Enantiomeric separation of metolachlor and its metabolites using LC-MS and CZE

The stereoisomers of metolachlor and its two polar metabolites [ethane sulfonic acid (ESA) and oxanilic acid (OXA)] were separated using liquid chromatography-mass spectrometry (LC-MS) and capillary zone electrophoresis (CZE), respectively. The separation of metolachlor enantiomers was achieved using a LC-MS equipped with a chiral stationary phase based on cellulose tris(3,5-dimethylphenyl carbamate) and an atmospheric pressure chemical ionization source operated under positive ion mode. The enantiomers of ESA and OXA were separated using CZE with gamma-cyclodextrin (gamma-CD) as chiral selector. Various CZE conditions were investigated to achieve the best resolution of the ESA and OXA enantiomers. The optimum background CZE electrolyte was found to consist of borate buffer (pH=9) containing 20% methanol (v/v) and 2.5% gamma-CD (w/v). Maximum resolution of ESA and OXA enantiomers was achieved using a capillary temperature of 15 degrees C and applied voltage of 30 kV. The applicability of the LC-MS and CZE methods was demonstrated successfully on the enantiomeric analysis of metolachlor and its metabolites in samples from a soil and water degradation study that was set up to probe the stereoselectivity of metolachlor biodegradation. These techniques allow the enantiomeric ratios of the target analytes to be followed over time during the degradation process and thus will prove useful in determining the role of chirality in pesticide degradation and metabolite formation.     

Microbial activity in an acid resin deposit: biodegradation potential and ecotoxicology in an extremely acidic hydrocarbon contamination

Acid resins are residues produced in a recycling process for used oils that was in use in the forties and fifties of the last century. The resin-like material is highly contaminated with mineral oil hydrocarbons, extremely acidic and co-contaminated with substituted and aromatic hydrocarbons, and heavy metals. To determine the potential for microbial biodegradation the acid resin deposit and its surroundings were screened for microbial activity by soil respiration measurements. No microbial activity was found in the core deposit. However, biodegradation of hydrocarbons was possible in zones with a lower degree of contamination surrounding the deposit. An extreme acidophilic microbial community was detected close to the core deposit. With a simple ecotoxicological approach it could be shown that the pure acid resin that formed the major part of the core deposit, was toxic to the indigenous microflora due to its extremely low pH of 0-1.     

Toxicity and repellency of hot pepper extracts to spider mite, Tetranychus urticae Koch

Increasing concern about persistence and environmental impact of synthetic pesticide residues require development of biodegradable and environmentally safe alternatives. The potential of using fruit extracts of hot pepper as alternatives to synthetic acaricides for controlling the two-spotted spider mite, Tetranychus urticae Koch, is explored in this study. Twenty-four Capsicum accessions (Solanaceae) were screened for their toxicity and repellency to the spider mites. Crude extracts from fruits of C. chinense, C. frutescens, C. baccatum, C. annuum, and C. pubescens were prepared in methanol and tested for their acaricidal properties. Spider mite mortality was greatest (45%) when fruit extract of accession Grif-9169 (C. annuum) was used. Results from diving board bioassays indicated that mites avoided filter paper strips treated with hot pepper extracts from accessions PI-596057 (C. baccatum), PI-195299 (C. annuum), and Grif- 9270 (C. annuum). This investigation suggests that methanolic extracts of these three accessions may have a great potential for repelling spider mites and should be field-tested on a large-scale to assess their value in managing populations of spider mites, which could reduce reliance on synthetic acaricides. An attempt was made to correlate repellency with chemical constituents of fruit extracts of the most repellent accessions to identify chemical sources of repellency. Capsaicin and dihydrocapsaicin, the pungent components of pepper fruit, were not correlated with toxicity or repellency, indicating that these are not likely related to the toxicity or repellency of the pepper fruit extracts. Other, unidentified chemicals are likely responsible for toxicity and repellency to the two-spotted spider mite.     

Effectiveness of Panama as an intercontinental land bridge for large mammals

Habitat fragmentation is a primary driver of wildlife loss, and establishment of biological corridors is a common strategy to mitigate this problem. A flagship example is the Mesoamerican Biological Corridor (MBC), which aims to connect protected forest areas between Mexico and Panama to allow dispersal and gene flow of forest organisms. Because forests across Central America have continued to degrade, the functioning of the MBC has been questioned, but reliable estimates of species occurrence were unavailable. Large mammals are suitable indicators of forest functioning, so we assessed their conservation status across the Isthmus of Panama, the narrowest section of the MBC. We used large-scale camera-trap surveys and hierarchical multispecies occupancy models in a Bayesian framework to estimate the occupancy of 9 medium to large mammals and developed an occupancy-weighted connectivity metric to evaluate species-specific functional connectivity. White-lipped peccary (Tayassu pecari), jaguar (Panthera onca), giant anteater (Myrmecophaga tridactyla), white-tailed deer (Odocoileus virginianus), and tapir (Tapirus bairdii) had low expected occupancy along the MBC in Panama. Puma (Puma concolor), red brocket deer (Mazama temama), ocelot (Leopardus pardalis), and collared peccary (Pecari tajacu), which are more adaptable, had higher occupancy, even in areas with low forest cover near infrastructure. However, the majority of species were subject to ≥1 gap that was larger than their known dispersal distances, suggesting poor connectivity along the MBC in Panama. Based on our results, forests in Darien, Donoso–Santa Fe, and La Amistad International Park are critical for survival of large terrestrial mammals in Panama and 2 areas need restoration.     

Optimization of capture–recapture monitoring of elusive species illustrated with a threatened grasshopper

Information on population sizes and trends of threatened species is essential for their conservation, but obtaining reliable estimates can be challenging. We devised a method to improve the precision of estimates of population size obtained from capture–recapture studies for species with low capture and recapture probabilities and short seasonal activity, illustrated with population data of an elusive grasshopper (Prionotropis rhodanica). We used data from 5 capture–recapture studies to identify methodological and environmental factors affecting capture and recapture probabilities and estimates of population size. In a simulation, we used the population size and capture and recapture probability estimates obtained from the field studies to identify the minimum number of sampling occasions needed to obtain unbiased and robust estimates of population size. Based on these results we optimized the capture–recapture design, implemented it in 2 additional studies, and compared their precision with those of the nonoptimized studies. Additionally, we simulated scenarios based on thresholds of population size in criteria C and D of the International Union for Conservation of Nature (IUCN) Red List to investigate whether estimates of population size for elusive species can reliably inform red-list assessments. Identifying parameters that affect capture and recapture probabilities (for the grasshopper time since emergence of first adults) and optimizing field protocols based on this information reduced study effort (−6% to −27% sampling occasions) and provided more precise estimates of population size (reduced coefficient of variation) compared with nonoptimized studies. Estimates of population size from the scenarios based on the IUCN thresholds were mostly unbiased and robust (only the combination of very small populations and little study effort produced unreliable estimates), suggesting capture–recapture can be considered reliable for informing red-list assessments. Although capture–recapture remains difficult and costly for elusive species, our optimization procedure can help determine efficient protocols to increase data quality and minimize monitoring effort.     

Applying the zoo model to conservation of threatened exceptional plant species

Maintaining a living plant collection is the most common method of ex situ conservation for plant species that cannot be seed banked (i.e., exceptional species). Viability of living collections, and their value for future conservation efforts, can be limited without coordinated efforts to track and manage individuals across institutions. Using a pedigree-focused approach, the zoological community has established an inter-institutional infrastructure to support long-term viability of captive animal populations. We assessed the ability of this coordinated metacollection infrastructure to support the conservation of 4 plant species curated in living collections at multiple botanic gardens around the world. Limitations in current practices include the inability to compile, share, and analyze plant collections data at the individual level, as well as difficulty in tracking original provenance of ex situ material. The coordinated metacollection framework used by zoos can be adopted by the botanical community to improve conservation outcomes by minimizing the loss of genetic diversity in collections. We suggest actions to improve ex situ conservation of exceptional plant species, including developing a central database to aggregate data and track unique individuals of priority threatened species among institutions and adapting a pedigree-based population management tool that incorporates life-history aspects unique to plants. If approached collaboratively across regional, national, and global scales, these actions could transform ex situ conservation of threatened plant species.