. In addition, it emerged that GI tract microbial communities are host and GI tract location-specific. This complicates establishing relevant links between the host’s health and the presence or abundance of specific microbial populations and argues for the implementation of novel high-throughput technologies in studying the diversity and functionality of the GI tract microbiota. Here, we focus on the recent developments and applications of phylogenetic microarrays based on SSU rRNA sequences and metagenomics approaches exploiting rapid sequencing technologies in unravelling the secrets of our GI tract microbiota.

Are functional traits good predictors of demographic rates? evidence from five neotropical forests

Abstract: A central goal of comparative plant ecology is to understand how functional traits vary among species and to what extent this variation has adaptive value. Here we evaluate relationships between four functional traits (seed volume, specific leaf area, wood density, and adult stature) and two demographic attributes (diameter growth and tree mortality) for large trees of 240 tree species from five Neotropical forests. We evaluate how these key functional traits are related to survival and growth and whether similar relationships between traits and demography hold across different tropical forests. There was a tendency for a trade-off between growth and survival across rain forest tree species. Wood density, seed volume, and adult stature were significant predictors of growth and/or mortality. Both growth and mortality rates declined with an increase in wood density. This is consistent with greater construction costs and greater resistance to stem damage for denser wood. Growth and mortality rates also declined as seed volume increased. This is consistent with an adaptive syndrome in which species tolerant of low resource availability (in this case shade-tolerant species) have large seeds to establish successfully and low inherent growth and mortality rates. Growth increased and mortality decreased with an increase in adult stature, because taller species have a greater access to light and longer life spans. Specific leaf area was, surprisingly, only modestly informative for the performance of large trees and had ambiguous relationships with growth and survival. Single traits accounted for 9-55% of the interspecific variation in growth and mortality rates at individual sites. Significant correlations with demographic rates tended to be similar across forests and for phylogenetically independent contrasts as well as for cross-species analyses that treated each species as an independent observation. In combination, the morphological traits explained 41% of the variation in growth rate and 54% of the variation in mortality rate, with wood density being the best predictor of growth and mortality. Relationships between functional traits and demographic rates were statistically similar across a wide range of Neotropical forests. The consistency of these results strongly suggests that tropical rain forest species face similar trade-offs in different sites and converge on similar sets of solutions.

Promoting coherent minimum reporting guidelines for biological and biomedical investigations: the MIBBI project

Abstract: The Minimum Information for Biological and Biomedical Investigations (MIBBI) project aims to foster the coordinated development of minimum-information checklists and provide a resource for those exploring the range of extant checklists.

Biological control and sustainable food production

Abstract: The use of biological control for the management of pest insects pre-dates the modern pesticide era. The first major successes in biological control occurred with exotic pests controlled by natural enemy species collected from the country or area of origin of the pest (classical control). Augmentative control has been successfully applied against a range of open-field and greenhouse pests, and conservation biological control schemes have been developed with indigenous predators and parasitoids. The cost–benefit ratio for classical biological control is highly favourable (1 : 250) and for augmentative control is similar to that of insecticides (1 : 2–1 : 5), with much lower development costs. Over the past 120 years, more than 5000 introductions of approximately 2000 non-native control agents have been made against arthropod pests in 196 countries or islands with remarkably few environmental problems. Biological control is a key component of a ‘systems approach’ to integrated pest management, to counteract insecticide-resistant pests, withdrawal of chemicals and minimize the usage of pesticides. Current studies indicate that genetically modified insect-resistant Bt crops may have no adverse effects on the activity or function of predators or parasitoids used in biological control. The introduction of rational approaches for the environmental risk assessment of non-native control agents is an essential step in the wider application of biological control, but future success is strongly dependent on a greater level of investment in research and development by governments and related organizations that are committed to a reduced reliance on chemical control.

S layer protein A of Lactobacillus acidophilus NCFM regulates immature dendritic cell and T cell functions

Abstract: Dendritic cells (DCs) are antigen-presenting cells that play an essential role in mucosal tolerance. They regularly encounter beneficial intestinal bacteria, but the nature of these cellular contacts and the immune responses elicited by the bacteria are not entirely elucidated. Here, we examined the interactions of Lactobacillus acidophilus NCFM and its cell surface compounds with DCs. L. acidophilus NCFM attached to DCs and induced a concentration-dependent production of IL-10, and low IL-12p70. We further demonstrated that the bacterium binds to DC-specific ICAM-3-grabbing nonintegrin (DC-SIGN), a DC- specific receptor. To identify the DC-SIGN ligand present on the bacterium, we took advantage of a generated array of L. acidophilus NCFM mutants. A knockout mutant of L. acidophilus NCFM lacking the surface (S) layer A protein (SlpA) was significantly reduced in binding to DC-SIGN. This mutant incurred a chromosomal inversion leading to dominant expression of a second S layer protein, SlpB. In the SlpB-dominant strain, the nature of the interaction of this bacterium with DCs changed dramatically. Higher concentrations of proinflammatory cytokines such as IL-12p70, TNFα, and IL-1β were produced by DCs interacting with the SlpB-dominant strain compared with the parent NCFM strain. Unlike the SlpA-knockout mutant, T cells primed with L. acidophilus NCFM stimulated DCs produced more IL-4. The SlpA–DC-SIGN interaction was further confirmed as purified SlpA protein ligated directly to the DC-SIGN. In conclusion, the major S layer protein, SlpA, of L. acidophilus NCFM is the first probiotic bacterial DC-SIGN ligand identified that is functionally involved in the modulation of DCs and T cells functions.

Long-term organic farming fosters below and aboveground biota: Implications for soil quality, biological control and productivity

Abstract: Organic farming may contribute substantially to future agricultural production worldwide by improving soil quality and pest control, thereby reducing environmental impacts of conventional farming. We investigated in a comprehensive way soil chemical, as well as below and aboveground biological parameters of two organic and two conventional wheat farming systems that primarily differed in fertilization and weed management strategies. Contrast analyses identified management related differences between “herbicide-free” bioorganic (BIOORG) and biodynamic (BIODYN) systems and conventional systems with (CONFYM) or without manure (CONMIN) and herbicide application within a long-term agricultural experiment (DOK trial, Switzerland). Soil carbon content was significantly higher in systems receiving farmyard manure and concomitantly microbial biomass (fungi and bacteria) was increased. Microbial activity parameters, such as microbial basal respiration and nitrogen mineralization, showed an opposite pattern, suggesting that soil carbon in the conventional system (CONFYM) was more easily accessible to microorganisms than in organic systems. Bacterivorous nematodes and earthworms were most abundant in systems that received farmyard manure, which is in line with the responses of their potential food sources (microbes and organic matter). Mineral fertilizer application detrimentally affected enchytraeids and Diptera larvae, whereas aphids benefited. Spider abundance was favoured by organic management, most likely a response to increased prey availability from the belowground subsystem or increased weed coverage. In contrast to most soil-based, bottom-up controlled interactions, the twofold higher abundance of this generalist predator group in organic systems likely contributed to the significantly lower abundance of aboveground herbivore pests (aphids) in these systems. Long-term organic farming and the application of farmyard manure promoted soil quality, microbial biomass and fostered natural enemies and ecosystem engineers, suggesting enhanced nutrient cycling and pest control. Mineral fertilizers and herbicide application, in contrast, affected the potential for top-down control of aboveground pests negatively and reduced the organic carbon levels. Our study indicates that the use of synthetic fertilizers and herbicide application changes interactions within and between below and aboveground components, ultimately promoting negative environmental impacts of agriculture by reducing internal biological cycles and pest control. On the contrary, organic farming fosters microbial and faunal decomposers and this propagates into the aboveground system via generalist predators thereby increasing conservation biological control. However, grain and straw yields were 23% higher in systems receiving mineral fertilizers and herbicides reflecting the trade-off between productivity and environmental responsibility.

Sewage Sludge as a Biomass Resource for the Production of Energy: Overview and Assessment of the Various Options†

Abstract: Treatment of municipal wastewater results worldwide in the production of large amounts of sewage sludge. The major part of the dry matter content of this sludge consists of nontoxic organic compounds, in general a combination of primary sludge and secondary (microbiological) sludge. The sludge also contains a substantial amount of inorganic material and a small amount of toxic components. There are many sludge-management options in which production of energy (heat, electricity, or biofuel) is one of the key treatment steps. The most important options are anaerobic digestion, co-digestion, incineration in combination with energy recovery, co-incineration in coal-fired power plants, co-incineration in combination with organic waste focused on energy recovery, use as an energy source in the production of cement or building materials, pyrolysis, gasification, supercritical (wet) oxidation, hydrolysis at high temperature, production of hydrogen, acetone, butanol, or ethanol, and direct generation of electrical energy by means of specific micro-organisms. Incineration and co-incineration with energy recovery and use of sewage sludge in the production of Portland cement are applied on a large scale. In these processes, the toxic organics are destructed and the heavy metals are immobilized in the ash or cement. The energy efficiency of these processes strongly depends upon the dewatering and drying step. It is expected that these applications will strongly increase in the future. Supercritical wet oxidation is a promising innovative technology but is still in the development stage. With the exception of biogas production, the other biological methods to produce energy are still in the initial research phase. Production of biogas from sewage sludge is already applied worldwide on small, medium, and large scales. With this process, a substantial experience exists and it is expected that this application is getting more and more attention. Besides the increasing focus on the recovery and reuse of energy, inorganics, and phosphorous, there is also an increasing focus to solve completely the problem of the toxic organics and inorganic compounds in sludge. In the assessment and selection of options for energy recovery by means of biological methods, this aspect has to be taken into account.

Social interactions in urban parks: stimulating social cohesion?

Abstract: People from all ethnic backgrounds spend some of their leisure time in green areas. This study found that urban parks are more inclusive green places than non-urban green areas, and that urban parks can promote social cohesion. The objective of the research was to establish the extent to which urban parks facilitate social cohesion and how social interaction and place attachment can contribute to such cohesion. Quantitative research (a survey) and qualitative research (observations and interviews) carried out in five urban parks in the Netherlands revealed that there are many similarities in the ways that ethnic groups use urban parks and in the meanings of such parks to these groups. Urban parks are sites where different ethnic groups mingle and where informal and cursory interactions can stimulate social cohesion. Furthermore, being involved and concerned with parks can facilitate attachment to these places. Urban parks can provide a vital locality where everyday experiences are shared and negotiated with a variety of people. The design of a park, its location and people's image of the park in combination with the cultural characteristics of various ethnic groups inform the opportunities for intercultural interactions.

Kinetic Modeling of Food Quality: A Critical Review

Abstract: This article discusses the possibilities to study relevant quality aspects of food, such as color, nutrient content, and safety, in a quantitative way via mathematical models. These quality parameters are governed by chemical, biochemical, microbial, and physical changes. It is argued that the modeling of such quality aspects is in fact kinetic modeling. Therefore, attention is paid to chemical kinetics, and its possibilities and limitations are discussed when applied to changes occurring in foods. The discussion is illustrated with examples from the literature. A major difficulty is that principles from chemical kinetics are strictly speaking only valid for simple elementary reactions, and foods are all but simple. Interactions in the food matrix and variability are 2 complicating factors. It is discussed how this difficulty can be tackled, and research priorities are suggested to come to better models in food science, and thereby to a better control of food quality.

Integrating Agricultural Landscapes with Biodiversity Conservation in the Mesoamerican Hotspot

Abstract: CELIA A. HARVEY,∗‡‡‡ OLIVER KOMAR,† ROBIN CHAZDON,‡ BRUCE G. FERGUSON,§ BRYAN FINEGAN,∗∗ DANIEL M. GRIFFITH,†† MIGUEL MARTINEZ-RAMOS,‡‡ HELDA MORALES,§ RONALD NIGH,§§ LORENA SOTO-PINTO,§ MICHIEL VAN BREUGEL,∗∗∗ AND MARK WISHNIE††† ∗Department of Agriculture and Agroforestry, CATIE, Apdo 7170, Turrialba, Costa Rica †Programa de Ciencias para la Conservacion, SalvaNATURA, Colonia Flor Blanca, 33 Avenida Sur #640, San Salvador, El Salvador ‡Department of Ecology and Evolutionary Biology, University of Connecticut, Storrs, CT 06268-3043, U.S.A. §Departamento de Agroecoloǵia, El Colegio de la Frontera Sur, Carretera Panamericana y Periferico Sur s-n, San Cristobal de Las Casas, Chiapas, Mexico ∗∗Department of Natural Resources and Environment, CATIE, Apdo 7170, Turrialba, Costa Rica ††Biodiversity of BOSAWAS Biosphere Reserve, Saint Louis Zoo, Managua, Nicaragua ‡‡Centro de Investigaciones en Ecosistemas, UNAM, AP 27-3 Santa Maŕia de Guido, CP 58089, Morelia, Michoacan, Mexico §§Centro de Investigaciones y Estudios Superiores en Antropoloǵia Social, San Cristobal de las Casas, Chiapas, Mexico ∗∗∗Centre for Ecosystem Studies, Wageningen University, P.O. Box 47, 6700 AA Wageningen, The Netherlands †††Equator Environmental, LLC, 250 Park Avenue South, New York, NY 10003, U.S.A.