Using Compound-Specific and Bulk Stable Isotope Analysis for Trophic Positioning of Bivalves in Contaminated Baltic Sea Sediments.

Stable nitrogen isotopes (δ15N) are used as indicators of trophic position (TP) of consumers. Deriving TP from δ15N of individual amino acids (AAs) is becoming popular in ecological studies, because of lower uncertainty than TP based on bulk δ15N (TPbulk). This method would also facilitate biomagnification studies provided that isotope fractionation is unaffected by toxic exposure. We compared TPAA and TPbulk estimates for a sediment-dwelling bivalve from two coastal sites, a pristine and a contaminated. Chemical analysis of PCB levels in mussels, sediments, and pore water confirmed the expected difference between sites. Both methods, but in particular the TPAA underestimated the actual TP of bivalves. Using error propagation, the total uncertainty related to the analytical precision and assumptions in the TP calculations was found to be similar between the two methods. Interestingly, the significantly higher intercept for the regression between TPAA and TPbulk in the contaminated site compared to the pristine site indicates a higher deamination rate due to detoxification as a result of chronic exposure and a higher 15N fractionation. Hence, there is a need for controlled experiments on assumptions underlying amino acid-specific stable isotope methods in food web and bimagnification studies.

Growth Retardation and Altered Isotope Composition As Delayed Effects of PCB Exposure in Daphnia magna.

Trophic magnification factor (TMF) analysis employs stable isotope signatures to derive biomagnification potential for environmental contaminants. This approach relies on species δ(15)N values aligning with their trophic position (TP). This, however, may not always be true, because toxic exposure can alter growth and isotope allocation patterns. Here, effects of PCB exposure (mixture of PCB18, PCB40, PCB128, and PCB209) on δ(15)N and δ(13)C as well as processes driving these effects were explored using the cladoceran Daphnia magna. A two-part experiment assessed effects of toxic exposure during and after exposure; juvenile daphnids were exposed during 3 days (accumulation phase) and then allowed to depurate for 4 days (depuration phase). No effects on survival, growth, carbon and nitrogen content, and stable isotope composition were observed after the accumulation phase, whereas significant changes were detected in adults after the depuration phase. In particular, a significantly lower nitrogen content and a growth inhibition were observed, with a concomitant increase in δ(15)N (+0.1 ‰) and decrease in δ(13)C (-0.1 ‰). Although of low magnitude, these changes followed the predicted direction indicating that sublethal effects of contaminant exposure can lead to overestimation of TP and hence underestimated TMF.

Stable isotope composition in Daphnia is modulated by growth, temperature, and toxic exposure: implications for trophic magnification factor assessment.

The potential for using stable isotope analysis in risk assessment of environmental contaminants is crucially dependent on the predictability of the trophic transfer of isotopes in food webs. The relationship between contaminant levels and trophic position of consumers is widely used to assess biomagnification properties of various pollutants by establishing trophic magnification factors (TMF). However, contaminant-induced variability of the isotopic composition in biota is poorly understood. Here, we investigated effects of toxic exposure on δ(15)N and δ(13)C values in a consumer, with a main hypothesis that these effects would be largely mediated via growth rate and metabolic turnover of the test animals. The cladoceran Daphnia magna was used in two experiments that were conducted to manipulate growth and body condition (assayed as C:N ratio) by food availability and temperature (Experiment 1) and by toxic exposure to the pesticide lindane (Experiment 2). We found a significant negative effect of growth rate and a positive effect of temperature on the consumer-diet discrimination factor for δ(15)N and δ(13)C, with no effects on the C:N ratio (Experiment 1). In lindane-exposed daphnids, a significant growth inhibition was observed, with concomitant increase in metabolic costs and significantly elevated size-specific δ(15)N and δ(13)C values. Moreover, a significantly higher incorporation of carbon relative to nitrogen, yet a concomitant decrease in C:N ratio was observed in the exposed animals. Together, these results have methodological implications for determining trophic positions and TMF in polluted environments, where elevated δ(15)N values would translate into overestimated trophic positions and underestimated TMF. Furthermore, altered δ(13)C values may lead to erroneous food-chain assignment of the consumer in question.

Does pre-exposure to polluted sediment affect sub-cellular to population-level responses to contaminant exposure in a sentinel species?

Understanding how key-species respond to anthropogenic stress such as chemical pollution is critical for predicting ecosystem changes. Little is however known about the intra-specific variability in the physiological and biochemical traits involved in contaminant exposure responses. Here, we explored this idea by exposing the Baltic amphipod Monoporeia affinis from two sites, one moderately polluted and one more pristine, to a sediment spiked with PAHs and PCBs. We evaluated the amphipods responses related to feeding, growth, a stress biomarker (acetylcholinesterase [AChE] inhibition) and stable isotope (δ13C and δ15N) composition including isotope niche analyses. More adverse responses were expected in animals from the low-pollution site than those from the high-pollution site due to tolerance development in the latter. Amphipods from both populations showed a ∼30% AChE inhibition when exposed to the contaminant spiked sediment. However, both controls and exposed amphipods from the high-pollution site had higher survival, nutrient uptake and condition status than the amphipods from the low-pollution site, which did not feed on the added diatoms as indicated by their isotope values. We found no signs of population-specific responses in physiological adjustments to contaminants with regard to classic ecotoxicological biomarkers such as AChE inhibition and growth status. Instead, isotope niche analyses proved useful in assessing contaminant stress responses at the population level.

Resource partitioning of a Mexican clam in species-poor Baltic Sea sediments indicates the existence of a vacant trophic niche.

Invasive species are often generalists that can take advantage of formerly unexploited resources. The existence of such vacant niches is more likely in species-poor systems like the Baltic Sea. The suspension feeding wedge clam, Rangia cuneata, native to estuarine environments in the Gulf of Mexico, was sighted for the first time in the southeastern Baltic in 2010 and a few years later in the northern Baltic along the Swedish coast. To explore possible competition for food resources between R. cuneata and the three native clams inhabiting Baltic shallow soft bottoms, stable isotope and fatty acid analyses were conducted. There was no overlap between R. cuneata and any of the native species in either stable isotope or fatty acid niches. This suggests efficient partitioning of resources; multivariate analyses indicate that separation was driven mainly by δ13C and by fatty acids reflecting diatoms and cyanobacteria, respectively (e.g. 16:1ω7 and 18:3ω3). R. cuneata reflected seasonal variation in phytoplankton more than other clams reflecting higher trophic plasticity. In conclusion, the addition of R. cuneata to the Baltic shallow soft bottoms suggests the existence of a vacant trophic niche in these sediment habitats, however the long-term effects on other species and nutrient cycling requires further studies focusing on the population dynamics of R. cuneata and its impact on the Baltic Sea ecosystem.

Shrews Under-Represented in Fruit Farms and Homesteads.

Shrews are a less studied group of small mammals than rodents. Between 2018 and 2022, we surveyed 23 sites in Lithuania, including natural and anthropogenic habitats, with the aim to assess the proportion of Soricidae in small mammal communities and their diet based on stable isotope analysis. The average representation of Soricidae was 3.1%, about half the long-term average in other habitats in the country. The highest proportions were in meadows and farmsteads, at 4.9% and 5.0% respectively. Shrews were not trapped on farms or in young orchards, and their relative abundance was very low in intensively managed orchards (0.006 individuals per 100 trap days). Neomys fodiens and N. anomalus were unexpectedly found in homesteads, including in outbuildings. Sorex araneus and S. minutus had similar diets. The trophic carbon/nitrogen discrimination factor between invertebrates and shrew hair was 2.74‱/3.98‱ for S. araneus, 1.90‱/3.78‱ for S. minutus in the orchards. The diet of N. fodiens and N. anomalus at the homesteads requires further investigation. We propose that the under-abundance of shrews may be due to contamination by plant protection products and a lack of invertebrates under intensive agricultural practices.

Horses in Lithuania in the Late Roman-Medieval Period (3rd-14th C AD) Burial Sites: Updates on Size, Age and Dating.

The tradition of burying horses in Lithuania lasted from the Early Roman period until the late 14th C AD. It was the longest-lasting custom in Europe, which has left about 2000 known horse burials. This paper publishes the osteometric data and age of horses found in Lithuanian cemeteries and castles of the 3rd-14th C AD, over 200 individuals in total. These are the remains of all the horses still stored in Lithuanian institutions. The paper discusses the dynamics of horse body size in order to test previously suggested hypotheses regarding the relationship between large horse body size and its military use, possibly non-local breed, and high social status of the owner. Moreover, we are publishing the AMS 14C dates of 13 horses previously assigned to the Migration period. The research results corrected the existing chronology. The abundant data also allowed an assessment of the development of the size and age of the horses in Lithuania between the 3rd and 14th C AD. Osteometric analyses have shown that Late Roman-post-Migration-period horses were unusually large compared to the Viking and medieval horses in Lithuania. Meanwhile, we suggest that the semi-slender-legged 118-125-cm-tall horse, which predominated in the Viking period, is the most consistent with the local horse type. In general, the horses in Lithuania in the 3rd-11th C AD were small compared to those in Central and Western Europe or Scandinavia. More significant changes can be observed in the Middle Ages. In the 12-14th C AD, there was a much greater variety of horses and the expansion of taller (140-150 cm) individuals. However, the continued abundance of small horses in the medieval times, found buried with saddles and other equipment, allows one to renew the debate on the formation of the cavalry, the tactics of combat, and the social composition of horsemen in Lithuania.

The elemental composition of small mammals in a commercial orchard-meadow system.

We analyze concentrations of various elements in small mammals from commercial orchards and berry plantations and test differences between them depending on species, individual body mass, age, gender, type and age of crop and intensity of applied agricultural measures. Skinned front legs (muscle and bones) were used to register the presence and concentration of Ca, Cd, Mo, Ni, Se, Ag, As, Be, Bi, Co, Cr, Cu, Fe, Ga, Mg, Mn, Pb, Sr, U, V and Zn. The most abundant species were common vole (Microtus arvalis), yellow-necked (Apodemus flavicollis) and striped field (A. agrarius) mice. The maximum recorded concentrations exceeded the minimums by 1.7-7.7 times in Mg, Zn, Cu, Ga, Ni and Ca, and 11.0-23.2 times in Mn, Be, Mo, Co, Sr, V, Pb and As. The hypothesis that the use of fertilization and pesticides in commercial orchards should induce differences in the elemental concentrations between crop areas and control habitats was confirmed by the higher concentrations of Cu, Mn, Bi, Co, Cr, Fe, Ni, Sr and Pb in rodents from the crop areas. Spatially controlled generalized linear mixed model confirmed the cumulative influence of species and crop, explaining 30-80% of the distribution of Ca, Ni, Co, Cu, Ga, Mn, Pb and U. The effect of species and the age of the individual was significant for Ni, Co, Cu, Fe, Ga, Mn and Pb, while effect of gender was not expressed. Depletion of Cu in older individuals was found in all three species. With species as a grouping factor, the effect of crop type and the intensity of agricultural practices were significant factors in the accumulation of Ca, Ni, Co, Cu, Ga, Mn and Pb, while effect of crop age was not expressed. The obtained elemental concentrations in rodents indicated orchards to be cleaner than heavy polluted areas.

Isotopic niche size variability in an ecosystem engineer along a disturbance gradient in a South African lagoon.

A key challenge for natural resource management is how to detect effects of environmental stress on individuals and populations before declines in abundance occur. Variability in carbon and nitrogen isotope composition (δ13C and δ15N) among consumers can provide information on the population trophic niche and how it may change in response to environmental stress. We measured δ13C and δ15N values in primary producers and in an ecosystem engineer, the bioturbating sandprawn Kraussillichirus kraussi, in Langebaan Lagoon, South Africa, along a human disturbance gradient. Diet partitioning mixing models were coupled with isotope niche analyses and individual body condition data to investigate shifts in resource utilisation and diet plasticity from minimally to highly disturbed sites. The δ15N values of seagrass, Zostera capensis indicated a nutrient gradient, with the highest δ15N values at highly disturbed sites indicating either anthropogenic or marine nitrogen inputs. A decreasing δ15N signal with distance from human disturbance/mouth of lagoon was however not evident for sandprawns nor their presumed dietary sources (phytoplankton, microphytobenthos or sediment organic matter), likely because of faster isotope turnover time compared to seagrass and/or differential fractionation for sandprawns among the sites. Sandprawn isotope niche sizes varied among sites, with no trend along the disturbance gradient. The smallest niche coincided with uniform feeding on microphytobenthos according to mixing models. On an individual level, deviating isotope values from population means were correlated to better body condition, suggesting that a divergent feeding strategy is beneficial. Our results support a generalist feeding behavior of the sandprawns with no evidence of reduced physiological status at the site with most human disturbance.

Development of graphitization method for low carbon aerosol filter samples with Automated Graphitization System AGE-3.

The wide applications of the radiocarbon (14C) approach in environmental, archeological, and geological research often necessitates the analysis of microgram-sized samples. The ability to measure low carbon samples is particularly relevant for aerosol particle filters, especially for samples from pristine environments. For this purpose, we investigated the sample dilution method for graphitization of low-carbon samples (20-200 µg C) with an Automated Graphitization System (AGE-3), and applied a mass balance equation for the calculation of 14C values. Materials with known 14C values (standards NIST-OXII and IAEA-C7) were diluted with 14C-free phthalic anhydride (PhA) until sufficient mass (500 µg C) for graphitization with the AGE-3 system was acquired. Reliable 14C values were obtained for samples with carbon amount in the range of 40-200 µg. Next, we adapted the dilution method for estimation of aerosol sample 14C values. Using it, we attained a precision of 0.71 ± 0.83 pMC for 14C measurements of aerosol samples containing 40-200 µg C. A shift of radiocarbon values to 5.07 pMC (average 3.08 ± 1.7 pMC) was observed for samples with low carbon content (<20 µg C). We determined that a precision of 2-3 pMC is acceptable for aerosol particle source apportionment studies. Using the sample dilution method, graphitization with AGE-3 of aerosol samples with carbon content >40 µg becomes a viable and efficient way of sample preparation for 14C analysis.

High capacity for a dietary specialist consumer population to cope with increasing cyanobacterial blooms.

We present a common-garden experiment to examine the amphipod Monoporeia affinis, a key deposit-feeder in the Baltic Sea, a low diversity system offering a good model for studying local adaptations. In the northern part of this system, the seasonal development of phytoplankton is characterized by a single diatom bloom (high nutritional quality), whereas in the south, the diatom bloom is followed by a cyanobacteria bloom (low nutritional quality) during summer. Therefore, the nutrient input to the benthic system differs between the sea basins. Accordingly, the amphipod populations were expected to be dietary specialists in the north and generalists in the south. We tested this hypothesis using a combination of stable isotope tracers, trophic niche analyses, and various endpoints of growth and health status. We found that when mixed with diatomes, the toxin-producing cyanobacteria, were efficiently incorporated and used for growth by both populations. However, contrary to expectations, the feeding plasticity was more pronounced in the northern population, indicating genetically-based divergence and suggesting that these animals can develop ecological adaptations to the climate-induced northward cyanobacteria expansion in this system. These findings improve our understanding regarding possible adaptations of the deposit-feeders to increasing cyanobacteria under global warming world in both limnic and marine ecosystems. It is possible that the observed effects apply to other consumers facing altered food quality due to environmental changes.

Impacts of land-use change and urban development on carbon sequestration in tropical seagrass meadow sediments.

Seagrass meadows store significant carbon stocks at a global scale, but land-use change and other anthropogenic activities can alter the natural process of organic carbon (Corg) accumulation. Here, we assessed the carbon accumulation history of two seagrass meadows in Zanzibar (Tanzania) that have experienced different degrees of disturbance. The meadow at Stone Town has been highly exposed to urban development during the 20th century, while the Mbweni meadow is located in an area with relatively low impacts but historical clearing of adjacent mangroves. The results showed that the two sites had similar sedimentary Corg accumulation rates (22-25 g m-2 yr-1) since the 1940s, while during the last two decades (∼1998 until 2018) they exhibited 24-30% higher accumulation of Corg, which was linked to shifts in Corg sources. The increase in the δ13C isotopic signature of sedimentary Corg (towards a higher seagrass contribution) at the Stone Town site since 1998 points to improved seagrass meadow conditions and Corg accumulation capacity of the meadow after the relocation of a major sewage outlet in the mid-1990s. In contrast, the decrease in the δ13C signatures of sedimentary Corg in the Mbweni meadow since the early 2010s was likely linked to increased Corg run-off of mangrove/terrestrial material following mangrove deforestation. This study exemplifies two different pathways by which land-based human activities can alter the carbon storage capacity of seagrass meadows (i.e. sewage waste management and mangrove deforestation) and showcases opportunities for management of vegetated coastal Corg sinks.

Stable Isotopes Reveal the Dominant Species to Have the Widest Trophic Niche of Three Syntopic Microtus Voles.

Diets and trophic positions of co-occurring animals are fundamental issues in their ecology, and these issues in syntopic rodents have been studied insufficiently. Using carbon (δ13C) and nitrogen (δ15N) stable isotope ratios from hair samples, we analysed the trophic niches of common (Microtus arvalis), field (M. agrestis), and root (M. oeconomus) voles co-occurring in orchards, berry plantations, and nearby meadows (as control habitat to orchards and plantations). We tested if the niche of the dominant common vole was the widest, whether its width depended on the presence of other vole species, and whether there were intraspecific differences. Results suggest stability in the trophic niches of all three Microtus species, as season explained only 2% of the variance. The widest trophic niche was a characteristic of the dominant common vole, the range of δ13C values exceeding the other two species by 1.6, the range of δ15N values exceeding the other two species by 1.9, and the total area of niche exceeding that of the other voles by 2.3-3 times. In the meadows and apple orchards, co-occurring vole species were separated according to δ13C (highest values in the dominant common vole), but they maintained similar δ15N values. Results give new insights into the trophic ecology small herbivores, showing the impact of species co-occurrence.

Isotopic Niche of Syntopic Granivores in Commercial Orchards and Meadows.

In agricultural habitats, diets and trophic positions of syntopic granivorous small mammals are not known sufficiently. Agroecosystems may be quite complex isotopically and the most complex situation concerns the nitrogen-15 isotope as δ15N values are influenced by many internal and external fluxes. We analysed the isotopic niches of striped field (Apodemus agrarius), yellow-necked (Apodemus flavicollis), and harvest (Micromys minutus) mice living sympatrically and syntopically in apple and plum orchards, raspberry and currant plantations, and nearby meadows that were used as control habitats. Carbon (δ13C) and nitrogen (δ15N) stable isotope ratios from hair samples were used as a proxy for their diet. As the main hypothesis, we expected differences in the isotopic niches of these three species, minimising interspecific competition. All species were fully separated according to δ15N values, presuming different amounts of food of animal origin in their diet. The separation of species according to δ13C was not expressed in all habitats. The core dietary niches of these species were fully separated in the apple orchards and raspberry plantations. Intraspecific differences of the isotopic niche were not present in any of the three species: that is, resources were equally used by males and females, adults, subadults, and juveniles.

How Copepods Can Eat Toxins Without Getting Sick: Gut Bacteria Help Zooplankton to Feed in Cyanobacteria Blooms.

Toxin-producing cyanobacteria can be harmful to aquatic biota, although some grazers utilize them with often beneficial effects on their growth and reproduction. It is commonly assumed that gut microbiota facilitates host adaptation to the diet; however, the evidence for adaptation mechanisms is scarce. Here, we investigated the abundance of mlrA genes in the gut of the Baltic copepods Acartia bifilosa and Eurytemora affinis during cyanobacteria bloom season (August) and outside it (February). The mlrA genes are unique to microcystin and nodularin degraders, thus indicating the capacity to break down these toxins by the microbiota. The mlrA genes were expressed in the copepod gut year-round, being >10-fold higher in the summer than in the winter populations. Moreover, they were significantly more abundant in Eurytemora than Acartia. To understand the ecological implications of this variability, we conducted feeding experiments using summer- and winter-collected copepods to examine if/how the mlrA abundance in the microbiota affect: (1) uptake of toxic Nodularia spumigena, (2) uptake of a non-toxic algal food offered in mixtures with N. spumigena, and (3) concomitant growth potential in the copepods. The findings provide empirical evidence that the occurrence of mlrA genes in the copepod microbiome facilitates nutrient uptake and growth when feeding on phytoplankton mixtures containing nodularin-producing cyanobacteria; thus, providing an adaptation mechanism to the cyanobacteria blooms.

Nitrogen isotope composition of amino acids reveals trophic partitioning in two sympatric amphipods.

According to ecological theory, two species cannot occupy the same niche. Using nitrogen isotope analyses (δ15N) of amino acids, we tested the extent to which two sympatric deposit-feeding amphipods, Monoporeia affinis and Pontoporeia femorata, partition their trophic resources. We found that trophic position (TP) and resynthesis index (∑V; a proxy for degradation status of ingested material prior to assimilation by the consumer) differ between species. The surface-feeding M. affinis had higher TP and intermediate ∑V, both pointing to a large contribution of metazoans in its diet. P. femorata, which feeds in the subsurface layers, had lower TP and a bimodal distribution of the ∑V values, supporting previous experimental evidence of a larger feeding niche. We also evaluated whether TP and ∑V values have consequences for amphipod fecundity and embryo viability and found that embryo viability in M. affinis was negatively linked to TP. Our results indicate that the amino acid-δ15N data paired with information about reproductive status are useful for detecting differences in the trophic ecology of sympatric amphipods.

Suppression of phase transitions and glass phase signatures in mixed cation halide perovskites.

Cation engineering provides a route to control the structure and properties of hybrid halide perovskites, which has resulted in the highest performance solar cells based on mixtures of Cs, methylammonium, and formamidinium. Here, we present a multi-technique experimental and theoretical study of structural phase transitions, structural phases and dipolar dynamics in the mixed methylammonium/dimethylammonium MA1-xDMAxPbBr3 hybrid perovskites (0 ≤ x ≤ 1). Our results demonstrate a significant suppression of the structural phase transitions, enhanced disorder and stabilization of the cubic phase even for a small amount of dimethylammonium cations. As the dimethylammonium concentration approaches the solubility limit in MAPbBr3, we observe the disappearance of the structural phase transitions and indications of a glassy dipolar phase. We also reveal a significant tunability of the dielectric permittivity upon mixing of the molecular cations that arises from frustrated electric dipoles.

Seasonal Variation in Stable Isotope Ratios of Cow Milk in Vilnius Region, Lithuania.

Various studies have shown that stable isotope analysis has the potential to verify the geographic origin of foods and drinks. However, stable isotope composition is not always constant in the environment and can even change in the same area. Dairy products are of particular interest as a group of foods that play an important role in feeding the population. The composition of milk is fundamentally dependent on the feeding of the cows, and thereby on a particular environment. To better understand the amount of variation in δ18O, δ13C, and δ15N values in the milk from the same area, we measured stable isotope ratios in cow milk water, artesian water, and precipitation (δ18O) as well as in bulk milk samples (δ13C and δ15N) collected in 2014⁻2016. Different water and food sources were available during the winter (artesian water only and dry grass) and summer (artesian water and fresh grass), and spring and autumn seasons reflected transitional periods. Oxygen stable isotope ratios in milk water were relatively lower in winter and transitional seasons and higher in summer, showing the dependence on the main water source. δ13C values reflected particular food sources. This study shows the applicability of the stable isotope ratio method in linking cow milk to specific environments and reveals the amount of variation in stable isotope ratios in the same area. These results could be valuable for other studies on geographical origin determination of dairy products.

Insufficient evidence for BMAA transfer in the pelagic and benthic food webs in the Baltic Sea.

The evidence regarding BMAA occurrence in the Baltic Sea is contradictory, with benthic sources appearing to be more important than pelagic ones. The latter is counterintuitive considering that the identified sources of this compound in the food webs are pelagic primary producers, such as diatoms, dinoflagellates, and cyanobacteria. To elucidate BMAA distribution, we analyzed BMAA in the pelagic and benthic food webs in the Northern Baltic Proper. As potential sources, phytoplankton communities were used. Pelagic food chain was represented by zooplankton, mysids and zooplanktivorous fish, whereas benthic invertebrates and benthivorous fish comprised the benthic chain. The trophic structure of the system was confirmed by stable isotope analysis. Contrary to the reported ubiquitous occurrence of BMAA in the Baltic food webs, only phytoplankton, zooplankton and mysids tested positive, whereas no measurable levels of this compound occurred in the benthic invertebrates and any of the tested fish species. These findings do not support the widely assumed occurrence and transfer of BMAA to the top consumers in the Baltic food webs. More controlled experiments and field observations are needed to understand the transfer and possible transformation of BMAA in the food web under various environmental settings.