More stories

  • in

    Population genomics reveal distinct and diverging populations of An. minimus in Cambodia

    Population sampling and sequencingWe generated whole genome sequence data from 302 wild-caught individual An. minimus female mosquitoes collected from five different field sites in Cambodia using the Illumina HiSeq 2000 platform with 150 bp paired-end reads with a target coverage of 30X for each. Mosquito collections in Thmar Da, in Eastern Cambodia, were done in 2010. Longitudinal monthly collections were performed from February 2014 to January 2015 in two sites in each of the Preah Vihear, and Ratanakiri provinces. Quarterly collections were also done in 2016 in one site in Preah Vihear province, Cambodia.Variant discoveryThe methods for sequencing and variant calling closely follow those of the Anopheles gambiae 1000 Genomes project phase 2 (Ag1000G)27. Sequence reads were aligned to the An. minimus reference genome AminM128. We restricted our analysis to the largest 40 contigs, which cover 96.6% of the AminM1 reference genome, as many smaller-sized contigs can confound diversity and divergence calculations. We found that 138,161,075 (75.4%) of sites within these 40 largest contigs pass our site filters and thus were accessible to SNP calling. Of these, we discovered 38,000,285 segregating single nucleotide polymorphisms (SNPs) that passed all of our quality control filters of 55,307,039 total segregating SNPs. 13.4% of these SNPs were multiallelic, with 32,906,471 biallelic SNPs. There were 4,807,355 triallelic and 286,459 quadriallelic SNPs. A total of 100,160,790 sites were invariant. The median genome-wide coverage was 35X.Population structureA principal component analysis (PCA) over biallelic SNPs distributed over the genome of 302 individual field-collected mosquitoes showed that there is clear population structure of An. minimus in Cambodia. Samples collected from five sites in three provinces split into three distinct clusters; here, we report on 283 individuals that could be clearly assigned to these clusters (Fig. 1), excluding 9 anomalous and 10 outlying individuals. One cluster includes all samples from the western collection site Thmar Da and the northern collection sites in Preah Vihear province, with two further clusters with samples from Ratanakiri province in the northeast. These clusters split primarily along the first and second principal components. This was a surprising finding because this population structure did not correlate to the geographic sampling of these mosquitoes. Individuals collected from the western and northern sites cluster tightly together despite being hundreds of kilometers apart.Fig. 1: Population structure of An. minimus in Cambodia.The map indicates the five Cambodian collection sites. Principal component analysis (PCA) of whole genome sequences of 283 individual An. minimus s.s. collected in five villages in Cambodia shows that there is a distinct population structure and three populations. When performing the same PCA on a large X-chromosomal contig (KB664054), these individuals break into four populations: TD from the West, PV from the northern province in Preah Vihear, and RK1 and RK2, both collected in two sites in Ratanakiri province in the Northeast.Full size imageTo further explore this population structure, we performed the same PCA over individual contigs from different regions of the genome. Performing PCA over the largest X-chromosomal contig KB664054 resulted in a splitting of the western and northern samples, indicating four distinct populations of An. minimus in Cambodia (Fig. 1). PCA from this contig on a quickly evolving sex chromosome revealed more population structure compared to autosomal contigs. The populations defined by these PCA clusters are designated in this study as TD from Thmar Da, in Western Cambodia (n = 41), on the Thai-Cambodian border, PV from the Northern province Preah Vihear (n = 156), and the two distinct populations collected in Ratanakiri province in the Northeast, each including individuals collected at both collection sites, these are designated as populations RK1 (n = 58) and RK2 (n = 28).To confirm our results from PCA, we also performed an admixture analysis. We ran admixture on each of the largest 10 contigs for values of K between 2 and 6 (Supplemental Fig. 1). At K = 2, the samples from Northeastern Cambodia split from Northern and Western Cambodia samples. At K = 3, the two different groups in Ratanakiri were separated, consistent with the PCA results. At K = 4, there was some evidence for geographical population structure between the Western TD and Northern PV populations, but the admixture results did not perfectly correspond with geographic sampling, with some evidence of mixed ancestry in the PV samples. Again, this is consistent with the PCA groupings, with the generally weaker evidence of geographic population structure between TD and PV. A cross-validation analysis showed the lowest cross-validation error for K = 2 and K = 3, consistent with the strongest evidence for population structure between the two RK groups and other populations. Cross-validation error was higher at K = 4, consistent with the weaker differentiation between TD and PV. At no point was their an indication of admixture between RK1 and RK2.To examine population differentiation, we computed differences in allele frequencies between each population using Pairwise Fst. Pairwise Fst between all 4 populations over the largest contig, KB663610, representing 16% of the An. minimus genome, (Fig. 2) shows that differentiation was relatively low between populations of TD and PV with an average pairwise Fst of 0.003, while the difference between RK2 and the other three populations is tenfold higher, around 0.03. Pairwise Fst estimates comparing these populations over other large An. minimus contigs indicate a similar level of differentiation, with average pairwise Fst values over 0.03 (Supplementary Data 3). The two sympatric populations from the Ratanakiri collection sites are as differentiated from each other as they are from the northern and western clusters.Fig. 2: Population diversity and divergence.Nucleotide diversity (π), Watterson’s Theta (θW), and Tajima’s D statistics were calculated over fourfold degenerate sites on autosomal contigs. The error bars indicate 95% confidence intervals calculated over 100 bootstrap replicates over samples. An average pairwise Fst in the table here was calculated in 20 kb windows over the largest contig KB663610.Full size imageThis level of differentiation of RK2, even from the RK1 population, might indicate an emerging cryptic species within An. minimus A or a newly diverging clade. RK1 and RK2 are sympatric populations, both being collected in the same two sites in Northeastern Cambodia. The differences seen here between RK1 and RK2 populations are consistent with cryptic taxa in other anopheline groups. For example, in the An. gambiae complex, the level of differentiation between recently diverged sibling species An. coluzzii and An. gambiae in West Africa is approximately 0.0319.Population diversity and variationTo characterize population diversity among these populations, nucleotide diversity (π), Watterson’s Theta (θW), and Tajima’s D statistics were calculated over 4-fold degenerate sites on autosomal contigs larger than 2 megabases with 100 bootstrap replicates over samples. These 17 contigs represent 80% of the Anopheles minimus genome (Fig. 2). The populations were downsampled for these calculations to have sizes equal to that of the smallest population RK2 (n = 28).There are small but significant differences in the magnitude of the genetic diversity summary statistics between these four different populations. In particular, there were notable differences between the putatively cryptic taxa RK1 and RK2, two populations that were collected in the same sites in Northeastern Cambodia. RK1 had higher levels of nucleotide diversity and lower levels of Tajima’s D than RK2. These differences are consistent with different population size histories between these sympatric groups. Lower values of Tajima’s D suggest stronger population growth in RK1. Comparing all four populations, higher levels of genetic diversity indicate larger effective population sizes of TD and PV compared to RK1 and RK2.RK2 has a significantly reduced nucleotide diversity and Watterson’s Theta compared to the other three populations. This may indicate a smaller population size and a recent bottleneck of the RK2 population in Cambodia. All four An. minimus populations have a negative Tajima’s D, indicating an excess of rare variants, particularly in RK1. This suggests recent population expansions in all populations.Signals of evolutionary selectionWe used Fst to scan across the Anopheles minimus genome to look for regions of the genome with increased differentiation. When we scanned the genome using pairwise Fst, there were no apparent long signals of differentiation that might indicate a large inversion or other structural variants, known to be major drivers of adaptive evolution in other Anopheles groups. To investigate increased differentiation across large regions of the genome, we performed scans of nucleotide diversity (π), Watterson’s Theta (θW), and Tajima’s D over the largest 14 contigs (representing 80% of the An. minimus genome). As with the Fst scans, there were no large regions of higher differentiation in any of the populations that might indicate major structural variants or inversions (Supplementary Figs. 2–4).Whole-genome sequencing allowed us to identify pointed signals occurring across the entire genome using scans of average pairwise Fst. Isolated points of high differentiation were compared over single contigs with average pairwise Fst calculated over windows of 1000 SNPs each and plotted over whole contigs. The strongest signals, indicated by the highest Fst value at the peak of a strong signal of differentiation, were ranked and compared. The five top signals in each of the six comparisons between the four populations are listed in Table 1. These isolated points of high differentiation are one indication of a signal of evolutionary selection. The most differentiated regions by Fst occurred when comparing the RK2 population to the other three populations, with the highest selection peaks with pairwise Fst over 0.4. RK2 also had more distinct signals of selection when compared to the other populations than RK1. Since these signals of differentiation were highly localized, we could look to known gene annotations and gene predictions across the AminM1 reference genome to see which genes were within 100 kbp of the peaks of these signals. We have noted candidate genes of interest that were near the strongest Fst signal peaks and also had known or predicted gene functions (Table 1, Supplementary Fig. 6, Supplementary Fig. 8).Table 1 The top five Fst signals of high differentiation within each of six population comparisons are reported here.Full size tableThere is almost no indication of selection when comparing the Thmar Da population with Preah Vihear, with all but one signal with Fst values below 0.05. The one strong signal between TD and PV (Fst = 0.125) is near a Carbohydrate sulfotransferase, which is involved in detoxification processes. Comparing TD to RK1 and RK2 reveals multiple strong signals of selection, some which are present in both Northeastern populations, as well as many unique RK2-specific signals (Fig. 3, Supplementary Fig. 5).Fig. 3: Signals of selection over a single autosomal contig.Pairwise Fst was calculated in 1000 SNP windows over autosomal contig KB664266, comparing the Thmar Da population to the three other populations, Ratanakiri 2, Ratanakiri 1, and Preah Vihear. There is almost no indication of selection when comparing Thmar Da with Preah Vihear. There is a strongly supported signal of differentiation in both Ratanakiri 1 and Ratanakiri 2 populations at 7.5 Mbp, which is in the same location as a cluster of GSTe genes, including GSTe2, which are known to be involved in metabolic resistance to DDT and pyrethroids. The signal with the highest Fst peak here in RK2, at 6 Mbp is close to an Ecdysteroid UDP-glucosyltransferase gene, shown to confer pyrethroid insecticide resistance in other anophelines. These are a few of many selection signals identified in this study that may be associated with insecticide pressure on these An. minimus populations.Full size imageMany of the strongest signals identified in this study may be associated with insecticide pressure on these An. minimus populations. The strongest selection signals in every population comparison were close to genes that are involved in detoxification, signal transduction, and adaptations to oxidative stress, or have been functionally validated to have mutations that confer resistance to insecticides (Table 1). Some signals of interest include a strongly supported signal of selection in both RK1 and RK2 populations at 7.5 Mbp on the contig KB664266, which is in the same location as a cluster of glutathione-S-transferases, including GSTe2, which has been shown to be involved in the metabolism of DDT and pyrethroids, mutations in which mediate metabolic insecticide resistance29. The signal with the highest pairwise Fst peak on the same contig KB664266, at 6 Mbp is an RK2-specific signal and close to an Ecdysteroid UDP-glucosyltransferase gene, which has been shown to confer pyrethroid insecticide resistance in An. stephensi30.Another notable signal is between the RK1 and RK2 populations on the contig KB663610, a Peptidase S1 domain-containing protein AMIN002286, which has been shown to be involved in response to parasite pathogens in insects31. The signals of selection observed in this study are mostly distinct from the main selection signals seen in An. gambiae complex mosquitoes19, the primary vectors of Plasmodium falciparum in Africa.Insecticide resistanceWe report here variants at known insecticide resistance-associated alleles for each of the four An. minimus populations. Variants occurring at a frequency of more than 2% in at least one of the four populations are reported in the known insecticide-resistance-associated genes Ace1, Rdl, KDR, and GSTe2 (Supplementary Data 2). GSTe2 mutants are present in multiple populations, at a low rate, and there are a few individuals in Thmar Da and Preah Vihear with the Rdl resistance mutation, which is known to confer resistance to cyclodiene insecticides, despite evidence from other studies that species in this region lack this resistance mutation32. We did not investigate copy number variation, which is one mechanism by which GSTe2 confers insecticide resistance. These SNP variants indicate variation throughout these insecticide-resistance-associated genes, and though most of these populations do not currently have high rates of validated insecticide resistance-associated mutations, this underlying variation provides the potential for structural and transcriptional events resulting in greater levels of insecticide resistance in An. minimus populations. More

  • in

    Integrated taxonomy reveals new threatened freshwater mussels (Bivalvia: Hyriidae: Westralunio) from southwestern Australia

    Genetic variationThe best fitting substitution models for COI codons 1–3 were identified as TN + F + G4, F81 + F + I, and TN + F, respectively. The maximum likelihood (ML) and Bayesian inference (BI) trees showed similar topologies of the main nodes, although the BI tree displayed greater resolution of the ingroup branches (Fig. 1). Furthermore, the BI tree revealed three monophyletic clades, while two of those clades were merged in the ML tree. Two of the three molecular species delimitation methods (ASAP and TCS) recovered three groups in the BI tree as distinct taxa (Fig. 1), corresponding to the three previously described ESUs27,28. The third method (bPTP) recovered between 8 and 43 groups (mean = 28.03) suggesting that there is evidence of additional genetic differentiation within the three groups identified by ASAP and TCS. The outputs of the three methods are provided in the Supplementary information. The molecular diagnosis uncovered several fixed nucleotide differences COI characters for each taxon (Table 1: “W. carteri” I = 10; “W. carteri” II = 3; “W. carteri” III = 5). There were also 13 fixed nucleotide differences in W. carteri for the 16S gene. The remaining two taxa had no fixed nucleotide differences for the 16S gene.Figure 1Phylogenetic trees obtained by maximum likelihood (left) and Bayesian inference (right) analysis of “Westralunio carteri” mtDNA COI sequences, including support values for the major genetic clades [ultrafast bootstrap values (left) and Bayesian posterior probabilities (right)]. Colour coded bars show support for the three major clades by the species delimitation methods (ASAP = dark shade; TCS = lighter shade). Green = WcI = “W. carteri” I; blue = WcIII = “W. carteri” III; red = WcII = “W. carteri” II. Results of bPTP analysis not shown (see supplementary data). Haplotype names correspond to Benson et al.28. Outgroup taxa are Velesunio ambiguus (Philippi, 1847) (Hyriidae: Velesunioninae) and Cucumerunio novaehollandiae (Gray, 1834) (Hyriidae: Hyriinae: Hyridellini).Full size imageTable 1 Molecular diagnoses of “Westralunio carteri” Evolutionarily Significant Units (ESUs) from southwestern Australia (after Bolotov et al.122 with reanalysis of data from Klunzinger et al.27 and Benson et al.28).Full size tableVariation in shell morphologyBased on results from analyses of variances (ANOVAs), shells of “W. carteri” I were significantly larger (for size metrics total length (TL), maximum height (MH), beak height (BH) and beak length (BL)) and more elongated (i.e., had a lower maximum height index (MHI)) than shells of “W. carteri” II and “W. carteri” II + III combined (Table 2). However, there was no difference in size or shape metrics between “W. carteri” I and “W. carteri” III (Table 2). The lack of significant differences in beak height index (BHI) and beak length index (BLI) among any of the taxa (Table 2) indicates that wing and anterior shell development was not discernibly different between any of the ESUs.Table 2 Shell size metrics [mm], shape indices [%] and scores for the first two principal components (PC) obtained by Principal Component Analysis of shape indices and 18 Fourier coefficients generated by Fourier Shape Analysis for each “Westralunio carteri” species and subspecies-level Evolutionarily Significant Units (ESUs): n, number of specimens measured; minimum (min) to maximum (max) and mean (± standard error (SE)).Full size tableThis pattern was partly confirmed in the principal component analysis (PCA) of these three shell shape indices, where PC1, largely explained by variation in BLI (Fig. 2A), did not differ between the two species (i.e., “W. carteri” I vs. “W. carteri” II + III) or among the three taxa (Table 2). The PC2, largely explained by variation in MHI and BHI (Fig. 2A), differed significantly between “W. carteri” I and “W. carteri” II (Table 2). Accordingly, 70% (70% jack-knifed) of specimens were assigned to the correct species in the corresponding discriminant analysis (DA), whilst this was true for only 55% (54%) at the MOTU-level.Figure 2Scatterplots of the first two PC axes obtained by PCA on (A) calculated shape indices based on shell measurements, and (B) 18 Fourier coefficients for “Westralunio carteri” I, “W. carteri” II and “W. carteri” III. 95% Confidence Intervals are displayed at the species level, i.e., for “W. carteri” I (full line) and “W. carteri” II + III (dashed line). Extreme shell outlines in (B) are depicted to visualise trends in sagittal shell shape, along PC axes.Full size imageThe difference in shell elongation between “W. carteri” I and “W. carteri” II was confirmed by Fourier shape analysis. As visualised by synthetic outlines in Fig. 2B, shell elongation is expressed along the PC1 (explaining 15% of total variation in Fourier coefficients). The PC1 as well as PC2 scores differed significantly between the two species (i.e., “W. carteri” I vs. “W. carteri” II + III) as well as between “W. carteri” I and “W. carteri” II, respectively (Table 2). Combined with synthetic outlines, this indicated a tendency towards a more elongated, somewhat wedge-shaped shell in “W. carteri” I, whilst “W. carteri” II shells tended to be relatively high with a stout anterior margin (Fig. 2B). An analysis of similarities (ANOSIM) analysis on all Fourier coefficients revealed no significant difference between the two species (i.e., “W. carteri” I vs. “W. carteri” II + III; ANOSIM: R = − 0.018, p = 0.097), but did indicate a significant difference between the three ESUs (ANOSIM: R = 0.0625, p = 0.0051). Specifically, “W. carteri” I differed significantly from “W. carteri” II (Bonferroni-corrected p = 0.0009). Only 66% and 65% (62% and 62% jack-knifed) of specimens were assigned to the correct species and taxon in DAs on that dataset, respectively.Taxonomic accountsClass: Bivalvia Linnaeus, 175831.Subclass: Autobranchia Grobben, 189432.Infraclass: Heteroconchia Gray, 185433.Cohort: Palaeoheterodonta Newell, 196534.Order: Unionida Gray, 185433 in Bouchet & Rocroi, 201035.Superfamily: Unionoidea Rafinesque, 182036.Family: Hyriidae Parodiz & Bonetto 196337.Genus: Westralunio Iredale, 19349.Type species: Westralunio ambiguus carteri Iredale, 19349 (by original designation).Redescription: Westralunio carteri (Iredale, 1934)SynonymyUnio australis Lamarck38: Menke39, p. 38, specimen 219. (Non Unio australis Lamarck, 181938).Unio moretonicus Reeve40: Smith41, p. 3, pl. iv, Fig. 2. (misidentified reference to Unio moretonicus Reeve, 186540).Hyridella australis (Lam.38): Cotton & Gabriel42 (in part), p. 156. (misidentified reference to Unio australis Lamarck, 181938).Hyridella ambigua (Philippi26): Cotton & Gabriel42 (in part), p. 157. (misidentified reference to Unio ambiguus Philippi, 184726).Westralunio ambiguus carteri: Iredale, 19349, p. 62.Westralunio ambiguus (Philippi26): Iredale9, p. 62, pl. iii, Fig. 8, pl. iv, Fig. 8. (Non Unio ambiguus Phil. 184726), Iredale43, p. 190.Centralhyria angasi subjecta Iredale, 19349, p. 67 (in part), Iredale43, p. 190.Westralunio carteri Iredale9: McMichael & Hiscock10pl. viii, Figs. 1, 2, 3, 4, 5, 6 and 7, pl. xvii, Figs. 4, 5.Type materialLectotype: AMS C.61724 (Fig. 3A) Westralunio ambiguus carteri Iredale, 19349.Figure 3(A) Westralunio ambiguus carteri Iredale, 1934, Lectotype: Victoria Reservoir, Darling Range, 12 mi E of Perth, AMS C.061724. Detail of fusion in anterior muscle scars from either valve represented by dashed lines and black polygons. Bottom image showing detail of hinge teeth. Photos provided with permission by Dr Mandy Reid, AMS. (B) Valves and detail of sculptured umbo of a juvenile W. carteri from Yule Brook, Western Australia, UMZC 2013.2.9. Photo by Dr Michael W. Klunzinger. (C) Glochidia of W. carteri from Canning River, Western Australia. Photo by Dr Michael W. Klunzinger.Full size imageParalectotypes: AMS C.170635 Westralunio ambiguus carteri Iredale, 19349 (n = 4).Type locality: Victoria Reservoir, Darling Range, 12 miles east of Perth, Western Australia (Fig. 4A).Figure 4(A) Victoria Reservoir, Canning River, near Perth, Western Australia, type locality for W. carteri. Photo by Corey Whisson. (B) Goodga River, Western Australia, type locality for W. inbisi inbisi, at vertical slot fishway where holotype of W. inbisi inbisi was collected from. Photo provided with permission by Dr Stephen J. Beatty. (C) Margaret River, Western Australia, type locality for W. inbisi meridiemus, at Canebreak Pool. Photo by Dr Michael W. Klunzinger.Full size imageLectotype: BMNH 1840–10-21–29 Centralhyria angasi subjecta Iredale (selected by McMichael & Hiscock10).Type locality: Avon River, Western Australia.Material examined for redescription: For W. carteri (= “W. carteri” I), molecular data examined included 52 and 61 individual COI mtDNA and 16S rDNA sequences, respectively, for species delimitation. Additionally, Fourier shell shape outline analysis and traditional shell morphometric measurements were examined from 238 and 290 individuals, respectively. Complete details on all specimens examined are provided in Supplementary Table S1.ZooBank registration: urn:lsid:zoobank.org:act:6B740F4D-40C3-4D6A-8938-B0FD7FD1F6D7.Etymology: The species name carteri is most likely named after the surname of the collector who provided original type specimens to the Australian Museum, although Iredale9 did not specify this as the case. We have applied ICZN Articles 46.1 and 47.144, designating W. carteri as the nominotypical species.Revised diagnosis: Specimens of W. carteri are distinguished from other Australian Westralunio taxa by having shell series that are significantly larger and more elongated than Westralunio inbisi inbisi subsp. nov., but not different from Westralunio inbisi meridiemus subsp. nov. The species has 10 diagnostic nucleotides at COI (57 G, 117 T, 210 G, 249 T, 255 C, 345 G, 423 T, 447 T, 465 A, 499 T) and 13 at 16S (137 T, 155 C, 228 C, 229 T, 260 G, 290 A, 305 G, 307 T, 310 A, 311 C, 321 T, 330 A, 460 A), which differentiate it from its sister taxa, W. inbisi inbisi and W. inbisi meridiemus (each described below) using ASAP and TCS species delimitation models.RedescriptionThis species is of the ESU “W. carteri” I27,28.Shell morphology: Shells of relatively small to medium size, generally less than 70 mm in length, but to a maximum length of approximately 100 mm10,45, MHI 46–89%; anterior portion of shell with moderate development, BLI 22–49%; larger shells with abraded umbos scarcely winged; wing development variable, generally decreasing with size, BHI 76–104% (Table 2). Shell outline oblong-ovate to rounded; posterior end obliquely to squarely truncate, anterior end round; ventral edge slightly curved, nearly straight in larger specimens; hinge line curved, hinge strong. Umbos usually abraded in specimens  > 20 mm in length; unabraded umbos with distinctive v- or w-shaped plicated sculpturing (Fig. 3B and Zieritz et al.46). Shell substance typically thick; shells of medium width with pronounced posterior ridge; periostracum smooth, dark brown to reddish, with fine growth lines. Pallial line less developed in smaller specimens and prominent only in large specimens (e.g.,  > 60 mm TL). Lateral teeth longer and blade-like, slightly serrated to smooth and singular in left valve, fitting into deep groove in right valve; pseudocardinal tooth in right valve coarsely serrated, thick, and erect, fitting into deeply grooved socket in left valve. Anterior muscle scars well impressed and anchored deeply in larger specimens; anterior retractor pedis and protractor pedis scars both small and fused with adductor muscle scar; posterior muscle scars lightly impressed; dorsal muscle scars usually with two or three deep pits anchored to internal umbo region.Anatomy: Supra-anal opening absent, siphons of moderate size, not prominent but protrude beyond shell margin in actively filtering live specimens, pigmented dark brown with mottled lighter brown to orange splotches; inhalant siphon aperture about 1.5 times size of exhalant and bearing 2–4 rows of internal papillae (Fig. 5A); ctenidial diaphragm relatively long and perforated. Outer lamellae of outer ctenidia completely fused to mantle, inner lamellae of inner ctenidia fused to visceral mass then united to form diaphragm; palps relatively small, usually semilunar in shape; marsupium well developed as a distinctive swollen interlamellar space in the middle third of the inner ctenidium of females. Outer ctenidia in both sexes thin, with numerous, short intrafilamentary junctions and few, irregular interlamellar junctions; in females similar, but marsupium has numerous, tightly packed, well-developed interlamellar junctions. Thus, brooding in females is endobranchous.Figure 5Live specimens of actively filtering freshwater mussels in the burrowed position. (A) Westralunio carteri (Iredale, 1934), Canning River at Kelmscott, Western Australia, inhalant siphon with 2–4 rows of papillae oriented toward substrate. Photo by Dr Michael W. Klunzinger. (B) Westralunio inbisi meridiemus subsp. nov., Canebreak Pool, Margaret River, Western Australia; inhalant siphon edges lined with protruding papillae facing towards water surface, away from substrate. Photo by Dr Michael W. Klunzinger.Full size imageLife history: Sexes are separate in W. carteri, and hermaphroditism appears to be rare47,48,49. Males and females both produce gametes year-round but brooding of glochidia appears to be seasonal and ‘tachyticitc’ (i.e., as defined by Bauer & Wächtler19, fertilisation and embryonic development occurring in late winter/early spring and glochidia release in early summer)50. Glochidia are released within vitelline membranes, embedded in mucus which extrude from exhalant siphons of females (i.e., ‘amorphous mucus conglutinates’) during spring/summer. Glochidia attach to host fishes and live parasitically on fins, gills or body surfaces for 3–4 weeks while undergoing metamorphosis to the juvenile stage. Host fishes which have been shown to support glochidia metamorphosis to the juvenile stage in the laboratory include Afurcagobius suppositus (Sauvage, 188051), Gambusia holbrooki (Girard, 185952), Nannoperca vitttata (Castelnau, 187353), Pseudogobius olorum (Sauvage, 188051) and Tandanus bostocki Whitley, 194454 but not Carassisus auratus Linnaeus, 175831 or Geophagus brasiliensis (Quoy & Gaimard, 1824 More

  • in

    Amoxicillin and thiamphenicol treatments may influence the co-selection of resistance genes in the chicken gut microbiota

    General description of sequencesAfter the quality filtering step, removal of chimeric fragments, and read merging, a total of 3,378,323 reads with 3007 different features was obtained, with an average of 27,244 sequences per individual sample. After quality filtering, none of the samples was excluded from the analysis of microbial communities.Amoxicillin and thiamphenicol treatments influence microbial diversity and the abundance of specific taxaUsing 16S rRNA NGS, the gut microbial community composition of the chicks in each group was characterized at different time points. At phylum level, microbiota composition varied with age rather than with treatment (Supplementary Fig. S1). Proteobacteria were the most abundant phyla at 1 day of age (d.o.a.), Firmicutes became dominant at later stages, while Bacteroidota were highly abundant in caecum samples collected at 46 d.o.a. Similar dynamics were observed also at family level, since Enterobacteriaceae and Clostridiaceae were significantly more abundant at 1 d.o.a. in all groups, Lactobacillaceae, Lachnospiraceae, and Ruminococcaceae seemed to bloom at 8 d.o.a., and Rikenellaceae were the dominant family in the caecum samples collected at 46 d.o.a. (Fig. 1; Supplementary Fig. S2).Figure 1Heatmap representing the microbial community composition at family level. The heatmap was generated in R (version 4.2.1) (https://www.r-project.org/) using package pheatmap (version 1.0.12).Full size imageEarly-age administrationIn both α-diversity indices (Fig. 2A,B), there was a trend towards increasing diversity from early to late time points in all groups; however, the only significant differences were between the group treated with amoxicillin (AMX1) and the other groups on day 21 post treatment (p.t.), and within AMX1 group between day 21 p.t. and the other time points. PERMANOVA showed that the microbial community was significantly different between the group treated with thiamphenicol (THP1) and the other two groups (i.e. AMX1 and control) on day 1 p.t. (p  More

  • in

    Semi-field and surveillance data define the natural diapause timeline for Culex pipiens across the United States

    Way, M. J., Hopkins, B. & Smith, P. M. Photoperiodism and diapause in insects. Nature 164, 615 (1949).Article 
    PubMed 

    Google Scholar 
    Beck, S. Photoperiod induction of diapause in an insect. Biol. Bull. 122, 1–12 (1962).Article 

    Google Scholar 
    Denlinger, D. L. & Armbruster, P. A. Mosquito diapause. Annu. Rev. Entomol. 59, 73–93 (2014).Article 
    PubMed 

    Google Scholar 
    Readio, J., Chen, M. H. & Meola, R. Juvenile hormone biosynthesis in diapausing and nondiapausing Culex pipiens (Diptera: Culicidae). J. Med. Entomol. 36, 355–360 (1999).Article 
    PubMed 

    Google Scholar 
    Eldridge, B. F. & Bailey, C. L. Experimental hibernation studies in Culex pipiens (Diptera: Culicidae): reactivation of ovarian development and blood-feeding in prehibernating females. J. Med Entomol. 15, 462–467 (1979).Article 
    PubMed 

    Google Scholar 
    Spielman, A. & Wong, J. Environmental control of ovarian diapause in Culex pipiens. Ann. Entomol. Soc. Am. 66, 905–907 (1973).Article 

    Google Scholar 
    Sanburg, L. L. & Larsen, J. R. Effect of photoperiod and temperature on ovarian development in Culex pipiens pipiens. J. Insect Physiol. 19, 1173–1190 (1973).Article 
    PubMed 

    Google Scholar 
    Eldridge, B. F. The effect of temperature and photoperiod on blood-feeding and ovarian development in mosquitoes of the Culex pipiens complex. Am. J. Trop. Med. Hyg. 17, 133–140 (1968).Article 
    PubMed 

    Google Scholar 
    Bowen, M. F. Patterns of sugar feeding in diapausing and nondiapausing Culex pipiens (Diptera: Culicidae) females. J. Med. Entomol. 29, 843–849 (1992).Article 
    PubMed 

    Google Scholar 
    Robich, R. M. & Denlinger, D. L. Diapause in the mosquito Culex pipiens evokes a metabolic switch from blood feeding to sugar gluttony. Proc. Natl Acad. Sci. USA 102, 15912–15917 (2005).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Eldridge, B. F. Environmental control of ovarian development in mosquitoes of the Culex pipiens complex. Am. Assoc. Adv. Sci. 151, 826–828 (1966).
    Google Scholar 
    Vinogradova, A. B. Culex pipiens Pipiens Mosquitoes: Taxonomy, Distribution, Ecology, Physiology, Genetics, Applied Importance And Control (Pensoft, 2000).Benoit, J. B. & Denlinger, D. L. Suppression of water loss during adult diapause in the northern house mosquito, Culex pipiens. J. Exp. Biol. 210, 217–226 (2007).Article 
    PubMed 

    Google Scholar 
    Li, A. & Denlinger, D. L. Pupal cuticle protein is abundant during early adult diapause in the mosquito Culex pipiens. J. Med. Entomol. 46, 1382–1386 (2009).Article 
    PubMed 

    Google Scholar 
    Yang, L., Denlinger, D. L. & Piermarini, P. M. The diapause program impacts renal excretion and molecular expression of aquaporins in the northern house mosquito, Culex pipiens. J. Insect Physiol. 98, 141–148 (2017).Article 
    PubMed 

    Google Scholar 
    King, B., Li, S., Liu, C., Kim, S. J. & Sim, C. Suppression of glycogen synthase expression reduces glycogen and lipid storage during mosquito overwintering diapause. J. Insect Physiol. 120, 103971 (2020).Article 
    PubMed 

    Google Scholar 
    Sim, C. & Denlinger, D. L. Transcription profiling and regulation of fat metabolism genes in diapausing adults of the mosquito Culex pipiens. Physiol. Genomics 39, 202–209 (2009).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Sim, C. & Denlinger, D. L. Insulin signaling and FOXO regulate the overwintering diapause of the mosquito Culex pipiens. Proc. Natl Acad. Sci. USA 105, 6777–6781 (2008).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Zhou, G. & Miesfeld, R. L. Energy metabolism during diapause in Culex pipiens mosquitoes. J. Insect Physiol. 55, 40–46 (2009).Article 
    PubMed 

    Google Scholar 
    Chang, J. et al. Solid-state NMR reveals differential carbohydrate utilization in diapausing Culex pipiens. Sci. Rep. 6, 37350 (2016).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Madder, D. J., Surgeoner, G. A. & Helson, B. V. Induction of diapause in Culex pipiens and Culex restuans (Diptera: Culicidae) in Southern Ontario. Can. Entomol. 115, 877–883 (1983).Article 

    Google Scholar 
    Spielman, A. Effect of synthetic juvenile hormone on ovarian diapause of Culex pipiens mosquitoes. J. Med. Entomol. 11, 223–225 (1974).Article 
    PubMed 

    Google Scholar 
    Sim, C. & Denlinger, D. L. Insulin signaling and the regulation of insect diapause. Front. Physiol. 4, 189 (2013).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Robich, R. M., Rinehart, J. P., Kitchen, L. J. & Denlinger, D. L. Diapause-specific gene expression in the northern house mosquito, Culex pipiens L., identified by suppressive subtractive hybridization. J. Insect Physiol. 53, 235–245 (2007).Article 
    PubMed 

    Google Scholar 
    Sim, C., Kang, D. S., Kim, S., Bai, X. & Denlinger, D. L. Identification of FOXO targets that generate diverse features of the diapause phenotype in the mosquito Culex pipiens. Proc. Natl Acad. Sci. USA 112, 3811–3816 (2015).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Kang, D. S., Cotten, M. A., Denlinger, D. L. & Sim, C. Comparative transcriptomics reveals key gene expression differences between diapausing and non-diapausing adults of Culex pipiens. PLoS ONE 11, e0154892 (2016).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Spielman, A. Structure and seasonality of nearctic Culex pipiens populations. Ann. N. Y. Acad. Sci. 951, 220–234 (2001).Article 
    PubMed 

    Google Scholar 
    Wilton, D. P. & Smith, G. C. Ovarian diapause in three geographic strains of Culex pipiens (Diptera: Culicidae). J. Med. Entomol. 22, 524–528 (1985).Article 
    PubMed 

    Google Scholar 
    Eldridge, B. F. Diapause and related phenomena in Culex mosquitoes: their relation to arbovirus disease ecology. In: Current Topics in Vector Research (ed. Harris, K. F.) 1–28 (Springer, 1987).Meuti, M. E., Short, C. A. & Denlinger, D. L. Mom matters: diapause characteristics of Culex pipiens-Culex quinquefasciatus (Diptera: Culicidae) hybrid mosquitoes. J. Med. Entomol. 52, 131–137 (2015).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Zhang, C. et al. Understanding the regulation of overwintering diapause molecular mechanisms in Culex pipiens pallens through comparative proteomics. Sci. Rep. 9, 6845 (2019).PubMed 
    PubMed Central 

    Google Scholar 
    Dunphy, B. M. et al. Long-term surveillance defines spatial and temporal patterns implicating Culex tarsalis as the primary vector of West Nile virus. Sci. Rep. 9, 6637 (2019).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Dunphy, B. M., Rowley, W. A. & Bartholomay, L. C. A Taxonomic checklist of the mosquitoes of Iowa. J. Am. Mosq. Control Assoc. 30, 119–121 (2014).Article 
    PubMed 

    Google Scholar 
    Sucaet, Y., Van Hemert, J., Tucker, B. & Bartholomay, L. C. A web-based relational database for monitoring and analyzing mosquito population dynamics. J. Med. Entomol. 45, 775–784 (2008).Article 
    PubMed 

    Google Scholar 
    Ryan, S. F., Valella, P., Thivierge, G., Aardema, M. L. & Scriber, J. M. The role of latitudinal, genetic and temperature variation in the induction of diapause of Papilio glaucus (Lepidoptera: Papilionidae). Insect Sci. 25, 328–336 (2018).Article 
    PubMed 

    Google Scholar 
    Huang, L. et al. Diapause incidence and critical day length of Asian corn borer (Ostrinia furnacalis) populations exhibit a latitudinal cline in both pure and hybrid strains. J. Pest Sci. 93, 559–568 (2020).Article 

    Google Scholar 
    Bradshaw, W. E. Geography of photoperiodic response in diapausing mosquito. Nature 262, 384–386 (1976).Article 
    PubMed 

    Google Scholar 
    Bradshaw, W. E. & Lounibos, L. P. Evolution of dormancy and its photoperiodic control in pitcher-plant mosquitoes. Nature 31, 546–567 (1977).
    Google Scholar 
    Kothera, L., Zimmerman, E. M., Richards, C. M. & Savage, H. M. Microsatellite characterization of subspecies and their hybrids in Culex pipiens complex (Diptera: Culicidae) mosquitoes along a North-South transect in the central United States. J. Med. Entomol. 46, 236–248 (2009).Article 
    PubMed 

    Google Scholar 
    Darsie, R. F. R. & Ward, R. A. R. Identification and Geographical Distribution of the Mosquitoes of North America, North of Mexico (University Press of Florida, 2005).Huang, S., Molaei, G. & Andreadis, T. G. Reexamination of Culex pipiens hybridization zone in the eastern United States by ribosomal DNA-based single nucleotide polymorphism markers. Am. J. Trop. Med. Hyg. 85, 434–441 (2011).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Reisen, W. K. Overwintering studies on Culex tarsalis (Diptera: Culicidae) in Kern County, California: life stages sensitive to diapause induction cues. Ann. Entomol. Soc. Am. 79, 674–676 (1986).Article 

    Google Scholar 
    Haba, Y. & McBride, L. Origin and status of Culex pipiens mosquito ecotypes. Curr. Biol. 32, R237–R246 (2022).Article 
    PubMed 

    Google Scholar 
    Holzapfel, C. M. & Bradshaw, W. E. Geography of larval dormancy in the tree-hole mosquito, Aedes triseriatus (Say). Can. J. Zool. 59, 1014–1021 (1981).Article 

    Google Scholar 
    Rinehart, J. P., Robich, R. M. & Denlinger, D. L. Enhanced cold and desiccation tolerance in diapausing adults of Culex pipiens, and a role for Hsp70 in response to cold shock but not as a component of the diapause program. J. Med. Entomol. 43, 713–722 (2006).Article 
    PubMed 

    Google Scholar 
    Faraji, A. & Gaugler, R. Experimental host preference of diapause and non-diapause induced Culex pipiens pipiens (Diptera: Culicidae). Parasit. Vectors 8, 389 (2015).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Washino, R. K. The physiological ecology of gonotrophic dissociation and related phenomena in mosquitoes. J. Med. Entomol. 13, 381–388 (1977).Article 
    PubMed 

    Google Scholar 
    Christophers, S. The development of the egg follicle in Anophelines. Paludism 1, 73–88 (1911).
    Google Scholar 
    Nelms, B. M., Macedo, P. A., Kothera, L., Savage, H. M. & Reisen, W. K. Overwintering biology of Culex (Diptera: Culicidae) mosquitoes in the Sacramento Valley of California. J. Med. Entomol. 50, 773–790 (2013).Article 
    PubMed 

    Google Scholar 
    Diniz, D. F. A., De Albuquerque, C. M. R., Oliva, L. O., De Melo-Santos, M. A. V. & Ayres, C. F. J. Diapause and quiescence: dormancy mechanisms that contribute to the geographical expansion of mosquitoes and their evolutionary success. Parasites Vectors 10, 1–13 (2017).Article 

    Google Scholar 
    Kingsolver, J. G. & Nagle, A. Evolutionary divergence in thermal sensitivity and diapause of field and laboratory populations of Manduca sexta. Physiol. Biochem. Zool. 80, 473–479 (2007).Article 
    PubMed 

    Google Scholar 
    Brent, C. S. & Spurgeon, D. W. Diapause response of laboratory reared and native lygus hesperus knight (Hemiptera: Miridae). Environ. Entomol. 40, 455–461 (2011).Article 

    Google Scholar 
    Rinehart, J. P., Yocum, G. D., Leopold, R. A. & Robich, R. M. Cold storage of Culex pipiens in the absence of diapause. J. Med. Entomol. 47, 1071–1076 (2014).Article 

    Google Scholar 
    Arora, A. K., Sim, C., Severson, D. W. & Kang, D. S. Random forest analysis of impact of abiotic factors on Culex pipiens and Culex quinquefasciatus occurrence. Front. Ecol. Evol. 9, 773360 (2022).Article 

    Google Scholar 
    Focks, D. A., Linda, S. B., Craig Jnr, G. B., Hawley, W. A. & Pumpuni, C. B. Aedes albopictus (Diptera: Culicidae): a statistical model of the role of temperature, photoperiod, and geography in the induction of egg diapause. J. Med. Entomol. 31, 278–286 (1994).Article 
    PubMed 

    Google Scholar 
    Urbanski, J. et al. Rapid adaptive evolution of photoperiodic response during invasion and range expansion across a climatic gradient. Am. Nat. 179, 490–500 (2012).Article 
    PubMed 

    Google Scholar 
    Kothera, L., Godsey, M. S., Doyle, M. S. & Savage, H. M. Characterization of Culex pipiens complex (Diptera: Culicidae) populations in Colorado, USA using microsatellites. PLoS ONE 7, e0047602 (2012).Article 

    Google Scholar 
    Kothera, L., Nelms, B. M., Reisen, W. K. & Savage, H. M. Population genetic and admixture analyses of Culex pipiens complex (Diptera: Culicidae) populations in California, United States. Am. J. Trop. Med. Hyg. 89, 1154–1167 (2013).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Kothera, L. et al. Bloodmeal, Host selection, and genetic admixture analyses of Culex pipiens Complex (Diptera: Culicidae) mosquitoes in Chicago, IL. J. Med. Entomol. 57, 78–87 (2020).Article 
    PubMed 

    Google Scholar 
    Huang, S., Molaei, G. & Andreadis, T. G. Genetic insights into the population structure of Culex pipiens (Diptera: Culicidae) in the Northeastern United States by using microsatellite analysis. Am. J. Trop. Med Hyg. 79, 518–527 (2008).Article 
    PubMed 

    Google Scholar 
    Barr, A. R. The Distribution of Culex p. pipiens and Cp quinquefasciatus in North America. Am. J. Trop. Med. Hyg. 6, 153–165 (1957).Article 
    PubMed 

    Google Scholar 
    Iltis, W. G. Biosystematics of the Culex pipiens Complex in Northern California. Thesis, University of California, Davis. (1966).Urbanelli, S., Silvestrini, F., Reisen, W. K., De Vito, E. & Bullini, L. Californian hybrid zone between Culex pipiens pipiens and Cx. p. quinquefasciatus revisited (Diptera: Culicidae). J. Med. Entomol. 34, 116–127 (1997).Article 
    PubMed 

    Google Scholar 
    Nelms, B. M. et al. Phenotypic variation among Culex pipiens complex (Diptera: Culicidae) populations from the Sacramento Valley, California: Horizontal and vertical transmission of West Nile virus, diapause potential, autogeny, and host selection. Am. J. Trop. Med. Hyg. 89, 1168–1178 (2013).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Dodson, B. L., Kramer, L. D. & Rasgon, J. L. Effects of larval rearing temperature on immature development and West Nile virus vector competence of Culex tarsalis. Parasit. Vectors 5, 199 (2012).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Ciota, A. T., Matacchiero, A. C., Marm Kilpatrick, A. & Kramer, L. D. The effect of temperature on life history traits of Culex mosquitoes. J. Med Entomol. 51, 55–62 (2014).Article 
    PubMed 

    Google Scholar 
    Carrington, L. B., Seifert, S. N., Willits, N. H., Lambrechts, L. & Scott, T. W. Large diurnal temperature fluctuations negatively influence Aedes aegypti (Diptera: Culicidae) life-history traits. J. Med. Entomol. 50, 43–51 (2013).Article 
    PubMed 

    Google Scholar 
    Lambrechts, L. et al. Impact of daily temperature fluctuations on dengue virus transmission by Aedes aegypti. Proc. Natl Acad. Sci. USA 108, 7460–7465 (2011).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Karki, S., Brown, W. M., Uelmen, J., O’Hara Ruiz, M. & Smith, R. L. The drivers of West Nile virus human illness in the Chicago, Illinois, USA area: fine scale dynamic effects of weather, mosquito infection, social, and biological conditions. PLoS ONE 15, e0227160 (2020).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Andreadis, T. G., Anderson, J. F., Vossbrinck, C. R. & Main, A. J. Epidemiology of West Nile virus in Connecticut: a five-year analysis of mosquito data 1999–2003. Vector-Borne Zoonotic Dis. 4, 360–378 (2004).Article 
    PubMed 

    Google Scholar 
    Anderson, J. F. & Main, A. J. Importance of vertical and horizontal transmission of West Nile virus by Culex pipiens in the northeastern United States. J. Infect. Dis. 194, 1577–1579 (2006).Article 
    PubMed 

    Google Scholar 
    Nasci, R. S. et al. West Nile virus in overwintering Culex mosquitoes, New York City, 2000. Emerg. Infect. Dis. 7, 742–744 (2001).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Kampen, H., Tews, B. A. & Werner, D. First evidence of West Nile virus overwintering in mosquitoes in Germany. Viruses 13, 1–7 (2021).Article 

    Google Scholar 
    Farajollahi, A. et al. Detection of West Nile viral RNA from an overwintering pool of Culex pipens pipiens (Diptera: Culicidae) in New Jersey, 2003. J. Med. Entomol. 42, 490–494 (2005).Article 
    PubMed 

    Google Scholar 
    Baqar, S., Hayes, C. G., Murphy, J. R. & Watts, D. M. Vertical transmission of West Nile virus by Culex and Aedes species mosquitoes. Am. J. Trop. Med. Hyg. 48, 757–762 (1993).Article 
    PubMed 

    Google Scholar 
    Miller, B. R. et al. First field evidence for natural vertical transmission of West Nile virus in Culex univittatus complex mosquitoes from Rift Valley Province, Kenya. Am. J. Trop. Med. Hyg. 62, 240–246 (2000).Article 
    PubMed 

    Google Scholar 
    Peffers, C. S., Pomeroy, L. W. & Meuti, M. E. Critical photoperiod and its potential to predict mosquito distributions and control medically important pests. J. Med. Entomol. 58, 1610–1618 (2021).Article 
    PubMed 

    Google Scholar 
    Bradshaw, W. E. & Holzapfel, C. M. Genetic shift in photoperiodic response correlated with global warming. Proc. Natl Acad. Sci. USA 98, 14509–14511 (2001).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Reiter, P. Climate change and mosquito-borne disease. Environ. Health Perspect. 109, 141–161 (2001).PubMed 
    PubMed Central 

    Google Scholar 
    Colón-González, F. J. et al. Projecting the risk of mosquito-borne diseases in a warmer and more populated world: a multi-model, multi-scenario intercomparison modelling study. Lancet Planet. Heal. 5, e404–e414 (2021).Article 

    Google Scholar 
    Barreaux, A. M. G., Stone, C. M., Barreaux, P. & Koella, J. C. The relationship between size and longevity of the malaria vector Anopheles gambiae (s.s.) depends on the larval environment. Parasites Vectors 11, 485 (2018).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Van Handel, E. & Day, J. F. Correlation between wing length and protein content of mosquitoes. J. Am. Mosq. Control Assoc. 5, 180–182 (1989).PubMed 

    Google Scholar 
    Ferreira-De-Freitas, L., Thrun, N. B., Tucker, B. J., Melidosian, L. & Bartholomay, L. C. An evaluation of characters for the separation of two Culex species (Diptera: Culicidae) based on material from the Upper Midwest. J. Insect Sci. 20, 21 (2020).Harrington, L. C. & Poulson, R. L. Considerations for accurate identification of adult Culex restuans (Diptera: Culicidae) in field studies. J. Med. Entomol. 45, 1–8 (2008).Article 
    PubMed 

    Google Scholar  More

  • in

    Plant genetic diversity affects multiple trophic levels and trophic interactions

    Effects of plant genetic diversity on multiple trophic groupsWe found that plant genetic diversity (i.e. diversification of cropping or plant cultivation systems; see Methods and Supplementary Table 15) decreased the overall performance of plant antagonists (effect size = −0.539, t = −2.070, P = 0.039) and several of its components (i.e., herbivores (effect size = −0.606, t = −4.127, P  More

  • in

    Local-scale feedbacks influencing cold-water coral growth and subsequent reef formation

    Henry, L.-A. & Roberts, J. M. Biodiversity and ecological composition of macrobenthos on cold-water coral mounds and adjacent off-mound habitat in the bathyal Porcupine Seabight, NE Atlantic. Deep Sea Res. I(54), 654–672 (2007).
    Google Scholar 
    Buhl-Mortensen, L. et al. First observations of the structure and megafaunal community of a large Lophelia reef on the Ghanaian shelf (the Gulf of Guinea). Deep Sea Res. II(137), 148–156 (2017).
    Google Scholar 
    Price, D. M. et al. Using 3D photogrammetry from ROV video to quantify cold-water coral reef structural complexity and investigate its influence on biodiversity and community assemblage. Coral Reefs 38, 1007–1021 (2019).
    Google Scholar 
    Roberts, J. M., Wheeler, A. J. & Freiwald, A. Reefs of the deep: the biology and geology of cold-water coral ecosystems. Science 312, 543–547 (2006).CAS 
    PubMed 

    Google Scholar 
    Henry, L. A., Nizinski, M. S. & Ross, S. W. Occurrence and biogeography of hydroids (Cnidaria: Hydrozoa) from deep-water coral habitats off the southeastern United States. Deep. Res. I(55), 788–800 (2008).
    Google Scholar 
    Henry, L.-A. & Roberts, J. M. Global Biodiversity in Cold-Water Coral Reef Ecosystems. In Marine Animal Forests (eds Rossi, S. et al.) 1–21 (Springer, 2016). https://doi.org/10.1007/978-3-319-17001-5_6-1.Chapter 

    Google Scholar 
    De Mol, B. et al. Large deep-water coral banks in the Porcupine Basin, southwest of Ireland. Mar. Geol. 188, 193–231 (2002).
    Google Scholar 
    Dorschel, B., Hebbeln, D., Rüggeberg, A., Dullo, W. C. & Freiwald, A. Growth and erosion of a cold-water coral covered carbonate mound in the Northeast Atlantic during the Late Pleistocene and Holocene. Earth Planet. Sci. Lett. 233, 33–44 (2005).CAS 

    Google Scholar 
    Hebbeln, D., Van Rooij, D. & Wienberg, C. Good neighbours shaped by vigorous currents: Cold-water coral mounds and contourites in the North Atlantic. Mar. Geol. 378, 171–185 (2016).
    Google Scholar 
    Wheeler, A. J. et al. Morphology and environment of cold-water coral carbonate mounds on the NW European margin. Int. J. Earth Sci. 96, 37–56 (2007).CAS 

    Google Scholar 
    Lo Iacono, C., Savini, A. & Basso, D. Cold-water carbonate bioconstructions. in Submarine Geomorphology, 425–455 (Springer, 2018).Hebbeln, D. Highly variable submarine landscapes in the Alborán sea created by cold-water corals. In Mediterranean Cold-Water Corals: Past, Present and Future (eds Orejas, C. & Jiménez, C.) 61–65 (Springer, 2019). https://doi.org/10.1007/978-3-319-91608-8_8.Chapter 

    Google Scholar 
    Addamo, A. M. et al. Merging scleractinian genera: The overwhelming genetic similarity between solitary Desmophyllum and colonial Lophelia. BMC Evol. Biol. 16, 1–17 (2016).
    Google Scholar 
    Wienberg, C. & Titschack, J. Framework-forming scleractinian cold-water corals through space and time: A late quaternary north atlantic perspective. in Marine Animal Forests 1–34 (Springer, 2017). https://doi.org/10.1007/978-3-319-17001-5_16-1Maier, C., Weinbauer, M. G. & Gattuso, J.-P. Fate of mediterranean scleractinian cold-water corals as a result of global climate change: A synthesis. In Mediterranean Cold-Water Corals: Past, Present and Future (eds Orejas, C. & Jiménez, C.) 517–529 (Springer, 2019). https://doi.org/10.1007/978-3-319-91608-8_44.Chapter 

    Google Scholar 
    Reynaud, S. & Ferrier-Pagès, C. Biology and ecophysiology of mediterranean cold-water corals. In Mediterranean Cold-Water Corals: Past, Present and Future (eds Orejas, C. & Jiménez, C.) 391–404 (Springer, 2019). https://doi.org/10.1007/978-3-319-91608-8_35.Chapter 

    Google Scholar 
    Hennige, S. J. et al. Using the Goldilocks principle to model coral ecosystem engineering. Proc. R. Soc. B Biol. Sci. 288, 20211260 (2021).CAS 

    Google Scholar 
    LoIacono, C. et al. The West Melilla cold water coral mounds, Eastern Alboran Sea: Morphological characterization and environmental context. Deep Sea Res. II(99), 316–326 (2014).
    Google Scholar 
    Mortensen, P. B., Hovland, T., Fosså, J. H. & Furevik, D. M. Distribution, abundance and size of Lophelia pertusa coral reefs in mid-Norway in relation to seabed characteristics. J. Mar. Biol. Assoc. 81, 581–597 (2001).
    Google Scholar 
    Mienis, F. et al. Hydrodynamic controls on cold-water coral growth and carbonate-mound development at the SW and SE Rockall Trough Margin, NE Atlantic. Ocean. Deep. Res. I(54), 1655–1674 (2007).
    Google Scholar 
    Davies, A. J. et al. Downwelling and deep-water bottom currents as food supply mechanisms to the cold-water coral Lophelia pertusa (Scleractinia) at the Mingulay Reef Complex. Limnol. Oceanogr. 54, 620–629 (2009).
    Google Scholar 
    Mohn, C. et al. Linking benthic hydrodynamics and cold-water coral occurrences: A high-resolution model study at three cold-water coral provinces in the NE Atlantic. Prog. Oceanogr. 122, 92–104 (2014).
    Google Scholar 
    Mienis, F. et al. Cold-water coral growth under extreme environmental conditions, the Cape Lookout area, NW Atlantic. Biogeosciences 11, 2543–2560 (2014).
    Google Scholar 
    Grasmueck, M. et al. Autonomous underwater vehicle (AUV) mapping reveals coral mound distribution, morphology, and oceanography in deep water of the Straits of Florida. Geophys. Res. Lett. 33, L23616 (2006).
    Google Scholar 
    Correa, T. B. S., Eberli, G. P., Grasmueck, M., Reed, J. K. & Correa, A. M. S. Genesis and morphology of cold-water coral ridges in a unidirectional current regime. Mar. Geol. 326–328, 14–27 (2012).
    Google Scholar 
    Lavaleye, M. et al. Cold-water corals on the tisler reef: Preliminary observations on the dynamic reef environment. Oceanography 22, 76–84 (2009).
    Google Scholar 
    Mortensen, P. B. et al. Seascape description of anunusual coral reef area off Vesteraålen, Northern Norway. in 4th International Symposium on deep-sea corals. (2008).Cathalot, C. et al. Cold-water coral reefs and adjacent sponge grounds: Hotspots of benthic respiration and organic carbon cycling in the deep sea. Front. Mar. Sci. 2, 37 (2015).
    Google Scholar 
    Buhl-Mortensen, P. & Sundahl, H. Environmental control of cold-water coral reef morphology. in 7th International Symposium on deep-sea corals. (2019).van der Kaaden, A.-S., van Oevelen, D., Rietkerk, M., Soetaert, K. & van de Koppel, J. Spatial self-organization as a new perspective on cold-water coral mound development. Front. Mar. Sci. 7, 631 (2020).
    Google Scholar 
    Buhl-Mortensen, L. et al. Biological structures as a source of habitat heterogeneity and biodiversity on the deep ocean margins. Mar. Ecol. 31, 21–50 (2010).
    Google Scholar 
    Jones, C. G., Lawton, J. H. & Shachak, M. Organisms as ecosystem engineers. Oikos 69, 373–386 (1994).
    Google Scholar 
    Mienis, F., Bouma, T., Witbaard, R., van Oevelen, D. & Duineveld, G. Experimental assessment of the effects of coldwater coral patches on water flow. Mar. Ecol. Prog. Ser. 609, 101–117 (2019).CAS 

    Google Scholar 
    van der Kaaden, A.-S. et al. Feedbacks between hydrodynamics and cold-water coral mound development. Deep Sea Res. I 178, 103641 (2021).
    Google Scholar 
    Mortensen, P. B., Hovland, M., Brattegard, T. & Farestveit, R. Deep water bioherms of the scleractinian coral Lophelia pertusa (L.) at 64° n on the norwegian shelf: Structure and associated megafauna. Sarsia 80, 145–158 (1995).
    Google Scholar 
    Corbera, G. et al. Ecological characterisation of a Mediterranean cold-water coral reef: Cabliers Coral Mound Province (Alboran Sea, western Mediterranean). Prog. Oceanogr. 175, 245–262 (2019).
    Google Scholar 
    Kano, A. et al. Age constraints on the origin and growth history of a deep-water coral mound in the northeast Atlantic drilled during Integrated Ocean Drilling Program Expedition 307. Geology 35, 1051–1054 (2007).CAS 

    Google Scholar 
    Douarin, M. et al. Growth of north-east Atlantic cold-water coral reefs and mounds during the Holocene: A high resolution U-series and 14C chronology. Earth Planet. Sci. Lett. 375, 176–187 (2013).CAS 

    Google Scholar 
    Orejas, C., Gori, A. & Gili, J. M. Growth rates of live Lophelia pertusa and Madrepora oculata from the Mediterranean Sea maintained in aquaria. Coral Reefs 27, 255–255 (2008).
    Google Scholar 
    Orejas, C. et al. Long-term growth rates of four Mediterranean cold-water coral species maintained in aquaria. Mar. Ecol. Prog. Ser. 429, 57–65 (2011).
    Google Scholar 
    Lartaud, F., Mouchi, V., Chapron, L., Meistertzheim, A.-L. & Le Bris, N. Growth Patterns of Mediterranean Calcifying Cold-Water Corals. in Mediterranean Cold-Water Corals: Past, Present and Future 405–422 (2019). https://doi.org/10.1007/978-3-319-91608-8_36.Büscher, J. V. et al. In situ growth and bioerosion rates of Lophelia pertusa in a Norwegian fjord and open shelf cold-water coral habitat. PeerJ 2019, 1–10 (2019).
    Google Scholar 
    Form, A. U. & Riebesell, U. Acclimation to ocean acidification during long-term CO2 exposure in the cold-water coral Lophelia pertusa. Glob. Chang. Biol. 18, 843–853 (2012).
    Google Scholar 
    Maier, C., Watremez, P., Taviani, M., Weinbauer, M. G. & Gattuso, J. P. Calcification rates and the effect of ocean acidification on Mediterranean cold-water corals. Proc. R. Soc. B Biol. Sci. 279, 1716–1723 (2012).CAS 

    Google Scholar 
    Lunden, J. J., McNicholl, C. G., Sears, C. R., Morrison, C. L. & Cordes, E. E. Acute survivorship of the deep-sea coral Lophelia pertusa from the Gulf of Mexico under acidification, warming, and deoxygenation. Front. Mar. Sci. 1, 78 (2014).
    Google Scholar 
    Gori, A., Reynaud, S., Orejas, C., Gili, J. M. & Ferrier-Pagès, C. Physiological performance of the cold-water coral Dendrophyllia cornigera reveals its preference for temperate environments. Coral Reefs 33, 665–674 (2014).
    Google Scholar 
    Huvenne, V. A. I. et al. Sediment dynamics and palaeo-environmental context at key stages in the Challenger cold-water coral mound formation: Clues from sediment deposits at the mound base. Deep. Res. I(56), 2263–2280 (2009).
    Google Scholar 
    Bartzke, G. et al. Investigating the prevailing hydrodynamics around a cold-water coral colony using a physical and a numerical approach. Front. Mar. Sci. 8, 3304 (2021).
    Google Scholar 
    Downs, C. A. et al. Cellular diagnostics and coral health: Declining coral health in the Florida Keys. Mar. Pollut. Bull. 51, 558–569 (2005).CAS 
    PubMed 

    Google Scholar 
    Ayala, A., Muñoz, M. F. & Argüelles, S. Lipid peroxidation: Production, metabolism, and signaling mechanisms of malondialdehyde and 4-hydroxy-2-nonenal. Oxid. Med. Cell. Long. 2014, 1–10 (2014).CAS 

    Google Scholar 
    Oh, T. J., Kim, I. G., Park, S. Y., Kim, K. C. & Shim, H. W. NAD-dependent malate dehydrogenase protects against oxidative damage in Escherichia coli K-12 through the action of oxaloacetate. Environ. Toxicol. Pharmacol. 11, 9–14 (2002).CAS 
    PubMed 

    Google Scholar 
    Dade, L., Hogg, A. & Boudreau, B. Physics of Flow Above the Sediment-Water Interface (Oxford University Press, 2001).
    Google Scholar 
    Gass, S. E. & Roberts, J. M. The occurrence of the cold-water coral Lophelia pertusa (Scleractinia) on oil and gas platforms in the North Sea: Colony growth, recruitment and environmental controls on distribution. Mar. Pollut. Bull. 52, 549–559 (2006).CAS 
    PubMed 

    Google Scholar 
    Brooke, S. & Young, C. M. In situ measurement of survival and growth of Lophelia pertusa in the northern Gulf of Mexico. Mar. Ecol. Prog. Ser. 397, 153–161 (2009).
    Google Scholar 
    Lartaud, F. et al. A new approach for assessing cold-water coral growth in situ using fluorescent calcein staining. Aquat. Living Resour. 26, 187–196 (2013).
    Google Scholar 
    Sebens, K. P., Witting, J. & Helmuth, B. Effects of water flow and branch spacing on particle capture by the reef coral Madracis mirabilis (Duchassaing and Michelotti). J. Exp. Mar. Bio. Ecol. 211, 1–28 (1997).
    Google Scholar 
    Sebens, K. P., Grace, S. P., Helmuth, B., Maney, E. J. Jr. & Miles, J. S. Water flow and prey capture by three scleractinian corals, Madracis mirabilis, Montastrea cavernosa and Porites porites, in a field enclosure. Mar. Biol. 131, 347–360 (1998).
    Google Scholar 
    Purser, A., Larsson, A. I., Thomsen, L. & van Oevelen, D. The influence of flow velocity and food concentration on Lophelia pertusa (Scleractinia) zooplankton capture rates. J. Exp. Mar. Bio. Ecol. 395, 55–62 (2010).
    Google Scholar 
    Orejas, C. et al. The effect of flow speed and food size on the capture efficiency and feeding behaviour of the cold-water coral Lophelia pertusa. J. Exp. Mar. Bio. Ecol. 481, 34–40 (2016).
    Google Scholar 
    Duineveld, G. C. A. et al. Spatial and tidal variation in food supply to shallow cold-water coral reefs of the Mingulay Reef complex (Outer Hebrides, Scotland). Mar. Ecol. Prog. Ser. 444, 97–115 (2012).
    Google Scholar 
    De Clippele, L. H. et al. The effect of local hydrodynamics on the spatial extent and morphology of cold-water coral habitats at Tisler Reef, Norway. Coral Reefs 37, 253–266 (2018).PubMed 

    Google Scholar 
    Jokiel, P. L. Effects of water motion on reef corals. J. Exp. Mar. Biol. Ecol. 35, 87–97 (1978).
    Google Scholar 
    Shashar, N., Cohen, Y. & Loya, Y. Extreme diel fluctuations of oxygen in diffusive boundary layers surrounding stony corals. Biol. Bull. 185, 455–461 (1993).CAS 
    PubMed 

    Google Scholar 
    Finelli, C. M., Helmuth, B. S. T., Pentcheff, N. D. & Wethey, D. S. Water flow influences oxygen transport and photosynthetic efficiency in corals. Coral Reefs 25, 47–57 (2006).
    Google Scholar 
    Atkinson, M. J. & Bilger, R. W. Effects of water velocity on phosphate uptake in coral reef-hat communities. Limnol. Oceanogr. 37, 273–279 (1992).CAS 

    Google Scholar 
    Mass, T., Genin, A., Shavit, U., Grinstein, M. & Tchernov, D. Flow enhances photosynthesis in marine benthic autotrophs by increasing the efflux of oxygen from the organism to the water. Proc. Natl. Acad. Sci. 107, 2527–2531 (2010).CAS 
    PubMed 
    PubMed Central 

    Google Scholar 
    Comeau, S., Edmunds, P. J., Lantz, C. A. & Carpenter, R. C. Water flow modulates the response of coral reef communities to ocean acidification. Sci. Rep. 4, 6681 (2014).CAS 
    PubMed 
    PubMed Central 

    Google Scholar 
    Larsson, A., Lundälv, T. & van Oevelen, D. Skeletal growth, respiration rate and fatty acid composition in the cold-water coral Lophelia pertusa under varying food conditions. Mar. Ecol. Prog. Ser. 483, 169–184 (2013).
    Google Scholar 
    Baussant, T., Nilsen, M., Ravagnan, E., Westerlund, S. & Ramanand, S. Physiological responses and lipid storage of the coral Lophelia pertusa at varying food density. J. Toxicol. Environ. Health. A 80, 266–284 (2017).CAS 
    PubMed 

    Google Scholar 
    Bouma, T. J. et al. Spatial flow and sedimentation patterns within patches of epibenthic structures: Combining field, flume and modelling experiments. Cont. Shelf Res. 27, 1020–1045 (2007).
    Google Scholar 
    Brooke, S. D., Holmes, M. W. & Young, C. M. Sediment tolerance of two different morphotypes of the deep-sea coral Lophelia pertusa from the Gulf of Mexico. Mar. Ecol. Prog. Ser. 390, 137–144 (2009).
    Google Scholar 
    Bøe, R. et al. Giant sandwaves in the Hola glacial trough off Vesterålen, North Norway. Mar. Geol. 267, 36–54 (2009).
    Google Scholar 
    Huvenne, V. A. I. et al. The Magellan mound province in the Porcupine Basin. Int. J. Earth Sci. 96, 85–101 (2007).CAS 

    Google Scholar 
    De Haas, H. et al. Morphology and sedimentology of (clustered) cold-water coral mounds at the south Rockall Trough margins, NE Atlantic Ocean. Facies 55, 1–26 (2009).
    Google Scholar 
    Lim, A., Huvenne, V. A. I., Vertino, A., Spezzaferri, S. & Wheeler, A. J. New insights on coral mound development from groundtruthed high-resolution ROV-mounted multibeam imaging. Mar. Geol. 403, 225–237 (2018).
    Google Scholar 
    Olariaga, A., Gori, A., Orejas, C. & Gili, J. M. Development of an autonomous aquarium system for maintaining deep corals. Oceanography 22, 44–45 (2009).
    Google Scholar 
    Davies, A. J. et al. Short-term environmental variability in cold-water coral habitat at Viosca Knoll, Gulf of Mexico. Deep Sea Res. I(57), 199–212 (2010).
    Google Scholar 
    Mienis, F. et al. The influence of near-bed hydrodynamic conditions on cold-water corals in the Viosca Knoll area, Gulf of Mexico. Deep Sea Res. I(60), 32–45 (2012).
    Google Scholar 
    Flo, E., Garcés, E., Manzanera, M. & Camp, J. Coastal inshore waters in the NW Mediterranean: Physicochemical and biological characterization and management implications. Estuar. Coast. Shelf Sci. 93, 279–289 (2011).CAS 

    Google Scholar 
    Davies, P. S. Short-term growth measurements of corals using an accurate buoyant weighing technique. Mar. Biol. 101, 389–395 (1989).
    Google Scholar 
    R Core Team. R: A Language and Environment for Statistical Computing. (R Core Team, 2018).Bradford, M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254 (1976).CAS 
    PubMed 

    Google Scholar 
    Thérond, P., Auger, J., Legrand, A. & Jouannet, P. α-tocopherol in human spermatozoa and seminal plasma: Relationships with motility, antioxidant enzymes and leukocytes. Mol. Hum. Reprod. 2, 739–744 (1996).PubMed 

    Google Scholar 
    Beers, R. F. & Sizer, I. W. A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J. Biol. Chem. 195, 133–140 (1952).CAS 
    PubMed 

    Google Scholar 
    Kalghatgi, S. et al. Bactericidal antibiotics induce mitochondrial dysfunction and oxidative damage in mammalian cells. Sci. Transl. Med. 5, 1–10 (2013).
    Google Scholar  More

  • in

    Green synthesis of zinc oxide nanoparticles using Sea Lavender (Limonium pruinosum L. Chaz.) extract: characterization, evaluation of anti-skin cancer, antimicrobial and antioxidant potentials

    Becker, J., Manske, C. & Randl, S. Green chemistry and sustainability metrics in the pharmaceutical manufacturing sector. Curr. Opin. Green Sustain. Chem. https://doi.org/10.1016/j.cogsc.2021.100562 (2022).Article 

    Google Scholar 
    Rajasekharreddy, P., Rani, P. U. & Sreedhar, B. Qualitative assessment of silver and gold nanoparticle synthesis in various plants: A photobiological approach. J. Nanoparticle Res. 12, 25 (2010).Article 

    Google Scholar 
    Mahmoud, A. E. D. Eco-friendly reduction of graphene oxide via agricultural byproducts or aquatic macrophytes. Mater. Chem. Phys. 253, 123336 (2020).Article 
    CAS 

    Google Scholar 
    Mahmoud, A. E. D., Stolle, A. & Stelter, M. Sustainable synthesis of high-surface-area graphite oxide via dry ball milling. ACS Sustain. Chem. Eng. 6, 25 (2018).Article 

    Google Scholar 
    Mellinas, C., Jiménez, A. & del Carmen Garrigós, M. Microwave-assisted green synthesis and antioxidant activity of selenium nanoparticles using theobroma cacao. l. bean shell extract. Molecules 24, 25 (2019).Article 

    Google Scholar 
    Ahmed, S., Saifullah, A. M., Swami, B. L. & Ikram, S. Green synthesis of silver nanoparticles using Azadirachta indica aqueous leaf extract. J. Radiat. Res. Appl. Sci. 9, 25 (2016).
    Google Scholar 
    Ebadi, M. et al. A bio-inspired strategy for the synthesis of zinc oxide nanoparticles (ZnO NPs) using the cell extract of cyanobacterium: Nostoc sp EA03: From biological function to toxicity evaluation. RSC Adv. 9, 25 (2019).Article 

    Google Scholar 
    Mahmoud, A. E. D. & Fawzy, M. Nanosensors and nanobiosensors for monitoring the environmental pollutants. Top. Min. Metallurg. Mater. Eng. https://doi.org/10.1007/978-3-030-68031-2_9 (2021).Article 

    Google Scholar 
    Mousavi, S. M. et al. Green synthesis of silver nanoparticles toward bio and medical applications: Review study. Artif. Cells Nanomed. Biotechnol. 46, 3. https://doi.org/10.1080/21691401.2018.1517769 (2018).Article 
    CAS 

    Google Scholar 
    Hussain, I., Singh, N. B., Singh, A., Singh, H. & Singh, S. C. Green synthesis of nanoparticles and its potential application. Biotechnol. Lett. 38, 25. https://doi.org/10.1007/s10529-015-2026-7 (2016).Article 
    CAS 

    Google Scholar 
    Singh, P., Kim, Y. J., Zhang, D. & Yang, D. C. Biological synthesis of nanoparticles from plants and microorganisms. Trends Biotechnol. 34, 25. https://doi.org/10.1016/j.tibtech.2016.02.006 (2016).Article 
    CAS 

    Google Scholar 
    Nilavukkarasi, M., Vijayakumar, S. & Prathipkumar, S. Capparis zeylanica mediated bio-synthesized ZnO nanoparticles as antimicrobial, photocatalytic and anti-cancer applications. Mater. Sci. Energy Technol. 3, 25 (2020).
    Google Scholar 
    Hussain, A. et al. Biogenesis of ZnO nanoparticles using: Pandanus odorifer leaf extract: Anticancer and antimicrobial activities. RSC Adv. 9, 25 (2019).Article 

    Google Scholar 
    Mohamed Isa, E. D., Shameli, K., Che Jusoh, N. W., Mohamad Sukri, S. N. A. & Ismail, N. A. Variation of green synthesis techniques in fabrication of zinc oxide nanoparticles—a mini review. IOP Conf. Ser. Mater. Sci. Eng. 1051, 25 (2021).Article 

    Google Scholar 
    Loganathan, S., Shivakumar, M. S., Karthi, S., Nathan, S. S. & Selvam, K. Metal oxide nanoparticle synthesis (ZnO-NPs) of Knoxia sumatrensis (Retz.) DC. Aqueous leaf extract and It’s evaluation of their antioxidant, anti-proliferative and larvicidal activities. Toxicol. Rep. 8, 25 (2021).
    Google Scholar 
    Mahmoud, A. E. D., El-Maghrabi, N., Hosny, M. & Fawzy, M. Biogenic synthesis of reduced graphene oxide from Ziziphus spina-christi (Christ’s thorn jujube) extracts for catalytic, antimicrobial, and antioxidant potentialities. Environ. Sci. Pollut. Res. 20, 1–16 (2022).
    Google Scholar 
    Ahmar Rauf, M., Oves, M., Ur Rehman, F., Rauf Khan, A. & Husain, N. Bougainvillea flower extract mediated zinc oxide’s nanomaterials for antimicrobial and anticancer activity. Biomed. Pharmacother. 116, 25 (2019).Article 

    Google Scholar 
    Chabattula, S. C. et al. Anticancer therapeutic efficacy of biogenic Am-ZnO nanoparticles on 2D and 3D tumor models. Mater. Today Chem. 22, 25 (2021).
    Google Scholar 
    Berehu, H. M. et al. Cytotoxic potential of biogenic zinc oxide nanoparticles synthesized from swertia chirayita leaf extract on colorectal cancer cells. Front. Bioeng. Biotechnol. 9, 25 (2021).Article 

    Google Scholar 
    Khezri, K., Saeedi, M. & Maleki Dizaj, S. Application of nanoparticles in percutaneous delivery of active ingredients in cosmetic preparations. Biomed. Pharmacother. 106, 25. https://doi.org/10.1016/j.biopha.2018.07.084 (2018).Article 
    CAS 

    Google Scholar 
    Smijs, T. G. & Pavel, S. Titanium dioxide and zinc oxide nanoparticles in sunscreens: Focus on their safety and effectiveness. Nanotechnol. Sci. Appl. 4, 25. https://doi.org/10.2147/nsa.s19419 (2011).Article 

    Google Scholar 
    Nasrollahzadeh, M. S. et al. Zinc oxide nanoparticles as a potential agent for antiviral drug delivery development: A systematic literature review. Curr. Nanosci. 18, 25 (2021).
    Google Scholar 
    Perera, W. P. T. D. et al. Albumin grafted coaxial electrosparyed polycaprolactone-zinc oxide nanoparticle for sustained release and activity enhanced antibacterial drug delivery. RSC Adv. 12, 25 (2022).Article 

    Google Scholar 
    Shalaby, M. A., Anwar, M. M. & Saeed, H. Nanomaterials for application in wound healing: Current state-of-the-art and future perspectives. J. Polym. Res. 29, 25. https://doi.org/10.1007/s10965-021-02870-x (2022).Article 
    CAS 

    Google Scholar 
    Kaushik, M. et al. Investigations on the antimicrobial activity and wound healing potential of ZnO nanoparticles. Appl. Surf. Sci. 479, 25 (2019).Article 

    Google Scholar 
    Espitia, P. J. P., Otoni, C. G. & Soares, N. F. F. Zinc oxide nanoparticles for food packaging applications. Antimicrob. Food Packag. https://doi.org/10.1016/B978-0-12-800723-5.00034-6.4 (2016).Article 

    Google Scholar 
    Doan Thi, T. U. et al. Green synthesis of ZnO nanoparticles using orange fruit peel extract for antibacterial activities. RSC Adv. 10, 25 (2020).Article 

    Google Scholar 
    Shobha, N. et al. Synthesis and characterization of Zinc oxide nanoparticles utilizing seed source of Ricinus communis and study of its antioxidant, antifungal and anticancer activity. Mater. Sci. Eng. C 97, 25 (2019).Article 

    Google Scholar 
    Zahran, M. A. & Willis, A. J. The vegetation of Egypt. Plant Veget. 2, 25 (2009).
    Google Scholar 
    El-Borady, O. M., Fawzy, M. & Hosny, M. Antioxidant, anticancer and enhanced photocatalytic potentials of gold nanoparticles biosynthesized by common reed leaf extract. Appl. Nanosci. (Switzerland) https://doi.org/10.1007/s13204-021-01776-w (2021).Article 

    Google Scholar 
    Hosny, M., Fawzy, M., Abdelfatah, A. M., Fawzy, E. E. & Eltaweil, A. S. Comparative study on the potentialities of two halophytic species in the green synthesis of gold nanoparticles and their anticancer, antioxidant and catalytic efficiencies. Adv. Powder Technol. 32, 25 (2021).Article 

    Google Scholar 
    Vijayakumar, S. et al. Acalypha fruticosa L. Leaf extract mediated synthesis of ZnO nanoparticles: Characterization and antimicrobial activities. Mater. Today Proc. 23, 25 (2019).
    Google Scholar 
    Fatimah, I., Pradita, R. Y. & Nurfalinda, A. Plant extract mediated of ZnO nanoparticles by using ethanol extract of mimosa pudica leaves and coffee powder. Proced. Eng. 148, 25 (2016).Article 

    Google Scholar 
    Heneidy, S. Z. & Bidak, L. M. Potential uses of plant species of the coastal mediterranean region, Egypt. Pak. J. Biol. Sci. 7, 1010–1023 (2004).Article 

    Google Scholar 
    Manousaki, E. & Kalogerakis, N. Halophytes present new opportunities in phytoremediation of heavy metals and saline soils. Ind. Eng. Chem. Res. 50, 25 (2011).Article 

    Google Scholar 
    Zengin, G., Aumeeruddy-Elalfi, Z., Mollica, A., Yilmaz, M. A. & Mahomoodally, M. F. In vitro and in silico perspectives on biological and phytochemical profile of three halophyte species—a source of innovative phytopharmaceuticals from nature. Phytomedicine 38, 35–44 (2018).Article 
    CAS 
    PubMed 

    Google Scholar 
    Xin, P. et al. Surface water and groundwater interactions in salt marshes and their impact on plant ecology and coastal biogeochemistry. Rev. Geophys. 60, 5. https://doi.org/10.1029/2021RG000740 (2022).Article 

    Google Scholar 
    International Union for Conservation of Nature. International Union for Conservation of Nature Natural Resources IUCN Red List Categories and Criteria (IUCN, 2001).
    Google Scholar 
    Boulos, L. Flora of Egypt Vol 417 21–22 (Al Hadara Publishing, 1999).
    Google Scholar 
    Safawo, T., Sandeep, B. V., Pola, S. & Tadesse, A. Synthesis and characterization of zinc oxide nanoparticles using tuber extract of anchote (Coccinia abyssinica (Lam.) Cong.) for antimicrobial and antioxidant activity assessment. Open Nano 3, 25 (2018).
    Google Scholar 
    Soltanian, S. et al. Biosynthesis of zinc oxide nanoparticles using hertia intermedia and evaluation of its cytotoxic and antimicrobial activities. https://doi.org/10.1007/s12668-020-00816-z/Published.Ogbole, O. O., Segun, P. A. & Adeniji, A. J. In vitro cytotoxic activity of medicinal plants from Nigeria ethnomedicine on Rhabdomyosarcoma cancer cell line and HPLC analysis of active extracts. BMC Complement Altern. Med. 17, 25 (2017).Article 

    Google Scholar 
    Slater, T. F., Sawyer, B. & Sträuli, U. Studies on succinate-tetrazolium reductase systems. III Points of coupling of four different tetrazolium salts. Biochim. Biophys. Acta 77, 25 (1963).Article 

    Google Scholar 
    Alley, M. C. et al. Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Res. 48, 25 (1988).
    Google Scholar 
    van de Loosdrecht, A. A., Beelen, R. H. J., Ossenkoppele, G. J., Broekhoven, M. G. & Langenhuijsen, M. M. A. C. A tetrazolium-based colorimetric MTT assay to quantitate human monocyte mediated cytotoxicity against leukemic cells from cell lines and patients with acute myeloid leukemia. J. Immunol. Methods 174, 25 (1994).
    Google Scholar 
    Gonelimali, F. D. et al. Antimicrobial properties and mechanism of action of some plant extracts against food pathogens and spoilage microorganisms. Front. Microbiol. 9, 25 (2018).Article 

    Google Scholar 
    Aldalbahi, A. et al. Greener synthesis of zinc oxide nanoparticles: Characterization and multifaceted applications. Molecules 25, 25 (2020).Article 

    Google Scholar 
    López-Cuenca, S. et al. High-yield synthesis of zinc oxide nanoparticles from bicontinuous microemulsions. J. Nanomater. 2011, 25 (2011).Article 

    Google Scholar 
    Sajadi, S. M. et al. Natural iron ore as a novel substrate for the biosynthesis of bioactive-stable ZnO@CuO@iron ore NCs: A magnetically recyclable and reusable superior nanocatalyst for the degradation of organic dyes, reduction of Cr(vi) and adsorption of crude oil aromatic compounds, including PAHs. RSC Adv. 8, 62. https://doi.org/10.1039/c8ra06028b (2018).Article 
    CAS 

    Google Scholar 
    Meena, P. L., Poswal, K. & Surela, A. K. Facile synthesis of ZnO nanoparticles for the effective photodegradation of malachite green dye in aqueous solution. Water Environ. J. 36, 25 (2022).Article 

    Google Scholar 
    El-Belely, E. F. et al. Green synthesis of zinc oxide nanoparticles (Zno-nps) using arthrospira platensis (class: Cyanophyceae) and evaluation of their biomedical activities. Nanomaterials 11, 25 (2021).Article 

    Google Scholar 
    Faye, G., Jebessa, T. & Wubalem, T. Biosynthesis, characterisation and antimicrobial activity of zinc oxide and nickel doped zinc oxide nanoparticles using Euphorbia abyssinica bark extract (2021). https://doi.org/10.1049/nbt2.12072.Dulta, K., Koşarsoy Ağçeli, G., Chauhan, P., Jasrotia, R. & Chauhan, P. K. A novel approach of synthesis zinc oxide nanoparticles by Bergenia ciliata rhizome extract: Antibacterial and anticancer potential. J. Inorg. Organomet. Polym. Mater. 31, 25 (2021).Article 

    Google Scholar 
    Faisal, S. et al. Green synthesis of zinc oxide (ZnO) nanoparticles using aqueous fruit extracts of Myristica fragrans: Their characterizations and biological and environmental applications. ACS Omega 6, 25 (2021).Article 

    Google Scholar 
    Adams, R. P. Identification of essential oil components by gas chromatography/mass spectrometry. J. Am. Soc. Mass Spectrometry 8, 25 (2007).
    Google Scholar 
    VStein, S., Mirokhin, D., Tchekhovskoi, D., & Nist, G. M. The NIST Mass Spectral Search Program for the NIST/EPA/NIH Mass Spectra Library. Gaithersburg, MD: Standard Reference Data Program of the National Institute of Standards and Technology (2002).Mahmoud, A. E. D., Hosny, M., El-Maghrabi, N. & Fawzy, M. Facile synthesis of reduced graphene oxide by Tecoma stans extracts for efficient removal of Ni (II) from water: Batch experiments and response surface methodology. Sustain. Environ. Res. 32, 25 (2022).Article 

    Google Scholar 
    Balasubramani, G. et al. GC-MS analysis of bioactive components and synthesis of gold nanoparticle using Chloroxylon swietenia DC leaf extract and its larvicidal activity. J. Photochem. Photobiol. B 148, 25 (2015).Article 

    Google Scholar 
    Barzinjy, A. A. & Azeez, H. H. Green synthesis and characterization of zinc oxide nanoparticles using Eucalyptus globulus Labill. leaf extract and zinc nitrate hexahydrate salt. SN Appl. Sci. 2, 25 (2020).Article 

    Google Scholar 
    Anitha, R., Ramesh, K. V., Ravishankar, T. N., Sudheer Kumar, K. H. & Ramakrishnappa, T. Cytotoxicity, antibacterial and antifungal activities of ZnO nanoparticles prepared by the Artocarpus gomezianus fruit mediated facile green combustion method. J. Sci. Adv. Mater. Devices 3, 25 (2018).
    Google Scholar 
    Chandra, H., Patel, D., Kumari, P., Jangwan, J. S. & Yadav, S. Phyto-mediated synthesis of zinc oxide nanoparticles of Berberis aristata: Characterization, antioxidant activity and antibacterial activity with special reference to urinary tract pathogens. Mater. Sci. Eng. C 102, 25 (2019).Article 

    Google Scholar 
    Majeed, S., Danish, M., Ismail, M. H., Ansari, M. T. & Ibrahim, M. N. M. Anticancer and apoptotic activity of biologically synthesized zinc oxide nanoparticles against human colon cancer HCT-116 cell line- in vitro study. Sustain. Chem. Pharm. 14, 25 (2019).
    Google Scholar 
    Miri, A., Khatami, M., Ebrahimy, O. & Sarani, M. Cytotoxic and antifungal studies of biosynthesized zinc oxide nanoparticles using extract of Prosopis farcta fruit. Green Chem. Lett. Rev. 13, 25. https://doi.org/10.1080/17518253.2020.1717005 (2020).Article 
    CAS 

    Google Scholar 
    Ahamed, M., Akhtar, M. J., Khan, M. A. M. & Alhadlaq, H. A. Enhanced anticancer performance of eco-friendly-prepared Mo-ZnO/RGO nanocomposites: Role of oxidative stress and apoptosis. ACS Omega 7, 25 (2022).Article 

    Google Scholar 
    Al-Mohaimeed, A. M., Al-Onazi, W. A. & El-Tohamy, M. F. Multifunctional eco-friendly synthesis of ZnO nanoparticles in biomedical applications. Molecules 27, 25 (2022).Article 

    Google Scholar 
    Schreyer, M., Guo, L., Thirunahari, S., Gao, F. & Garland, M. Simultaneous determination of several crystal structures from powder mixtures: The combination of powder X-ray diffraction, band-target entropy minimization and Rietveld methods. J. Appl. Crystallogr. 47, 25 (2014).Article 

    Google Scholar 
    Pu, Y., Niu, Y., Wang, Y., Liu, S. & Zhang, B. Statistical morphological identification of low-dimensional nanomaterials by using TEM. Particuology 61, 11–17 (2022).Article 
    CAS 

    Google Scholar 
    Wu, C. M., Baltrusaitis, J., Gillan, E. G. & Grassian, V. H. Sulfur dioxide adsorption on ZnO nanoparticles and nanorods. J. Phys. Chem. C 115, 10164–10172 (2011).Article 
    CAS 

    Google Scholar 
    Saranya, S., Vijayaranai, K., Pavithra, S., Raihana, N. & Kumanan, K. In vitro cytotoxicity of zinc oxide, iron oxide and copper nanopowders prepared by green synthesis. Toxicol. Rep. 4, 25 (2017).
    Google Scholar 
    Chelladurai, M. et al. Anti-skin cancer activity of Alpinia calcarata ZnO nanoparticles: Characterization and potential antimicrobial effects. J Drug Deliv. Sci. Technol. 61, 102180 (2021).Article 
    CAS 

    Google Scholar 
    Lingaraju, K., Naika, H. R., Nagabhushana, H. & Nagaraju, G. Euphorbia heterophylla (L.) mediated fabrication of ZnO NPs: Characterization and evaluation of antibacterial and anticancer properties. Biocatal. Agric. Biotechnol. 18, 25 (2019).Article 

    Google Scholar 
    Sana, S. S. et al. Crotalaria verrucosa leaf extract mediated synthesis of zinc oxide nanoparticles: Assessment of antimicrobial and anticancer activity. Molecules 25, 25 (2020).Article 

    Google Scholar 
    Bisht, G. & Rayamajhi, S. ZnO nanoparticles: A promising anticancer agent. Nanobiomedicine https://doi.org/10.5772/63437 (2016).Article 
    PubMed 
    PubMed Central 

    Google Scholar 
    Bharath, B., Perinbam, K., Devanesan, S., AlSalhi, M. S. & Saravanan, M. Evaluation of the anticancer potential of Hexadecanoic acid from brown algae Turbinaria ornata on HT–29 colon cancer cells. J. Mol. Struct. 1235, 25 (2021).Article 

    Google Scholar 
    Selim, Y. A., Azb, M. A., Ragab, I., Abd El-Azim, H. M. & M.,. Green synthesis of zinc oxide nanoparticles using aqueous extract of Deverra tortuosa and their cytotoxic activities. Sci. Rep. 10, 25 (2020).Article 

    Google Scholar 
    Medini, F. et al. Phytochemical analysis, antioxidant, anti-inflammatory, and anticancer activities of the halophyte Limonium densiflorum extracts on human cell lines and murine macrophages. South Afr. J. Bot. 99, 25 (2015).Article 

    Google Scholar 
    Pan, M. H., Ghai, G. & Ho, C. T. Food bioactives, apoptosis, and cancer. Mol. Nutr. Food Res. 52, 20. https://doi.org/10.1002/mnfr.200700380 (2008).Article 
    CAS 

    Google Scholar 
    Abdallah, H. M. & Ezzat, S. M. Effect of the method of preparation on the composition and cytotoxic activity of the essential oil of Pituranthos tortuosus. Z. Nat. Sect. C J. Biosci. 66 C, 25 (2011).
    Google Scholar 
    Iqbal, J. et al. Green synthesis of zinc oxide nanoparticles using Elaeagnus angustifolia L. leaf extracts and their multiple in vitro biological applications. Sci. Rep. 11, 25 (2021).Article 

    Google Scholar 
    Norouzi Jobie, F., Ranjbar, M., Hajizadeh Moghaddam, A. & Kiani, M. Green synthesis of zinc oxide nanoparticles using Amygdalus scoparia Spach stem bark extract and their applications as an alternative antimicrobial, anticancer, and anti-diabetic agent. Adv. Powder Technol. 32, 21 (2021).Article 

    Google Scholar 
    Chen, F. C., Huang, C. M., Yu, X. W. & Chen, Y. Y. Effect of nano zinc oxide on proliferation and toxicity of human gingival cells. Hum. Exp. Toxicol. 41, 15 (2022).Article 

    Google Scholar 
    Sajjad, A. et al. Photoinduced fabrication of zinc oxide nanoparticles: Transformation of morphological and biological response on light irradiance. ACS Omega 6, 75 (2021).Article 

    Google Scholar 
    Sohail, M. F. et al. Green synthesis of zinc oxide nanoparticles by neem extract as multi-facet therapeutic agents. J. Drug Deliv. Sci. Technol. 59, 15 (2020).
    Google Scholar 
    Lopes, M., Sanches-Silva, A., Castilho, M., Cavaleiro, C. & Ramos, F. Halophytes as source of bioactive phenolic compounds and their potential applications. Crit. Rev. Food Sci. Nutr. 20, 20. https://doi.org/10.1080/10408398.2021.1959295 (2021).Article 
    CAS 

    Google Scholar 
    Bouarab-Chibane, L. et al. Antibacterial properties of polyphenols: Characterization and QSAR (quantitative structure-activity relationship) models. Front. Microbiol. 10, 77 (2019).Article 

    Google Scholar 
    Guimarães, A. C. et al. Antibacterial activity of terpenes and terpenoids present in essential oils. Molecules 24, 11 (2019).Article 

    Google Scholar 
    Sirelkhatim, A. et al. Review on zinc oxide nanoparticles: Antibacterial activity and toxicity mechanism. Nano-Micro Lett. 7, 219–242. https://doi.org/10.1007/s40820-015-0040-x (2015).Article 
    CAS 

    Google Scholar 
    Singh, T. A. et al. A state of the art review on the synthesis, antibacterial, antioxidant, antidiabetic and tissue regeneration activities of zinc oxide nanoparticles. Adv. Coll. Interface Sci. 295, 25. https://doi.org/10.1016/j.cis.2021.102495 (2021).Article 
    CAS 

    Google Scholar 
    Gao, Y. et al. Biofabrication of zinc oxide nanoparticles from Aspergillus niger, their antioxidant, antimicrobial and anticancer activity. J. Clust. Sci. 30, 11 (2019).Article 

    Google Scholar 
    Luo, Q. et al. Terpenoid composition and antioxidant activity of extracts from four chemotypes of Cinnamomum camphora and their main antioxidant agents. Biofuels Bioprod. Biorefin. 16, 510–522 (2022).Article 
    CAS 

    Google Scholar 
    Bose, J., Rodrigo-Moreno, A. & Shabala, S. ROS homeostasis in halophytes in the context of salinity stress tolerance. J. Exp. Bot. 65, 25. https://doi.org/10.1093/jxb/ert430 (2014).Article 
    CAS 

    Google Scholar  More

  • in

    Experimental climate change impacts on Baltic coastal wetland plant communities

    Kimmel, K., Kull, A., Salm, J. & Mander, Ü. The status, conservation and sustainable use of Estonian wetlands. Wetl. Ecol. Manag. 18, 375–395. https://doi.org/10.1007/s11273-008-9129-z (2008).Article 

    Google Scholar 
    Engle, V. Estimating the provision of ecosystem services by Gulf of Mexico coastal wetlands. Wetlands 31, 179–193. https://doi.org/10.1007/s13157-010-0132-9 (2011).Article 

    Google Scholar 
    Ward, R., Teasdale, P., Burnside, N., Joyce, C. & Sepp, K. Recent rates of sedimentation on irregularly flooded Boreal Baltic coastal wetlands: Responses to recent changes in sea level. Geomorphology 217, 61–72. https://doi.org/10.1016/j.geomorph.2014.03.045 (2014).Article 

    Google Scholar 
    Villoslada Peciña, M. et al. Country-scale mapping of ecosystem services provided by semi-natural grasslands. Sci. Total Environ. 661, 212–225. https://doi.org/10.1016/j.scitotenv.2019.01.174 (2019).Article 
    CAS 
    PubMed 

    Google Scholar 
    Lima, M., Ward, R. & Joyce, C. Environmental drivers of sediment carbon storage in temperate seagrass meadows. Hydrobiologia 847, 1773–1792. https://doi.org/10.1007/s10750-019-04153-5 (2019).Article 
    CAS 

    Google Scholar 
    Ward, R. Sedimentary response of Arctic coastal wetlands to sea level rise. Geomorphology 370, 107400. https://doi.org/10.1016/j.geomorph.2020.107400 (2020).Article 

    Google Scholar 
    Akumu, C., Pathirana, S., Baban, S. & Bucher, D. Examining the potential impacts of sea level rise on coastal wetlands in north-eastern NSW, Australia. J. Coast. Conserv. 15, 15–22. https://doi.org/10.1007/s11852-010-0114-3 (2010).Article 

    Google Scholar 
    Ward, R. Carbon sequestration and storage in Norwegian Arctic coastal wetlands: Impacts of climate change. Sci. Total Environ. 748, 141343. https://doi.org/10.1016/j.scitotenv.2020.141343 (2020).Article 
    CAS 
    PubMed 

    Google Scholar 
    Hossain, M., Hein, L., Rip, F. & Dearing, J. Integrating ecosystem services and climate change responses in coastal wetlands development plans for Bangladesh. Mitig. Adapt. Strateg. Glob. Chang. 20, 241–261. https://doi.org/10.1007/s11027-013-9489-4 (2015).Article 

    Google Scholar 
    Ward, R., Friess, D., Day, R. & Mackenzie, R. Impacts of climate change on global mangrove ecosystems: A regional comparison. Ecosyst. Health Sustain. 4, 1–25 (2016).
    Google Scholar 
    Graham, L. P. et al. Climate change. In The Baltic Sea Area Draft HELCOM Thematic Assessment. (Helsinki Commission, Baltic Marine Environmental Protection Commission, 2007).BACC. Assessment of Climate Change for the Baltic Sea Basin. (Springer Science & Business Media, 2008).
    Google Scholar 
    Rivis, R. et al. Trends in the development of Estonian coastal land cover and landscapes caused by natural changes and human impact. J. Coast. Conserv. 20, 199–209. https://doi.org/10.1007/s11852-016-0430-3 (2016).Article 

    Google Scholar 
    Cubasch, U. et al. Projections of future climate change. in IPCC Climate Change 2001: The Scientific Basis Contribution of Working Group I to the Third Assessment Report of the Intergovernmental Panel on Climate Change. (Cambridge University Press, 2001).Mafi-Gholami, D., Zenner, E., Jaafari, A. & Ward, R. Modeling multi-decadal mangrove leaf area index in response to drought along the semi-arid southern coasts of Iran. Sci. Total Environ. 656, 1326–1336. https://doi.org/10.1016/j.scitotenv.2018.11.462 (2019).Article 
    CAS 
    PubMed 

    Google Scholar 
    IPCC. Global Warming of 1.5 ºC. Ipcc.ch. https://www.ipcc.ch/sr15/ (2008).Omstedt, A., Pettersen, C., Rodhe, J. & Winsor, P. Baltic Sea climate: 200 yr of data on air temperature, sea level variation, ice cover, and atmospheric circulation. Clim. Res. 25, 205–216 (2004).Article 

    Google Scholar 
    Räisänen, J. Future climate change in the Baltic Sea Region and environmental impacts. Oxf. Res. Encycl. Clim. Sci. https://doi.org/10.1093/acrefore/9780190228620.013.634 (2017).Article 

    Google Scholar 
    Dippner, J. W. et al. Climate-related marine ecosystem change. in Team, B. A. Assessment of Climate Change for the Baltic Sea Basin. (SSBM, 2008).Ward, R., Burnside, N., Joyce, C., Sepp, K. & Teasdale, P. Improved modelling of the impacts of sea level rise on coastal wetland plant communities. Hydrobiologia 774, 203–216. https://doi.org/10.1007/s10750-015-2374-2 (2016).Article 

    Google Scholar 
    Vuorinen, I. Proportion of copepod biomass declines with decreasing salinity in the Baltic Sea. ICES Mar. Sci. 55, 767–774. https://doi.org/10.1006/jmsc.1998.0398 (1998).Article 

    Google Scholar 
    Berg, M., Joyce, C. & Burnside, N. Differential responses of abandoned wet grassland plant communities to reinstated cutting management. Hydrobiologia 692, 83–97. https://doi.org/10.1007/s10750-011-0826-x (2011).Article 

    Google Scholar 
    Short, F., Kosten, S., Morgan, P., Malone, S. & Moore, G. Impacts of climate change on submerged and emergent wetland plants. Aquat. Bot. 135, 3–17. https://doi.org/10.1016/j.aquabot.2016.06.006 (2016).Article 

    Google Scholar 
    Ward, R., Burnside, N., Joyce, C. & Sepp, K. Importance of microtopography in determining plant community distribution in Baltic coastal wetlands. J. Coast. Res. 321, 1062–1070. https://doi.org/10.2112/JCOASTRES-D-15-00065.1 (2016).Article 

    Google Scholar 
    Burnside, N., Joyce, C., Puurmann, E. & Scott, D. Use of vegetation classification and plant indicators to assess grazing abandonment in Estonian coastal wetlands. J. Veg. Sci. 18, 645–654. https://doi.org/10.1111/j.1654-1103.2007.tb02578.x (2007).Article 

    Google Scholar 
    Ward, R., Burnside, N., Joyce, C. & Sepp, K. The use of medium point density LiDAR elevation data to determine plant community types in Baltic coastal wetlands. Ecol. Indic. 33, 96–104. https://doi.org/10.1016/j.ecolind.2012.08.016 (2013).Article 

    Google Scholar 
    Goud, E., Watt, C. & Moore, T. Plant community composition along a peatland margin follows alternate successional pathways after hydrologic disturbance. Acta Oecol. 91, 65–72. https://doi.org/10.1016/j.actao.2018.06.006 (2018).Article 

    Google Scholar 
    Moreno, J., Terrones, A., Juan, A. & Alonso, M. Halophytic plant community patterns in Mediterranean saltmarshes: Shedding light on the connection between abiotic factors and the distribution of halophytes. Plant Soil 430, 185–204. https://doi.org/10.1007/s11104-018-3671-0 (2018).Article 
    CAS 

    Google Scholar 
    Sharpe, P. & Baldwin, A. Tidal marsh plant community response to sea-level rise: A mesocosm study. Aquat. Bot. 101, 34–40. https://doi.org/10.1016/j.aquabot.2012.03.015 (2012).Article 

    Google Scholar 
    Lindig-Cisneros, R. & Zedler, J. Phalaris arundinacea seedling establishment: Effects of canopy complexity in fen, mesocosm, and restoration experiments. J. Bot. 80, 617–624. https://doi.org/10.1139/b02-042 (2002).Article 

    Google Scholar 
    Ahn, C. & Mitsch, W. Scaling considerations of mesocosm wetlands in simulating large created freshwater marshes. Ecol. Eng. 18, 327–342. https://doi.org/10.1016/S0925-8574(01)00092-1 (2002).Article 

    Google Scholar 
    Brotherton, S. & Joyce, C. Extreme climate events and wet grasslands: Plant traits for ecological resilience. Hydrobiologia 750, 229–243. https://doi.org/10.1007/s10750-014-2129-5 (2015).Article 

    Google Scholar 
    Stewart, R. I. et al. Mesocosm experiments as a tool for ecological climate-change research. Adv. Ecol. Res. AP. 48, 71–181 (2013).Article 

    Google Scholar 
    Kont, A., Ratas, U. & Puurmann, E. Sea-level rise impact on coastal areas of Estonia. Clim. Change 36, 175–184. https://doi.org/10.1023/A:1005352715752 (1997).Article 

    Google Scholar 
    Short, F. & Neckles, H. The effects of global climate change on seagrasses. Aquat. Bot. 63, 169–196. https://doi.org/10.1016/S0304-3770(98)00117-X (1999).Article 

    Google Scholar 
    Engels, J., Rink, F. & Jensen, K. Stress tolerance and biotic interactions determine plant zonation patterns in estuarine marshes during seedling emergence and early establishment. J. Ecol. 99, 277–287. https://doi.org/10.1111/j.1365-2745.2010.01745.x (2010).Article 

    Google Scholar 
    Rayner, D. et al. Intertidal wetland vegetation dynamics under rising sea levels. Sci. Total Environ. 766, 144237. https://doi.org/10.1016/j.scitotenv.2020.144237 (2021).Article 
    CAS 
    PubMed 

    Google Scholar 
    Toogood, S. & Joyce, C. Effects of raised water levels on wet grassland plant communities. Appl. Veg. Sci. 12, 283–294. https://doi.org/10.1111/j.1654-109X.2009.01028.x (2009).Article 

    Google Scholar 
    Humphreys, A., Gorsky, A., Bilkovic, D. & Chambers, R. Changes in plant communities of low-salinity tidal marshes in response to sea-level rise. Ecosphere https://doi.org/10.1002/ecs2.3630 (2021).Article 

    Google Scholar 
    Jarvis, J. C., McKenna, S. A. & Rasheed, M. A. Seagrass seed bank spatial structure and function following a large-scale decline. Mar. Ecol. Prog. Ser. 665, 75–87. https://doi.org/10.3354/meps13668 (2021).Article 

    Google Scholar 
    Elsey-Quirk, T. & Leck, M. Patterns of seed bank and vegetation diversity along a tidal freshwater river. Am. J. Bot. 102, 1996–2012. https://doi.org/10.3732/ajb.1500314 (2015).Article 
    PubMed 

    Google Scholar 
    Jutila, H. Germination in Baltic coastal wetland meadows: Similarities and differences between vegetation and seed bank. Plant Ecol. 166, 275–293 (2003).Article 

    Google Scholar 
    Ellenberg, H. Zeigerwerte der Gefässpflanzen Mitteleuropas. 42–111. (Scr. Geobot., 1979).Joshi, R. et al. Salt adaptation mechanisms of halophytes: Improvement of salt tolerance in crop plants. in Elucidation of Abiotic Stress Signaling in Plants. (Springer, 2015).Tessier, M., Gloaguen, J. & Lefeuvre, J. Factors affecting the population dynamics of Suaeda maritima at initial stages of development. Plant Ecol. 147, 193–203 (2000).Article 

    Google Scholar 
    Hanslin, H. & Eggen, T. Salinity tolerance during germination of seashore halophytes and salt-tolerant grass cultivars. Seed Sci. Res. 15, 43–50. https://doi.org/10.1079/SSR2004196 (2005).Article 

    Google Scholar 
    Köster, T. et al. The management of the coastal grasslands of Estonia. WIT Trans. Ecol. Environ. https://doi.org/10.2495/CENV040051 (2004).Article 

    Google Scholar 
    Spencer, T. et al. Global coastal wetland change under sea-level rise and related stresses: The DIVA wetland change model. Glob. Planet. Change 139, 15–30. https://doi.org/10.1016/j.gloplacha.2015.12.018 (2016).Article 

    Google Scholar 
    Marani, M., D’Alpaos, A., Lanzoni, S., Carniello, L. & Rinaldo, A. Biologically-controlled multiple equilibria of tidal landforms and the fate of the Venice lagoon. Geophys. Res. Lett. https://doi.org/10.1029/2007GL030178 (2007).Article 

    Google Scholar 
    Petersen, K., Frank, H., Paytan, A. & Bar-Zeev, E. Impacts of seawater desalination on coastal environments. Sustain. Desalin. Handb. https://doi.org/10.1016/B978-0-12-809240-8.00011-3 (2018).Article 

    Google Scholar 
    Rannap, R. et al. Coastal meadow management for threatened waders has a strong supporting impact on meadow plants and amphibians. J. Nat. Conserv. 35, 77–91. https://doi.org/10.1016/j.jnc.2016.12.004 (2017).Article 

    Google Scholar 
    Krauss, K. et al. How mangrove forests adjust to rising sea level. New Phytol. 202, 19–34. https://doi.org/10.1111/nph.12605 (2014).Article 
    PubMed 

    Google Scholar 
    Kirwan, M. et al. Limits on the adaptability of coastal marshes to rising sea level. Geophys. Res. Lett. https://doi.org/10.1029/2010GL045489 (2010).Article 

    Google Scholar 
    Burnside, N., Joyce, C., Berg, M. & Puurman, E. The relationship between microtopography and vegetation in Estonian coastal wetlands: Implications for climate change. Publ. Inst. Geogr. Univ. Tartu. 106, 19–23 (2008).
    Google Scholar 
    Hulisz, P., Piernik, A., Mantilla-Contreras, J. & Elvisto, T. Main driving factors for seacoast vegetation in the southern and eastern Baltic. Wetlands 36, 909–919. https://doi.org/10.1007/s13157-016-0803-2 (2016).Article 

    Google Scholar 
    Gough, L. & Grace, J. Effects of flooding, salinity and herbivory on coastal plant communities, Louisiana, United States. Oecologia 117, 527–535. https://doi.org/10.1007/s004420050689 (1998).Article 
    PubMed 

    Google Scholar 
    Hannerz, F. & Destouni, G. Spatial characterization of the Baltic sea drainage basin and its unmonitored catchments. Ambio 35, 214–219. https://doi.org/10.1579/05-A-022R.1 (2006).Article 
    PubMed 

    Google Scholar 
    Kont, A., Jaagus, J. & Aunap, R. Climate change scenarios and the effect of sea-level rise for Estonia. Glob. Planet. Change 36, 1–15. https://doi.org/10.1016/S0921-8181(02)00149-2 (2003).Article 

    Google Scholar 
    von Storch, H. & Omstedt, A. Introduction and summary. in Team, B. A. Assessment of Climate Change for the Baltic Sea Basin. (SSBM, 2008).Stigebrandt, A. Physical oceanography of the Baltic Sea. in A Systems Analysis of the Baltic Sea. 19–74 (Springer, 2001).Ingerpuu, N. & Sarv, M. Effect of grazing on plant diversity of coastal meadows in Estonia. Ann. Bot. Fenn. 52, 84–92. https://doi.org/10.5735/085.052.0210 (2015).Article 

    Google Scholar 
    Moinardeau, C., Mesléard, F., Ramone, H. & Dutoit, T. Short-term effects on diversity and biomass on grasslands from artificial dykes under grazing and mowing treatments. Environ. Conserv. 46, 132–139. https://doi.org/10.1017/S0376892918000346 (2019).Article 

    Google Scholar 
    Tardella, F. M., Bricca, A., Goia, I. G. & Catorci, A. How mowing restores montane Mediterranean grasslands following cessation of traditional livestock grazing. Agric. Ecosyst. Environ. 295, 1158. https://doi.org/10.1016/j.agee.2020.106880 (2020).Article 

    Google Scholar 
    Lindborg, R. & Eriksson, O. Historical landscape connectivity affects present plant species diversity. Ecology 85, 1840–1845. https://doi.org/10.1890/04-0367 (2004).Article 

    Google Scholar 
    Villoslada Peciña, M., Bergamo, T., Ward, R., Joyce, C. & Sepp, K. A novel UAV-based approach for biomass prediction and grassland structure assessment in coastal meadows. Ecol. Indic. 122, 107227. https://doi.org/10.1016/j.ecolind.2020.107227 (2021).Article 

    Google Scholar 
    Villoslada, M. et al. Fine scale plant community assessment in coastal meadows using UAV based multispectral data. Ecol. Indic. 111, 105979. https://doi.org/10.1016/j.ecolind.2019.105979 (2020).Article 

    Google Scholar 
    Araya, Y., Gowing, D. & Dise, N. A controlled water-table depth system to study the influence of fine-scale differences in water regime for plant growth. Aquat. Bot. 92, 70–74. https://doi.org/10.1016/j.aquabot.2009.10.004 (2010).Article 

    Google Scholar 
    Koch, E. et al. Non-linearity in ecosystem services: Temporal and spatial variability in coastal protection. Front. Ecol. Environ. 7, 29–37. https://doi.org/10.1890/080126 (2009).Article 

    Google Scholar 
    Church, J. & White, N. Sea-level rise from the late 19th to the early 21st century. Surv. Geophys. 32, 585–602. https://doi.org/10.1007/s10712-011-9119-1 (2011).Article 

    Google Scholar 
    Goodwillie, C., McCoy, M. & Peralta, A. Long-term nutrient enrichment, mowing, and ditch drainage interact in the dynamics of a wetland plant community. Ecosphere. https://doi.org/10.1002/ecs2.3252 (2020).Article 

    Google Scholar 
    Kindt, R. & Coe, R. Tree diversity analysis. A manual and software for common statistical methods for ecological and biodiversity studies. in World Agroforestry | Transforming Lives and Landscapes with Trees. http://www.worldagroforestry.org/output/tree-diversity-analysis (2005).Oksanen, J. et al. CRAN—Package Vegan. Cran.r-project.org. https://CRAN.R-project.org/package=vegan. (2022).Wickham, H. Create Elegant Data Visualisations Using the Grammar of Graphics. Ggplot2.tidyverse.org. https://ggplot2.tidyverse.org (2016).Avolio, M. et al. A comprehensive approach to analyzing community dynamics using rank abundance curves. Ecosphere. https://doi.org/10.1002/ecs2.2881 (2019).Article 

    Google Scholar 
    Curtis, J. & McIntosh, R. The interrelations of certain analytic and synthetic phytosociological characters. Ecology 31, 434–455. https://doi.org/10.2307/1931497 (1950).Article 

    Google Scholar 
    Porto, A. B., do Prado, M. A., Rodrigues, L. D. S. & Overbeck, G. E. Restoration of subtropical grasslands degraded by non-native pine plantations: Effects of litter removal and hay transfer. Restor. Ecol. https://doi.org/10.1111/rec.13773 (2022).Article 

    Google Scholar 
    Cáceres, M. D. & Legendre, P. Associations between species and groups of sites: Indices and statistical inference. Ecol. 90, 3566–3574. https://doi.org/10.1890/08-1823.1 (2009).Article 

    Google Scholar 
    Wickham, H., François, R., Henry, L. & Müller, K. dplyr: A Grammar of Data Manipulation. https://dplyr.tidyverse.org; https://github.com/tidyverse/dplyr (2022). More