In the EURAPMON programme it was identified that differences between studies in sampling and processing strategies for contaminant monitoring hampered direct comparison of study results and an integrated interpretation (Movalli et al. 2008). The use of appropriate sample containers and storage conditions was also identified as an essential step to avoid contamination of samples or degradation of the compound of interest, and information on this needed to be easily accessible to personnel collecting and sending samples to ecotoxicological laboratories. Although toxicology specialists have a deep understanding on these issues, other groups of professionals (e.g. field researchers) or volunteers (e.g. ringers) collecting samples need guidance that ensures appropriate collection of samples without compromising the analytical quality of the sample. Indeed, raptor population monitoring activities have the potential to enhance a widescale availability of raptor samples that are interlinked with key contextual data (e.g. breeding success, population trends, survival, diet, etc.), since some of the biological matrices needed for analysis (e.g. feathers, addled/deserted eggs, blood) are routinely collected as part of surveys of raptor breeding populations (Espín et al. 2016; Derlink et al. 2018). Consequently, it is essential to provide protocols on appropriate sampling methods for contaminant monitoring, for field ornithologists, personnel at museums and Wildlife Rehabilitation Centres, and other people involved in raptor sample collection.
A detailed best-practice sampling protocol was previously prepared by EURAPMON (Espín et al. 2014), as well as a publication of the potential widescale availability of raptor samples and the relative merits of each matrix type (Espín et al. 2016). However, through more recent meetings and workshops under ERBFacility involving researchers across 27 countries, the need to elaborate a protocol in a more schematic and clear format was identified. This schematic protocol should provide essential information on the practicalities of sampling and storage that were not earlier reported by Espín et al. 2014 (e.g. type of containers to conserve matrices, storage and transport conditions, and how these differ depending on the sample matrices and the contaminant to be analysed). Twenty-three researchers involved in the three arenas of the ERBFacility COST Action and representing 11 countries (Denmark, Germany, Iceland, Norway, Portugal, Slovenia, Spain, Sweden, Switzerland, The Netherlands and United Kingdom), have participated in the preparation of this schematic sampling protocol based upon their expertise in sampling and contaminant monitoring in raptors.
The protocol presented in this perspective (see Electronic Supplementary Material) provides guidance on sample collection for contaminant monitoring in raptors with a view to increasing sampling capacity across countries and facilitating harmonization of procedures to maximize the reliability, comparability and interoperability of biomonitoring data. While the protocol has been prepared under a Europe COST Action, the guidance is applicable to raptor monitoring worldwide. The protocol is presented as Supplementary Material to this communication, and has been prepared following an easy-to-follow style, with hyperlinks to redirect the reader to the relevant information elsewhere in the protocol.
The schematic protocol starts with each matrix type, including sample types that are collected both during active sampling (samples taken from captured live birds and monitored nests) and passive monitoring (samples taken from dead birds and deserted nests) (Fig. 1). These sample types include whole blood, plasma, serum, deserted or addled eggs, feathers, preen oil, regurgitated pellets, prey remains, gastric content and internal tissues. The reader clicks on the sample type of interest and is redirected to the specific protocol for each matrix (e.g. see protocol for blood sampling in Fig. 2), which in turn offers further hyperlinks to additional and more detailed information. Some important general guidelines are also given regarding sampling and ethical permits, personal safety and wildlife health, animal welfare, labelling samples, and essential guidance to avoid contamination and to record basic data (date, location, etc.) in the sampling report. Moreover, new information is provided on the volume/mass of sample needed for contaminant monitoring, the most suitable container type to conserve the sample, and the necessary conditions required for transportation and storage.
Different sample matrices provide different information about exposure and effects and not all are suitable for biomonitoring (Espín et al. 2016). Thus, information on the type of contaminants and biomarkers most frequently analysed in the different matrices in live and dead birds is also shown. Photographs and links to web-based videos are also provided to illustrate the proper materials and methods needed for sample collection, taking measurements (such as measuring egg size, eggshell thickness, body fat tissue during necropsy, etc.) and identifying gonads and internal tissues.
We recommend the schematic protocol for use by professionals and volunteers as a standard guide to ensure harmonised sampling methods and appropriate quality of samples collected for contaminant monitoring in raptors. Contaminant issues often occur across national and continental boundaries, and therefore require harmonised methods to study their occurrence, impact and any effect of legal or voluntary mitigation, whether of legacy, current or emerging contaminants.