High Altitude Lake and Hydrochemistry: A Study of Lam Dal and Six Consecutive Lakes of Dhauladhar, Himachal Himalaya, India

Increased human influence has warmed the atmosphere, ocean and land thereby resulting in widespread and rapid changes in the atmosphere, ocean, cryosphere and biosphere (IPCC, 2021). High altitude lakes are generally small and quite sensitive to natural and anthropogenic perturbations. The present work is a preliminary work to investigate different hydro chemical processes and factors that controls the geochemistry of a high altitude lake, Lam Lake (dal) and its consecutive six lakes flowing through the Chamba district, Himachal Pradesh. Two hundred and eighty (n=280) water samples were collected in the year 2017 during the pre-monsoon and post-monsoon season. The anion concentration for all the seven lakes followed the trend HCO3- > NO3- > Cl- > SO42- > PO43- whereas the order of cation concentrations was Ca2+> Mg2+> Na+> K+ for both the seasons. Less EC of the water samples shows its serene nature. Components of bicarbonate (HCO3-) were found to be the major anion whereas calcium (Ca2+) was found to be the major cation present in the lakes. Piper Plot and Durov plot indicated Ca2+ – HCO3- as the major hydrogeochemical facies with comparatively less contribution from Mg2+–HCO3- type. The dominance of Ca2+ – HCO3- over Mg2+– HCO3- reflects the possibility of the natural order of dominance in the geology of the catchment area. The low Na+ + K+/TZ+ (total cations) ratios and the high Ca2+ + Mg2+/TZ+ (total cations) and (Ca2+ + Mg2+)/(Na+ + K+) ratios showed dominance of carbonate weathering. The average carbon ratios during pre-monsoon and post-monsoon were found to be 0.97 and 0.98, respectively, suggesting that proton is primarily derived from the oxidation of sulphide involving carbonate dissolution. The baseline data generated for a high-altitude lake shows that weathering and erosion during monsoonal precipitation and snow melt runoff during ablation season are the main sources of the chemical composition of lake water. Further to trace the imprints of climate change and seasonal variations in the high-altitude lakes, long term monitoring is recommended along with isotopic tracer techniques.

Integrating multiple chemical tracers to elucidate the diet and habitat of Cookiecutter Sharks

The Cookiecutter shark (Isistius brasiliensis) is an ectoparasitic, mesopelagic shark that is known for removing plugs of tissue from larger prey, including teleosts, chondrichthyans, cephalopods, and marine mammals. Although this species is widely distributed throughout the world’s tropical and subtropical oceanic waters, like many deep-water species, it remains very poorly understood due to its mesopelagic distribution. We used a suite of biochemical tracers, including stable isotope analysis (SIA), fatty acid analysis (FAA), and environmental DNA (eDNA), to investigate the trophic ecology of this species in the Central Pacific around Hawaii. We found that large epipelagic prey constituted a relatively minor part of the overall diet. Surprisingly, small micronektonic and forage species (meso- and epipelagic) are the most important prey group for Cookiecutter sharks across the studied size range (17–43 cm total length), with larger mesopelagic species or species that exhibit diel vertical migration also being important prey. These results were consistent across all the tracer techniques employed. Our results indicate that Cookiecutter sharks play a unique role in pelagic food webs, feeding on prey ranging from the largest apex predators to small, low trophic level species, in particular those that overlap with the depth distribution of the sharks throughout the diel cycle. We also found evidence of a potential shift in diet and/or habitat with size and season. Environmental DNA metabarcoding revealed new prey items for Cookiecutter sharks while also demonstrating that eDNA can be used to identify recent prey in stomachs frozen for extended periods. Integrating across chemical tracers is a powerful tool for investigating the ecology of elusive and difficult to study species, such as meso- and bathypelagic chondrichthyans, and can increase the amount of information gained from small sample sizes. Better resolving the foraging ecology of these mesopelagic predators is critical for effective conservation and management of these taxa and ecosystems, which are intrinsically vulnerable to overfishing and exploitation.

Magnetic nanoparticle technique versus radioisotope technique in detection of sentinel lymph node in early breast cancer: a systematic review and meta-analysis

Sentinel lymph node biopsy is the standard technique to stage the axilla in early breast cancer. The gold standard is the dual technique of radioisotope and blue dye injection. The drawbacks of dual technique include handling and disposal of radioactive material. Equally reliable, user and patient friendly, magnetic tracer super paramagnetic iron oxide, was compared with the radioisotope tracer in detection of Sentinel lymph node in a clinically node negative axilla in early breast cancer in this meta-analysis, with emphasis on the utility and safety of both techniques. PubMed, Medline were searched from April 2015 to October 2019. 1395 patients’ data was included from seven homogenous studies in this meta-analysis. A statistical analysis was performed using STATA 16.1 version for sentinel lymph node detection rate using standard technique, magnetic tracer technique and both techniques. Ratio of successes and failures of the outcomes was measured and analysed. A paired two sample ‘Z’ test was performed to compare between the standard technique and magnetic tracer techniques. The standardised mean difference of ratio of success rate between two techniques was; 1.013334 with a p value of 0.3136.The standardised mean difference of ratio of failure rate between two techniques was 1.016667, with a p value of 0.3132. Success and failure rate showed statistically no significant difference between the two techniques. This two-way analysis with paired two sample ‘Z’ test confirms that neither standard technique nor magnetic tracer techniques are superior or inferior to each other.

CARD-FISH in the Sequencing Era: Opening a New Universe of Protistan Ecology

Phagotrophic protists are key players in aquatic food webs. Although sequencing-based studies have revealed their enormous diversity, ecological information on in situ abundance, feeding modes, grazing preferences, and growth rates of specific lineages can be reliably obtained only using microscopy-based molecular methods, such as Catalyzed Reporter Deposition-Fluorescence in situ Hybridization (CARD-FISH). CARD-FISH is commonly applied to study prokaryotes, but less so to microbial eukaryotes. Application of this technique revealed that Paraphysomonas or Spumella-like chrysophytes, considered to be among the most prominent members of protistan communities in pelagic environments, are omnipresent but actually less abundant than expected, in contrast to little known groups such as heterotrophic cryptophyte lineages (e.g., CRY1), cercozoans, katablepharids, or the MAST lineages. Combination of CARD-FISH with tracer techniques and application of double CARD-FISH allow visualization of food vacuole contents of specific flagellate groups, thus considerably challenging our current, simplistic view that they are predominantly bacterivores. Experimental manipulations with natural communities revealed that larger flagellates are actually omnivores ingesting both prokaryotes and other protists. These new findings justify our proposition of an updated model of microbial food webs in pelagic environments, reflecting more authentically the complex trophic interactions and specific roles of flagellated protists, with inclusion of at least two additional trophic levels in the nanoplankton size fraction. Moreover, we provide a detailed CARD-FISH protocol for protists, exemplified on mixo- and heterotrophic nanoplanktonic flagellates, together with tips on probe design, a troubleshooting guide addressing most frequent obstacles, and an exhaustive list of published probes targeting protists.

Effects of warming and oxalic acid addition on plant–microbial competition in Picea brachytyla

The importance of oxalic acid for tree seedling growth and the competition for inorganic nitrogen (N) by plants and soil microorganisms under warming was investigated using 15N tracer techniques in Picea brachytyla (Franch.) E. Pritz. Results showed that warming combined with oxalic acid application induced growth enhancements in seedlings primarily through increases in fine root length and fine root surface area. Moreover, soil NH4 +, NO3 –, PO4 3–, N mineralization, microbial biomass carbon (MBC), and microbial biomass nitrogen (MBN) were significantly higher or tended to be higher with oxalic acid application. However, warming with oxalic acid application altered the partitioning of N between plants and soil microorganisms by increasing microbial 15N recovery to a lesser extent than it increased plant 15N recovery. While plants showed no specific preferences between N forms under normal conditions or warming alone, under warming and oxalic acid, plants showed a preference for 15NO3. Microorganisms showed a stronger preference for 15NH4 especially under warming and oxalic acid treatments. These findings suggest that plasticity in resource use could be an important mechanism in alleviating competition for soil N between plants and microbes under warming and oxalic acid addition.

Subcellular view of host–microbiome nutrient exchange in sponges: insights into the ecological success of an early metazoan–microbe symbiosis

Background Sponges are increasingly recognised as key ecosystem engineers in many aquatic habitats. They play an important role in nutrient cycling due to their unrivalled capacity for processing both dissolved and particulate organic matter (DOM and POM) and the exceptional metabolic repertoire of their diverse and abundant microbial communities. Functional studies determining the role of host and microbiome in organic nutrient uptake and exchange, however, are limited. Therefore, we coupled pulse-chase isotopic tracer techniques with nanoscale secondary ion mass spectrometry (NanoSIMS) to visualise the uptake and translocation of 13C- and 15N-labelled dissolved and particulate organic food at subcellular level in the high microbial abundance sponge Plakortis angulospiculatus and the low microbial abundance sponge Halisarca caerulea. Results The two sponge species showed significant enrichment of DOM- and POM-derived 13C and 15N into their tissue over time. Microbial symbionts were actively involved in the assimilation of DOM, but host filtering cells (choanocytes) appeared to be the primary site of DOM and POM uptake in both sponge species overall, via pinocytosis and phagocytosis, respectively. Translocation of carbon and nitrogen from choanocytes to microbial symbionts occurred over time, irrespective of microbial abundance, reflecting recycling of host waste products by the microbiome. Conclusions Here, we provide empirical evidence indicating that the prokaryotic communities of a high and a low microbial abundance sponge obtain nutritional benefits from their host-associated lifestyle. The metabolic interaction between the highly efficient filter-feeding host and its microbial symbionts likely provides a competitive advantage to the sponge holobiont in the oligotrophic environments in which they thrive, by retaining and recycling limiting nutrients. Sponges present a unique model to link nutritional symbiotic interactions to holobiont function, and, via cascading effects, ecosystem functioning, in one of the earliest metazoan–microbe symbioses.

Quantifying Potential N Turnover Rates in Hypersaline Microbial Mats by Using 15N Tracer Techniques

ABSTRACT Microbial mats, due to stratification of the redox zones, have the potential to include a complete N cycle; however, an attempt to evaluate a complete N cycle in these ecosystems has not been yet made. In this study, the occurrence and rates of major N cycle processes were evaluated in intact microbial mats from Elkhorn Slough, Monterey Bay, CA, USA, and Baja California Sur, Mexico, under oxic and anoxic conditions using 15N-labeling techniques. All the major N transformation pathways, with the exception of anammox, were detected in both microbial mats. Nitrification rates were found to be low at both sites for both seasons investigated. The highest rates of ammonium assimilation were measured in Elkhorn Slough mats in April and corresponded to high in situ ammonium concentrations in the overlying water. Baja mats featured higher ammonification than ammonium assimilation rates, and this, along with their higher affinity for nitrate compared to ammonium and low dissimilatory nitrate reduction to ammonium rates, characterized their differences from Elkhorn Slough mats. Nitrogen fixation rates in Elkhorn Slough microbial mats were found to be low, implying that other processes, such as recycling and assimilation from water, are the main sources of N for these mats at the times sampled. Denitrification in all the mats was incomplete, with nitrous oxide as the end product and not dinitrogen. Our findings highlight N cycling features not previously quantified in microbial mats and indicate a need for further investigations of these microbial ecosystems. IMPORTANCE Nitrogen is essential for life. The nitrogen cycle on Earth is mediated by microbial activity and has had a profound impact on both the atmosphere and the biosphere throughout geologic time. Microbial mats, present in many modern environments, have been regarded as living records of the organisms, genes, and phylogenies of microbes, as they are one of the most ancient ecosystems on Earth. While rates of major nitrogen metabolic pathways have been evaluated in a number of ecosystems, they remain elusive in microbial mats. In particular, it is unclear what factors affect nitrogen cycling in these ecosystems and how morphological differences between mats impact nitrogen transformations. In this study, we investigate nitrogen cycling in two microbial mats having morphological differences. Our findings provide insight for further understanding of biogeochemistry and microbial ecology of microbial mats.

Optimizing Dose and Timing in Magnetic Tracer Techniques for Sentinel Lymph Node Detection in Early Breast Cancers: The Prospective Multicenter SentiDose Trial

Superparamagnetic iron oxide nanoparticles (SPIO) are non-inferior to radioisotope and blue dye (RI + BD) for sentinel lymph node (SLN) detection. Previously, 2 mL SPIO (Sienna+®) in 3 mL NaCl was used. In this dose-optimizing study, lower doses of a new refined SPIO solution (Magtrace®) (1.5 vs. 1.0 mL) were tested in different timeframes (0–24 h perioperative vs. 1–7 days preoperative) and injections sites (subareolar vs. peritumoral). Two consecutive breast cancer cohorts (n = 328) scheduled for SLN-biopsy were included from 2017 to 2019. All patients received isotope ± blue dye as back-up. SLNs were identified primarily with the SentiMag® probe and thereafter a gamma-probe. The primary endpoint was SLN detection rate with SPIO. Analyses were performed as a one-step individual patient-level meta-analysis using patient-level data from the previously published Nordic Trial (n = 206) as a third, reference cohort. In 534 patients, the SPIO SLN detection rates were similar (97.5% vs. 100% vs. 97.6%, p = 0.11) and non-inferior to the dual technique. Significantly more SLNs were retrieved in the preoperative 1.0 mL cohort compared with 1.5 and the 2.0 mL cohorts (2.18 vs. 1.85 vs. 1.83, p = 0.003). Lower SPIO volumes injected up to 7 days before the operation have comparable efficacy to standard SPIO dose and RI + BD for SLN detection.