N = 20) and non-musician (N = 20) participants were required to judge musical stimuli in terms of “liking” and “emotions”. No significant differences between groups were found, although musicians were slower than non-musicians in both tasks, likely indicating a more analytic judgment, due to musical expertise. Experiment #2 investigated the putative causal role of the left dorsolateral pre-frontal cortex (DLPFC) in the esthetic appreciation of music, by means of transcranial direct current stimulation (tDCS). Unlike previous findings in visual art, no significant effects of tDCS were found, suggesting that stimulating the left DLPFC is not enough to affect the esthetic appreciation of music, although this conclusion is based on negative evidence,.Humans are the only species capable of experiencing pleasure from esthetic stimuli, such as art and music. Neuroimaging evidence suggests that the left dorsolateral prefrontal cortex (DLPFC) plays a critical role in esthetic judgments, both in music and in visual art. In the last decade, non-invasive brain stimulation (NIBS) has been increasingly employed to shed light on the causal role of different brain regions contributing to esthetic appreciation. In Experiment #1, musician ( Inucleus accumbens (NAcc) and the right caudate nucleus) [Across the years, research in the field of esthetic appreciation moved from a behavioral to a neurofunctional level of investigation. As for its neural correlates, studies investigating the neuroscience of art assign an important role to the left dorsolateral prefrontal cortex (DLFPC) ,4,37. Thnucleus) ,44. Acconucleus) . Changesnucleus) . In sum,nucleus) ,46. Expenucleus) . In factnucleus) . Overallnucleus) , in thatnucleus) ,39. Forty participants, divided in two groups, volunteered to participate in this study. The non-musician group included 20 students from the University of Milano-Bicocca  who reported no previous training or special interest in music . The musician group included 20 musicians  with musical education . Exclusion criteria were uncorrected hearing or visual impairments. All participants were right-handed  and had https://www.audacityteam.org/, accessed on 1 February 2020).Stimuli consisted of 45 excerpts cut out from musical pieces of different cultures and different musical genres. None of them had been used for soundtracks of movies or television spots. The excerpts never corresponded to the beginning of the track or containing vocal parts, to rule out the possibility of recognizing the track, or of introducing other confounding variables, due to the presence of language. Excerpts performed in various formations  were selected; a third of them had a tonal structure, a third was, instead, atonal and the remaining tracks had a mixed structure. The instruments used were both canonical traditional  and electronic . Furthermore, stimuli that did not contain a melodic line, but had just a rhythmic structure, were also included. The tracks were cut, so that each of them lasted 7 s, and, subsequently, were equalized for intensity. Finally, a fade-out effect in the last 1500 msec was applied. All adjustments were made through the Audacity© software : “beautiful”, “ugly”, “unsure judgment”  through a pilot study in which five western professional musicians, graduated at the Conservatory, were enrolled, and asked to classify the tracks into the three categories proposed, according to their professional judgment.As a result, the “beautiful” category was mostly composed of excerpts with a tonal framework, a regular rhythmic structure, or easily perceptible and consonant sounds. On the contrary, the “ugly” category was mostly composed of excerpts with an atonal framework, an irregular rhythmic structure, or hardly perceptible and dissonant sounds. The “medium” category included excerpts with mixed characteristics, as compared to the other two groups. Participants, seated in front of a 15.500 PC (1280 × 800 pixels) screen at an approximate distance of 57 cm, in a normal-lightened and silent room, were asked to perform a computerized rating task. Before starting the experiment, participants were informed that they would be performing two computer tasks, in which they would be listening to a few short musical excerpts through earphones (sound-proof over-ear headphones). The first task required an esthetic judgment (“How much do you like this excerpt?”); while the second task required an expression of the emotional valence evoked by the excerpt (“Positive or negative emotion?”). https://jasp-stats.org, accessed on 1 March 2021). A Bayesian ANOVA with an analysis of the effects [incl), which can be interpreted as evidence in the data for including a predictor, either a main effect or an interaction. BFsincl between 1 and 3, 3 and 10, and larger than 10, are considered respectively as anecdotal, moderate, and strong evidence for including a predictor; conversely, BFsincl between 1 and 1/3, 1/3 and 1/10, and smaller than 1/10 indicate anecdotal, moderate, or strong evidence for excluding a predictor. A BFincl = 1 indicates no evidence in favor of including or excluding a predictor [10) compare the alternative hypothesis that two levels of a factor differ from each other with the null hypothesis of no difference, the thresholds for supporting the alternative vs. null hypotheses being the same as the one illustrated above for including or excluding a factor using BFincl.The position of the mouse cursor along the bar was automatically converted by the software to percentage rating scores, where a 0% score corresponded to the mouse cursor positioned at the lower end (red) of the rating bar and a 100% score corresponded to the mouse cursor positioned at the upper end (green) of the rating bar. Analyses were run using SPSS© 19. Bayesian analyses were performed using the JASP software (Version 0.16.0) , divided in two groups, who reported no previous training or special interest in music volunteered to participate in this study . None of them had taken part in Experiment #1. Furthermore, as no significant differences were found between musicians and laypersons neither in the esthetical nor in the emotional judgment, we decided not to include a second group of professional musicians. Participants were tested all right-handed  and had t-tested between Group #1 and Group #2 on the pre-tDCS stimulation (to ensure no conditioning on judgments) scores obtained by participants in Set #1 vs. Set #2. No differences were found between each category of stimuli .Two different sets of musical excerpts were set up, each including 42 musical excerpts. Set 1 was the same set used in Experiment #1. In the esthetic judgment task, three out of 45 tracks, which had obtained, on average, the most extreme judgment scores, were excluded, to adopt only the excerpts with a certain degree of modulation in their esthetic judgment. Set #2, instead, was created ad hoc for this experiment. Within the same songs the tracks used for Set #1, new excerpts, alike to the ones chosen for Set #1, with reference to the criteria of harmonic, rhythmic and melodic structure, were selected, to obtain two largely superimposable, but not identical, sets. This was verified through independent samples Participants were seated in front of a 15.500 PC (1280 × 800 pixels) screen at an approximate distance of 57 cm, in a lightened and silent room, and were asked to perform the computerized rating task of Experiment #1. Both groups performed the same task, but with a reversed presentation order of the two sets of stimuli: (a) Group #1 listened to set #2, pre-stimulation and to set #1, post-stimulation; (b) Group #2 listened to set #1, pre-stimulation and to set #2, post-stimulation.Before starting the experiment, participants were informed that they would be listening a few short musical excerpts through earphones (sound-proof over-ear headphones). The task required an esthetic judgment  kept firm by elastic bands. The excitability-enhancing anodal electrode was placed over the left DLPFC localized as the middle point between F3 and F5 in the 10–20 electroencephalography (EEG) system [Transcranial direct current stimulation (tDCS) was delivered by a battery driven, constant current stimulator  through a pair of saline-soaked sponge electrodes (7 × 5 cm: 35 cm) system , whereas) system ,53,54. E) system . Moreove) system . The stuBefore starting, instructions were repeated to participants. Excerpts were presented in a random fixed order. The whole experimental session lasted approximately 45 min. The software E-prime 2.0  was used for stimuli presentation and data recording.Group (musicians and non-musicians) and Judgement . A repeated-measures analysis of variance (ANOVA), with Judgment  as a within-subjects main factor and Group (musicians and non-musicians) as a between-subjects main factor, was carried out on the mean percentage rating scores for the esthetic evaluation  no significant main effect of Group (musicians vs. non-musicians); (iii) a trend towards significance for the interaction group × judgment, indicating that the effect of group varied at different levels of judgment. As for the main effect, the differences among the three levels of judgment were then analyzed by multiple comparisons applying the Bonferroni correction. Significant differences between “beautiful” vs. “ugly” (p < 0.001), “beautiful” vs. “medium” (p < 0.001), and “medium” vs. “ugly” (p < 0.001) were found. Regarding the interaction comparisons, musicians reported a lower score for “beautiful” , and a higher score for “medium”  and for “ugly”  judgments.tion see . The anaJudgment as a predictor, BFincl = 8.75 × 1016, but only very weak evidence for excluding the main effect of Group (BFincl = 0.47) or including the interaction effect (BFincl = 1.05). Post hoc comparisons for the Judgment effect provided evidence indicating different rating scores between the “beautiful” and “ugly” (BF10 = 2.99 × 1011), “beautiful” and “medium” , and “ugly” and “medium” (BF10 = 1.045 × 106) judgment levels.Bayesian analysis indicated that data provided very strong support for the inclusion of Group  and Judgment . A repeated-measures ANOVA, with Judgment as a within-subjects main factor, and Group as a between-subjects main factor, was performed  but no significant interaction effect Judgment × Group. As for the Judgment factor, post hoc analyses (Bonferroni correction applied) indicated only a significant difference between “beautiful” and “ugly”, p = 0.04 , with “beautiful” excerpts obtaining the fastest times (M ± SD = 1465 ± 67 msec), followed by “medium” (M ± SD = 1559 ± 86 msec), and “ugly” (M ± SD = 1617 ± 99 msec) tracks. Regarding the Group factor, musicians proved to be slower (M ± SD = 1704 ± 607 msec), as compared to non-musicians (M ± SD = 1389 ± 338 msec), in making an esthetic evaluation of musical excerpts.rmed see . The anaJudgment, BFincl = 1.35, and Group, BFincl = 1.69 as predictors, and moderate evidence for excluding the interaction effect (BFincl = 0.12). Post hoc comparisons for the Judgment effect provided evidence indicating that reaction times were faster in the “beautiful” than in the “ugly” condition (BF10 = 3.36), but not when the “beautiful” and “medium” (BF10 = 0.80), and the “ugly” and “medium” Judgment (BF10 = 0.24) were compared.Bayesian analysis indicated that data provided anecdotal evidence for the inclusion of In sum, the results demonstrated significant differences for all participants for all three categories: “beautiful”, “ugly”, and “medium”. The “beautiful” excerpts obtained the highest scores, the “ugly” ones the lowest scores, and the “medium” ones were placed at an intermediate level. As for musicians and non-musicians, the present data did not show significant differences in the judgements of the two groups about the excerpts. However, musicians were significantly slower than non-musicians.Group (musicians and non-musicians) and Judgment . In line with the results obtained in the Esthetic evaluation, the “beautiful” category obtained the highest rating, followed by the “medium” and lastly by the “ugly”.Judgment  as the within-subjects main factor and Group (musicians and non-musicians) as the between-subjects main factor was carried out  no significant main effect of Group; and (iii) no significant interaction effect of Judgment by Group. As for the main effect of Judgment, Bonferroni-corrected multiple comparisons demonstrated significant differences between the three categories . A repeated-measures ANOVA with  out see . The anaJudgment as a predictor, BFincl = 2.67 × 1028, but only very weak evidence for excluding the main effect of Group (BFincl = 0.46), and moderate evidence for excluding the interaction effect Judgment by Group (BFincl = 0.18). Post hoc comparisons for the Judgment effect provided evidence indicating that the emotional mean rating score was higher for the “beautiful” compared to both the “ugly” (BF10 = 3.05 × 1018) and the “medium” judgment (BF10 = 2.96 × 1011), and for the “medium” compared to the “ugly” judgment .Bayesian analysis indicated that data provided very strong support for the inclusion of Group (musicians and non-musicians) and Judgment . A repeated-measures ANOVA with Judgment as a within-subjects main factor and Group as a between-subjects main factor revealed a significant main effect of Judgment, and a main effect of Group. The Judgment by Group interaction was not significant. As for the main effect of Judgment, multiple comparisons, with the Bonferroni correction applied, showed significant differences between the “beautiful” vs. “ugly” (p < 0.001) and between the “medium” vs. “ugly” (p = 0.04) judgements; the difference between the “beautiful” vs. “medium” judgements was not significant (p = 0.15). “Beautiful” excerpts obtained the fastest reaction times (M ± SD = 1584 ± 94 msec), followed by “medium” (M ± SD = 1684 ± 105 msec) and “ugly” tracks (M ± SD = 1826 ± 125 msec). As for the differences between the groups, musicians were slower in judging the emotional valence of musical excerpts (M ± SD = 1989 ± 863.9 msec), compared to non-musicians (M ± SD = 1407 ± 353 msec).Judgment as a predictor, BFincl = 242.25, and moderate evidence for including the main effect of Group, BFincl = 4.63. The BF for the Judgment by Group interaction was 0.19, indicating moderate evidence for excluding the interaction effect as the predictor. Post hoc comparisons for the Judgment effect provided evidence indicating faster reaction times for the “beautiful” compared to the “ugly” judgment (BF10 = 306.3), and for the “medium” compared to the “ugly” judgment (BF10 = 3.09). Results indicated evidence that reaction times of “beautiful” and “medium” judgment were the same (BF10 = 1.12).Bayesian analysis indicated that data provided very strong support for the inclusion of Overall, Experiment #1 had a double aim: firstly, to assess whether there was any difference in the esthetic and emotional evaluation of musical excerpts between musicians and non-musicians; secondly, to set up a task, providing a useful tool, to assess “liking” and emotions in music, both for music experts and naïve individuals. As for the emotional and esthetic experience, the results demonstrated significant differences for all participants in both tasks, and for all three categories: “beautiful”, “ugly”, and “medium”. For both the esthetic and the emotional judgments, the “beautiful” excerpts obtained the highest scores, the “ugly” ones the lowest scores, and the “medium” ones were placed at an intermediate level.As for the differences between musicians and non-musicians, the present data do not show significant differences in the judgements of the two groups about the excerpts, both emotionally and esthetically: the recorded judgements are then independent of musical expertise. On the other hand, musicians are significantly slower than non-musicians.Overall, the results highlight a correlation between esthetic and emotional evaluations in music after a short listen , demonstrating that higher levels of beauty correspond to more positive emotions. Furthermore, this correlation is independent of a previous musical training, which, in contrast, seems to influence only the time needed to express the judgment, suggesting a more analytic process for higher levels of expertise.t-tests were carried out between the two groups, for each category of stimuli: “beautiful”, “medium”, and “ugly”. No significant differences were found .To evaluate the robustness of the results of Experiment #1 about esthetic judgments, the esthetic scores obtained by non-musicians in Experiment #1 were compared with those obtained in Experiment #2 by the participants who scored stimuli of Set #1 in the pre-stimulation stage. Three independent sample After each stimulation, a questionnaire was administered to all participants, to detect the presence of any side effect or discomfort sensations. Transient discomfort sensations as itch or light burning on the scalp were sometimes  reported during the initial phases of either stimulation. As preliminary analyses demonstrated no significant differences between the evaluations of set #1 and set #2, in all subsequent analyses, the participants were considered as a single group (N = 22). The position of the mouse cursor along the bar was automatically converted by the software to percentage rating scores, where a 0% score corresponded to the mouse cursor positioned at the lower end (red) of the rating bar, and a 100% score to the mouse cursor positioned at the upper end (green) of the rating bar. Analyses were performed on mean rating scores.tDCS Condition , Session (pre- and post-tDCS) and Judgment  as within-subject main factors, demonstrated no significant main effect of the tDCS Condition  to post-tDCS (M ± SD = 49.82 ± 11.65%). Furthermore, the main effect of Judgement and the Session by the Judgment interaction were also significant. Nevertheless, the tDCS Condition by Session, the tDCS Condition by Judgment, and the tDCS Condition by Session * Judgment interactions were not significant. As for the main effect of Judgment, the Bonferroni-corrected multiple comparisons showed significant differences between the three categories , with a higher mean score for the “beautiful” category (M ± SD = 64.93 ± 12.57%), as compared to the “medium” (M ± SD = 54.52 ± 12.96%) and the “ugly” (M ± SD = 33.25 ± 13.79%) one, which obtained the lowest rating. As for the Session × Judgment interaction, mean scores indicate that the “beautiful”  and the “medium”  categories, but not the “ugly”  category, obtained higher scores during the pre-tDCS condition as compared to the post one.tion see . On the Judgment as a predictor, BFincl = 4.91 × 1067. The post hoc tests indicated overwhelming evidence that the “liking” rating score differed between “beautiful” and “medium” judgment (BF10 = 2.98 × 1011), between “beautiful” and “ugly” judgment (BF10 = 3.10 × 1032), and between “medium” and “ugly” judgment (BF10 = 6.61 × 1027). The data also indicated weak evidence for the inclusion of Session as a predictor, BFincl = 1.12. Finally, the data indicated moderate evidence for excluding the tDCS Condition as predictor, BFincl = 0.21, as well as all the interactions effects .Bayesian analysis indicated that the data provided very strong support for the inclusion of In line with the procedure followed for the rating scores, participants were considered as a single group including 22 subjects.tDCS Condition , Session (pre- and post-tDCS) and Judgment  as within-subject main factors, demonstrated no significant main effect of Condition  than during the post-tDCS task, (M ± SD = 1322 ± 518 msec). Moreover, the main effect of Judgment was significant. All interaction effects were not significant: Condition × Session, Condition × Judgment, Session × Judgment and Condition × Session × Judgment. The differences among the three levels of Judgment were analyzed by Bonferroni-corrected multiple comparisons; significant differences were found between the “beautiful” vs. “ugly” (p = 0.01) and the “medium” vs. “ugly” (p = 0.02), with the “beautiful” vs. “medium” comparison being not significant (p = 0.89). Particularly, participants were faster for “beautiful” excerpts (M ± SD = 1310 ± 104 msec), as compared to the “medium” tracks (M ± SD = 1360 ± 109 msec) and to the “ugly” ones (M ± SD = 1453 ± 106 msec), which required the longest times.tion see . The maiSession, BFincl = 4.80 and Judgment, BFincl = 3.36, as predictors. Post hoc tests provided evidence that latencies differed between the “beautiful” and the “ugly” judgments (BF10 = 153.45) and between the “medium” and the “ugly” ones BF10 = 8.37). Conversely, the latencies of “beautiful” and “medium” judgment did not differ from each other (BF10 = 0.24). Analyses also indicated moderate evidence for excluding the tDCS Condition as predictor, BFincl = 0.29, as well as the interaction effects Session × tDCS Condition: BF incl = 0.35, and Session × tDCS Condition × Judgment: BF incl = 0.17. Finally, data indicated strong evidence for excluding as predictors both the Session × Judgment, BF incl = 0.07, and the tDCS Condition × Judgment, BF incl = 0.07, interactions. Bayesian analysis indicated moderate evidence for including both Experiment #2 investigated the role of the left DLPFC in the esthetical judgment of musical pieces, using the tDCS. Contrary to the expectations and previous findings in the field of visual art , the resConcerning the ratings, the results show a significant difference between pre- and post-tDCS scores, independent of the type of stimulation , with rates being lower after tDCS. This result may be accounted for in terms of a generalized effect due to the participants’ fatigue and the repetition of the task.Finally, the pattern of response latencies, with faster responses in the second (post-tDCS) part, with respect to the first (pre-tDCS) part of the study, would likely reflect a stabilization of the response criterion used and a task familiarization effect, resulting in faster responses.Experiment #1 highlighted how esthetic judgment and emotional valence follow the same trend in music, both reporting higher scores for excerpts previously classified as “beautiful” and lower scores for the “ugly” tracks.The classification of the stimuli made before the experiments, together with the results of the present study, support the hypothesis that esthetics judgment and emotional valence generally correlate in music. In fact, previous findings show that the music preferences of both musicians and non-musicians vary only in a few music genres, such as classical and jazz music, which are preferred by musicians . HoweverFurthermore, esthetic judgment and emotional valence in music seem to be independent of the level of musical training, adding more evidence to previous findings showing a homogeneity of the answers of musicians and non-musicians about esthetic judgment of chord sequences, with no differences between expert and naïve participants . As for Experiment #2 does not demonstrate any significant effect of tDCS stimulation. Cattaneo and colleagues  performeFurthermore, music preferences may vary widely among individuals, depending on several modulating factors, including: (i) person-specific features or the “internal context”, including expertise, internal state, mood, personality, and attitude; (ii) factors related to the listening situation, also described as “external context”, including the physical and social environment, such as being at home or elsewhere, as in a concert hall; being alone or with others) ,69. TherRatings, independent of the type of stimulation , are lower after tDCS. This result may be accounted for in terms of a generalized effect due to the participants’ fatigue and the repetition of the task. Fatigue may be an interfering variable in the evaluation of pleasantness, as observed in other areas, such as food pleasantness ratings .Finally, the pattern of response latencies, with faster responses in the second (post-tDCS) part, with respect to the first (pre-tDCS) part of the study, may reflect a stabilization of the response criterion used and a task familiarization effect, resulting in faster responses, as observed in previous studies .On the other hand, overall, the results obtained in Experiments #1 and #2 have to be considered along with a few limitations aspects: first, the present samples included only relatively young participants; second, music excerpts were not chosen by the participants themselves. Finally, future studies should also specifically explore the influence of different music genres on the esthetic judgment of music.In summary, the present findings show that esthetic and emotional judgments agree when people  are asked to rate their first impression about music tracks, independent of their musical expertise. Furthermore, musicians tend to be slower than laypersons, possibly indicating that they allocate comparatively more resources and effort when required to express an esthetic judgment about music. The present results, unlike previous findings in visual art , demonst


One type, mangroves, utilizes less freshwater during transpiration than adjacent terrestrial ecoregions, equating to only 43% (average) to 57%  of evapotranspiration ( Competition for water between humans and natural ecosystems is critical in driving outcomes for natural ecosystem health, and water supply is projected to increasingly shift toward human use3. Selection of “nature-based solutions” to sequester carbon or enhance climate resilience could simultaneously consider the water economy of the ecosystem doing the work. To that end, “blue carbon” wetlands may have emergent but yet unrecognized ecological value.Water will be among the most limiting natural resource of the future, influencing ecological flows among coastal environments and soon affecting at least 80% of the world’s population directly– but also include upper estuarine tidal wetlands, and adjacent ecosystems such as salt flats and macroalgal communities5. As a group, they provide tremendous value to human societies7. Among these, blue carbon wetlands store large amounts of carbon in aboveground biomass and in soils as their roots trap sediments and expand soil volumes9. As a result, blue carbon wetlands have increasingly become important for environmental management of coasts and for use as self-sustaining nature-based solutions in adapting coastlines to rising seas10.Blue carbon wetlands comprise a variety of coastal ecosystems – most notably mangroves, saltmarshes and seagrasses11 and tolerates but does not always require saline water12 (but see ref.13). Mangroves extract freshwater from ultrafiltration of seawater through root tissue15 and consume freshwater differentially when it is readily available 17. Once water is absorbed by roots and the metabolic costs of excluding salt are incurred, bulk water transport from roots-to-leaves is driven by pressure gradients created as water is transpired18. Especially under saline soil conditions, mangroves have been recognized as having high plasticity in adjusting their rates of leaf-level water use efficiency to accommodate osmotic gradients19.Mangrove vegetation is located within the intertidal zone20, that intrinsic leaf-level photosynthetic water use efficiency  of leaf-level water-use efficiency published over the last three decades (source: Scopus/Web of Science), most confirm what was originally presented from a 1989 field studyses Fig. . This re15. Mangroves can, in many locations, grow with limited freshwater availability17, their roots extracting freshwater from seawater14, and they can obtain some freshwater directly through foliage25. We explore three questions. First, are leaf-level and tree-level water use strategies in mangroves similarly conservative? Second, how much water does mangrove vegetation use? Third, is mangrove vegetation water use low compared to water use from non-wetland ecosystems  or from vegetation that arises from the conversion of mangroves to alternative land uses ? We answer these questions by exploring a pulse of recent studies and model development that allow insight across scales.In this study, we assess whether one type of blue carbon wetland – mangroves – may be particularly efficient in the amount of water they use at leaf, tree, and ecosystem scales during the process of atmospheric carbon capture2 (mol H2O)−1 from dry sub-humid environments to 88–95 μmol CO2/(mol H2O) in arid and semi-arid environments −1  which is only slightly higher than shrubs and trees from dry sub-humid environments .Mangrove water use efficiency is reported over a range of scales; as long-term, instantaneous, or intrinsic, and at the leaf, plant, or ecosystem level, each necessitating a range of assumptions. This 3 × 3 matrix of scale is dominated in the mangrove literature by leaf-level mentclass2pt{minim2O day−1 cm−1 of dbh  while mangrove trees used only 1.4 L H2O day−1 cm−1 of dbh . Therefore, from sap flow-derived individual tree water flux data, we can begin to understand organizational scales at which transpirational water loss is most limited from mangrove vegetation.However, lack of differentiation among comparative water use efficiencies at the leaf-level did not scale. Instead, upland forest trees used, on average, 3.5 L Hp < 0.001; r2 ≥ 0.955) suggesting the potential to use NPP of focal mangrove forests to estimate 2O year−1 and NPP is in units of kg C m-2 year−1. Note that 1 mm H2O is equivalent to 1 L H2O per square meter of ground area.Scaling further to the canopy, we discovered that slopes of relationships relating canopy transpiration to net primary productivity  and the Asia–Pacific Region (N = 47) that had sufficient data to derive approximate NPP values, we used Eq.  during periodic (or even sustained) reductions in freshwater to the coast29, at least to a point30.Surface or groundwater seeping through mangroves and out to the ocean is not necessarily available for direct human consumption, but unconsumed freshwater that flows to marine and estuarine environments has inherently high value because of its importance in supporting wetland and coastal ocean productivity, and thus, sustained ecological processes within adjacent estuariesElaeis guineensis Jacq.) plantations, have replaced 18,467 ha of mangroves in Indonesia, Myanmar, Malaysia, and Thailand between the years 2000–201231. These plantations use more water through 32 to 70%33 of 2O year−1 toward transpirational water loss that would simultaneously reduce water flows toward estuaries. As oil palm plantations often need irrigation in dry years, the additional amount of freshwater needed to sustain oil palm 34. Furthermore, freshwater use by mangroves would not compete directly with humans under most land use or climate change scenarios.Mangroves also have lower water use than alternative land uses to which they have been converted. For example, oil palm  that ensure optimal water potentials and sap flow with increasing environmental stress. Thus, in addition to inherently lower whole-tree water use in mangroves compared to upland trees, saline conditions exert additional constraints to whole-tree mangrove water use. For example, water use can become theoretically high (> 150 L H2O day −1), and far greater than the literature has documented to date . This dynamic xylem sap osmolality control increases the water available to the transpiration stream without actively increasing transpiration, further reducing the risk of embolisms under salt stress46.Plasticity in water use along environmental gradients has also been studied from anatomical and physiological perspectives in mangroves2 from the MODIS satellite product (MOD16-A3), which is an analytical approach used by Schlesinger & Jasechko48 to permit direct comparisons of sap flow-derived transpiration rates and For our analysis focused at the broader canopy scale, we report average 2. We assume that plots are representative of the larger stand. Second, our approximations of 49. Third, our sap flow generated determinations of At least four assumptions apply to this 2 ha−1) and average canopy heights (10–11 m), regional 2O year−1 while dominant canopy 2O year−1, or 34–50% of 50. Short mangrove forests  on Hainan Island, China, used only 269–357 mm H2O year−1 versus an 2O year−1, or 23–31% of 51. Mangrove stands did use as much as 872 mm H2O year−1 versus an −1 (or 63% of 2 ha−1), and taller trees (19 m)50. 52. Unlike the direct linear increases in 20, mangrove 52. Thus, empirical estimations of Empirical assessment of sed Fig. 20, mangr2O year−1 if comparable areas to mangrove are assumed (Table 2O year−1). Likewise, our predictive variable (NPP) for approximating 53. Thus, mangroves receiving higher rainfall benefit from reduced salinities and increased nutrients, which in turn illicit productivity gains requiring more freshwater use by trees.By aligning global mangrove area with the distribution of adjacent ecoregions to mangroves, water use differences from our analysis scale to 34.7 Teraliters (TL) Hed Table . We add 54, due to changes in diets, increased demand for freshwater resources to support fossil fuel extraction55, and competition between water used for agricultural production of biofuels and food56. Placing potential water use reductions by mangroves within this food-energy-water nexus requires understanding of the multiple ecosystem services that mangrove wetlands provide, which encompass food resources, building materials and energy utilization. The quantification and comparison of freshwater consumption in the production of 268 goods and services can be determined from estimates of 57.Finally, there is a growing concern for the ability to feed the Planet’s increasing population with limited renewable freshwater resources58, we estimate that the value of food production per hectare for a tropical rainforest (e.g.), in USD, is approximately $ 0.27 mm−1 of H2O but for mangroves it is $ 2.20 mm−1 of H2O 59. High productivity of mangroves for their low usage of freshwater compared to land uses that have replaced them provides evidence of additional benefits to the ecology of associated estuaries that are dependent on freshwater flows. The water economy of ecosystems may require greater worldwide consideration as humans look for options to ameliorate, limit, and adapt to future water shortages while sustaining coastal productivity and ecological connections. The conservative water use of blue carbon ecosystems may add to their value and function as nature-based solutions, and to coastal resilience as freshwater availability is reduced.Efficient and conservative use of freshwater at the individual tree and stand levels by mangroves equate to global water use differentials in the trillions of liters annually compared to adjacent ecoregions and alternate land use area. We can gain additional information on levels of water use from other blue carbon wetlands through expanding eco-hydrologic studies, especially as these wetlands may also be efficient in maintaining their functions with reduced freshwater availability. Low freshwater use by mangroves could significantly augment the global blue carbon wealth of nations, recently estimated to be $190.67 ± 30 billion year5; all but three of these would be considered wetlands, with salt flats and mud flats being examples of non-emergent (plant) blue carbon wetlands. Herein, we focus on mangroves.At least 11 coastal ecosystems have been considered based on a minimum of actionably defined criteria to be “blue carbon ecosystems”. These include mangrove wetlands, tidal marshes , seagrasses, salt flats, freshwater (upper estuarine) tidal forests, macroalgae, phytoplankton, coral reef, marine fauna (fish), oyster reefs, and mud flats20, which presented leaf-scale net photosynthesis (2 m−2 s−1), stomatal conductance (−2 s−1), leaf-intercellular CO2 concentrations (−1), and intrinsic photosynthetic water use efficiency (−2 s−1). Soil salinity at the time of data collection ranged from 10 to 49 psu, and median long-term atmospheric temperature and relative humidity among sites ranged from 19.9 to 27.4 °C and 35.1 to 92.2%, respectively , including arid, semi-arid, dry semi-humid, and humid locations.We then accessed an existing database  that reported raw 61 that provides maximum individual tree water use data (L H2O day−1) from 52 published studies representing 67 species of upland trees from around the world. Of those studies, dbh values (8 to 134 cm) were provided alongside 47 individual tree water use values. Maximum individual tree water use and dbh (4.1 to 45.3 cm) were available from the original source for 8 mangrove studies representing 7 species from French Guiana, Mayotte Island (Indian Ocean), China, Florida (USA), and Louisiana (USA) ; e.g., we could not extract specific individual tree water use versus dbh from a Moreton Bay  study site62, south Florida study site63, or from five additional study sites in China52. However, regressions for two of the Chinese study sites provided over two years51 indicated that mangrove trees from a suite of species ranging in dbh from 8 to 24 cm used approximately 0.76 and 9.31 L H2O day−1, or 0.53 L H2O day−1 cm−1 of dbh. These apparent rates were even lower than what was reported as average for mangroves in Fig. 2O day−1 cm−1. The mangrove species included in this analysis were Avicennia germinans (L.) L., Laguncularia racemosa (L.) C.F. Gaertn., Rhizophora mangle L., Ceriops tagal (Perr.) C.B. Rob., Rhizophora mucronata Lam., Sonneratia apetala Buch.-Ham, and Sonneratia caseolaris (L.) Engl.. Additional comparative mangrove species reported by B. Leng & K.-F. Cao51 included Bruguiera sexangula (Lour.) Poir., Bruguiera sexangula var. rhynchopetala W.C. Ko, Excoecaria agallocha L., Rhizophora apiculata Blume, Sonneratia alba Sm., and Xylocarpus granatum J. Koenig.For Fig. A) Table . These r2 can provide insight into the water use requirement for that uptake of carbon64. We used leaf-level instantaneous water use efficiency  for determining mangrove net primary productivity (NPP) values from A. germinans, L. racemosa, and R. mangle forest components, we used light-saturated, leaf-level 2 (mol H2O)−1 [± 1 SE], respectively, from mangrove saplings and small trees of south Florida65. 20 , NPP becomes 0.96 kg C m−2 for 2004 and 0.85 kg C m−2 for January to August of 2005  and was nearly identical for 2005 .This predictive relationship was validated in two independent ways. First, for one of the calibration sites , we modeled 69. The relationships of predicted NPP versus p = 0.643).Second, we wanted to determine whether 70; and reveals unit conversions.We reviewed data from 26 published records that report mangrove NPP, or enough data to estimate NPP, from 71 study sites located in the Florida-Caribbean Region (25 sites) and Asia–Pacific Region (46 sites) Table . Table Spredicted curve in Fig. 2 grid in MOD16, and 2) was combined with the average of up to 8 additional neighboring cells to provide comparative 2 for each location from 2000 to 2013 to compare sensitivity among suites of the specific MODIS16-A3 cells selected over land. When neighboring cells were completely over water, they were excluded since component mangrove forest 2O year−1 . Therefore, we use We then convert NPP to mentclass2pt{minim48, and this ratio difference was assumed to represent net water use strategy affecting differences by the dominant vegetation between ecosystem types. We were also mindful that salinity reductions can affect 20. Standard deviation was 32% of mean The average ves Fig. 20. Stand2O year−1), temperate deciduous forest (131.76 mm H2O year−1), tropical grassland (110.77 mm H2O year−1), temperate grassland (46.48 mm H2O year−1), temperate coniferous forest (54.96 mm H2O year−1), desert (22.99 mm H2O year−1), and Mediterranean shrubland (12.08 mm H2O year−1). To convert potential water use differences to kL H2O ha−1 year−1 (as presented in the abstract), the following calculation is used :Table Elaeis guineensis Jacq.) plantations, 32 to 70%33, for a water use difference of 1170 and 3160 kL H2O ha−1 year−1, respectively, relative to annual global mangrove 31 to attain potential water use differences of 21.6 to 58.4 GL H2O year−1 from avoided conversion of mangrove to oil palm in this region.For comparisons made to mature (> 12 years) oil palm :In order to determine how much global mangrove area is adjacent to each ecoregion, we conducted a cross-walk between terrestrial ecoregionsas Table . In othe41. In the model, an individual tree is described by four geometric measures, including stem radius, stem height, crown radius and root radius; attributing functional relevance in terms of resource uptake. Aiming to maximize resource uptake, new biomass is allocated to increase these measures in an optimal but not constant proportion. Water uptake of the tree is driven by the water potential gradient between the soil and the leaves. Thus, porewater salinity is part of what determines the water availability for plants.The BETTINA model simulates the growth of mangrove trees as a response to above- and below-ground resources, i.e. light and water42. We can show that the ratio of the actual transpiration to the potential transpiration decreases with increasing salinity; plants use less water. Potential transpiration was the transpiration of a given tree without a simulated reduction in water availability due to porewater salinity. These parameter details are presented graphically for mangroves , food production. If a net kilogram of a food is grown on a hectare of land where water is abundant and that kilogram of food requires 400 mm of water to be produced, the export of that food to an area of low water availability provides an ecosystem service in the amount of 1 kg of food, plus 400 mm of “virtual” water not actually needed at the destination but used at the source. This water is defined as the product’s “virtual water content”dbh for both terrestrial and mangrove trees, were not normally distributed. We used a Kruskal–Wallis ANOVA based on ranks, and the Dunn’s Method for difference tests. Individual tree water use by dbh for both terrestrial and mangrove trees were determined using linear regression, mostly applied to mean values. For a couple of mangrove studies, only median values were extractable from minimum and maximum values. Likewise, all other data relationships were best fit with linear models, including the calibration curves between Data for leaf-level Supplementary Information.


Explorations of indefinite nanocavities have attracted surging interest in the past few years as such cavities enable light confinement to exceptionally small dimensions, relying on the hyperbolic dispersion of their consisting medium. Here, we propose and study indefinite graphene nanocavities, which support ultra-compressed mode volumes with confinement factors up to  Optical nanocavities provide a indispensable platform to study various sorts of light–matter interactions, including light emission ,2,3, optRecently, hyperbolic metamaterials (HMMs) and metasurfaces ,21,22,23In this article, we demonstrated a novel type of graphene indefinite nanocavity consisting of alternating graphene and silicon layers. The hyperbolic dispersion of such graphene-silicon HMM allows for propagating waves with large wave vectors and high effective indexes. This leads to ultra-confined modes with volume confinement factors up to troscopy , biosenstroscopy ,48, and troscopy ,50, overd = 9 nm and its relative permittivity is set to be T is temperature; e is elementary electric charge; and ℏ is the reduced Planck constant. In this research, T = 300 K, The scheme for an indefinite graphene nanocavity is presented in ectively :(1)σintrx-y plane [z coskz-y plane : ϵx=ϵy=ϵx, y, and z-directions, respectively. We assume that Cavity modes in the graphene indefinite cavity and the iso-frequency contour of HMM are both shown in ties see b and lx=z reach almost 300. However, the quantum effect should then be taken into consideration if the distance between neighbouring graphene sheets goes into the sub-nanometer regime.The spatial distributions of the electric field for a 2D indefinite cavity see b are preon modes ,54. Inderelation :(5)kx2/ϵAs a next step, we perform simulations of a 2D nanocavity array see b with 50As a last step, we study the mode volume of graphene indefinite cavities by means of quasi-normal mode theory . Figure  Formula . In the 0−5∼10−4  and simi0−5∼10−4 . Althoug to Ref. ), the obcavities . The extcavities .In conclusion, we propose and demonstrate graphene indefinite nanocavities with ultra-compressed mode volumes and extraordinary optical confinement at mid-infrared and THz spectral regimes. The normalized mode volume can reach approximately


When a twist angleis applied between two layered materials (LMs),the registry of the layers and the associated change in their functionalproperties are spatially modulated, and a moiré superlatticearises. Several works explored the optical, electric, and electromechanicalmoiré-dependent properties of such twisted LMs but, to thebest of our knowledge, no direct visualization and quantificationof van der Waals (vdW) interlayer interactions has been presented,so far. Here, we use tapping mode atomic force microscopy phase-imagingto probe the spatial modulation of the vdW potential in twisted hexagonalboron nitride. We find a moiré superlattice in the phase channelonly when noncontact (long-range) forces are probed, revealing themodulation of the vdW potential at the sample surface, following ABand BA stacking domains. The creation of scalable electrostatic domains,modulating the vdW potential at the interface with the environmentby means of layer twisting, could be used for local adhesion engineeringand surface functionalization by affecting the deposition of moleculesor nanoparticles. For example, scattering near-field optical microscopy (s-SNOM)uncovered the variation of the in-plane optical phonon frequenciesfor different stacking in the moiré superlattice of a twistedhBN (t-hBN).24 Piezo force microscopy revealedstrain gradients along moiré stacking domain boundaries, throughpiezoelectric coupling to an electric field applied between atomicforce microscope (AFM) tip and hBN sample.19 Electrostatic force microscopy (EFM) and kelvin probe force microscopy(KPFM) were performed on t-hBN (1–20L-BN on top of a thicker>30L flake20), addressing the existenceof two opposite permanent out-of-plane polarizations emerging fromthe moiré pattern.22 However, the impact of moirésuperlattices on local vdW interactions in twisted LMs has not beenexplored so far, to the best of our knowledge.hBN isa wide-bandgap (∼6 eV)25 In tapping mode AFM, the sine of the phase channelis proportional to the energy dissipated in the tip–sampleinteraction.33 This depends on the tip–sample distance in a way that isspecific to the probed force,26 allowingnoncontact (or long-range) vdW forces to be distinguished from otherlocal interactions, such as capillary, surface energy hysteresis,and viscoelasticity forces.34 By tuning the phasechannel to the local vdW dissipation, we quantify the dissipated energyand visualize the modulated vdW potential at the top layer–airinterface, resulting from the t-hBN moiré superlattices. Weprovide a physical interpretation of the nanoscale origin of the vdWdissipation contrast based on analysis of the tip–sample interaction,showing that the Debye force between the neutral tip and interlayerpermanent electric dipoles is the principal source of the imagingcontrast. We explain this Debye interaction for the two main stackingdomains involved in the t-hBN structure, i.e., AB and BA.Here, we investigatethe moiré interlayer modulation ofthe vdW potential of t-hBN by using tapping mode AFM phase-imaging,a widely used tool for nanoscale force characterization.AFMphase imaging is a simpler and more reliable way to visualizemoiré patterns in t-LMs. Unlike electric force microscopy techniques,such as EFM and KPFM, it does not require any specific sample or tipbiasing. This simplifies sample preparation and reduces the possibilityof damage.35 to tribology,36 both at the macroscopic and microscopic level.36 We find a modulation on the vdW potential atthe sample surface in t-hBN and quantify the related energy dissipation,after calibration of the AFM parameters. The fact that such potentialcan be patterned in scalable domains engineered by twisting providesa tool for functionalization of surfaces. Locally engineered adhesion,periodically spaced anchoring sites for molecules and nanoparticlesdeposition, and electrostatically patterned substrates for controlledcells stimulation are a few applications that could benefit from ourfindings.Weak electrostatic potentials at the interface withthe environmentare at the origin of numerous phenomena in fields ranging from fluiddynamics2, as describedin twist, is defined as the angle between the lattice vectors of the topand bottom hBN flakes.38 We control this by first identifyingneighboring flakes cleaved from the same bulk hBN, as determined byinspection of the relative orientation of their faceted edges, andthen picking one flake up using the other.37 θtwist may be tailored by rotating the transferstage between picking up the first and second flakes. The accuracyof θtwist is limited by the resolution and wobbleof the transfer stage (±0.01° and ±0.008°), monitoredby tracking the relative orientations of the faceted edges of topand bottom hBN using optical microscopy and AFM.We use a t-hBN sample consistingof a 2 nm (∼5L) top hBNlayer and an 8 nm (∼20L) bottom hBN, k ∼ 0.7 N·m–1, v ∼ 150 kHz), Scanasyst Air HR , 240AC-NG ,and ASYELEC.01-R2 cantilevers. To avoid damaging the tips, calibration procedures areperformed at the end of the experiments. The deflection sensitivityis obtained by recording 10 force–distance curves on mica (withoutchanging the laser spot position onto the cantilever) and calculatingthe average inverse slope of the contact region. The cantilevers springconstant is then obtained using the standard thermal tune method.39 All the AFM images are obtained in tapping modeat ∼0.5–1 Hz scan rate. These are all postprocessedusing Gwyddion.40 Phase imaging theory25 states that phase contrast is inversely relatedto the cantilever spring constant. This points to the need of a softcantilever. Thus, phase images are taken with k ∼1 N·m–1. No phase moiré contrast isobtained for k ∼ 30 N·m–1 . KPFM maps are also takenwith soft cantilevers with the sample holder connected to ground.AFM/phase/KPFM measurements are taken at ∼25 °C (RH∼ 40%), using a Multimode 8 (Bruker) AFM microscope, with ScanasystFluid  to 0°, forcing the attractive regime (AR)to correspond to negative phase values and the repulsive regime (RR)to positive ones. Instead, Asylum Research AFM microscopes set thefree phase to 90°, with AR (RR) phase values higher (lower) thanthis. AFM tapping mode force spectroscopy can be performed to verifythese definitions. All phase values in the rest of this paper arerenormalized from Bruker to Asylum Research convention.AFM phase values tend to follow different conventions dependingon the AFM microscope brand. Ref  summariz42 appliedto the dynamics of the AFM cantilever. Adrive sin(ωt) , the tiposcillation corresponds to a delayed sinusoidal motion, i.e., A sin(ωt – φ) . Adrive is related to the free oscillation amplitude A0  as Q the quality factorof the cantilever resonance.42We simultaneouslyrecord topography and phase channels in tappingmode AFM. The phase signal can be described in terms of a forced anddamped harmonic oscillator model22 In 25 The AR and RR concepts in AFM phase-imaging can be describedin terms of the nonlinear AFM cantilever dynamics.43 Assuming the tip–sample interaction to be describedby a Lennard-Jones force curve, the tip experiences, over the oscillationperiod T, either attractive (force F < 0) or repulsive (F > 0) interactions, dependingon the instantaneous tip–sample distance  dt. Approachingthe tip to the sample, i.e., decreasing the distance zc of the cantilever chip from the sample; 2) RR,for zc ≪ A0, F̅ > 0. Thus, inAR(RR) the AFM cantilever experiences an average negative (positive)deflection, 43 It is possible to move from one to the otherby modifying A0 and A. In our case, to visualizethe moiré superlattice via the phase channel, it is necessaryto operate in AR.An average force, e sample2b, two pA while keeping constant A0, the topography is unaltered  and scan size, and on a differentt-hBN; see To test the generalapplicability of our methodology, we imagemoiré superlattices in different regions of the same sample,with different cantilevers  and sin φ are linked by:26Fts is the total tip–sample interactionand dz/dt the tip speed along the z-axisat time t. Q(<10) environment.31 In this case, contributions from higher cantilevermodes should be considered and a sinusoidal oscillation cannot beaccepted.31 Our Q ∼150 (derived by fitting the resonance curve)25 allows us to use Further insightscan be obtained by introducing the local dissipationenergy of the tip–sample interaction. As discussed in refs , 44, 45,k, Q, A, and A0 is done. Once a phase image is acquired, a dissipation map can bethen reconstructedthrough 27 In termsof local interactions at the nanoscale,3 main dissipative mechanisms at the tip–sample junction canbe considered:26 (1) long-range  vdW-like forces; (2) short-rangesurface energy hysteresis; and (3) short-range viscoelasticity. Ultimately,all are characterized by a different force expression when the tipapproaches (forward movement) or withdraws (backward movement) to/fromthe sample surface, resulting in what is usually called a force–distancehysteretic behavior.48 These differ fortheir dependence on the minimum tip–sample distance, dmin, that can be controlled by adjusting A/A0.28 Plotting the dissipation energy as a function of A/A0 enables the identification of themain dissipation channel responsible for the moiré superlatticecontrast of A0 constant and decreasing, ateach image, A; see Ediss as a function of A/A0. The trend in 28 Since surface energy hysteresisand viscoelasticity emerge from a tip–sample interaction typicalof the repulsive regime, we consider them negligible in the operatingattractive regime.Dissipation in AFM measurementscan have different origins.34 In this case, the contributionof capillary forces would result from the presence of an uncontrolledwater layer on the sample, due to ambient humidity .Even though the energydissipation trend excludes short-range forcesas the moiré imaging contrast mechanism, besides vdW forces,capillary forces could, in-principle, contribute to a similar dissipationbehavior.n = 300 force–distance curves are collected at the center ofboth “dark” and “bright” domains of apreviously acquired phase map, k.In order to distinguish betweencapillary and vdW forces as dissipativemechanisms, we perform AFM force-spectroscopy. In this case, the tipis not oscillated, but approached and withdrawn to/from the sample,while recording the deflection of the cantilever. As discussed in F–d curvefor an AB domain  that can be ascribed to the formationof a capillary bridge between tip and sample.51 At such distances,attractive vdW forces can affect the tip–sample interaction.4848 At this point, the tip contacts the sample enteringa repulsive regime, with a force increasing until set-point deflectionis reached.When the gradient of the total attractiveforce overcomes the springconstant of the cantilever, a sudden collapse of the tip toward thesample takes place, caused by the so-called snap-in mechanical instability.don to doff, where doff corresponds to the distance of rupture of the capillarybridge;51 the other from the adhesion point  to the end of thesnap-out48 . In this region, the nanoscalewater bridge is not broken yet (since d < doff); therefore, any dissipative contributiononly results from vdW forces; see Retracting the tip from the sample (red), in RR,two separatedregions can be distinguished, where approach and withdraw curves arenot overlapping, signature of hysteresis. In such regions, the dissipationcan be calculated as the enclosed area between the approach and withdrawcurves. One hysteresis region extends from A0 is ∼16 nm, A0) is spanned,covering both vdW and capillary interaction regions. These results,for both AB and BA stacking . Debye forces56 affect a neutral molecule interacting with a polar molecule.Keesom forces57 emerge from the interactionbetween two polar molecules. All have an attractive energy UvdW ∝ 1/d6, where d is the distance between the two parts.52VdW forces emerge from the quantum mechanical interaction betweenpermanent or transient electric dipoles between molecules,twist between thetwo crystal structures. Hence, our moiré phase-image contrastemerges from the Debye dissipative vdW interaction between tip andsample.Refs −22 suggeEdissvdW) are in The values of dissipation energy related only to thevdW contribution emerging fromthe moiré superlattice. We quantified the vdW interactionson both AB and BA regions, through the proportionality between phasesignal and dissipative tip–sample forces, indicating the BAregions as the most dissipative. We discussed the origin of this nanoscalevdW dissipation and related the interaction between tip and interlayerelectric dipoles to a Debye vdW force.We observed the spatial modulation of the vdW potential inducedby the moiré superlattice of t-hBN and t-WSe2),59 while maintaining a nm resolution. Nanopatterning is an importantand diverse research topic continuously enriched by different approaches.60 Of particular relevance is high-spatial resolutioncombined with large scale patterning . LM twi63 onto Si + 90 nm SiO2 by micromechanicalcleavage (MC). In order to control θtwist, eitherlarge flakes (>50 μm) selectively torn during transfer37 or neighboring hBN flakes cleaved from the samebulk crystal during MC20 are identifiedby studying the orientation of their faceted edges using optical microscopy.64 t-hBN samples with controlled interlayer rotation are then fabricated using polycarbonate (PC) stamps.65 First, a PC film on polydimethylsiloxane (PDMS)is brought into contact with the substrate with hBN flakes at 40 °Cusing a micromanipulator, so that the contact front between stampand substrate covers part of one flake or one of two adjacent flakesexfoliated from the same crystal on the tape. Stamps are then retracted,and the material in contact with the PC is picked up from the substrate.After picking up the first flake, a controlled θtwist  can be applied by rotating the sample stage, beforethe flake on PC is aligned to the second one and brought into contactat 40 °C. The stamp is then retracted and the resulting t-hBNis picked up by PC. t-hBN is then transferred onto Si + 285 nm SiO2 at 180 °C, before the PC residue is removed by immersionin chloroform and then ethanol for 30 min. While Si + 90 nm SiO2 is used to facilitate the identification of hBN flakes,66 Si + 285 nm SiO2 is chosen for furthercharacterization, such as gate dependent electrical measurements.t-hBNsamples are prepared by first exfoliating bulk hBN (B-hBN) crystals,grown at high pressure and temperature in a barium boron nitride solvent,N, of hBN by measuringthe position of the C mode,68 Pos(C). For N > 5, the shift in Pos(C), ΔPos(C), can be smallerthan the spectral resolution  ∼ 0.15 cm–1 between N = 10 and N = 11 vs a resolution of ∼0.6 cm–1, correspondingto the wavenumber interval between detector pixels for the combinationof diffraction grating and CCD used in the measurements). However,as the Raman peaks are represented by multiple data points even forspectrally narrow ULF modes ,it is possible to extract their position with accuracy exceeding thespectral resolution of the experimental setup, via spectral fitting.In general, the error of the peak position extracted via fitting isdetermined by the fitting error, statistical errors arising from spatialvariation, CCD noise and errors associated with the registry of pixelsrelative to the position of peaks.Ultralow frequency (ULF) Raman spectroscopy may be used in orderto estimate the number of layers, N > 15 l-hBN using aHoribaLabRAM Evolution at 514 nm, with an 1800 l/mm grating and volume Braggfilters with a ∼5 cm–1 cutoff frequency anda 100× objective (NA: 0.9). –1, expected to be negligible compared to statisticalerrors and pixel registry. In order to evaluate the error due to detectornoise, lateral variations across the sample surface and other statisticalvariations, a series of spectra are acquired at different positionson the same hBN flake. A histogram of Pos(C), from 64 different locationsis shown in –1, witha standard deviation ∼0.05 cm–1 and a variationrange ∼0.25 cm–1, which compares favorablywith the spectral resolution of the system (∼0.6 cm–1).In order to extract the errorof our measurements for Pos(C) dueto fitting and statistical variations, a series of ULF Raman spectraare measured on –1, such that the C peak is depicted by <10 pixels, the registryof the CCD pixels is expected to contribute an additional error. Toevaluate this, Pos(C) is extracted by fitting spectra acquired fromthe same position of a N > 15 l-hBN flake,with grating position offset from −3 to +3 cm–1 in 0.5 cm–1 increments, As the spectral resolution of the system used is comparabletothe full width half-maximum, FWHM(C) ∼ 1.1 cm–1, with a variation range ∼0.27 cm–1, less than the spectral resolution of the system. The values ofthe main fitting errors are in A range of grating registriesare used so that Pos(C) is at thecenter of two adjacent pixels or between them. The standard deviationof Pos(C), extracted from Lorentzian fitting, is ∼0.06 cmN,68 for the hBN flakes used herethe change in Pos(C) ∼ 0.15 cm–1 between N = 10 and 11 is comparable with the total fitting error∼ ± 0.15, allowing N to be determined±1 layer for N < 11.As the relative change of Pos(C) reduces with increasing 2g mode ∼1366 cm–1.71 The errors associated with fitting it to a Lorentzian,from statistical variation, and pixel registry are summarized in 2g mode relative to C.2. Pos(C) = 52.5± 0.14 cm–1 for the 8 nm flake, t-hBN and B-hBN,with FWHM(C) = 1 ± 0.2 cm–1, whereas Pos(C)= 50.1 ± 0.14 cm–1 for the 2 nm flake. Pos(C)can be used to determine N, for N > 2 as73c the speed of lightin cm s –1, μ = 6.9 × 10–27 kg Å–2 the mass of one layer per unit areaand α⊥ the interlayer coupling.73–1. Fromthis we can derive α⊥ = 1.69 × 1018 N m–3. We then use it in N = 5± 1 for the 2 nm thick flake and N > 10 forthe 8 nm one. 2g) = 1366 ± 0.2 cm–1 with FWHM(E2g) = 8.1 ± 0.2 cm–1 for 8 nm, t-hBN, and B-hBN, whereas FWHM(E2g) = 9.8 ±0.2 cm–1 for the 2 nm flake. The peak broadening∼1.7 cm–1 in the 2 nm flake can be attributedto strain variations within the laser spot, as thinner flakes conformmore closely to the roughness of the underlying SiO2. Thisis also confirmed by the higher RMS roughness of the 2 nm flake (∼0.6nm) as measured by AFM, compared to ∼ 0.2 nm for the 8 nm flakeand t-hBN.In B-hBN, 75 a higher (lower) surface potential corresponds to an upward (downward)polarization, a feature of BA (AB) stacking domains in t-hBN.22 Hence, bright (dark) phase domains  corresp2pz orbitals, AA has a zeronet electric dipole . Thephase image shows a moiré pattern over the whole 8 μm× 8 μm scan size.twist  than the one discussedin the main text. While the topography maps  of the tip from the consideredinterlayers dipoles.26 Therefore, a thickertop-hBN will necessarily increase dmax and dmin, decreasing Ediss. The thickness of the top layer can affect the formationof the domains itself.20 Such effect wouldcomplicate the possibility to set a reference for experimentally derivingthe trend of the dissipation energy with respect to the increasingdistance due to a thicker top layer.The dissipated energy doesnot only depend on the tip–sampledistance, but also on the hysteresis coefficient α. The physicalorigin of this parameter is not unique, since several phenomena cancontribute to increase the adhesion in the withdraw curves. In ref , an exteSee 78Long-range dissipative forcesact upon the tip in the noncontact attractive regime, and are typicallyrepresented by a vdW-like distance-dependent expressionHeff, represents themagnitude of the vdW interactionbetweenan AFM tip with radius R and the sample at a distance d.78 α ≥ 0 distinguishesbetween forward and backward movements of the tip with respect tothe sample during one oscillation.78 Ifthe two tip–sample regimes are equal in magnitude (α= 0), a conservative interaction arises providing no dissipation.The existence of a magnitude difference (α > 0), instead,yieldsa dissipation ∝ α.28Heff in effective parameter, takinginto account all 3 main interactions between tip and sample-substrate.Since thicknesses are in the few nm range, Heff can be identified for tip-ambient-top hBN, tip-ambient-bottomhBN, and tip-ambient-substrate systems.80In See In 2/1L-WSe2 on Si + 285 nm SiO2. While the morphology (rphology 16a does rphology 16b.F–d curves (out of 300) measuredon the center of both BA (twist = 0.2°) sample of –1 in –1 for  both BA 17a and A both BA 17b domai


We aim to reveal the impact of socio-economic circumstances on early childhood nutrition, which affects the physical development and overall survival of this susceptible population group. Bone collagen samples from 176 individuals between the fetal and the 7–15 age categories from four urban/rural South-Estonian cemeteries were cross-sectionally analyzed via EA-IRMS  for δ13C and δ15N. Results suggest that South-Estonian children had a staple terrestrial C3 diet integrated with animal proteins. Significant divergences were observed between urban and rural sites and slight variation occurred among rural subgroups, possibly resulting from a wider food choice available in towns, different consumption of C4 foods, and/or secular changes. This study provides the first data regarding infant feeding practices in medieval and early modern Livonia. These practices were similar among the different contexts, indicating comparable cultural traditions in child rearing. Breastfeeding was likely practiced for 1–2 years, with supplementary foods introduced around 1 year of age. The weaning process was probably concluded around the age of 3. The δ13C and δ15N values of older children are comparable to those of the adults from the same sites, indicating their diets became similar after weaning, when they started working and obtained a more mature social status.Studying infant diet and feeding practices through stable isotope analysis provides direct insight into the life and health of vulnerable population groups in the past. Although the general diet in medieval and early modern Livonia has been reconstructed from written sources, little is known about childhood diet during this tumultuous period of Eastern European history. This study presents a comparative investigation of the staple non-adult diet in urban/rural communities during the 13 From a socio-cultural point of view, understanding these practices helps reveal the complex interactions between childcare in different socio-economic contexts, treatment of minorities, and local traditions in dietary selection , 2. On tl stress –10. The l stress , 8.The study of breastfeeding and weaning patterns through stable isotope analysis has become an increasingly important line of research in bioarchaeology –15. StabThe study of infant diet and feeding practices has received little attention in the Baltics. Biomolecular analyses of weaning patterns were published for Swedish and Latvian prehistoric populations –24. The th-17th centuries AD), were analyzed to answer the following research questions:The aim of this paper is to investigate non-adult diet in medieval and early modern Livonia, a historic region that mostly corresponds to present-day Estonia and Latvia, through carbon and nitrogen stable isotope analysis, with a special focus on early childhood breastfeeding and weaning practices. Bone samples from 176 individuals between the fetal and the 7–15 age ranges, recovered from four urban and rural cemeteries from Southern-Estonia  and nitrogen (δ15N) stable isotope analyses have been frequently used to study breastfeeding and weaning patterns in archeological samples. The basic principle is that a breastfeeding infant is consuming the mother’s tissues, showing a rise in δ15N values of about one trophic level (2–3‰) compared to the mother [15N values dropping to a level similar to those of the mother [13C values, with a trophic level shift of around 1‰ [Carbon  and early modern (16th–18th centuries AD) Livonia. Historical records testify to a series of socio-cultural processes such as foreign conquests and supplanting of local politico-economic powers by several non-local stakeholder groups , and development of urban settlements with typically “European” features  and the local lower class  has been reported among the urban populations [Southern-Estonia formed a central part of medieval ..th–16th We can take a glance into the general dietary habits of medieval and early modern Estonian populations through a plethora of archaeological and historical sources –49. The The medieval city dweller typically obtained the necessary food from the household, since the preparation of bread, mead, beer, and meat/fish was mainly carried out at home . Small gUnfortunately, few historical studies regarding childhood in medieval and early modern Livonia exist, and no specific information concerning the duration of breastfeeding or weaning is available , 53. Thith century [th/early 17th centuries. The burials were mostly simple graves with no coffin or grave goods, and the dead were buried with their heads turned westwards. The proportion of non-adults in St Jacob’s cemetery  is located southeast of Tartu. Rescue excavations were carried out in 2011, unearthing the ancient settlement and the village cemetery [th and to the 17th century. However, one burial contained a 14th–16th century needle-sheath, suggesting that the cemetery was already in use in the medieval period [Three village cemeteries were selected from the rural surroundings of Tartu: Lohkva, Mäletjärve, and Makita. The medieval/early modern settlement of Lohkva , but most graves are from the 16th century [The village cemetery of Mäletjärve, excavated in 1984, is located southeast of Tartu. It was in use between the 14 century . Due to th and the 17th centuries [th–15th and the 15th–17th centuries AD. Non-adults constituted 60.8% of the recovered burials  date the cemetery between the 13enturies . The burenturies . A hundr burials . The demfetus (under 28 gestation weeks), perinate I , perinate II , and neonate (from 42 weeks to around a month old). The term “perinate” defines the individuals who died around the time of birth, with the perinate II category including possible full term and post-term births. Those aged younger than 37 weeks (perinate I) were possibly either aborted (death in utero), stillborn (death after 28 weeks of gestation), or dead immediately after birth, in the case of pre-terms or babies that were small for gestational age [Ethical approval was granted by the Department of Archaeology, Institute of History and Archaeology, University of Tartu (UT). According to the research procedures in place at the institute, no permits were required for the described study, which complied with all relevant regulations. The specimens studied are stored among the UT Institute of History and Archaeology archaeological collections, Tartu, Estonia. A detailed anthropological study was carried out on the non-adult individuals. Age at death was estimated using standards based on tooth formation and eruption –67, boneonal age .A hundred and seventy-six non-adults were sampled from the urban and rural contexts . ExceptiThe principal limitation of this dataset is represented by the age-at-death composition of the urban versus the rural samples. The funerary record of the urban sample is skewed towards the youngest individuals, while the rural samples tend to represent older children, lowering the informative power of the statistical comparisons between sites. This is an unfortunate consequence of performing cross-sectional studies on archaeological populations, which mostly make a biased proxy of the original human communities . IntrinsRegarding our dataset, the large number of perinates from St Jacob’s cemetery come from mass graves outside of the cemetery walls specifically dedicated to perinates, indicating the choice of a specific burial area for particular population groups . The samThe samples were processed at the Archemy Laboratory, UT. Collagen extraction was performed using the modified Longin method . Bone sa13C) and nitrogen (δ15N) isotopic compositions were measured using an automated carbon and nitrogen elemental analyzer isotope ratio mass spectrometer (EA-IRMS) in the Department of Geology, UT. Bone collagen samples were weighed into tin capsules (~1.0–1.2 mg) and combusted in a Thermo FlashTM HT EA with the introduction of separated N2 and CO2 to a Delta V plus via a ConFlo IV interface. The analysis was run in duplicates , and the final values averaged. The data were calibrated against international standards from IAEA . The results are expressed using the delta notation in parts per thousand (per mil or ‰) [Stable carbon (δil or ‰)  relativeAs references, the mean values of the adult population from these cemeteries are used. The adult individuals (n = 25) were sampled and analyzed by the first author. The local and temporally coeval faunal stable isotope results were provided by Aguraiuja-Lätti , and Malve .U test and a Kruskall–Wallis test were used for comparison of two or more than two samples (ex. among cemeteries/age ranges). Possible statistical outliers were identified using the IQR (Tukey’s Interquartile Range) criterion.Statistical analysis was performed using PAST 4.09 software . A ShapiOut of all 176 samples, 140 (79.55%) presented sufficient collagen preservation according to standard quality control methods: C:N atomic ratio between 3.07 and 3.59 (mean 3.31 ± 0.09); weight percent collagen yield ≥1%; weight percent carbon (%C) >13%; weight percent nitrogen (%N) >4% –87. The 13C values of the whole sample of 140 non-adults range from -22.42‰ to -19.23‰ (mean ± sd = -21.10 ± 0.50‰), while the δ15N values range from 7.72‰ to 16.05‰ (mean ± sd = 11.79 ± 1.72‰).The δ13C values of the urban non-adults range from -22.19‰ to -20.26‰ (mean ± sd = -21.06 ± 0.43‰) and the δ15N values range from 9.96‰ to 16.05‰ (mean ± sd = 12.82 ± 1.32‰). For the rural non-adults, the δ13C results range from -22.42‰ to -19.23‰ (mean ± sd = -21.15 ± 0.56‰) and the δ15N values range from 7.72‰ to 15.63‰ (mean ± sd = 10.82 ± 1.48‰). Comparing the contexts, the nitrogen values show a notable variability reflecting the differences in the age range distribution between sites, as well as the presence of outliers , but a significant difference was observed for the δ15N values .The isotopic values for the urban sample were higher compared to the rural sample by 0.09‰ for δfor δ15N . Since t15N and δ13C values between children from different sites. No significant difference between sample medians was observed for δ13C. The Mann–Whitney tests between group pairs showed a significant difference between Mäletjärve and the other sites , Mäletjärve (2.04‰), and Makita (2.11‰), confirming the results of the Mann-Whitney test on the urban and rural whole samples.For δ13C values of the urban women range from -21.47‰ to -20.48‰ (mean ± sd = -20.02 ± 0.29‰) and the δ15N values range from 10.01‰ to 12.67‰ (mean ± sd = 11.18 ± 0.80‰). The δ13C values of the rural women range from -22.39‰ to -20.63‰ (mean ± sd = -21.66 ± 0.45‰) and the δ15N values range from 10.39‰ to 12.31‰ (mean ± sd = 11.17 ± 0.63‰)  reach the expected ca. 2–3‰ increase indicative of a breastfeeding signal, touching the highest values of ca. 3.7‰ when urban infants and women are compared . This suggests an overall similar diet for children living in different contexts. The small difference in δcontexts , 49. Fros millet . Althougs millet . Althoug13C values in non-adults (mean ± sd = -21.10 ± 0.50‰) are comparable with the adult values (mean ± sd = -21.31 ± 0.47‰), especially with regard to those in the 1–7 and 7–15 age categories  from which it is inferable that children were put to work quite early, already at 5–6 years of age, and given simpler tasks in the household. For instance, one of the most common tasks of child laborers was herding. The common principle was that hard workers could not be under-nourished and required more food, so it is reasonable to assume similar diets for children and adult workers. Yet, it was also stated that children were usually given less meat [15N values of older children and adolescents  are similar to the adult values (δ15N mean ± sd = 11.26 ± 0.69‰), although they show greater variability due to the wide age range selection. As can be inferred from the comparison with the animal baseline, these results imply that adults and children in South-Estonia likely had wide access to animal products, possibly relying on small amounts of higher trophic level proteins, such as fish. In particular, human individuals show δ15N values elevated by 4.94‰ (δ13C = 0.28‰) compared to cattle, by 5.81‰ (δ13C = 0.45‰) compared to horses, by 5.33‰ (δ13C = 0.00‰) related to ovicaprines, and between 0.41 and 2.93‰ (δ13C between 0.28 and 0.86‰) in respect to omnivores, with the dog showing the greatest similarities to humans . These values, together with the similarity with omnivores, therefore seem to show a trophic effect that is compatible with consumption of protein sources not only from plants, but also of animal origin. This would also be in line with historical sources reporting a consistent presence of animal products in the diet of Estonians, including fish [15N values are generally above 10‰, fish consumption may explain higher human δ15N values [The δing fish –50, 90. N values , 90. SimN values , 91, 92;N values , 88, 90.N values , 89, 90.N values , 48. FurN values .15N values [The reliance on manured plants  may be undetectable from isotopic values. Using animal values as proxies, the herbivore nitrogen values tend to be quite low (between 4–6‰), while the increased human (and omnivore) values are compatible with a standard trophic effect –96, withN values .15N values (1.99‰ higher in the urban sample), a general trend detectable in both babies  and MK-74  show the lowest δ15Ν values of the whole sample, comparable to the rural female means  and one for nitrogen (LK-73), all among the rural samples. However, the δle means . Accordi15Ν values around 10.80‰, much lower than the other urban perinates . These individuals have been sampled for aDNA analyses in order to investigate kinship and search for evidence of infectious neonatal diseases.On the urban counterpart, three babies clearly stood out. Individuals JK-16, JK-17 and JK-18  exhibited δerinates . These b15Ν (11.20‰), while the other three lie within 1sd from the female means  with very low δ15Ν values [A somewhat unexpected result concerns the neonates, where the mean values are closer to the perinate I category (ca. 1–2‰) than to what is expected in a nursing context. However, there are only four neonates in this sample and they show quite varied values, which complicates the task of inferring possible patterns and influences their mean values. In particular, individual JK-50 (around 42 weeks) shows quite low δle means . Since tΝ values , 13, 36.15Ν values. In particular, individuals JK-43 (around 6 months), JK-59 (ca. 9 months) and LK-73 (about 1 year), show the highest δ15Ν values in the whole dataset   . This ma15Ν values ranging from 12.61‰ to 13.81‰ in the urban cemetery and from 9.83‰ to 15.63‰ in the rural areas, possibly reflecting different individual choices or local customs in the weaning practice. A striking outlier is MK-155 (ca. 4 years), showing high δ13C and δ15Ν values  . These values may be reflecting the dietary choices of the caregivers and, given the estimated age, perhaps the supplementary foods selected for weaning. However, the individual displayed porotic bone production on the orbits as well as abnormal porosity on the palate and around the infraorbital foramen. Bone production was also observed on the superior surface of the sphenoid lesser wings, on the pterygoid processes and mostly around the rotundus and ovale foramina. The individual also showed signs of endocranial lesions in the form of patchy capillary defects on the frontal bones above the orbital floor and on the greater wings of the sphenoid, as well as hair-on-end lesions on the temporal bone squama and inflammatory pitting on the occipital squama. Abnormal porosity of the postcranium was also detected. This pattern is compatible with a case of scurvy [13C, which in MK-155 is also associated with high δ15N. These recorded values therefore likely reflect physiological stress. Given the estimated age, the chosen weaning strategy may have played a role in the development or worsening of a systemic metabolic disease.The results of this cross-sectional analysis seem to reflect an effective breastfeeding signal, with a successive drop between 1 and 3 years of age. The children in this category show δ 13.12‰) . This chf scurvy , 102. Itth–17th century written sources, the families of German citizens and manor owners often hired Estonian wet-nurses and nannies. The 18th century publicist Peter Ernst Wilde wrote that a good wet-nurse must not be a drinker, wanton, or angry woman [th century, when more German women began to breastfeed their own children [The high variability encountered in this study may be explained by the historical context and social composition of this dataset, which included townspeople from different social statuses, foreign elites, and countryside migrants. People involved in childcare, including mothers and perhaps wet-nurses, likely came from different backgrounds. Hence, in addition to rural and urban food availability, childcare practices at the local or family level also ought to be considered as possible factors of diversity. For instance, according to the few available 16ry woman . Hiring ry woman . This trchildren . It is uth to early 18th centuries). The sampling of adult bone and dentine formed during childhood revealed that individuals had higher δ15N values during the first years of life as compared to their last years of life. The authors interpreted this as a result of an incorporated breastfeeding signature, and/or of an access to terrestrial or aquatic protein sources even at very young ages at Alytus. Skipitytė et al. [th–early 20th centuries) located near the Curonian Lagoon and the Baltic Sea in Lithuania. Their results are comparable to those of this study, and the paper reported that δ15N values were slightly elevated in non-adults as compared to adults, probably indicating nursing. The authors argue that breastfeeding was practiced until the age of 3–4 years, suggesting that early weaning was not the cause of death in these populations [15Ν values at younger ages to physiological stress in utero [Our results seem to place the duration of breastfeeding between 1 and 2 years, with the introduction of supplementary foods apparently starting around 1 year and ending close to or after 3 years of age. This is in line with isotopic data from other medieval/early modern contexts in Latvia and Lithuania –29. For ulations . The resin utero .15Ν values respectively 1.98‰ and 1.65‰ higher compared to rural individuals. This is likely reflecting the general dietary differences discussed above, especially considering that the female baseline follows a similar trend. The women in Tartu may have had access to a wider range of foodstuffs, including imported terrestrial and marine products. This would have reasonably resulted in higher δ15Ν values in their breastmilk, reflected in the overall values of their nursing and weaning children. It can also be hypothesized that 1–3 year old children were breastfed longer in urban areas compared to the countryside, but this cannot be confirmed in the absence of written sources.Observing the Infant 0–1 and Child 1–3 categories , a strikth century publicist August Wilhelm Hupel wrote that Estonian mothers let their babies crawl on the ground as soon as possible. This could have meant that lower class mothers needed their hands to participate in the housework and had to leave their babies free to crawl, but also that caregivers allowed them to be quite independent very early. Their older siblings (5–6 years old) used to look after the younger ones [Unfortunately, no historical data regarding the length of breastfeeding and the age of weaning in Estonia has been recovered. Entering the realm of speculation, indirect inferences can be attempted on the basis of written sources: for instance, the 18ger ones . Hence, ger ones . It cannger ones . Becominth–20th century sources mention that barley-water, breadcrumbs in milk, and probably gruel were used as supplementary foods for infants [The introduction of supplementary foods may have therefore started before one year of age, as reported for more recent contexts , and giv infants , 106. ThThis paper reports on the first isotopic data of non-adults in Southern Estonia, providing new insights into infant diet and feeding practices in medieval and early modern Livonia. Carbon and nitrogen stable isotope analyses were performed with a cross-sectional approach, allowing reconstruction of the staple diet, detection of dietary differences between urban and rural contexts, and identification of breastfeeding and weaning patterns.3 sources and was supplemented by animal proteins. Significant differences were observed between urban and rural cemeteries, possibly linked to a wider choice of different foodstuffs (including imported marine products) accessible in urban contexts like Tartu as compared to a countryside diet with less variety of foods. A significant difference was also observed among rural subgroups, possibly due to different consumption of small amounts of C4 foods, but also to secular dietary changes.The results of this study indicate that the diet of Livonian children was mainly from terrestrial COur results indicate that breastfeeding may have been practiced for 1–2 years, with the introduction of supplementary foods beginning around 1 year. The weaning process appears to have been concluded around 3 years of age, in line with isotopic data from other coeval Baltic samples. The isotopic values of older children and adolescents are comparable to the adult means, indicating that their diets were quite similar, although children may have consumed slightly fewer protein sources. This is supported by historical sources reporting that child labor began around 5–6 years of age, at which time youngsters likely began to consume an adult diet. For perinates and neonates, the isotopic values were discussed in the light of physiological stress and maternal nutrition.This work provides a large and unique sample of well-documented material, and thus contributes to the wider picture of non-adult bioarchaeology in Eastern Europe. Despite the limitations of cross-sectional studies, initial data regarding infant dietary practices in the Eastern Baltics is presented, forming a solid baseline to understand the non-survivors; it will support future high-resolution analyses, such as incremental dentine sampling, in order to reduce the amount of speculation in the interpretation of these results. The integration of incremental data from individual survivors will allow for the creation of a finer timescale and longitudinal approach, confirming or adjusting the results presented here. Finally, future inclusion of additional individuals from other coeval rural Southern Estonian sites will contribute to obtain a more complete and homogeneous sample with fair representation of all age ranges.S1 FileThe file includes the results of the Shapiro-Wilk tests of normality and Mann-Whitney tests, the raw data for the carbon and nitrogen isotope analyses performed on the non-adult samples from the four urban and rural cemeteries studied, and the descriptive statistical analysis of the urban and rural female individuals categorized by age.(DOCX)Click here for additional data file.


Following discussions with colleagues and peer reviewers on further work extending the model introduced in the manuscript, we have identified a technical issue in the way we mathematically phrased the phage-bacterium interaction. This has been corrected with an improved equation which restricts the maximum rate of phage predation at higher phage concentrations, exactly as our original frequency-dependent approach did, but with greater biological and mathematical clarity. Refitting the model with this improved “saturated phage predation” term does not substantially change our parameter values, figures, results, or conclusions.Volume 7, no. 2, e00135-22, 2022, All mentions of “frequency-dependent” phage predation should be replaced by “saturated” phage predation, and all mentions of “density-dependent” phage predation should be replaced by “linear” phage predation.P50 corresponds to the phage concentration at half-saturation, i.e., where the adsorption rate is equal to half the maximum.Page 6: The two equations used for the frequency-dependent interaction (equations 2 and 3) should be replaced by this single improved equation for saturated predation:Page 11: Figure 5 should appear as shown below, with the results from fitting the model with the improved “saturated phage predation” term.Page 12: Table 1 should appear as shown below.Pages 18 and 19: Equations 34 to 39 should read as follows:Page 19: The two equations used for the density-dependent interaction (equations 40 and 41) should be replaced by this single equation, to be consistent with the model equations above:Page 19: The two equations used for the frequency-dependent interaction (equations 42 and 43) should be replaced by this single improved equation for saturated predation:Page 22: Reference 36 should read as follows: Roach DR, Leung CY, Henry M, Morello E, Singh D, Di Santo JP, Weitz J S, Debarbieux L. 2017. Synergy between the host immune system and bacteriophage is essential for successful phage therapy against an acute respiratory pathogen. Cell Host Microbe 22:38–47.e4.