The sea surface microlayer (SML) of marine environments is hostile, exposing bacteria to extreme conditions such as temperatures and salinity fluctuations, solar radiation, and the presence of toxic substances such as heavy metals, microplastics, and pharmaceutical compounds. Our study focused on investigating the ecotoxicological effects of varying ciprofloxacin concentrations (0, 10, 50, and 100 ng/ml) on bacterial abundance and enrichment in natural SML and underlying water (ULW) samples obtained from the southern North Sea. In addition, we analyzed the samples for the presence of ciprofloxacin and 25 other antibiotics, including lincomycin, clindamycin, clarithromycin, erythromycin, azithromycin, ofloxacin and novobiocin. Furthermore, we carried out antibiotic susceptibility tests on marine bacterial isolates cultivated in the presence of 100 ng mL ⁻¹, utilizing antibiotics commonly found in their natural habitats. We observed a decrease in bacterial abundance and enrichment in the SML and ULW when exposed to 50 and 100 ng/ml of ciprofloxacin. However, over time, the abundance and enrichment of bacteria increased at these concentrations, indicating resistance. The presence of 100 ng/ml of ciprofloxacin also exerted selective pressure on bacterial members in the SML and ULW, and we cultivated 42 marine and 55 nonmarine bacteria in the presence of 100 ng/ml of ciprofloxacin. Although ciprofloxacin was not detected, we found 11 other antibiotics, particularly in the SML. When we exposed marine bacteria to antibiotics such as novobiocin, ofloxacin, clarithromycin, erythromycin, and clindamycin found in the environment, we observed that some marine bacteria are resistant to these antibiotics. Our findings suggest that resistance in marine bacteria can be acquired through exposure to antibiotics released into coastal water bodies.
Due to its location at the air-sea interface, the sea surface microlayer is prone to accumulating anthropogenic pollutants. We analyzed the presence of antibiotic compounds, including ciprofloxacin, in this area using ultra-performance liquid chromatography—triple quadrupole mass spectrometry in tandem with ionization spray (UPLC-QqQMS/MS) (Bruker EVOQ Elite system, Billerica MA). We compared the numbers and concentrations of these compounds in the microlayer to those found in the underlying water column.
The effects of a phytoplankton bloom and photobleaching on colored dissolved organic matter (CDOM) in the sea-surface microlayer (SML) and the underlying water (ULW) were studied in a month-long mesocosm study, in May and June of 2023, at the Institute for Chemistry and Biology of the Marine Environment (ICBM) in Wilhelmshaven, Germany. The mesocosm study was conducted by the DFG research group BASS (Biogeochemical processes and Air–sea exchange in the Sea-Surface microlayer, Bibi et al., 2025) in the Sea Surface Facility (SURF) of the ICBM. The facility contains an 8 m × 1.5 m × 0.8 m large outdoor basin with a retractable roof, which was closed at night and during rain events. The basin was filled with North Sea water from the adjacent Jade Bay. Homogeneity of the ULW in the basin was achieved by constant mixing of the water column. The daily SML and ULW samples were collected alternating in the morning, about 1 h after sunrise, and in the afternoon, about 10 h after sunrise. The alternation of sampling times intended to capture a potential effect of sun-exposure duration on DOM transformations and elucidated the day and night variability of the layers. The SML was collected via glass plate sampling (Cunliffe and Wurl, 2014). The ULW was sampled via a submerged tube and a connected syringe suction system in 0.4 m depth. The removed sample volume was refilled with Jade Bay water every day. SML and ULW samples were filtered through pre-flushed 0.7 µm Whatman GF/F and 0.2 nucleopore filters into brown bottles and were stored dark and at 4 °C until measurement within weeks of the study. The brown bottles were previously combusted at 500 °C. CDOM was measured with three liquid waveguide capillary cells (LWCC, WPI, USA) of different pathlengths (10 cm, 50 cm, 250 cm) to increase the measurement sensitivity following the protocols of Röttgers et al. (2024) using a spectral detector (Avantes, Netherlands) for a total spectral range from 230 to 750 nm. A sodium chloride (NaCl) solution was used for the salinity correction. The blank-corrected absorbance spectra were then converted into Napierian absorption coefficients (Bricaud et al., 1981).
The effects of a phytoplankton bloom and photobleaching on colored dissolved organic matter (CDOM) in the sea-surface microlayer (SML) and the underlying water (ULW) were studied in a month-long mesocosm study, in May and June of 2023, at the Institute for Chemistry and Biology of the Marine Environment (ICBM) in Wilhelmshaven, Germany. The mesocosm study was conducted by the DFG research group BASS (Biogeochemical processes and Air–sea exchange in the Sea-Surface microlayer, Bibi et al., 2025) in the Sea Surface Facility (SURF) of the ICBM. The facility contains an 8 m × 1.5 m × 0.8 m large outdoor basin with a retractable roof, which was closed at night and during rain events. The basin was filled with North Sea water from the adjacent Jade Bay. Homogeneity of the ULW in the basin was achieved by constant mixing of the water column. The daily SML and ULW samples were collected alternating in the morning, about 1 h after sunrise, and in the afternoon, about 10 h after sunrise. The alternation of sampling times intended to capture a potential effect of sun-exposure duration on DOM transformations and elucidated the day and night variability of the layers. The SML was collected via glass plate sampling (Cunliffe and Wurl, 2014). The ULW was sampled via a submerged tube and a connected syringe suction system in 0.4 m depth. The removed sample volume was refilled with Jade Bay water every day. SML and ULW samples were filtered through pre-flushed 0.7 µm Whatman GF/F and 0.2 nucleopore filters into clear 40 ml SUPELCO bottles. These bottles were acid-washed twice and combusted at 500 °C for 5 h. The samples were stored dark and at 4 °C and measured within a few months of the study. FDOM was measured using a Aqualog fluorescence spectrometer (Horiba Scientific, Japan) with 10 seconds integration time and high gain of the CCD (charge-coupled device) sensor within an excitation range from 240 to 500 nm, and an emission range from 209.15 to 618.53 nm. The Aqualog measures fluorescence as well as absorption. The resulting data includes an excitation-emission-matrix (EEM) of the blank (MilliQ Starna cuvette), an EEM of the sample, and the absorption values of the sample. The raw exported Aqualog data was corrected for errors and lamp shifts. The corrected EEM data is then decomposed by PARAFAC (Murphy et al., 2013) for its underlying fluorophore components. Before running the PARAFAC routine, the corrected data needed to undergo a correction process by subtracting the blank from the sample EEM and canceling the influences of the inner-filter effect (IFE, Parker & Rees, 1962; Kothawala et al., 2013). The fluorescence intensity of the IFE-corrected EEM is calibrated by using the Raman scatter peak of water (Lawaetz & Stedmon, 2009). For PARAFAC the corrected data was processed using the drEEM and NWAY toolbox (version 0.6.5; Murphy et al., 2013) in MATLAB (R2020b). A 4-component model was validated with the validation style S4C6T3 for the split half analysis with nonnegativity constraints and 1-8e as the convergence criteria with 50 random starts and a maximum number of 2500 iterations. The resulting final model had a core consistency of 82.04 and the explained percentage was 99.54%. Furthermore, four fluorescence indices were calculated from the corrected EEM data (HIX – Humification index, Zsolnay et al., 1999; BIX – Biological index, Huguet et al., 2009; REPIX – Recently produced index, Parlanti et al., 2000, Drozdowska et al., 2015; ARIX, Murphy, 2025).