Versatile simulations of admixture and accurate local ancestry inference with mixnmatch and ancestryinfer

It has become clear that hybridization between species is much more common than previously recognized. As a result, we now know that the genomes of many modern species, including our own, are a patchwork of regions derived from past hybridization events. Increasingly researchers are interested in disentangling which regions of the genome originated from each parental species using local ancestry inference methods. Due to the diverse effects of admixture, this interest is shared across disparate fields, from human genetics to research in ecology and evolutionary biology. However, local ancestry inference methods are sensitive to a range of biological and technical parameters which can impact accuracy. Here we present paired simulation and ancestry inference pipelines, mixnmatch and ancestryinfer, to help researchers plan and execute local ancestry inference studies. mixnmatch can simulate arbitrarily complex demographic histories in the parental and hybrid populations, selection on hybrids, and technical variables such as coverage and contamination. ancestryinfer takes as input sequencing reads from simulated or real individuals, and implements an efficient local ancestry inference pipeline. We perform a series of simulations with mixnmatch to pinpoint factors that influence accuracy in local ancestry inference and highlight useful features of the two pipelines. mixnmatch is a powerful tool for simulations of hybridization while ancestryinfer facilitates local ancestry inference on real or simulated data.     

Gaidropsarus (Gadidae,Teleostei) of the North Atlantic Ocean: a brief phylogenetic review

The phylogenetic relationships among the North Atlantic Gaidropsarus and between the three Gaidropsarinae genera Gaidropsarus, Ciliata and Enchelyopus are reviewed with the hitherto most comprehensive taxonomic sampling of this group. Phylogenetic results (maximum parsimony, maximum likelihood and Bayesian inference) based on nuclear (rhodopsin) and concatenated mitochondrial (12s, 16s and cytb) markers clearly support this subfamily. For the north‐eastern Atlantic species of Gaidropsarus, two previously unreported clades were strongly supported, clarifying the relationships within the genus, and revealing fewer distinct taxa in the north Atlantic Gaidropsarus than previously stipulated. The data challenge the specific status of Gaidropsarus mediterraneus and Gaidropsarus guttatus and raise doubts concerning the distinctiveness of other species. A taxonomic revision of the genus is suggested.     

The anatomy of choice: dopamine and decision-making

This paper considers goal-directed decision-making in terms of embodied or active inference. We associate bounded rationality with approximate Bayesian inference that optimizes a free energy bound on model evidence. Several constructs such as expected utility, exploration or novelty bonuses, softmax choice rules and optimism bias emerge as natural consequences of free energy minimization. Previous accounts of active inference have focused on predictive coding. In this paper, we consider variational Bayes as a scheme that the brain might use for approximate Bayesian inference. This scheme provides formal constraints on the computational anatomy of inference and action, which appear to be remarkably consistent with neuroanatomy. Active inference contextualizes optimal decision theory within embodied inference, where goals become prior beliefs. For example, expected utility theory emerges as a special case of free energy minimization, where the sensitivity or inverse temperature (associated with softmax functions and quantal response equilibria) has a unique and Bayes-optimal solution. Crucially, this sensitivity corresponds to the precision of beliefs about behaviour. The changes in precision during variational updates are remarkably reminiscent of empirical dopaminergic responses—and they may provide a new perspective on the role of dopamine in assimilating reward prediction errors to optimize decision-making.     

A standardized reanalysis of molecular phylogenetic hypotheses of Gobioidei

Gobioidei is a suborder of perciform fishes with about 2000 species distributed worldwide. Despite the evolutionary and ecological importance of gobioids, their phylogenetic inter- and intrarelationships are still poorly understood. Only a few studies (either morphological or molecular) have tackled the phylogeny of Gobioidei as a whole. Of these, only six studies thus far have addressed gobioid intrarelationships based on molecular data (each using different taxon sampling, genes, outgroups and method of phylogenetic inference), yielding contrasting results regarding the phylogenetic relationships among major lineages. In this study, we have reanalysed data from four of these molecular phylogenetic studies of Gobioidei under standardization criteria (same outgroup and methods of phylogenetic inference) in order to assess the robustness of their results, as well as to identify which parts of the gobioid tree are least resolved. Results from all datasets reanalysed in this study are generally similar to those of the respective original studies, and suggest broad patterns of phylogenetic relationships among gobioid lineages. However, there are numerous topological discrepancies among the four studies, support is low for many phylogenetic relationships and topology tests are unable to reject the vast majority of alternative topologies tested. The concatenation of datasets yields a relatively robust phylogeny of major lineages of Gobioidei, but there are some issues of overlap and missing data, which are ameliorated with the inclusion of additional homologous sequences from GenBank that increase dataset completeness. Because both monophyly of major gobioid groups and phylogenetic relationships among them cannot be fully resolved, it is clear that further phylogenetic research is needed, and this should be accompanied by a major taxonomic revision of the Gobioidei. Nevertheless, even with the relatively unstable nature of the available molecular phylogenies, there are some monophyletic units that can be identified, and a basic structure of the gobioid tree appears evident.     

PHYLOGENY OF PHAGOTROPHIC EUGLENIDS (EUGLENOZOA): A MOLECULAR APPROACH BASED ON CULTURE MATERIAL AND ENVIRONMENTAL SAMPLES1

Molecular studies based on small subunit (SSU) rDNA sequences addressing euglenid phylogeny hitherto suffered from the lack of available data about phagotrophic species. To extend the taxon sampling, SSU rRNA genes from species of seven genera of phagotrophic euglenids were investigated. Sequence analyses revealed an increasing genetic diversity among euglenid SSU rDNA sequences compared with other well‐known eukaryotic groups, reflecting an equally broad diversity of morphological characters among euglenid phagotrophs. Phylogenetic inference using standard parsimony and likelihood approaches as well as Bayesian inference and spectral analyses revealed no clear support for euglenid monophyly. Among phagotrophs, monophyly of Petalomonas cantuscygni and Notosolenus ostium, both comprising simple ingestion apparatuses, is strongly supported. A moderately supported clade comprises phototrophic euglenids and primary osmotrophic euglenids together with phagotrophs, exhibiting a primarily flexible pellicle composed of numerous helically arranged strips and a complex ingestion apparatus with two supporting rods and four curved vanes. Comparison of molecular and morphological data is used to demonstrate the difficulties to formulate a hypothesis about how the ingestion apparatus evolved in this group.     

Fourteenth research symposium University of Durham 1974

Abstract

Background: Few studies analysing lichen diversity have simultaneously considered interactions among drivers that operate at different spatial and temporal scales.

Aims: The aims of this study were to evaluate the relative importance of host tree, and local, landscape and historical factors in explaining lichen diversity in managed temperate forests, and to test the potential interactions among factors acting at different spatial scales.

Methods: Thirty-five stands were selected in the ?rség region of western Hungary. Linear models and multi-model inference within an information-theory framework were used to evaluate the role of different variables on lichen species richness.

Results: Drivers at multiple spatial scales contributed to shaping lichen species richness both at the tree and plot levels. Tree-level species richness was related to both tree- and plot-level factors. With increasing relative diffuse light lichen species richness increased; this effect was stronger on the higher than on the lower part of the trunks. At the plot scale, species richness was affected by local drivers. Landscape and historical factors had no, or only a marginal, effect.

Conclusions: Lichen conservation in temperate managed forests could be improved if the complex interactions among host tree quality and availability, micro-climatic conditions, and management were taken into consideration.     

Haplotype Inference for Population Data with Genotyping Errors

Inference of haplotypes is important in genetic epidemiology studies. However, all large genotype data sets have errors due to the use of inexpensive genotyping machines that are fallible and shortcomings in genotyping scoring softwares, which can have an enormous impact on haplotype inference. In this article, we propose two novel strategies to reduce the impact induced by genotyping errors in haplotype inference. The first method makes use of double sampling. For each individual, the “GenoSpectrum” that consists of all possible genotypes and their corresponding likelihoods are computed. The second method is a genotype clustering algorithm based on multi‐genotyping data, which also assigns a “GenoSpectrum” for each individual. We then describe two hybrid EM algorithms (called DS‐EM and MG‐EM) that perform haplotype inference based on “GenoSpectrum” of each individual obtained by double sampling and multi‐genotyping data. Both simulated data sets and a quasi real‐data set demonstrate that our proposed methods perform well in different situations and outperform the conventional EM algorithm and the HMM algorithm proposed by Sun, Greenwood, and Neal (2007, Genetic Epidemiology 31 , 937–948) when the genotype data sets have errors.     

Simulations inform design of regional occupancy‐based monitoring for a sparsely distributed,territorial species

Sparsely distributed species attract conservation concern, but insufficient information on population trends challenges conservation and funding prioritization. Occupancy‐based monitoring is attractive for these species, but appropriate sampling design and inference depend on particulars of the study system. We employed spatially explicit simulations to identify minimum levels of sampling effort for a regional occupancy monitoring study design, using white‐headed woodpeckers (Picoides albolvartus), a sparsely distributed, territorial species threatened by habitat decline and degradation, as a case study. We compared the original design with commonly proposed alternatives with varying targets of inference (i.e., species range, space use, or abundance) and spatial extent of sampling. Sampling effort needed to achieve adequate power to observe a long‐term population trend (≥80% chance to observe a 2% yearly decline over 20 years) with the previously used study design consisted of annually monitoring ≥120 transects using a single‐survey approach or ≥90 transects surveyed twice per year using a repeat‐survey approach. Designs that shifted inference toward finer‐resolution trends in abundance and extended the spatial extent of sampling by shortening transects, employing a single‐survey approach to monitoring, and incorporating a panel design (33% of units surveyed per year) improved power and reduced error in estimating abundance trends. In contrast, efforts to monitor coarse‐scale trends in species range or space use with repeat surveys provided extremely limited statistical power. Synthesis and applications. Sampling resolutions that approximate home range size, spatially extensive sampling, and designs that target inference of abundance trends rather than range dynamics are probably best suited and most feasible for broad‐scale occupancy‐based monitoring of sparsely distributed territorial animal species.     

Genomic data detect corresponding signatures of population size change on an ecological time scale in two salamander species

Understanding the demography of species over recent history (e.g. <100 years) is critical in studies of ecology and evolution, but records of population history are rarely available. Surveying genetic variation is a potential alternative to census‐based estimates of population size, and can yield insight into the demography of a population. However, to assess the performance of genetic methods, it is important to compare their estimates of population history to known demography. Here, we leveraged the exceptional resources from a wetland with 37 years of amphibian mark–recapture data to study the utility of genetically based demographic inference on salamander species with documented population declines (Ambystoma talpoideum) and expansions (A. opacum), patterns that have been shown to be correlated with changes in wetland hydroperiod. We generated ddRAD data from two temporally sampled populations of A. opacum (1993, 2013) and A. talpoideum (1984, 2011) and used coalescent‐based demographic inference to compare alternate evolutionary models. For both species, demographic model inference supported population size changes that corroborated mark–recapture data. Parameter estimation in A. talpoideum was robust to our variations in analytical approach, while estimates for A. opacum were highly inconsistent, tempering our confidence in detecting a demographic trend in this species. Overall, our robust results in A. talpoideum suggest that genome‐based demographic inference has utility on an ecological scale, but researchers should also be cognizant that these methods may not work in all systems and evolutionary scenarios. Demographic inference may be an important tool for population monitoring and conservation management planning.     

In and out: Effects of shoot- vs. rooted-presence sampling methods on plant diversity measures in mountain grasslands

Plant diversity measures (e.g., alpha- and beta-diversity) provide the basis for a number of ecological indication and monitoring methods. These measures are based on species counts in sampling units (plots or quadrats). However, there are two alternative conventions for defining a vascular plant species as “present” in a plot, i.e. “shoot presence” (a species is recorded if the vertical projection of any above-ground part falls within the plot) and “rooted presence” (a species is recorded only when an individual is rooted inside the plot). Very few studies addressed the effects of the two sampling conventions on species richness and diversity indices. We sampled mountain dry grasslands in Italy across different plot sizes and vegetation types to assess how large is the difference in alpha- and beta-diversity values and in sample-based rarefaction curves between the two methods. We found that the difference is greatly dependent on plot size, being more relevant, both in absolute and percentage values, at smaller grain; it is also dependent on habitat type, being larger in shallow-soil communities, as they have a sparser vegetation structure and host life-form types with a larger lateral spread. At fine spatial scales (<1 m²) the difference is large enough to bias statistical inference, and we conclude that at such scales one should not attempt to compare plant diversity indices if they were not obtained with the same sampling convention.     

The aggregate site frequency spectrum for comparative population genomic inference

Understanding how assemblages of species responded to past climate change is a central goal of comparative phylogeography and comparative population genomics, an endeavour that has increasing potential to integrate with community ecology. New sequencing technology now provides the potential to perform complex demographic inference at unprecedented resolution across assemblages of nonmodel species. To this end, we introduce the aggregate site frequency spectrum (aSFS), an expansion of the site frequency spectrum to use single nucleotide polymorphism (SNP) data sets collected from multiple, co‐distributed species for assemblage‐level demographic inference. We describe how the aSFS is constructed over an arbitrary number of independent population samples and then demonstrate how the aSFS can differentiate various multispecies demographic histories under a wide range of sampling configurations while allowing effective population sizes and expansion magnitudes to vary independently. We subsequently couple the aSFS with a hierarchical approximate Bayesian computation (hABC) framework to estimate degree of temporal synchronicity in expansion times across taxa, including an empirical demonstration with a data set consisting of five populations of the threespine stickleback (Gasterosteus aculeatus). Corroborating what is generally understood about the recent postglacial origins of these populations, the joint aSFS/hABC analysis strongly suggests that the stickleback data are most consistent with synchronous expansion after the Last Glacial Maximum (posterior probability = 0.99). The aSFS will have general application for multilevel statistical frameworks to test models involving assemblages and/or communities, and as large‐scale SNP data from nonmodel species become routine, the aSFS expands the potential for powerful next‐generation comparative population genomic inference.     

基于高密度单核苷酸多态性的共祖远亲缘关系预测算法准确性研究

目的 研究构建基于共祖（identity-by-descent，IBD）片段算法预测远亲缘关系分析流程并评估预测准确性。方法 采用高密度单核苷酸多态性（single nucleotide polymorphism，SNP）芯片对253份家系样本进行检测，研究基于IBD片段算法的分析流程进行两两个体间亲缘关系预测，评估预测准确性。随机减少SNP位点，评估位点数对算法预测准确性的影响。结果 IBD片段算法预测1~7级亲缘关系平均置信区间准确率为94.72%，预测可信度为99.77%，6级及以上亲缘关系预测时出现假阴性。随着SNP数量减少，预测准确性会出现一定程度的下降。结论 IBD片段算法可用于7级以内亲缘关系的预测，该算法在群体遗传学、法医遗传学等领域有重要应用价值。     

The efficacy of respondent-driven sampling for the health assessment of minority populations

BackgroundRespondent driven sampling (RDS) is a relatively new network sampling technique typically employed for hard-to-reach populations. Like snowball sampling, initial respondents or “seeds” recruit additional respondents from their network of friends. Under certain assumptions, the method promises to produce a sample independent from the biases that may have been introduced by the non-random choice of “seeds.” We conducted a survey on health communication in Guam’s general population using the RDS method, the first survey that has utilized this methodology in Guam. It was conducted in hopes of identifying a cost-efficient non-probability sampling strategy that could generate reasonable population estimates for both minority and general populations.MethodsRDS data was collected in Guam in 2013 (n = 511) and population estimates were compared with 2012 BRFSS data (n = 2031) and the 2010 census data. The estimates were calculated using the unweighted RDS sample and the weighted sample using RDS inference methods and compared with known population characteristics.ResultsThe sample size was reached in 23 days, providing evidence that the RDS method is a viable, cost-effective data collection method, which can provide reasonable population estimates. However, the results also suggest that the RDS inference methods used to reduce bias, based on self-reported estimates of network sizes, may not always work. Caution is needed when interpreting RDS study findings.ConclusionsFor a more diverse sample, data collection should not be conducted in just one location. Fewer questions about network estimates should be asked, and more careful consideration should be given to the kind of incentives offered to participants.     

A new genus of the tribe Acratini from the continental Neotropical region (Coleoptera: Curculionoidea,Brentidae)

A group of 10 species belonging to the Neotropical tribe Acratini Alonso-Zarazaga, Lyal, Bartolozzi & Sforzi 1999, is shown to be monophyletic by a phylogenetic analysis based on 48 morphological characters. A new genus, Pertusius n. gen., is described, based on three derived characters of external and genital morphology: venter of prorostrum of males with fine median longitudinal carina, base of elytra with deep pit at the place of insertion on mesonotum, and proximal sclerite of endophallus more or less horseshoe-shaped. Eight species were previously known and, according to former authors, belonged to the genera Acratus Lacordaire 1865 [Pertusius apicalis (Sharp 1895), n. comb., P. errabundus (Kleine 1927), n. comb., P. extrarius (Kleine 1927), n. comb., P. fidus (Kleine 1927), n. comb. and P. telesi (Soares & Meyer 1959), n. comb.] and Proteramocerus Kleine 1921 [Pertusius chalcites (Perty 1832), n. comb., P. filum (Sharp 1895), n. comb., and P. laevis (Germar 1824), n. comb.]. Four new synonymies are proposed: Proteramocerus disparilis Soares & Dias 1971, n. syn. for Acratus apicalis Sharp 1895; Teramocerus laevigatus Boheman 1840, n. syn. for Arrhenodes chalcites Perty 1832; Acratus extraordinarius Kleine 1927, n. syn. for Acratus errabundus Kleine 1927; Proteramocerus diringshofeni Soares & Dias 1971, n. syn. for Acratus filum Sharp 1895. Two new species are described: Pertusius guyanensis n. sp. from French Guiana, and P. mexicanus n. sp. from Mexico (Yucatán Peninsula). Pertusius apicalis is newly cited from Bolivia and Peru, P. chalcites from Argentina and Paraguay, P. extrarius from Ecuador, P. fidus from Peru and P. telesi from Trinidad and Tobago and Venezuela. An identification key to species of the genus is provided.     

Phylogenetic relationships in Bulbostylis (Abildgaardieae: Cyperaceae) inferred from nuclear and plastid DNA sequence data

Previous molecular phylogenetic analyses of the family Cyperaceae based on rbcL sequences showed Bulbostylis as paraphyletic, with B. atrosanguinea and B. hispidula forming a clade with Nemum spadiceum. On the contrary, phylogenetic analyses of the tribe Abildgaardieae based on nuclear (ITS ribosomal region) and plastid sequences (trnL-F region) showed Bulbostylis as monophyletic, although they only incorporated four species of Bulbostylis and none of Nemum. In this work, we presented a phylogenetic hypothesis of Bulbostylis based on a comprehensive sampling, including species from different continents for the first time. New sequences of Abildgaardia, Crosslandia, Fimbristylis, and Nemum were included to test the monophyly of Bulbostylis. In total, 84 sequences of both ITS and trnL regions were generated. Analyses were performed using Bayesian inference, maximum likelihood, and parsimony. Ancestral state reconstruction was performed using ML, MCMC, and parsimony methods. In all analyses, Bulbostylis resulted paraphyletic as Nemum atracuminatum is nested within it. Most American species of Bulbostylis grouped together, but relationships amongst them appeared poorly resolved. Ancestral state reconstructions of native distribution suggest an African ancestor of Bulbostylis, with at least three introduction independent events of the species in America. Morphological diagnostic characters such as the ‘style base permanence or detachment from the ripe achene’, and the ‘micromorphological patterns of the achene surface’ are homoplastic in this phylogenetic context, and therefore unsuitable to propose infrageneric groupings within the Bulbostylis.     

Do species of Neacratus with anchor-shaped sclerite of endophallus really belong to Neacratus (Coleoptera: Brentidae: Acratini)?

A small group of nine nominal species belonging to the genus Neacratus Alonso-Zarazaga, Lyal, Sforzi & Bartolozzi, 1999 is studied in detail from a morphological point of view. Lectotypes are designated for Brentus obtusus Lund, 1800 and Nemocephalus fulgidus Kleine, 1928. Three new synonymies are proposed: Nemocephalus brevicostatus Kleine, 1922 n. syn. for Brenthus obtusus Lund, 1800, Nemocephalus longiceps Perroud, 1853 n. syn. for Brenthus famulus Boheman, 1840, and Nemocephalus fulgidus Kleine, 1928 n. syn. for B. famulus Boheman, 1840. A new species, Neacratus pascali n. sp., is described from French Guiana. New country records are provided for Neacratus obtusus (Bolivia, Ecuador, French Guiana, Grenada, Peru, Trinidad and Tobago, Venezuela), N. puncticeps (Sharp, 1895) (Colombia, Ecuador, Venezuela), N. guatemalensis (Senna, 1893) (Belize, Colombia, El Salvador), N. deplanatus (Sharp, 1895) (Colombia, Costa Rica, French Guiana, Mexico, Panama, Venezuela) and N. famulus (Paraguay). A phylogenetic analysis carried out on this group shows it forms a monophyletic lineage included in a clade containing most of other species of Neacratus, of which it is the type species; it is therefore not justified to create a new generic name. The possible polyphyletism of the genus Neacratus as a whole and the development of an excessively long rostrum in some male Acratini are discussed.     

Variation of plant diversity components in different scales in relation to grazing and climatic conditions

Background: Understanding the role of livestock grazing on plant diversity can be improved by an accurate measurement of diversity at all hierarchical scales due to the changeability of diversity components in space.

Aims: We evaluated the effects of grazing on plant species diversity at different scales of all common and rare species in two regions that have different climatic conditions (arid vs. semi-arid).

Methods: In each region, we collected abundant data of plant species from a nested sampling design that consisted of local (80 plots) and regional (16 sites) scales. We partitioned total species diversity (γ) into within plots (α_l), among plots (β_l) and among sites (β₂) using the additive partitioning.

Results: Diversity among sites contributed the most to total diversity for all and rare plant species in both regions. In addition, α₁ and β₁ diversities in ungrazed areas were greater than those in grazed areas for all and common species in both climates.

Conclusion: Abandonment of grazing after 10 years resulted in significant regeneration of common species at the local scale, with no change in rare species. We conclude that low grazing intensity is likely to be an important tool for conservation of plant diversity in which all scales should be considered.     

Exact Bayesian bin classification: a fast alternative to Bayesian classification and its application to neural response analysis

We investigate the general problem of signal classification and, in particular, that of assigning stimulus labels to neural spike trains recorded from single cortical neurons. Finding efficient ways of classifying neural responses is especially important in experiments involving rapid presentation of stimuli. We introduce a fast, exact alternative to Bayesian classification. Instead of estimating the class-conditional densities p(x|y) (where x is a scalar function of the feature[s], y the class label) and converting them to P(y|x) via Bayes’ theorem, this probability is evaluated directly and without the need for approximations. This is achieved by integrating over all possible binnings of x with an upper limit on the number of bins. Computational time is quadratic in both the number of observed data points and the number of bins. The algorithm also allows for the computation of feedback signals, which can be used as input to subsequent stages of inference, e.g. neural network training. Responses of single neurons from high-level visual cortex (area STSa) to rapid sequences of complex visual stimuli are analysed. Information latency and response duration increase nonlinearly with presentation duration, suggesting that neural processing speeds adapt to presentation speeds. Action Editor: Alexander Borst     

The ‘requirement of total evidence’ and its role in phylogenetic systematics

The question of whether or not to partition data for the purposes of inferring phylogenetic hypotheses remains controversial. Opinions have been especially divided since Kluge's (1989, Systematic Zoology 38, 7–25) claim that data partitioning violates the requirement of total evidence (RTE). Unfortunately, advocacy for or against the RTE has not been based on accurate portrayals of the requirement. The RTE is a basic maxim for non-deductive inference, stipulating that evidence must be considered if it has relevance to an inference. Evidence is relevant if it has a positive or negative effect on a given conclusion. In the case of ℈partitioned’ phylogenetic inferences, the RTE is violated, and the basis for rational belief in any conclusion is compromised, unless it is shown that the partitions are evidentially irrelevant to one another. The goal of phylogenetic systematics is to hypothesize past causal conditions to account for observed shared similarities among two or more species. Such inferences are non-deductive, necessitating consideration of the RTE. Some phylogeneticists claim the parsimony criterion as justification for the RTE. There is no relation between the two – parsimony is a relation between a hypothesis and causal question(s). Parsimony does not dictate the content of premises prior to an inference. ℈Taxonomic congruence,’ ℈supertrees,’ and ℈conditional combination’ methods violate the RTE. Taxonomic congruence and supertree methods also fail to achieve the intended goal of phylogenetic inference, such that ℈consensus trees’ and ℈supertrees’ lack an empirical basis. ℈Conditional combination’ is problematic because hypotheses derived from partitioned data cannot be compared – a causal hypothesis inferred to account for a set of effects only has relevance to those effects, not any comparative relevance to other causal hypotheses. A similar problem arises in the comparisons of hypotheses derived from different causal theories.