Biogeochemical particulate and microplastics analysis in Antarctic penguin guano samples, collected 2021-2022
GB/NERC/BAS/PDC/02205
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Summary
Abstract:
This dataset presents the results of the quantification of biogeochemical particulate components (carbon, nitrogen, and biogenic silica) and the identification of microplastics in penguin guano samples collected from Deception and Livingston Islands (South Shetland Islands) during the 2020/2021 and 2021/2022 austral summers. Both species (Chinstrap and Gentoo) showed similar values for natural particulate fractions. Adapted and refined FTIR techniques enabled the detection of the smaller fraction of microplastics up to 25 micrometers in penguin guano for the first time. Microplastics were detected in 91 percent of samples, dominated by small particles (46 percent 25-50 micrometers). Chinstrap guano contained the highest amount of microplastics. Polypropylene was the predominant polymer (34 percent in Chinstrap, 75 percent in Gentoo) followed by Polyethylene (37 percent in Chinstrap, not found in Gentoo). This study provides the first survey of the smallest microplastic fraction (down to 25 micrometers) in penguin guano, offering new insight into how a shift in the partitioning from natural to microplastic particles, previously overlooked, may influence guano-mediated carbon pathways.
This research falls under the framework of the Spanish Government project PIMETAN (RTI2018-098048-B-I00) and the UKRI-FLF project CUPIDO (MR/T020962/1). E. Sparaventi was supported by the Spanish predoctoral FPI grant (PRE2019-089679) and a bursary from Antarctic Science Ltd.
Keywords:
Antarctica, biogeochemical cycles, carbon, guano, microplastics, penguins
Citation
Sparaventi, E., Rowlands, E., Giglio, F., Rodriguez-Romero, A., Tovar-Sanchez, A., & Manno, C. (2026). Biogeochemical particulate and microplastics analysis in Antarctic penguin guano samples, collected 2021-2022 (Version 1.0) [Data set]. NERC EDS UK Polar Data Centre. https://doi.org/10.5285/88c79a01-d3c0-4fca-835c-481930948a5e
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REFERENCE MATERIALS
- https://doi.org/10.1016/j.scitotenv.2021.147698
- https://doi.org/10.1021/acs.estlett.1c00851
- https://doi.org/10.1021/es2031505
- https://doi.org/10.1039/c9ay00126c
REFERENCE MATERIALS
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Constraints
| Access Constraints: | No restrictions apply |
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| Use Constraints: | This data is governed by the NERC data policy (http://www.nerc.ac.uk/research/sites/data/policy/) and supplied under Open Government Licence v.3 (http://www.nationalarchives.gov.uk/doc/open-government-licence/version/3/). |
Basic Information
| Creation Date: | 2026-05-14 |
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| Dataset Progress: | Complete |
| Dataset Language: | English |
| ISO Topic Categories: |
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| Parameters: |
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| Personnel: | |
| Name | UK PDC |
| Role(s) | Metadata Author |
| Organisation | British Antarctic Survey |
| Name | Erica Sparaventi |
| Role(s) | Investigator, Technical Contact |
| Organisation | British Antarctic Survey |
| Name | Emily Rowlands |
| Role(s) | Investigator |
| Organisation | British Antarctic Survey |
| Name | Federico Giglio |
| Role(s) | Investigator |
| Organisation | Istituto di Scienze Polari |
| Name | Araceli Rodriguez-Romero |
| Role(s) | Investigator |
| Organisation | Institute of Marine Sciences of Andalusia |
| Name | Antonio Tovar-Sanchez |
| Role(s) | Investigator |
| Organisation | Institute of Marine Sciences of Andalusia |
| Name | Clara Manno |
| Role(s) | Investigator |
| Organisation | British Antarctic Survey |
| Parent Dataset: | N/A |
Additional Information
| Reference: | Fragao, J.; Bessa, F.; Otero, V.; Barbosa, A.; Sobral, P.; Waluda, C. M.; Guimaro, H. R.; Xavier, J. C. Microplastics and Other Anthropogenic Particles in Antarctica: Using Penguins as Biological Samplers. Sci. Total Environ. 2021, 788, 147698. https://doi.org/10.1016/j.scitotenv.2021.147698 Hidalgo-Ruz, V.; Gutow, L.; Thompson, R. C.; Thiel, M. Microplastics in the Marine Environment: A Review of the Methods Used for Identification and Quantification. Environ. Sci. Technol. 2012, 46 (6), 3060-3075. https://doi.org/10.1021/es2031505 Hufnagl, B.; Stibi, M.; Martirosyan, H.; Wilczek, U.; Moller, J. N.; Loder, M. G. J.; Laforsch, C.; Lohninger, H. Computer-Assisted Analysis of Microplastics in Environmental Samples Based on uFTIR Imaging in Combination with Machine Learning. Environ. Sci. Technol. Lett. 2022, 9 (1), 90-95. https://doi.org/10.1021/acs.estlett.1c00851 Primpke, S., Dias, P.A., Gerdts, G., 2019. Automated identification and quantification of microfibres and microplastics. https://doi.org/10.1039/c9ay00126c Roscher, L. et al. (2021) 'Microplastic pollution in the Weser estuary and the German North Sea', Environmental Pollution, 288, p. 117681. doi:10.1016/j.envpol.2021.117681. |
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| Quality: | To avoid potential contamination, all handling steps were performed under controlled clean conditions. Guano samples were collected with PS spoons and stored using falcon tubes, zip lock bags, however, all storage material were new and the spoon was cleaned after each use. Samples were handled to minimize external contamination prior to frozen condition storage, with exposure kept to a minimum. Once in the lab, for the microplastic particles extraction procedures all reagents were first filtered through a 2 micrometer polycarbonate filter. All glass and beakers were cleaned with 95 percent ethanol and ultrapure water before use. Furthermore, all the steps included in the protocol were performed in a laminar flow hood in a restricted access laboratory and only cotton lab coats and nitrile gloves were used. To check for contamination, three types of blanks were performed: i) procedural blanks (3 replicates); ii) lab contamination air blanks (2 replicates); and iii) storage blanks, for zip lock bags (2 replicates) and falcon tubes (2 replicates). In particular, a total of 3 procedural blanks were performed for the entire analysis and processed with the same protocol as the samples but using only filtered milli-Q. In addition, lab contamination air blanks used polycarbonate filters (47 mm diameter and 0.2 micrometer pore size) placed in glass petri dishes on the laboratory bench and simulate all the steps and procedures that have been carried out. Furthermore, storage blanks, consisting of 2 empty zip-lock bags and 2 falcon tubes stored for over a year, were analyzed to check potential contamination during storage. No field blanks were taken to evaluate background contamination, since guano sampling was not initially designed for investigating plastic pollution. | |
| Lineage/Methodology: | A total of 32 fresh penguin guano samples were collected during two Antarctic campaigns in the South Shetland Islands (SSI). In the austral summers of 2020/2021 and 2021/2022, samples were collected from Chinstrap penguin breeding colonies at Vapour Col (62 deg 59' S, 60 deg 44' W) (20,000 breeding pairs) and Baily Head (62 deg 58' S, 60 deg 30' W) (50,000 breeding pairs) on Deception Island (n=25). In the austral summer of 2021/2022, samples were collected from Gentoo penguin breeding colonies at Hannah Point (62 deg 39' S, 60 deg 36' W), Sally Rock (62 deg 42' S, 60 deg 25' W) and Argentina Cove (62 deg 40' S, 60 deg 24' W) on Livingston Island (n=7). Single colony census is not available for Gentoo penguins, since they are distributed across multiple smaller colonies, but total abundance is estimated to be on the order of several thousand breeding pairs (10^3-10^4), based on site-specific monitoring data on Livingston Island). Wet guano was carefully collected by hand or with a plastic (polystyrene, PS) spoon from snow, ice, or rocks, avoiding collecting ground remains, placed in a conical centrifuge tube or in a zip lock bag and frozen at -20 deg C prior to processing in the laboratory. Biogeochemical analysis: The 32 guano samples were freeze-dried (LABCONCO, model 798,030). Subsamples of dried guano (3.5 mg) were used for analyses of particulate biogeochemical matter. Total particulate carbon (TC), particulate organic carbon (POC) and total nitrogen (TN) were measured in triplicate by combustion in an elemental analyser (CHN, Exeter Analytical Inc. CE440 elemental analyser, accuracy +/- 0.15 percent). For POC determination, samples were pre-treated with 1 N hydrochloric acid (HCl, analytical grade, >= 37 percent, then diluted, Sigma-Aldrich). Particulate inorganic carbon (PIC) was obtained by determining the difference between TC and POC. Three blank filters (GF/F filters, 25 mm, 0.45 micrometer pore size) were interspersed regularly between samples (run every 25 samples), to measure and correct for drift. The amount of particulate biogenic silica (BSi) in guano was quantified by colorimetric analysis (Jasco UVIDEC accuracy +/- 1 percent). For the silicate extraction, approximately 10-20 mg of guano sample was suspended in 50 mL of 0.5 M sodium hydroxide (NaOH, pellets, Carlo Erba, Milan, Italy) at 85 deg C for 5 h. Following a progressive dissolution method, an aliquot of each sample was taken for analysis every hour and the relative silica data were extrapolated back to time zero to correct for the silica originating from coexisting clay minerals. The dissolved silicate was then reacted with Ammonium Molybdate Tetrahydrate (99+ percent, Thermo Scientific). To shift the solution toward blue (measured at 810 nm), a reducing solution was added, composed of: Metol sulfite (Sodium Sulfite anhydrous, Thermo Scientific), 4-Methylaminophenol sulfate (99 percent extra pure, Thermo Scientific), Oxalic acid dihydrate (VWR), and sulfuric acid (H2SO4, diluted 50 percent, 96 percent purity, Panreac AppliChem). Microplastics extraction: We adapted and further improved a methodology used for the extraction and identification of microplastics in penguin guano (Fragao et al., 2021), to enable the analyses of smaller microplastics (>25 micrometers). This utilized alkaline digestion, which has been proposed as an effective and cheaper option to enzymatic digestion, as it is also less damaging for plastic particles or fibres. An initial optimisation step was performed to determine the appropriate sample mass required to ensure complete digestion of the guano matrix. Different sample weights were tested, after which a standardised mass of approximately 1.5 g, was selected and used for subsequent analyses. The 32 freeze-dried samples were weighed (1-3 g), transferred into clean glass beakers with a 10 percent potassium hydroxide (KOH) solution, placed on a shaker for 72 h at room temperature. The volume of KOH added was 3 times the volume of biological material. After 72 h, the floating phase was separated from the settled part, by filtration onto separate 10 micrometer metal meshes using a glass filtration set-up (45 mm) with a vacuum pump. Due to the large amount of organic matter (mainly due to the presence of a large volume of krill cuticles) the filters from both the floating and settled parts were placed in clean beakers and a further 10 percent hydrogen peroxide (H2O2) solution was added for 24 h. The volume of the solution depended on the amount of sample, up to a maximum of 100 mL. The digested floating phase filters were then vacuum filtered on Anodiscs (Anopore Inorganic Membrane, 0.2 micrometers, 25 mm, Whatman), to allow a full scan of the filter surface for the analysis of the smaller plastics, (detection limit down to 11 micrometers), using a Focal Plane Array (FPA)-based imaging. The settled parts were vacuum filtered onto another 10 micrometer metal mesh. The meshes from the settled parts were systematically examined using a stereomicroscope LEICA M80 (Leica Microsystems GmbH), to identify potential microplastic particles and/or fibres candidates, which were then hand-picked using forceps and placed onto the Anodiscs for the further analysis. For the selection of possible microplastics, the criteria proposed by Hidalgo-Ruz et al. (2012) have been followed as guidelines (i.e. choosing fibers with equal thickness and homogeneous colors). Microplastics characterisation: A total of eleven digested floating phase Anodiscs, were randomly selected from the 32 digested samples, five of Chinstrap penguin guano and six of Gentoo penguin samples, and analysed for the identification of microplastic particles using FPA-based imaging. To identify the chemical composition of the particles/fibres, the Anodisc filters were analysed using the Agilent 670 micro-Fourier Transform Infrared Spectroscopy (u-FTIR, Agilent Techologies, Santa Clara, CA, USA). The spectrometer was coupled to an Agilent 620 microscope with an automated XYZ-stage and 128 x 128 focal plane array (FPA) detector, cooled with liquid nitrogen, in transmission mode. FTIR spectral analysis was conducted in the spectral range of 3600 - 1250 cm-1 (spectral resolution: 8 cm-1). Each filter was fully analysed by sequentially scanning one half at a time to ensure accurate FTIR measurement. Before each sample scan, a background scan was collected on a clean Anodiscs in the same spectral range. Spectra were further analysed via Purency Microplastics Finder (MPF) (Purency GmbH, Austria) running with machine learning model (by Hufnagl et al. 2022) which supports the identification of 21 polymers. This post-hoc spectral analysis in Purency used a relevance tolerance limit of 0.6 (the relevance value represents the degree of similarity between the measured FTIR spectrum of a particle and the reference spectra contained in the spectral library; higher relevance values indicate a better spectral match and therefore a higher confidence in the polymer identification) and spectral matches were filtered to match the lower size threshold of the analytical technique of 11 micrometers. |
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Locality
| Temporal Coverage: | |
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| Start Date | 2021-01-28 |
| End Date | 2022-03-01 |
| Spatial Coverage: | |
| Latitude | |
| Southernmost | -62.98333 |
| Northernmost | -62.65 |
| Longitude | |
| Westernmost | -60.73333 |
| Easternmost | -60.41667 |
| Altitude | |
| Min Altitude | N/A |
| Max Altitude | N/A |
| Depth | |
| Min Depth | N/A |
| Max Depth | N/A |
| Location: | |
| Location | Antarctica |
| Detailed Location | Livingstone Island, South Shetland Islands |
| Location | Antarctica |
| Detailed Location | Deception Island, South Shetland Islands |
Instrumentation
| Data Collection: | Biogeochemical particulate instrumentation: Elemental analyser (CHN, Exeter Analytical Inc. CE440 elemental analyser, accuracy +/- 0.15 percent). Microplastic data collection: All measurements were carried out using the Agilent 670 Fourier Transform Infrared (FTIR) spectrometer (Agilent Technologies, Santa Clara, CA), USA, with a cryogenically cooled mercury cadmium telluride (MCT) detector. The spectrometer was coupled to an Agilent 620 microscope with an automated XYZ-stage and 128 x 128 focal plane array (FPA) detector, cooled with liquid nitrogen. The FTIR system was continuously purged using a dry air generator (FGSR). The FTIR microscope was equipped with a 15 x IR objective lens and a 15-x visual objective lens. This stage held the sample in a bespoke filter holder enabling the transmission of infrared through the lower Cassegrain, the base of an Anodisc filter where the sample was held, and through the Barium fluoride slide of the same shape and diameter as the filter. This setup facilitated FTIR imaging of both fibres and particles (microplastics of all other morphologies) (Primpke et al., 2019, Roscher et al., 2021). Filters were scanned in halves, with each half measuring approximately 25mm x 14mm to allow a small overlap between scans when stitching together the data. The FPA detector enabled each of the halves to be scanned, acquiring a mosaic of spectra (Primpke et al.,2017, 2020a). This was calibrated with an XYZ stage allowing the exact coordinates at the end of the scan to be used to accurately identify the start of the next scanning area, covering the complete area of each filter (diameter of 25mm, area 625mm2). The collection of multiple mosaics meant each sample took approximately 13 scannable hours spread over 2.5 days. |
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Storage
| Data Storage: | 2x .csv files Biogeochemical.csv contains biogeochemical analysis of 32 penguin guano samples, measured in triplicate. Microplastics.csv contains data on microplastics detected in 11 of the guano samples. |
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