ALBERT

Description: An increase in human Vibrio spp. infections has been linked to climate change related events, in particular to seawater warming and heatwaves. However, there is a distinct lack of research of pathogenic Vibrio spp. occurrences in the temperate North Sea, one of the fastest warming seas globally. Particularly in the German Bight, Vibrio investigations are still scarce. This study focuses on the spatio-temporal quantification and pathogenic characterization of V. parahaemolyticus, V. vulnificus and V. cholerae over the course of 14 months. Species-specific MPN-PCR (Most probable number – polymerase chain reaction) conducted on selectively enriched surface water samples revealed seasonal patterns of all three species with increased abundances during summer months. The extended period of warm seawater coincided with prolonged Vibrio spp. occurrences in the German Bight. Temperature and nitrite were the factors explaining variations in Vibrio spp. abundances after generalized additive mixed models. The specific detection of pathogenic markers via PCR revealed trh-positive V. parahaemolyticus, pathogenic V. vulnificus (nanA, manIIA, PRXII) and V. cholerae serotype O139 presence. Additionally, spatio-temporally varying virulence profiles of V. cholerae with multiple accessory virulence-associated genes, such as the El Tor variant hemolysin (hlyAET), acyltransferase of the repeats-in-toxin cluster (rtxC), Vibrio 7th pandemic island II (VSP-II), Type III Secretion System (TTSS) and the Cholix Toxin (chxA) were detected. Overall, this study highlights that environmental human pathogenic Vibrio spp. comprise a reservoir of virulence-associated genes in the German Bight, especially in estuarine regions. Due to their known vast genetic plasticity, we point to the possible emergence of highly pathogenic V. cholerae strains. Particularly, the presence of V. cholerae serotype O139 is unusual and needs urgent continuous surveillance. Given the predictions of further warming and more frequent heatwave events, human pathogenic Vibrio spp. should be seriously considered as a developing risk to human health in the German Bight.

Description: Protoceratium reticulatum as a producer of yessotoxin (YTX) and its analogues is common in several coastal environments. The YTX-producing strain of P. reticulatum, isolated from the German Bight (North Sea), was analysed to study toxin production under various autecological conditions. Experiments were carried out to investigate the influence of N/P ratio (2.44 (1/10 N), 24.36 (f/2) and 243.65 (1/10 P)), temperature (15 and 20 °C), salinity (5, 10, 15, 20, 25 and 30) and growth phase on YTX formation, cell size and chlorophyll a concentration. P. reticulatum showed the highest growth at 15 °C and higher salinities (25 and 30). In particular, higher temperature led to a reduced growth. The total YTX concentrations were higher at lower temperature. However, a clear correlation between salinity and YTX production was not observed at lower temperature. Furthermore, 1/10 P and f/2 cultures exhibited the highest cell quota of YTX at the end of the stationary phase; a dramatic effect occurred at 15 °C in 1/10 P media, when the toxicity increased to ten fold higher values. Slight variations of the composition of the YTX analogues under nutrient limitation were observable. In addition, the results indicate that N-limitation cause a lower cell size, whereas P-limitation leads to a higher cell size; an influence of the salinity on cell size was also observable.

Description: Microplastics (MP) have been recorded in various environments around the globe. For a better understanding of distribution patterns and for providing a basis for risk assessments, detailed data on MP concentrations and polymer compositions are required. This study investigated the effluents of two German wastewater treatment plants (WWTP) monthly over one year, in order to better understand their temporal input of MP into the receiving river systems. MP item data down to 11 μm were obtained by means of Fourier Transform Infrared (FTIR) spectroscopy under the application of an improved polymer database. Complementary mass data were obtained by pyrolysis gas chromatography–mass spectrometry (Py-GC/MS) (for one WWTP). Both FTIR and Py-GC/MS analysis revealed a homogeneous polymer composition over the year, with a general dominance of polyolefins. Elevated MP item and mass concentrations (maximum: 3 × 104 items m−3 and 3.8 × 103 μg m−3) were observed during winter months and were accompanied by either heavy rainfall (increased discharge and total organic carbon) or elevated turbidity values. These observations emphasize the need for the assessment of background parameters in future MP monitoring studies. By providing monthly data over one year on MP items and masses in WWTP effluents, this study helps enhancing the understanding of temporal MP dynamics and can act as a valuable reference point for future assessments.

Description: The prevalence of multidrug-resistant Gram-negative bacteria in aquatic environments has been a long withstanding health concern, namely extended-spectrumbeta-lactamase (ESBL) producing Escherichia coli. Given increasing reports on microplastic (MP) pollution in these environments, it has become crucial to better understand the role of MP particles as transport vectors for such multidrug-resistant bacteria. In this study, an incubation experiment was designed where particles of both synthetic and natural material (HDPE, tyre wear, and wood) were sequentially incubated atmultiple sites along a salinity gradient from the Lower Weser estuary (Germany) to the offshore island Helgoland (German Bight, North Sea). Following each incubation period, particle biofilms andwater samples were assessed for ESBL-producing E. coli, first by the enrichment and detection of E. coli using Fluorocult® LMX Broth followed by cultivation on CHROMAgar™ ESBL media to select for ESBLproducers. Results showed that general E. coli populations were present on the surfaces of wood particles across all sites but nonewere found to produce ESBLs. Additionally, neither HDPE nor tyrewear particles were found to harbour any E. coli. Conversely, ESBL-producing E. coli were present in surrounding waters from all sites, 64% of which conferred resistances against up to 3 other antibiotic groups, additional to the beta-lactam resistances intrinsic to ESBL-producers. This study provides a first look into the potential of MP to harbour and transport multidrug-resistant E. coli across different environments and the approach serves as an important precursor to further studies on other potentially harmful MP-colonizing species.