Deep-sea meiofaunal communities in the south-eastern Levantine basin and their shaping factors - Morphological-taxonomy-free metabarcoding approach.

The <3% dissimilar Amplicon Sequence Variant (ASV) clusters of the 18S-V4 barcode were used as species-proxies for the evaluation of ASV composition and ASV diversity indices characterizing the hitherto poorly investigated meiofaunal communities of the south-eastern part of the Levantine basin. Accompanied by abundance measurements, the relationships of these characteristics with sedimentary and bottom terrain parameters were interpreted. The construction of community composition profiles, namely ASVs' list and their estimated abundances, was done using our previously established procedure (Harbuzov et al., 2022, Marine Genomics 65, 100980), combining metabarcoding with sample reads normalization by the abundance of hard-bodied meiofaunal taxa. The study province included the 54-1418 m depth range, across vertical sub-bottom horizons ranging 0-17 cm. Oxygen, hydrogen sulfide and methane concentrations in the pore water, as well as sediment grain size spectra and sedimentary protein and carbohydrate levels, were measured, followed by an evaluation of their involvement in the shaping of the meiofaunal communities' characteristics. Community composition was generally site-and-horizon dependent and its abundance decreased with increasing bottom depth and across sub-bottom horizons, typical to benthic habitats which are nourished by organic carbon from the euphotic zone. The relatively sharply inclined continental slope bottom located in the northern part of the Israeli coast was an exception. Its meiofaunal community characteristics were speculated to be affected by intensive sediment mixing and lateral transport of food from the shelf, in addition to the effect of the euphotic zone-originated food sources.

A broad-taxa approach as an important concept in ecotoxicological studies and pollution monitoring.

Aquatic invertebrates play a pivotal role in (eco)toxicological assessments because they offer ethical, cost-effective and repeatable testing options. Additionally, their significance in the food chain and their ability to represent diverse aquatic ecosystems make them valuable subjects for (eco)toxicological studies. To ensure consistency and comparability across studies, international (eco)toxicology guidelines have been used to establish standardised methods and protocols for data collection, analysis and interpretation. However, the current standardised protocols primarily focus on a limited number of aquatic invertebrate species, mainly from Arthropoda, Mollusca and Annelida. These protocols are suitable for basic toxicity screening, effectively assessing the immediate and severe effects of toxic substances on organisms. For more comprehensive and ecologically relevant assessments, particularly those addressing long-term effects and ecosystem-wide impacts, we recommended the use of a broader diversity of species, since the present choice of taxa exacerbates the limited scope of basic ecotoxicological studies. This review provides a comprehensive overview of (eco)toxicological studies, focusing on major aquatic invertebrate taxa and how they are used to assess the impact of chemicals in diverse aquatic environments. The present work supports the use of a broad-taxa approach in basic environmental assessments, as it better represents the natural populations inhabiting various ecosystems. Advances in omics and other biochemical and computational techniques make the broad-taxa approach more feasible, enabling mechanistic studies on non-model organisms. By combining these approaches with in vitro techniques together with the broad-taxa approach, researchers can gain insights into less-explored impacts of pollution, such as changes in population diversity, the development of tolerance and transgenerational inheritance of pollution responses, the impact on organism phenotypic plasticity, biological invasion outcomes, social behaviour changes, metabolome changes, regeneration phenomena, disease susceptibility and tissue pathologies. This review also emphasises the need for harmonised data-reporting standards and minimum annotation checklists to ensure that research results are findable, accessible, interoperable and reusable (FAIR), maximising the use and reusability of data. The ultimate goal is to encourage integrated and holistic problem-focused collaboration between diverse scientific disciplines, international standardisation organisations and decision-making bodies, with a focus on transdisciplinary knowledge co-production for the One-Health approach.

Anthropogenic and natural disturbances along a river and its estuary alter the diversity of pathogens and antibiotic resistance mechanisms.

Antibiotic resistance (AMR) in pathogens threatens human health worldwide, and antibiotic-resistant bacteria (ARB) are widespread in the environment. In particular, anthropogenically-disturbed rivers became reservoirs of ARBs and hotspots of antibiotic resistance gene (ARG) transmission. However, the diversity and sources of ARB, and the mechanisms of ARG transmission are not fully known. Here, we used deep metagenomic sequencing to study the dynamics of pathogens and their antibiotic resistance mechanisms along the Alexander River (Israel), affected by sewage and animal farm runoffs. Putative pathogens such as Aeromicrobium marinum and Mycobacterium massilipolynesiensis were enriched in western stations, following the inputs of polluted Nablus River. Aeromonas veronii was dominant in eastern stations in Spring. Several AMR mechanisms showed distinct patterns in Summer-Spring (dry season) and Winter (rainy season). We found low abundance beta-lactamases conferring carbapenem resistance: e.g., OXA-912 was linked to A. veronii in Spring; OXA-119 and OXA-205 to Xanthomonadaceae in Winter. We classified 33 % of ARG-containing contigs as putative plasmid sequences, indicating the high potential for resistome transmission. A limited number of ARGs were linked to putative phages. Our results suggest that this model river is a hotspot for AMR activity and transmission, and highlight the merit of deep sequencing for AMR discovery.

Amplicon sequence variant-based meiofaunal community composition revealed by DADA2 tool is compatible with species composition.

The present study is aimed at implementing the morphological identification-free amplicon sequence variant (ASV) concept for describing meiofaunal species composition, while strongly indicating reasonable compatibility with the underlying species. A primer pair was constructed and demonstrated to PCR amplify a 470-490 bp 18S barcode from a variety of meiofaunal taxa, high throughput sequenced using the Illumina 300 × 2 bps platform. Sixteen 18S multi-species HTS assemblies were created from meiofaunal samples and merged to one assembly of ~2,150,000 reads. Five quality scores (q = 35, 30, 25, 20, 15) were implemented to filter five 18S barcode assemblies, which served as inputs for the DADA2 software, ending with five reference ASV libraries. Each of these libraries was clustered, applying 3% dissimilarity threshold, revealed an average number of 1.38 ±â€¯0.078 ASVs / cluster. Hence, demonstrating high level of ASV uniqueness. The libraries which were based on q ≤ 25 reached a near-asymptote number of ASVs which together with the low average number of ASVs / cluster, strongly indicated fair representation of the actual number of the underlying species. Hence, the q = 25 library was selected to be used as metabarcoding reference library. It contained 461 ASVs and 342-3% clusters with average number of 1.34 ±â€¯1.036 ASV / cluster and their BLASTN annotation elucidated a variety of expected meiofaunal taxa. The sixteen assemblies of sample-specific paired reads were mapped to this reference library and sample ASV profiles, namely the list of ASVs and their proportional copy numbers were created and clustered.

Developing novel microsatellite markers by NGS technology for Rhopilema nomadica, an invasive jellyfish.

Twelve microsatellite loci, obtained by whole genome sequencing approach, were developed and validated for the rhizostomatid jellyfish Rhopilema nomadica, the most pernicious invasive species in the Mediterranean Sea. A sample of 40 specimens collected at six locations along the Mediterranean coast of Israel were genotyped and all loci presented suitable outcomes to population genetic studies, revealing 5-19 alleles/locus with clean and reproducible amplifications. Observed and expected heterozygosity ranged 0.0.353 to 0.971 and 0.335 to 0.870, respectively, and the fixation index (inbreeding coefficient) and the polymorphic information content (PIC) ranged between - 0.190 and 0.240 and 0.32 to 0.858, respectively. The new set of microsatellite loci will be used to study long-term changes in the population genetic parameters of this invasive species.