Functional similarity affects similarity in partner composition in flea-mammal networks.

Functional signal in an interaction network is a phenomenon in which species resembling each other in their traits interact with similar partners. We tested the functional signal concept in realm-specific and regional flea-host networks from four biogeographic realms and asked whether the species composition of (a) host spectra and (b) flea assemblages is similar between functionally similar flea and host species, respectively. Analogously to testing for phylogenetic signal, we applied Mantel tests to investigate the correlation between flea or host functional distances calculated from functional dendrograms and dissimilarities in sets of interacting partners. In all realm-specific networks, functionally similar fleas tended to exploit similar hosts often belonging to the same genus, whereas functionally similar hosts tended to harbour similar fleas, again often belonging to the same genus. The strength of realm-specific functional signals and the frequency of detecting a significant functional signal in the regional networks differed between realms. The frequency of detecting a significant functional signal in the regional networks correlated positively with the network size for fleas and with the number of hosts in a network for hosts. A functional signal in the regional networks was more frequently found for hosts than for fleas. We discuss the mechanisms behind the functional signal in both fleas and their hosts, relate geographic functional signal patterns to the historic biogeography of fleas and conclude that functional signals in the species composition of host spectra for fleas and of flea assemblages for hosts result from the interplay of evolutionary and ecological processes.

[Changes in Soil Nitrogen Components and Their Relationship with Environmental Factors with Different Tea Plantation Ages].

Changes in soil nitrogen components in tea gardens affect the soil nitrogen supply capacity and nitrogen cycle. In this study, soil samples were collected from forest land, cultivated land, and tea gardens with different plantation ages (30, 50, and 70 years) to explore the changes in soil nitrogen components and their relationship with physicochemical properties and enzyme activities. The results showed that:① with the increase in tea plantation age, the silt, total phosphorus, and urease and catalase activities gradually increased, whereas the sand, clay, pH, electrical conductivity, soil organic carbon, and the activities of invertase gradually decreased. The alkaline phosphatase activity increased first and then decreased with the increase in tea plantation age, and no significant differences were observed in soil water content and acid phosphatase activity. ② With the increase in tea plantation age, the contents of acid ammonia nitrogen, amino acid nitrogen, and nitrate nitrogen (NO3--N) increased significantly, and the contents of total nitrogen, acid ammonia nitrogen, hydrolyzable unknown nitrogen, and non-hydrolyzable nitrogen in tea gardens were significantly higher than those in forest land. ③ The total phosphorus, alkaline phosphatase, and urease were the main factors affecting soil nitrogen components. Among them, organic nitrogen components were significantly correlated with total phosphorus and alkaline phosphatase, and inorganic nitrogen components were significantly correlated with alkaline phosphatase, whereas total nitrogen had significant correlations with sand, silt, total phosphorus, urease, and alkaline phosphatase.

Combining Genetic and Phenotypic Analyses for Detecting Bread Wheat Genotypes of Drought Tolerance through Multivariate Analysis Techniques.

Successfully promoting drought tolerance in wheat genotypes will require several procedures, such as field experimentations, measuring relevant traits, using analysis tools of high precision and efficiency, and taking a complementary approach that combines analyses of phenotyping and genotyping at once. The aim of this study is to assess the genetic diversity of 60 genotypes using SSR (simple sequence repeat) markers collected from several regions of the world and select 13 of them as more genetically diverse to be re-evaluated under field conditions to study drought stress by estimating 30 agro-physio-biochemical traits. Genetic parameters and multivariate analysis were used to compare genotype traits and identify which traits are increasingly efficient at detecting wheat genotypes of drought tolerance. Hierarchical cluster (HC) analysis of SSR markers divided the genotypes into five main categories of drought tolerance: four high tolerant (HT), eight tolerant (T), nine moderate tolerant (MT), six sensitive (S), and 33 high sensitive (HS). Six traits exhibit a combination of high heritability (>60%) and genetic gain (>20%). Analyses of principal components and stepwise multiple linear regression together identified nine traits (grain yield, flag leaf area, stomatal conductance, plant height, relative turgidity, glycine betaine, polyphenol oxidase, chlorophyll content, and grain-filling duration) as a screening tool that effectively detects the variation among the 13 genotypes used. HC analysis of the nine traits divided genotypes into three main categories: T, MT, and S, representing three, five, and five genotypes, respectively, and were completely identical in linear discriminant analysis. But in the case of SSR markers, they were classified into three main categories: T, MT, and S, representing five, three, and five genotypes, respectively, which are both significantly correlated as per the Mantel test. The SSR markers were associated with nine traits, which are considered an assistance tool in the selection process for drought tolerance. So, this study is useful and has successfully detected several agro-physio-biochemical traits, associated SSR markers, and some drought-tolerant genotypes, coupled with our knowledge of the phenotypic and genotypic basis of wheat genotypes.

Benchmarking the Mantel test and derived methods for testing association between distance matrices.

Testing the association between objects is central in ecology, evolution, and quantitative sciences in general. Two types of variables can describe the relationships between objects: point variables (measured on individual objects), and distance variables (measured between pairs of objects). The Mantel test and derived methods have been extensively used for distance variables. Yet, these methods have been criticized due to low statistical power and inflated type I error when spatial autocorrelation is present. Here, we assessed the statistical power between different types of tested variables and the type I error rate over a wider range of autocorrelation intensities than previously assessed, both on univariate and multivariate data. We also illustrated the performance of distance matrix statistics through computational simulations of genetic diversity. We show that the Mantel test and derived methods are not affected by inflated type I error when spatial autocorrelation affects only one variable when investigating correlations, or when either the response or the explanatory variable(s) is affected by spatial autocorrelation while investigating causal relationships. As previously noted, with autocorrelation affecting more variables, inflated type I error could be reduced by modifying the significance threshold. Additionally, the Mantel test has no problem of statistical power when the hypothesis is formulated in terms of distance variables. We highlight that transformation of variable types should be avoided because of the potential information loss and modification of the tested hypothesis. We propose a set of guidelines to help choose the appropriate method according to the type of variables and defined hypothesis.

The Effects of Genotype × Environment on Physicochemical and Sensory Properties and Differences of Volatile Organic Compounds of Three Rice Types (Oryza sativa L.).

Understanding the effects of genotype, environment and their interactions on rice quality is of great importance for rice breeding and cultivation. In this study, six rice varieties with two indica, two japonica and two indica-japonica types of rice were selected and planted at ten locations in Zhejiang Province to investigate the genotype (G) × environment (E) on physicochemical and sensory properties and the differences of volatile organic compounds (VOCs) among the three types of rice. Analysis of variances showed that apparent amylose content (AC), total protein content (PC), alkali spreading value (ASV), RVA profiles, and appearance (ACR), palatability (PCR), and sensory evaluation value (SEV) of cooked rice and texture of cooled cooked rice (TCCR) were mainly affected by genotypic variation, whereas the smell of cooked rice (SCR) was mainly affected by environment (p < 0.05). The G × E effect was significant for most parameters. The weather in the middle and late periods of filling had important effects on the formation of rice quality, especially on setback (SB) and pasting temperature (PT) (p < 0.01). They were negatively correlated with the texture of cooked rice (TCR) and SEV (p < 0.05). Peak viscosity (PV) and breakdown (BD) were positively related to the sensory evaluation parameters (p < 0.01) and could be used to predict cooked rice quality. A total of 59 VOCs were detected, and indica, japonica and indica-japonica had 9, 6 and 19 characteristic compounds, respectively. The principal component analysis showed that the physicochemical and sensory properties and VOCs of indica-japonica rice were more stable than those of indica and japonica rice at ten locations in Zhejiang Province. It is helpful for rice breeders to understand how the environment affects the physicochemical, sensory properties and VOCs of the three rice types, and it is also important for food enterprises to provide rice products with stable quality.

Novel insight into the evolution of volatile compounds during dynamic freeze-drying of Ziziphus jujuba cv. Huizao based on GC-MS combined with multivariate data analysis.

To understand the evolution of aroma in jujubes during dynamic freeze drying (FD), the relationship between aroma compounds, precursors, and related enzyme activities were analyzed. Fifty-three volatiles were identified during FD processing. After FD, the total aroma contents were increased from 11,004 to 14,603 µg/kg, ketones content was significantly decreased by 54.11 %, resulted in the loss of creamy note in freeze-dried jujube (FDJ). Through the network analysis, serine, glycine, proline, valine, cysteine, arginine, glutamic acid, lysine and leucine had the significant correlation with pyrazines, dominated the roasty note of FDJ. Linoleic acid, α-linolenic acid and oleic acid with lipoxygenase had important effects on the increase of esters (from 412 to 9,486 µg/kg), contributed fruity and sweet notes of FDJ. Besides, through the Mantel test, the influence degree of factors on the formation of FDJ aroma was ranked as temperature > enzyme activity > fatty acids > amino acids.

Effects of additives on the co-composting of forest residues with cattle manure.

Co-composting of forest residues (FR) with cattle manure (CM) results in a low nitrogen (N) conversion efficiency, a low organic matter (OM) degradation rate, and a low quality compost product. This study evaluated the effects of addition of bone charcoal (BC), pumice (PM), or straw biochar (SB) at a ratio of 10 % (w/w) on the co-composting of FR with CM. The highest quality compost was obtained with addition of 10 % PM. Compared with the control (without any additive), PM addition increased the OM degradation rate, the nitrate-N, the available phosphorus, and the available potassium by 25 %, 110 %, 24 %, and 9 %, respectively, and increased the relative abundance of bacteria (Planomicrobium, Flavobacterium, and Pseudomonas) involved in lignocellulose degradation and N transformation. With the addition of PM, the co-composting of FR with CM generated a high quality, useful product in only 39 days.

Flavor improving effects of cysteine in xylose-glycine-fish waste protein hydrolysates (FPHs) Maillard reaction system.

A promising way to utilize fish by-products is to develop hydrolysis of fish proteins with enzymes. The obtained fish protein hydrolysates (FPHs) are rich in peptides and amino acids, but bitterness and aroma defects impede further utilization of FPHs. The present study adopted Maillard reaction to improve FPHs' flavor and illustrated the role of cysteine in this system. We investigated the impact of cysteine (0, 0.25%, 0.5%, 0.75%, and 1%) on the browning intensity, free amino acids (FAAs), molecular weight distribution, structure of MRPs, volatile compounds changes and organoleptic characteristics of xylose-glycine-FPHs Maillard reaction systems. Results showed that the addition of cysteine lowered the browning degree of Maillard reaction products (MRPs) by inhibiting the cross-linking of small peptides and reducing the production of melanin. GC-MS and GC-IMS analysis indicated that cysteine inhibited the formation of furans and nitrogen-containing compounds and facilitated the formation of sulfur-containing compounds contributing to the meaty flavor. Sensory analysis revealed that 0.25-0.75% range of cysteine increased the meaty, caramel, umami, mouthfulness and salty notes, and caused a decrease in bitter taste of the MRPs as confirmed by GC-MS. A highly significant correlation between the organoleptic characteristics and physicochemical indicators of MRPs was found by Mantel test. These results elucidated the influence of cysteine on the formation of Maillard reaction products and will help improve the flavor profile of meat flavorings.

Phytochemicals and Their Correlation with Molecular Data in Micromeria and Clinopodium (Lamiaceae) Taxa.

A study of the phytochemical and molecular characteristics of ten Micromeria and six Clinopodium taxa (family Lamiaceae) distributed in the Balkan Peninsula was carried out. The phytochemicals detected in essential oils by gas chromatography, mass spectrometry, and molecular data amplified fragment length polymorphism were used to study the taxonomic relationships among the taxa and the correlations between phytochemical and molecular data. STRUCTURE analysis revealed three genetic groups, while Bayesian Analysis of Population Structure grouped the studied taxa into 11 clusters nested in the groups obtained by STRUCTURE. Principal components analysis performed with the 21 most represented compounds in the essential oils yielded results that were partly consistent with those obtained by STRUCTURE and neighbour-joining analyses. However, their geographic distributions did not support the genetic grouping of the studied taxa and populations. The Mantel test showed a significant correlation between the phytochemical and genetic data (r = 0.421, p < 0.001). Genetic distance explained 17.8% of the phytochemical distance between populations. The current taxonomic position of several of the studied taxa is yet to be satisfactorily resolved, and further studies are needed. Such future research should include nuclear and plastid DNA sequences from a larger sample of populations and individuals.

Effect of a chitosan coating incorporating epigallocatechin gallate on the quality and shelf life of bighead carp (Aristichthys nobilis) fillets during chilled storage.

The objective of this study was to investigate the potential application of chitosan coatings incorporating epigallocatechin gallate (EGCG) for preserving fillets of bighead carp during chilled storage. The fillets were coated with acetic acid and glycerol, chitosan, and chitosan-EGCG, respectively, and the changes in their physicochemical, microbiological, and sensory characteristics during storage at 4 °C were determined. Notably, total volatile basic nitrogen, thiobarbituric-acid-reactive substances, and K value of chitosan-EGCG coated fillets sampled on day 15 were 48.04 %, 60.19 %, and 32.91 % lower than untreated fillets, respectively. Microbial enumeration suggested that the inclusion of EGCG significantly improved the inhibitory effect of pure chitosan coating on the proliferation of microorganisms. Furthermore, the chitosan-EGCG coated fillets also performed the best in terms of color, texture, and sensory analysis, and extended the shelf-life of the fillets for at least 6 days. A principal component analysis further confirmed the preserving effect of the chitosan-EGCG coating. Mantel test results suggested that the fillets' organoleptic characteristics strongly correlated with physicochemical and microbiological indicators. Overall, this work provides an effective protocol for food quality control and the extension of shelf life during chilled storage, and it clarifies the relationships between organoleptic characteristics and physicochemical and microbiological indexes.

Population genetic structure and morphological diversity of Cruzia tentaculata (Nematoda: Ascaridida), a parasite of marsupials (Didelphinae), along the Atlantic Forest on the eastern coast of South America.

Cruzia tentaculata is a helminth parasite of marsupials and has a wide geographic distribution from Mexico to Argentina. The aim of this study was to analyse the genetic population structure of this nematode along the Atlantic Forest biome. Cruzia tentaculata specimens were recovered from Didelphis aurita, Didelphis albiventris and Philander quica in 9 localities. Morphological and morphometric data were investigated for phenotypic diversity among localities and hosts using multivariate discriminant analysis of principal components. Phylogenetic relationships of C. tentaculata were determined using maximum likelihood and Bayesian inference. The population structure was analysed by fixation indices, molecular variance analysis, Tajima's D and Fu's Fs neutrality tests, Mantel tests and Bayesian clustering analysis. A higher significant morphometric difference for males was observed between localities. In the haplogroup networks, 2 groups were recovered, separating locations from the north and from the south/southeast. The morphometric variation in C. tentaculata between different localities was compatible with this north and southeast/south pattern, suggesting adaptation to different ecological conditions. Population genetic analyses suggested a pattern of evolutionary processes driven by Pleistocene glacial refugia in the northeast and southeast of the Atlantic Forest based on the distribution of genetic diversity.

Putting the Mantel test back together again.

The Mantel test has been widely used in ecology and evolution, but over the last two decades it has been frequently critiqued because results were inconsistent with expectations and there were issues with Type I (false-positive) and Type II (false-negative) error rates. Three-matrix extensions of the Mantel test have been challenged for similar reasons. Even the null hypotheses underlying the Mantel test have been questioned. As a result, use of the Mantel test and its variants has been discouraged or limited to special situations. Here, we examine Mantel test criticisms including the lack of agreement between traditional variable-based Pearson correlations (r) and observation-based Mantel correlations (rm ), and the unusual Type I and Type II error rates. We propose an alternate proximity measure that resolves these issues. We use simulations and examples to contrast Mantel results based on Euclidean distance, squared Euclidean distance, and the simple difference (Diff) with traditional bivariate Pearson correlations. We demonstrate that use of the simple difference in Mantel tests can resolve the underlying problems with poor agreement between bivariate Pearson and Mantel correlations, as well as appropriate Type I and Type II errors (i.e., where r = cor(x,y) and rm = cor(dx ,dy ), if dx = Diff(x) and dy = Diff(y), r = rm ). We also show that the simple difference can provide solutions to issues with partial Mantel tests and distance-based MANOVA. Because our results resolve many of the issues with Mantel tests, we hope that these findings will restore the popularity of the Mantel test.

Population genetic structure and phenotypic diversity of Aspidodera raillieti (Nematoda: Heterakoidea), a parasite of Didelphini marsupials in Brazil's South and Southeast Atlantic Forest.

BACKGROUND: The population genetics of parasites may be influenced by host specificity, life cycle, host geographical range, evolutionary history, and host population structure. The nematode Aspidodera raillieti infects different marsupial and rodent hosts in the Nearctic and Neotropical regions, implying a gene flow among populations. However, niche diversification of the main hosts of A. raillieti in superimposed areas may provide conditions for population genetic structuring within this parasite species. We examined the genetic structuring of A. raillieti infecting three marsupial species co-occurring along the South and Southeast Brazilian Atlantic Forest, a hotspot of biodiversity. METHODS: We employed morphometric analyses and partial mitochondrial cytochrome c oxidase I gene sequences (MT-CO1) to characterize populations via phylogenetic and phylogeographic analyses. RESULTS: Among 175 A. raillieti specimens recovered from the marsupial hosts Didelphis aurita, D. albiventris, and Philander quica, we identified 99 MT-CO1 haplotypes forming four haplogroups and four clades in networks and phylogenetic trees, respectively. Clades I and II encompassed parasites of D. albiventris from the South region, clade III comprised parasites of D. aurita from the South and Southeast regions, and clade IV encompassed parasites of D. aurita and D. albiventris from the South and Southeast regions and parasites of P. quica from the South region. High genetic differentiation between clades, with a high fixation index and greater genetic variation in the analysis of molecular variance (AMOVA), indicated low gene flow between clades. Haplotypes shared among host species revealed a lack of host specificity. A significant correlation in the Mantel test suggested parasite isolation by distance, while there was no evidence of geographical structure between populations. Negative neutrality test values for clades III and IV suggested recent population expansion. Morphometric differentiation between A. raillieti specimens recovered from different host species, as well as from different localities, was more evident in males. CONCLUSION: The genetic structure of A. raillieti populations in the South and Southeast Atlantic Forest resulted from historical events rather than from current geographical distribution or host specificity. We also demonstrate morphometric variation associated with host species and localities, suggesting phenotypic plasticity to host attributes and to spatial variables.

Dynamics and key drivers of antibiotic resistance genes during aerobic composting amended with plant-derived and animal manure-derived biochars.

Plant-derived and animal manure-derived biochars have been used to improve the quality of compost but the differences in their effects on antibiotic resistance genes (ARGs) during composting are unclear. This study selected two types of biochar (RB and PB) produced from abundant agricultural waste to be added to the compost. Adding plant-derived RB performed better in ARGs, mobile genetic elements, and human pathogenic bacteria removal during aerobic composting, whereas adding manure-derived PB even increased ARGs abundance. Vertical gene transfer was possibly the key mechanism for persistent ARGs, and easily removed ARGs were regulated by horizontal and vertical gene transfer. Adding plant-derived RB reduced the abundances of persistent ARG hosts (e.g., Pseudomonas and Longispora) and ARG-related metabolic pathways and genes. The higher nitrogen content of manure-derived PB may have promoted the proliferation of ARG hosts. Overall, adding manure-derived biochar during composting may not be the optimal option for eliminating ARGs.

Combination of linear and nonlinear multivariate approaches effectively uncover responses of phytoplankton communities to environmental changes at regional scale.

The response of a community to environmental changes is either linear or non-linear, so that they can be investigated approximately by linear or nonlinear models. At community level, redundancy analysis (RDA) and canonical correspondence analysis (CCA), and Mantel test and Generalized Dissimilarity Modelling (GDM) are two pairs of fundamental multivariate approaches. Thus, it is necessary to determine how they are used for a given group of communities or a metacommunity. In the present study, we explored the applications of the two pairs of commonly used multivariate methods for the analysis of tropical phytoplankton communities. Phytoplankton were collected from 60 tropical reservoirs in southern China at two distinct regions and two hydrological seasons. Because of a short environmental gradient, response of phytoplankton communities to the environmental gradients was first explored with linear models: distance-based redundancy analysis (db-RDA) and Mantel test. Then, CCA and GDM were further applied to recognize the nonlinear relationship between phytoplankton community variation and environmental changes, and to detect the significant environmental and/or spatial variables. Our results strongly suggest that the combination of db-RDA and GDM provides a highly effective tool to uncover the linearity and nonlinearity in community responses and the important associated environmental and spatial variables, which were significantly different between flooding and dry seasons.

Diversity, Dispersal and Mode of Reproduction of Amanita exitialis in Southern China.

Amanita exitialis is a poisonous mushroom and has caused many deaths in southern China. In this study, we collected 118 fruiting bodies of A. exitialis from seven different sites in Guangdong Province in southern China and investigated their genetic relationships using 14 polymorphic molecular markers. These 14 markers grouped the 118 fruiting bodies into 20 multilocus genotypes. Among these 20 genotypes, eight were each found only once while the remaining 12 were each represented by two to 54 fruiting bodies. Interestingly, among the 12 shared genotypes, four were shared between/among local populations that were separated by as far as over 80 km, a result consistent with secondary homothallic reproduction and long-distance spore dispersal. Despite the observed gene flow, significant genetic differentiations were found among the local populations, primarily due to the over-representation of certain genotypes within individual local populations. STRUCTURE analyses revealed that the 118 fruiting bodies belonged to three genetic clusters, consistent with divergence within this species in this geographic region. Interestingly, we found an excess of heterozygous individuals at both the local and the total sample level, suggesting potential inbreeding depression and heterozygous advantage in these populations of A. exitialis. We discuss the implications of our results for understanding the life cycle, dispersal, and evolution of this poisonous mushroom.

Extracting consistent biological information from functional results of metabolomic pathway analysis using the Mantel's test.

Extraction of meaningful biological information from the vast array of data that metabolomics analyses generate is a major challenge in the field. A variety of computational and visual tools that help to identify changes in metabolic pathways have been proposed including functional analysis and pathway analysis. Meta-analysis of metabolomic data has emerged as a powerful source of information. In this work, the applicability of the Mantel's test for the correlation of functional results from metabolic pathway analysis is shown using experimental and simulated data sets as evaluation examples. The statistical significance of the correlation coefficient can be assessed by permutation testing requiring practically no computation time. The use of the Mantel's test can assist the critical comparison of different phenotypes, studies, methods, platforms, or data preprocessing strategies, as well as help to identify inconsistencies between metabolomic study outcomes, making this algorithm attractive for data interpretation and meta-analysis on a routine basis.

Assessing the correlations and selection criteria between different traits in wheat salt-tolerant genotypes.

Salinity is one of the largest stresses blocking horizontal and vertical expansion in agricultural lands. Establishing salt-tolerant genotypes is a promising method to benefit from poor water quality and salinized lands. An integrated method was developed for accomplishing reliable and effective evaluation of traits stability of salt-tolerant wheat. The study aims were to estimate the genetic relationships between explanatory traits and shoot dry matter (SDM), and determine the traits stability under three salinity levels. Morphophysiological and biochemical traits were evaluated as selection criteria for SDM improvement in wheat for salinity tolerance. Three cultivars and three high-yielding doubled haploid lines (DHLs) were used. Three salt (NaCl) levels (control (washed sand), 7 and 14 dS m-1) were applied for 45 days (at the first signs of death in the sensitive genotypes). All morphophysiological traits gradually decreased as salinity levels increased, excluding the number of roots. Decreases were more visible in sensitive genotypes than in tolerant genotypes. All biochemical traits increased as salinity levels increased. Variance inflation factors (VIFs) and condition number exhibited multicollinearity for membrane stability index and polyphenol oxidase activity. After their removal, all VIFs were <10, thereby increasing path coefficient accuracy. Total chlorophyll content (CHL) and catalase (CAT) provided significant direct effects regarding genetic and phenotypic correlations for the three salinity levels and their interactions in path analysis on SDM, indicating their stability. CHL and CAT had high heritability (>0.60%) and genetic gain (>20%) and highly significant genetic correlation, co-heritability, and selection efficiencies for SDM. CHL and CAT could be used as selection criteria for salinity tolerance in wheat-breeding programs. The tolerated line (DHL21) with the check cultivar (Sakha 93) can be also recommended as novel genetic resource for improving salinity tolerance of wheat.

Comparative population genetic structure of two ixodid tick species (Acari:Ixodidae) (Ixodes ovatus and Haemaphysalis flava) in Niigata prefecture, Japan.

Ixodid ticks (Acari:Ixodidae) are essential vectors of tick-borne diseases in Japan. In this study, we characterized the population genetic structure and inferred genetic divergence in two widespread and abundant ixodid species, Ixodes ovatus and Haemaphysalis flava. Our hypothesis was that genetic divergence would be high in I. ovatus because of the low mobility of their small rodent hosts of immature I. ovatus would limit their gene flow compared to more mobile avian hosts of immature H. flava. We collected 320 adult I. ovatus from 29 locations and 223 adult H. flava from 17 locations across Niigata Prefecture, Japan, and investigated their genetic structure using DNA sequences from fragments of two mitochondrial gene regions, cox1 and the 16S rRNA gene. For I. ovatus, pairwise FST and analysis of molecular variance (AMOVA) analyses of cox1 and 16S sequences indicated significant genetic variation among populations, whereas both markers showed non-significant genetic variation among locations for H. flava. A cox1 gene tree and haplotype network revealed three genetic groups of I. ovatus. One of these groups consisted of haplotypes distributed at lower altitudes (251-471 m.a.s.l.). The cox1 sequences of I. ovatus from Japan clustered separately from I. ovatus sequences reported from China, suggesting the potential for cryptic species in Japan. Our results support our hypothesis and suggest that the host preference of ticks at the immature stage may influence the genetic structure of the ticks. This information may be important for understanding the tick-host interactions in the field to better understand the tick-borne disease transmission and in designing an effective tick control program.

Sensitivity of migratory connectivity estimates to spatial sampling design.

BACKGROUND: The use of statistical methods to quantify the strength of migratory connectivity is commonplace. However, little attention has been given to their sensitivity to spatial sampling designs and scales of inference. METHODS: We examine sources of bias and imprecision in the most widely used methodology, Mantel correlations, under a range of plausible sampling regimes using simulated migratory populations. RESULTS: As Mantel correlations depend fundamentally on the spatial scale and configuration of sampling, unbiased inferences about population-scale connectivity can only be made under certain sampling regimes. Within a contiguous population, samples drawn from smaller spatial subsets of the range generate lower connectivity metrics than samples drawn from the range as a whole, even when the underlying migratory ecology of the population is constant across the population. Random sampling of individuals from contiguous subsets of species ranges can therefore underestimate population-scale connectivity. Where multiple discrete sampling sites are used, by contrast, overestimation of connectivity can arise due to samples being biased towards larger between-individual pairwise distances in the seasonal range where sampling occurs (typically breeding). Severity of all biases was greater for populations with lower levels of true connectivity. When plausible sampling regimes were applied to realistic simulated populations, accuracy of connectivity measures was maximised by increasing the number of discrete sampling sites and ensuring an even spread of sites across the full range. CONCLUSIONS: These results suggest strong potential for bias and imprecision when making quantitative inferences about migratory connectivity using Mantel statistics. Researchers wishing to apply these methods should limit inference to the spatial extent of their sampling, maximise their number of sampling sites, and avoid drawing strong conclusions based on small sample sizes.