With the development of neural network technology and the rapid growth of China's tourism economic income at this stage, the research on the comprehensive evaluation of tourism resources has gradually emerged. Based on this, this paper studies the neural network comprehensive evaluation model based on multispecies evolutionary genetic algorithm and designs the neural network analysis system of influencing factors of tourism resources based on multispecies evolutionary genetic algorithm. The collection and acquisition of data information are realized from the aspects of resource income status, tourism development investment, and sustainability evaluation in the tourism area. The multispecies evolutionary genetic algorithm is used for comprehensive analysis and evaluation. The algorithm can realize the complex analysis and comprehensive evaluation of the core influencing factors of neural network. Accurate analysis and evaluation were carried out according to the different characteristics of tourism resources and the current situation of tourism income. The results show that the neural network comprehensive evaluation model based on multispecies evolutionary genetic algorithm has the advantages of high practicability, good sorting effect of variable ratio, and good data integration. It can effectively analyze and compare the comprehensive evaluation factors affecting tourism resources in different ratios. Tourism resources are the premise and foundation of tourism development. Tourism resources mainly include natural landscape tourism resources and cultural landscape tourism resources. Tourism resources are the basis of tourism development. China's tourism resources are very rich and have broad development prospects. They are widely used and paid more and more attention to tourism research, regional development, resource protection, and so on. At present, in terms of the comprehensive evaluation of tourism resources in China, the comprehensive evaluation model of input and recovery is mostly adopted, and the recovery analysis is rarely carried out from the perspective of sustainability . Since eThis paper studies the influencing factors of neural network and puts forward the Manchester comprehensive evaluation model of the ratio of variables, which is mainly divided into three parts. The first part introduces and summarizes the research status of influencing factors of tourism resources at home and abroad. The second part constructs a neural network-influencing factor ranking model based on multispecies evolutionary genetic algorithm and objective evaluation and constructs the impact evaluation index system of different types of tourism resources by using the multispecies evolutionary method. The third part takes multiregional tourism resources as the experimental object, tests and verifies the variable-ratio ranking model of influencing factors of comprehensive evaluation constructed in this paper, draws conclusions, and makes corresponding analysis. The innovation of this paper is that the multispecies evolutionary genetic algorithm is applied to the comprehensive evaluation model of influencing factors of neural network. On this basis, it can make full use of the development characteristics of each tourism resource and the difference information of comprehensive evaluation, realize the full type simulation at the simulation level, and quantitatively describe different comprehensive evaluation modes with multitransformed genetic factors. The similarity of sustainability research strategies is consistent with the expected evaluation indicators, and the degree of influence of weight ranking of the comprehensive evaluation is completed with quantitative indicators, which can efficiently rank the factors affecting tourism resources according to different sustainability orientation.To sum up, it can be seen that most of the current comprehensive evaluation models of regional tourism resources do not involve intelligent algorithms based on the difference of regional tourist group characteristics. On the other hand, although China has performed a lot of basic research in the comprehensive evaluation of regional tourism resources, there are relatively few research results in the specific quantitative dynamic neural network evaluation system and the quality evaluation of the comprehensive evaluation of regional tourism resources. There is no research on the objectivity of the comprehensive evaluation quality of regional tourism resources and the construction of relevant models.China is relatively backward in the comprehensive evaluation of tourism resources, and some foreign developed countries are more advanced in the research of the comprehensive evaluation of tourism resources . Bollt fMultispecies evolutionary genetic algorithm and neural network algorithm are one of successful intelligent optimization algorithms. Internationally, data analysis optimization and quality evaluation of different types of problems have become a hot research field . Aiming Based on this, in the process of neural network research on the comprehensive evaluation model of tourism resources based on multispecies evolutionary genetic algorithm, this paper first designs a multispecies evolutionary genetic algorithm based on the influence degree of multidimensional index factors; that is, according to the level differences of different internal capacities and different technical requirements in different regions of China's tourism field, and the differences in the evaluation difficulty of different scenic spots in the same conditions, the highly targeted multispecies evolutionary genetic algorithm is used to realize the comprehensive evaluation and global random search of different scenes, and the differentiation analysis is carried out. Second, through the neural network based on multispecies evolutionary genetic algorithm, a series of data information expressed in different regions are accurately divided in the process of comprehensive analysis of tourism resources, so as to realize the high classification of different quality of different types of tourism resources in the process of comprehensive analysis of tourism resources, The goal with strong synergy and relevance  is pushed to the process to be optimized in the next stage. Combined with the guidance of sustainability, the neural network model is used to realize the goal-oriented and high-quality recovery of special objectives in different types of tourism resources and accurately improve the stability of the comprehensive evaluation and quality assessment system.In the process of neural network research on the comprehensive evaluation model of tourism resources, this part first adopts the neural network based on multispecies evolutionary genetic algorithm based on multivariate transformation factors and selects three characteristic parameters related to the tourism innovation quality and tourism evaluation of tourism resources. The data processing process of neural network based on multispecies evolutionary genetic algorithm is shown in Through the research on the common innovation types of tourism resources and the evaluation and incentive rules for regions, this paper clearly defines the hierarchical framework and classification subordination of the whole tourism resource system. Finally, the comprehensive evaluation of the impact of multiple genetic factors can be established from the perspective of the comprehensive evaluation of the impact of genetic resources, so as to provide an objective reference for the comprehensive evaluation of the impact of multiple genetic factors. In the construction of tourism resource optimization model and evaluation link based on multispecies evolutionary genetic algorithm, we will use a neural network based on multispecies evolutionary genetic algorithm to classify the comprehensive evaluation methods of tourism resources in different stages according to the similarity and collaborative similarity of regional innovation ability and then divide the data information of regional innovation methods in tourism work. Through the selection of multispecies evolutionary genetic algorithm, the divided comprehensive evaluation scheme can be divided and updated twice, so as to ensure the stratification and updation of technological innovation in the process of regional sustainability research and comprehensive analysis. The simulation results of neural network based on the multispecies evolutionary genetic algorithm are shown in As can be seen from It can be seen from m= (tn+1 is t1) is taken as an individual. The inverse function of the sum of distances between adjacent data in this sequence can be used as the fitness of the corresponding individual sustainability m, so the fitness function is as follows:As can be seen from m= is coded. However, it is difficult to directly encode such individuals [U, V, W, X, Y, and Z to represent the corresponding sustainability, The sequence of these six symbols is used to represent the possible solution, that is, the chromosome in the neural network based on multispecies evolutionary genetic algorithm [In this process, when we interpret each sustainability, the corresponding individual ividuals . Becauselgorithm . Then, tAs can be seen from Then, we perform conventional crossover or mutation operations in neural network based on multispecies evolutionary genetic algorithm, such as exchanging the last three bits as follows:X at the fifth position of chromosome t2 into W and get formula (or change  formula :(9)t2″=Ut1′, t2′, and t2″ obtained above are illegal sequences.From this, we can see that The idea of sustainable development originated in the field of ecological environment and is truly a classic definition of sustainable development recognized by the international community. It not only meets the needs of contemporary people but also does not harm the development of the ability of the future generations to meet their needs, such as economic sustainable development, ecological sustainable development, and social sustainable development. The relationship between population, resources, and environment should be correctly handled, so as to continuously enhance China's sustainable development ability, continuously improve the ecological environment, and significantly improve the efficiency of resource utilization, and so as to promote the whole society to embark on the road of ecological good and harmonious development.In order to solve this problem, we carry out set operation (intersection and union operation) and chromosome-pairing regularization on the tourism resource evaluation data of the tourism resource sustainability group with the number P in the same spatial location stage and carry out comparative analysis for many times. When the visitor object of each region belongs to the set corresponding to the nearest genetic node center, the iterative processing is ended.W and the target genetic crossover space as U.t represents the location of the  tourism resource scheme, andt  represents the variation rate of t.We set the population scale of tourism resource sustainability as Finally, through the database information of the computer and the preset automatic judgment program, some data information is restored, so as to reprocess and process the secondary data information and then cycle back and forth to form three clusters . The datIt can be seen from In the process of evaluating the influencing factors of the comprehensive evaluation of tourism resources from the neural network based on the multispecies evolutionary genetic algorithm, in order to maximize the innovation level of regions with different backgrounds according to their existing comprehensive evaluation incentive level and characteristic information status, the neural network based on multispecies evolutionary genetic algorithm used in this paper will improve the existing resource level of tourism resources, determine the appropriate genetic strategy according to the corresponding eigenvalues and fitness, including selecting population size, selecting and crossing the mutation methods, and determining genetic parameters such as crossover probability and mutation probability, and further randomly generate the initial population . When thWhen the neural network of multispecies evolutionary genetic algorithm deeply excavates the corresponding comprehensive evaluation level for different tourism resources, it will produce different similarities of innovation quality levels for the groups corresponding to tourism resources .Therefore, in the research process of the influencing factor model of tourism resources based on neural network based on multispecies evolutionary genetic algorithm, when a certain index is met, this research generally means that when the fitness of the optimal individual reaches a given threshold, or the fitness of the optimal individual and population does not rise, the iterative process of the algorithm converges and the algorithm ends. A neural network based on the multispecies evolutionary genetic algorithm is used to classify and analyze the sustainability of the same innovation focus . In the Finally, for the key requirements of different sustainability on tourism innovation, this model will be divided according to the current internal comprehensive evaluation scheme and implementation effect of tourism and conduct specific index evaluation. In the comprehensive evaluation model of influencing factors of tourism resources based on multispecies evolutionary genetic algorithm, it will combine the distribution and ranking of different influencing factors of tourism resources, so as to efficiently help tourism resources quickly improve the internal comprehensive evaluation of tourism according to their own development needs, which can be used as the evaluation index of tourism resources.In order to combine the sustainability and actual development needs of China's tourism resources, this study establishes a comprehensive evaluation model of influencing factor ranking based on the efficient interaction between sustainability and tourism resource scheme, which can realize the scientific evaluation of the actual ratio of influencing factors of tourism resources. Based on the above evaluation rules of neural network based on the multispecies evolutionary genetic algorithm, we first establish a fuzzy comprehensive evaluation model based on the incentive orientation and focus of sustainability on tourism. The model takes the characteristic indicators of factors affecting tourism resources as the center and takes the existing tourism resource level and different sustainability-oriented information as the core evaluation indicators to evaluate the innovative achievements and quality of tourism areas.The experimental process takes the comprehensive evaluation difference degree of different tourism resource types  as the experimental object. By using the optimized neural network based on multispecies evolutionary genetic algorithm based on deep mining, three-dimensional simulation process, and multilevel population division, it is the test data source for the comprehensive evaluation of different tourism resources under different sustainability incentives and carries out the impact analysis according to the innovation achievements and quality of different tourism as the standard, so as to realize the differential evaluation of different tourism resources and the nondifferential evaluation within tourism, and so as to sort and analyze different influencing factors.The experiment is divided into two parts: the control group and the experimental group. The experimental results of the population and its fitness of the first three generations are shown in For the analysis of influencing factors of the comprehensive evaluation of different types of tourism resources in China, the comparison between the analysis of influencing factors based on multispecies evolutionary genetic algorithm and not based on multispecies evolutionary genetic algorithm is shown in It can be seen from the above table and In order to better analyze the influencing factors of the neural network, we need to focus on the analysis and research from the perspective of sustainability. On this basis, the neural network comprehensive evaluation model based on multispecies evolutionary genetic algorithm is studied, and the neural network analysis system of influencing factors of tourism resources based on multispecies evolutionary genetic algorithm is designed. The innovation of this paper is to apply a multipopulation evolutionary genetic algorithm to the comprehensive evaluation model of influencing factors of neural network. First, three characteristic parameters related to the influencing factors of tourism resources are selected, and an evaluation system of influencing factors of tourism resources based on multispecies evolutionary genetic algorithm is proposed. Through the research on the status of tourism resources, tourism development investment, and evaluation, this paper clearly defines the hierarchical framework and index relationship of the whole tourism resources evaluation system. Second, the evaluation methods of the influencing factors of this comprehensive evaluation are evaluated from multiple angles, which provide a reference scientific research case for establishing an intelligent comprehensive evaluation system. Experiments show that based on multispecies evolutionary genetic algorithm, the comprehensive evaluation of the influencing factors of tourism resources can be realized by using fuzzy evaluation, which is conducive to improving the sustainability of tourism resources and the possibility of obtaining tourist support. However, this paper only focuses on the construction of the tourism resource system and does not take the influencing factors of the comprehensive evaluation of different types of tourism into account. Therefore, the comprehensive analysis of the influencing factors of the comprehensive evaluation system needs to be further studied.


The use of genetic test results in risk-rated insurance is a significant concern internationally, with many countries banning or restricting the use of genetic test results in underwriting. In Australia, life insurers’ use of genetic test results is legal and self-regulated by the insurance industry ). In 2018, an Australian Parliamentary Inquiry recommended that insurers’ use of genetic test results in underwriting should be prohibited. In 2019, the FSC introduced an industry self-regulated moratorium on the use of genetic test results. In the absence of government oversight, it is critical that the impact, effectiveness and appropriateness of the moratorium is monitored. Here we describe the protocol of our government-funded research project, which will serve that critical function between 2020 and 2023.A realist evaluation framework was developed for the project, using a context-mechanism-outcome (CMO) approach, to systematically assess the impact of the moratorium for a range of stakeholders. Outcomes which need to be achieved for the moratorium to accomplish its intended aims were identified, and specific data collection measures methods were developed to gather the evidence from relevant stakeholder groups  to determine if aims are achieved. Results from each arm of the study will be analysed and published in peer-reviewed journals as they become available.The A-GLIMMER project will provide essential monitoring of the impact and effectiveness of the self-regulated insurance moratorium. On completion of the study (3 years) a Stakeholder Report will be compiled. The Stakeholder Report will synthesise the evidence gathered in each arm of the study and use the CMO framework to evaluate the extent to which each of the outcomes have been achieved, and make evidence-based recommendations to the Australian federal government, life insurance industry and other stakeholders.The online version contains supplementary material available at 10.1186/s12910-021-00634-2. Genetic Nondiscrimination Act (2017) prohibits insurers  from using genetic test results without an individual’s express consent. In the US, the Genetic Information Nondiscrimination Act (2008) (GINA) limits the use of genetic information only in health insurance underwriting (and employment contexts). It does not apply to life insurance, although some individual states have legislated to limit genetic discrimination in life insurance [The use of genetic test results in risk-rated insurance is a significant concern internationally –4. A majnsurance .Other jurisdictions have implemented alternative mechanisms, such as a moratorium in the UK (now the Code on Genetic Testing and Insurance ) which wDisability Discrimination Act 1992 (Cth), life insurers are legally permitted to use genetic test results to discriminate against all applicants [In Australia, risk-rated insurance is provided by life insurers, not health insurers. Under the plicants , 17, 18.Additionally, GD in this context has been identified as one of the most significant ethical, legal and social issues (ELSI) in genomics currently facing Australia, both in terms of policy development and its impact on genetic research and clinical services . In 2018Disability Discrimination Act 1992 (Cth)—that is, insurers are still legally allowed to use genetic test results to discriminate against all applicants [Although the federal government has not yet responded to the recommendations, in July 2019 the FSC independently introduced an industry-led moratorium [RESEARCH QUESTION: To what extent does the self-regulated FSC moratorium achieve the critical policy aims identified by the Parliamentary Joint Committee (PJC)?The A-GLIMMER study, funded by the Australian government’s Genomic Health Futures Mission, will evaluate the current Australian response to GD in life insurance:To reduce consumer fears related to insurance, which deter the uptake of clinical genetic testing and/or research participation (s9.98)To eliminate genetic discrimination in the Australian life insurance industry (ss9.84 &9.86)To remove a barrier currently compromising the success of genetic medicine in Australia (s9.89)To ensure Australian government oversight and monitoring to combat concerns with industry self-regulation (ss 9.94 & 9.96)The aims of the recommended policy change in this area, as discussed in the PJC Report  are:To rOur research project will assess whether the moratorium is effective in achieving these aims. This research will serve a critical role in increasing the evidence base internationally and helping Australia achieve appropriate long-term regulation for this important issue, taking into consideration the perspective of all key stakeholders .Internationally, various measures have been introduced to address GD. Research has been conducted into the effectiveness of the regulatory mechanisms used in European countries, such as ethnographic fieldwork within insurance companies  and postWe have identified four major stakeholder groups, whose perspectives must be considered in order to rigorously assess whether the current Australian moratorium is an appropriate and effective long-term regulatory solution. Some research has previously been conducted internationally on these stakeholder groups to gauge experiences and perceptions of genetic discrimination, views on regulation of genetic discrimination and knowledge of relevant local non-discrimination instruments, as set out below. While these studies represent findings at various timepoints across a variety of regulatory contexts, which may differ from those currently in Australia, they demonstrate the research which has been conducted in this area.Since the 1990s, numerous studies in North America, the United Kingdom, Europe and Australia have described concerns regarding GD. These concerns were voiced by at-risk clinical patients –39, suppHealth professionals (HPs) —both genetic and non-genetic clinicians—are key to ensuring adequate communication of information about GD to patients. In a survey undertaken before the recent introduction of Canadian non-discrimination legislation , all CanVarious studies have also surveyed health professionals without a genetics qualification about their views and experiences regarding genetic testing and insurance discrimination. In one US study of over 1000 physicians and nurse practitioners , 96% of Studies have also tested health professionals’ knowledge and understanding of legal non-discrimination provisions. In one US study, > 90% of participants (n = 1110) had an inaccurate knowledge of current legal protections , and in Genetic Non-Discrimination Act) showed that more than a third of families with “very sick children”, declined to participate in a free research study because of such fears [International researchers have described the deterrent effect of GD fears on research participation . Evidencch fears . Less thch fears . Similarch fears .Research has been conducted internationally (primarily in the USA) directly with insurance companies to understand their practices and perspectives regarding the use of genetic test results in underwriting. This research includes both life and health insurance providers, but more recently has been focused on health insurers with the introduction of GINA legislation  in 2008. In 1993, medical directors of US life insurance companies were surveyed  using a It is critical that the impact, effectiveness and appropriateness of the FSC moratorium is monitored, taking into account these different stakeholder perspectives , to ensure that the proposed FSC review in 2022 is informed by rigorous and evidence-based submissions. Currently, there are no other mechanisms in place to do this, and this project addresses that critical gap. Our project, funded by an Australian government grant, will utilise a nationally coordinated effort to collect data from different stakeholder perspectives, to build a complete picture of the impact of the moratorium.To systematically assess the impact of the moratorium, a realist evaluation paradigm was employed in constructing the evaluation framework. “Realist evaluations asks not, ‘What works?’ or, ‘Does this program work?’ but asks instead, ‘What works for whom in what circumstances and in what respects, and how?’” , p.2. ThWidespread and accurate awareness of the existence of the moratorium and its terms among consumer groups, health professionals, genetic researchers and research participants, ethics committees, financial industry members and regulators.Confidence among consumers, health professionals, researchers and the insurance industry that the moratorium terms are strictly adhered to, and that breaches are rectified.Timely and regular updates to policy, practice and processes in health care, industry and research to reflect the moratorium  guidelines).Adherence to the terms of the moratorium in the collection and use of genetic test results by all insurance companies, in practice.For the moratorium to accomplish its intended aims (see above), the following outcomes must be achieved:Inattention to any of these areas will reduce the ability of the moratorium to achieve its intended outcomes.As indicated in Table In order to assess the outcome measures in Table When considering genetic testing, a consumer is any individual who has had, or may have in the future, a genetic test. Consumers include those with a personal and family history of genetic or medical conditions, as well as ostensibly healthy individuals who may consider genetic testing for potential preventative health benefit or may be offered population genetic testing or genetic testing as part of a research study. With respect to genetic testing and life insurance, individuals fall into a range of different categories see Fig. .Fig. 4CaOBJECTIVE 1.1 Assess levels of awareness and understanding of the moratorium in the general population, genetic testers, pre-testers and declinersOBJECTIVE 1.2 Assess the self-described impact of the moratorium on the decision-making of pre-testers and declinersOBJECTIVE 1.3 Assess the impact of the moratorium on genetic testers’ ability to access insurance products compared to pre-moratoriumPart 1 of A-GLIMMER is designed to address the following objectives  was held to seek input regarding the proposed methodology for gathering consumer views.Prior to the commencement of the moratorium, research was conducted with Australian consumers to assist with identifying experiences in access to life insurance products following genetic testing . This rePost-moratorium, this research methodology will be repeated and expanded. Individuals with variants that increase their risk of disease, as well as favourable results that negate a family history of disease, will be surveyed to capture levels of understanding of the moratorium, impact of the moratorium on decision-making and experiences with accessing life insurance products. Recruitment will be through support groups and other consumer groups, but the reach will be expanded considerably to groups supporting consumers with a range of genetic conditions, including but not limited to LSA, Pink Hope, Mito Foundation, Familial Hypercholesterolemia Network Australia, Rare Cancers Australia, Genetic Undiagnosed and Rare Disease Network, Rare Voices Australia, and Cancer Council Victoria. With an expanded reach, we expect that the number of participants will exceed the number from the previous survey.Unaffected individuals who are considering having predictive genetic testing will be surveyed to capture levels of understanding of the moratorium and the impact of the moratorium on decision-making. Decliners will be surveyed to understand reasons for their decision to not have testing.The PRiMo (using Polygenic Risk Modification to improve breast cancer prevention) trial is recruiting female participants who will be offered genetic testing through Australian Familial Cancer Clinics (FCCs) for breast and ovarian cancer-predisposing genetic variants. Questions regarding knowledge of the moratorium, influence of insurance implications on decisions regarding genetic testing, and experiences with accessing life insurance will be included in the questionnaires received by participants soon after receiving results. Follow-up questions about experiences with accessing life insurance will be included in questionnaires administered at subsequent 6-12 month intervals.Males attending an FCC and considering predictive genetic testing for adult-onset autosomal dominant conditions, and females considering predictive genetic testing for adult-onset autosomal dominant conditions who are not eligible for PRiMo, will be invited separately to answer questions regarding knowledge of the moratorium, influence of insurance implications on decisions regarding genetic testing, and experiences with accessing life insurance.Each year, the Australian Consortium for Social and Political Research Incorporated administers the Australian Survey of Social Attitudes (AuSSA) to representative sections of the general public. The AuSSA is “Australia’s main source of data for the scientific study of the social attitudes, beliefs and opinions of Australians, how they change over time, and how they compare with other societies” . We prevFor the purposes of the A-GLIMMER project, health professionals (HPs) include any qualified health professional who has direct contact with patients who are considering genetic testing. This includes HPs working in genetics services, such as genetic counsellors and clinical geneticists, as well as other non-genetic HPs who discuss genetic testing with patients, such as nurses and oncologists.OBJECTIVE 2.1 Assess the level of understanding of the moratorium by health professionals.OBJECTIVE 2.2 Describe experiences of health professionals regarding the impact of the moratorium on patients.OBJECTIVE 2.3 Describe health professionals’ views on regulation and the moratorium’s effectivenessPart 2 is designed to address the following objectives:In Australia, some opportunistic data collection from interviewing health professionals occurred as part of a project which aimed to verify reports of GD by consumers , but didDuring the data collection period for the pre-moratorium survey, some feedback was obtained regarding the questions asked and the process of completing the survey. Prior to finalising this protocol, the proposed follow-up survey questions were piloted on several genetics professionals in different roles, who provided feedback about content, clarity and flow.Following commencement of the moratorium, health professionals who discuss genetic testing with patients will be invited to participate in an online survey  impact of the moratorium on the Financial Services IndustryOBJECTIVE 4.3 Assess the level of adherence to the moratorium by life insurance companiesPart 4 is designed to address the following objectives:Before the protocol was finalised, a meeting was held with key underwriting representatives from several of the large Australian life insurance providers to seek feedback regarding the proposed methodology, target groups, and subject matter of interviews.The Australian government publishes a list  of registered Australian financial advisers. Financial advisers will be randomly selected  and invited to complete a short anonymous telephone survey, to assess the understanding of financial industry personnel who are not part of a life insurance company. Participants will be asked questions relating to their knowledge and understanding of the existence and terms of the moratorium.Application forms  will be collected from all underwriters offering risk-rated life insurance in Australia. Content analysis will be conducted to determine whether the forms comply with the terms of the moratorium. Specifically, fields considered will include those seeking information from applicants about past or future genetic testing, and explanation (if any) of the terms of the moratorium. Previous research conducted in 2003  collecteUnderwriting representatives from FSC member life insurance companies will be invited to participate in semi-structured interviews or focus groups to explore their views on the moratorium, changes to practice, benefits and limitations, and adherence to terms. Focus groups and interviews will be conducted by videoconference and facilitated by members of the research team. Sixteen life insurance companies are currently members of FSC and it is expected that approximately 10–15 underwriters will attend either a focus group or take part in an interview.The FSC requires its member companies to record in a dedicated database de-identified information regarding all applications for a life insurance product where a genetic test result has been disclosed, either voluntarily or inadvertently . PreviouResults from each arm of the study will be analysed and published in peer-reviewed journals as they become available.At the end of the study term (3 years) a Regulatory Evaluation Report will be prepared. The Regulatory Evaluation Report will synthesise the evidence gathered in each arm of the study and use the CMO framework to evaluate the extent to which the moratorium, as implemented by the FSC Standard, has achieved the outcomes intended by the PJC recommendations. The Regulatory Evaluation Report will identify any outcomes that have not been achieved and will draw on the collected data to provide possible reasons why this has occurred. The Report will make recommendations to rectify any failings in relation to the moratorium and to enhance its operation in the future. Consequently, this research project and the Regulatory Evaluation Report will provide valuable evidence toward, although it will not replace, the FSC’s review of the moratorium . The rep—to what extent does the self-regulated FSC moratorium achieve the aims of addressing concerns with GD as identified by the Parliamentary Joint Committee?Our project brings together Australia’s leading researchers, clinicians, patient groups, policy experts and industry representatives to answer an over-arching research questionStrengths of the study include an experienced and diverse investigator group from across Australia that has published extensively together in the area , 73–78, The study has limitations and risks which must be acknowledged. The diverse methods of data collection being undertaken across the four stakeholder groups could be challenging to synthesise in a final report. The study may be more likely to collect data from highly motivated or vocal stakeholders, rather than a truly representative cross-section of the community. Further, there is a risk of investigator team bias, given individual views on the issue of GD. We have taken deliberate steps to mitigate against these risks, to ensure rigour and objectivity in our study.The study’s limited timeline presents another challenge, given the broad and diverse scope of work to be completed. Various challenges or delays could prevent key milestones from being achieved. For example, difficulties in recruiting participants, or obtaining necessary ethics approvals, could influence the planned timeline and milestones. Further disruptions caused by the Covid-19 pandemic may also create challenges for recruitment and data collection. Other risks for the study include the availability of industry-collected data. As study partners, FSC has pledged to provide access to certain industry data, but the research team does not have primary access to this data, and so it is possible that access to this data could be delayed or inconsistent.In conclusion, the findings of this study will provide valuable evidence to inform the FSC review of the moratorium in 2022, and future policy regarding the use of genetic information in life insurance.Additional file 1. Title of data: Health professional questionnaire. Description of data: questionnaire to be administered to health professionals after the introduction of the moratorium


Olea europaea ‘Galega vulgar’ variety is a blend of West and Central Mediterranean germplasm with cultivated-wild admixture characteristics. ‘Galega vulgar’ is known for its high rusticity and superior-quality olive oil, being the main Portuguese variety with high impact for bioeconomy. Nevertheless, it has been replaced by higher-yielding and more adapted to intensive production foreign varieties. To clarify the potential ancestral origin, genetic diversity evolution, and existing genetic relationships within the national heritage of ‘Galega vulgar’, 595 trees, belonging to ancient and centenary age groups and prospected among ten traditional production regions, were characterized using 14 SSR markers after variety validation by endocarp measurements. Ninety-five distinguishable genets were identified, revealing the presence of a reasonable amount of intra-genetic and morphological variability. A minimum spanning tree, depicting the complete genealogy of all identified genets, represented the ‘Galega vulgar’ intra-varietal diversity, with 94% of the trees showing only a two-allele difference from the most frequent genet (C001). Strong correlations between the number of differentiating alleles from C001, the clonal size, and their net divergence suggested an ancestral monoclonal origin of the ‘Galega vulgar’, with the most frequent genet identified as the most likely origin of all the genets and phenotypic diversification occurring through somatic mutations. Genetic erosion was detected through the loss of some allele combinations across time. This work highlights the need to recover the lost diversity in this traditional olive variety by including ancient private genets  in future breeding programs and investing in the protection of these valuable resources in situ by safeguarding the defined region of origin and dispersion of ‘Galega vulgar’. Furthermore, this approach proved useful on a highly diverse olive variety and thus applicable to other diverse varieties due either to their intermediate nature between different gene pools or to the presence of a mixture of cultivated and wild traits . Olea europaea L. subsp. europaea var. europaea  , and it is the fourth-largest olive producer in Europe, after Spain, Greece, and Italy IOC, 2020, with thNowadays, molecular markers are fundamental tools used in the characterization and understanding of diversity, due to their independence from environmental effects , to trace back any eventual loss of allelic diversity that might have occurred across time. This study will allow us to track and recover the genetic diversity still existing in the country but that might be already lost in the present ‘Galega vulgar’ orchards; broaden the available genetic basis for breeding purposes; and unravel the ancestral origin of this variety. All these efforts will contribute to the efficient conservation of this variety and to promote its use.To assess the diversity of the ‘Galega vulgar’ variety present in the Portuguese orchards or kept on isolated trees, a collection was established with individual trees prospected from different traditional regions of olive production. Prospections were performed taking into consideration the age of the olive trees provided by farmers or conservation sites . Within each orchard, the prospected olive trees (about ten trees/orchard) were randomly selected and tagged with a unique identifier, and the tree global position system (GPS) coordinates recorded. The same procedure was used for isolated individuals. This resulted in 629 prospected trees, 356 of which were centennial and 273 ancient, from ten geographical districts . A complper tree were collected to perform endocarp validation of the ‘Galega vulgar’ variety; and during spring, ten young leaves per tree were collected for DNA isolation and genetic diversity characterization. The fruits were kept on ice at harvest and then stored at 4 ± 2°C until variety validation, while leaves were kept on ice when collected and then frozen in liquid nitrogen and stored at −80 ± 2°C until genomic DNA extraction.For each tree, leaf and fruit samples were collected in two different seasons: during autumn, 50 fruits Representative sample of each ‘Galega vulgar’, ‘Arbequina’, and ‘Picual’ varieties from the World Olive Germplasm Bank (WOGB) were also included in the SSR analysis.per each of the 629 prospected trees, were characterized using nine different qualitative parameters and one quantitative parameter: shape in the position of greatest eccentricity, symmetry in position A, symmetry in position B, number of grooves on basal end, distribution of grooves on basal end, shape of apex in position A, mucron, shape of the base in position A, rugosity of surface, and weight , stained with SYBR® Safe , and visualized using a GEL-DOC1000 System . Quantification and extra quality evaluation were performed using a Nanodrop spectrophotometer, Nanodrop ND-2000C .DNA was extracted from the sampled young leaves using the CTAB procedure developed by Doyle and Doyle . The quaThe SSR analysis consisted of 14 SSRs, selected from the consensus list of the best olive SSRs  of PCR products were estimated using SagaGT software from LI-COR and data compiled in a matrix for further analysis. All alleles were double scored and manually reviewed by two independent researchers to prevent scoring errors. Samples with small differences in allele sizes were additionally checked by re-amplifications to exclude possible genotyping and allele sizing errors. The ‘Galega vulgar’, ‘Arbequina’, and ‘Picual’ representative samples from the WOGB were also included in the allele sizes estimation.a) and genotypes (Ng) per locus as well as the Polymorphic Information Content (PIC)  was plotted on a log-log scale. The Pareto's parameter β was calculated by regression from the Pareto distribution as −1 multiplied by the regression slope  in the set of ‘Galega vulgar’ tree samples was identified using GenClone 2.0  germplasm groups [centennial trees (<100 years old) vs. ancient trees (>400 years old)], (2) age groups , (3) groups according to endocarp profile (E1–E4), (4) geographical groups (10 regions of origin), and (5) groups according to tree location (orchard vs. isolated tree).per locus (Nav), the allelic richness (Nar) as a measure of the number of alleles per locus independent of sample size, and the number of private alleles (Npr), using FSTAT v. 2.9.3.2 software , clonal richness (Ncr), and genotypic richness [R; Dorken and Eckert  is dependent on sample size, the clonal richness (Ncr) was calculated according with the modified formula proposed by El Mousadik and Petit (Nar), based on the rarefaction method of Hurlbert  and expected (HE) heterozygosity within each group.Intra-varietal genetic diversity was analyzed after identifying the tree samples that shared an identical genet and removing the duplicates from the data set. GENEPOP 4.0  the net divergence of the genets and the logarithm of the number of ramets or clonal size, (B) the number of different alleles of each genet to the most frequent genet (C001) and the net divergence of the genet, and (C) the number of different alleles of each genet to the most frequent genet (C001) and the number of ramets in each genet SAS, .https://www.graphviz.org/). The tree was rooted using the most frequent genet (C001) as outgroup.The minimum spanning tree based on the matrix of genetic distances computed as the number of different alleles among genets was constructed using a custom script in Python version 3.6 (available on request) and the tree was visualized using Graphviz  to 9 (GAPU103A), with an average of 5.50. The most informative SSRs were ssrOeUA-DCA11 and GAPU103A, showing a PIC value of 0.61 and 0.57, respectively  was identified in the panel of the 595 trees analyzed. The distribution of genets according to the number of genetically identical samples  was highly skewed, with two large genets and many small ones resulting in a shallow regression line of Pareto's plot . The larpsa) based on the 12 polymorphic microsatellites was calculated among the 95 genets and ranged from 0.04  to 0.33   . The twoalleles) . When coalleles) .A principal coordinate analysis (PCoA) based on the proportion-of-shared-alleles distance was performed to graphically depict the genetic relationships among genets . The firr = −0.00) (as shown in r = −0.03)  . Indeed,Nar, HO, HE) (Nc) identified in the centenary and ancient germplasm groups was also similar (57 vs. 58), but the clonal richness (Ncr) was higher in ancient trees (48.94 vs. 58.00). Among age groups, the clonal richness was higher in A1 and A3 groups . The number of private clones (Npc) in the two germplasm groups (centenary vs. ancient) was similar (37 vs. 38) but the private clonal richness was higher in ancient trees (30.07 vs. 38.00), with the highest value in A1 group. Regarding the genotypic richness , it was higher in ancient trees (0.17 vs. 0.22), assuming the highest value in the oldest A3 group. Consequently, the probability of encountering distinct genets when randomly taking two trees of an age group (D*) was also slightly higher in ancient trees (0.77 vs. 0.79). In conclusion, some specific alleles or genets of each age group were different , with the number of allele combinations higher on ancient genotypes, which translates into the loss of some allele combinations across time.Regarding the comparison of overall centennial and ancient trees and between age groups , no group differed significantly in any of the standard genetic diversity parameters  . HoweverHO and HE), no significant differences were found between centenary and ancient germplasm groups , neither among age groups. In all the cases, the observed heterozygosity was higher than the expected heterozygosity . Even so, the unique ancient group presented the highest values for allelic richness (4.33 vs. 4.95), clonal richness (34.25 vs. 38.00), private clonal richness (34.25 vs. 38.00), R (0.68 vs. 0.76), D* (0.96 vs. 0.99) and V (0.72 vs. 0.88). Regarding the observed and expected heterozygosity, both were higher in the centennial group.Out of the 95 identified genets, only 20 (21.05%) were identified in both germplasm groups. Nevertheless, the six most frequent genets (C001-C006), represented by 426 (71.60%) out of 595 sampled trees, were found in both germplasm groups. By analyzing the genets with unique age individuals  , no signIn this study, we did not find any evidence for cases of homonymy , nor for sexual propagation  that could be explained as a result of out-crossing events, or by a selection pressure made by olive growers to maintain a ‘Galega vulgar’ morphotype.P < 0.05) among the four endocarp profiles. A Tukey's HSD test  revealed that the E1 and E4 profiles were significantly different from each other but not from E2 and E3 profiles. For clonal richness (Ncr), private clonal richness (Npcr) and expected heterozygosity (HE), comparing all the endocarp profiles, E2 presented the highest values , and E4 the lowest values   .Nar, HO, HE among districts .From the comparison of the ten prospected geographical districts , no signcr, Npcr, D*, and V), Portalegre, Santarém, Lisboa, and Évora showed the highest values (exception for Lisboa Npcr) (cr) and therefore the highest probability of encountering two different genets , while Portalegre had the highest number of private genets . Finally, the region having the most equitable clonal distribution, i.e., all the genets had almost the same frequency of tested trees , was Santarém  . More spSantarém .Nar, HO, HE  .Regarding the comparison between orchard and isolated trees, no significant differences were found for ctively) . NeverthAnalysis of molecular variance (AMOVA) was performed to test the existence of genetic structure among and within groups: germplasm groups (centenary vs. ancient), age groups , endocarp profile groups , prospected geographical districts groups, and tree location groups (orchard vs. isolated tree). Negative values of ϕ-statistics were observed in both tests, suggesting the absence of any genetic structure .Values for the correlation coefficients calculated to analyze the ancestral origin of this variety after identifying the presence of one main genet (C001), by clarifying the relationship between the amount of clonal divergence and clonal size (number of ramets), can be found in r = −0.49; P < 0.0001) was observed between the net divergence of the genets and their number of ramets  was observed between the number of differentiating alleles of genets in comparison to the most frequent genet (C001) and their net divergence  was observed between the number of differentiating alleles of genets in comparison to the most frequent genet (C001) and their number of ramets. The other most frequent genets (C002–C008) had a single allele difference from C001  had the lowest net divergence (198), followed by the second most frequent genet (C002) with 221. A negative correlation  results based on the proportion-of-shared-alleles distance matrix, we also observed that the majority of the individual trees were clustered in a few genets, with a mixture of undistinguishable ancient and centennial germplasm. Similar findings were observed for other Portuguese varieties, namely ‘Verdeal-Transmontana’ and ‘Cobrançosa’, that display a wide intra-varietal genetic variability  very frequent and genetically similar genets. The same results were already described for other Portuguese varieties, as ‘Verdeal-Transmontana’, where the analyzed trees fell into two major genotypic groups  was higher than the expected heterozygosity (HE) in both germplasm groups (centenary and ancient), and also considering the different age groups . Using SSR markers, several authors also reported the same trend in different olive germplasm , being the most frequent genets present in centennial orchards and therefore still conserved under production. On the other hand, the ancient trees showed a higher clonal richness and private clonal richness  compared with the centennial trees, meaning that some allele combinations were lost across time, representing a case of genetic erosion. Indeed, Maxted and Guarino  describeHowever, it is important to mention that estimating the age of live olive trees represented a very challenging task. According to Lavee , the innin situ in particular regions presenting higher diversity could be a way to recover from this genetic erosion.The identified genetic erosion highlights the need to recover the lost diversity at the more recently established orchards. In fact, isolated ancient trees showed a higher clonal richness compared to orchard trees. Therefore, the inclusion of ancient private genets in breeding programs, broadening the programs' genetic basis is a key aspect, since these ancient genets might have interesting traits for survival adaptation due to their longevity . Indeed, according to Belaj et al. , the WOGThe morphological diversity with the simultaneous consideration of rounded to acute endocarp shapes and medium to small endocarp weights in the ‘Galega vulgar’ variety could be a sign of an ongoing domestication process or the effect of a long-term clonal propagation process. According to Fuller , in the in situ in regions with the highest diversity, and eventually ex situ, which might allow genetic studies under the same environmental conditions. As far as we know, this was the most representative study made so far in Portugal regarding the ‘Galega vulgar’ diversity and is a conscious first effort to efficiently preserve and use this national high-quality genetic resource. Moreover, this approach has the potential to be replicated in other diverse varieties, either due to their intermediate nature between different gene pools or due to the presence of a mixture of cultivated and wild traits, as is the case of ‘Galega vulgar’. Thus, it will be possible to accurately access and counteract the risk of further genetic erosion of these varieties, toward their efficient use in breeding programs.This study allowed us to track, for an eventual recovery, the genetic diversity still existing in Portugal but that might already be lost in the present ‘Galega vulgar’ orchards and unraveled the ancestral origin of this variety. The inclusion of the ancient private genets  in breeding programs will broaden their present genetic basis. Moreover, investment should be made in the protection of these resources https://figshare.com/, doi: 10.6084/m9.figshare.12666446.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: HS performed the prospection of the individual trees, morphological endocarp characterization, DNA isolation, the SSR genotyping, participated in the analysis of the molecular data, and drafted the manuscript. ZS performed the statistical analysis of the molecular data, and critically participated in the manuscript revision. MLA participate in the analysis of the molecular data and revised the manuscript critically. PF was involved in the prospection phase of this study and revised the manuscript critically. JN revised the manuscript critically. MCVP designed and coordinated the study and participated in the drafting and revising of the manuscript. All authors read and approved the final manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Procyon lotor) is an invasive carnivore that invaded various areas of the world. Although controlling feral raccoon populations is important to reduce serious threats to local ecosystems, raccoons are not under rigid population control in Europe and Japan. We examined the D-loop and nuclear microsatellite regions to identify spatially explicit and feasible management units for effective population control and further range expansion retardation. Through the identification of five mitochondrial DNA haplotypes and three nuclear genetic groups, we identified at least three independent introductions, range expansion, and subsequent genetic admixture in the Boso Peninsula. The management unit considered that two were appropriate because two populations have already genetic exchange. Furthermore, when taking management, we think that it is important to monitor DNA at the same time as capture measures for feasible management. This makes it possible to determine whether there is a invasion that has a significant impact on population growth from out of the unit, and enables adaptive management.The raccoon ( Invasion may ultimately lead to serious human health4 and economic6 problems.Controlling invasive species is important for reducing threats to biodiversity and natural resources on a global scale7. For example, a systematic measure includes early detection and rapid response to eradicate newly established populations; monitoring population dynamics and range expansion are important components of adaptive management approaches. Not only distribution data but also molecular analyses are effective tools for understanding the process of range expansion by understanding the degree of genetic exchange under various environmental conditions8. Moreover, an eradication program can be adopted by identifying the marginal populations to limit range expansion and avoid genetic admixture9, whereas the core populations must be kept at low density to prevent negative impacts10. Asada11 proposed “lag-phase management” as an effective management technique for mammals, considering the sexual difference in dispersal distance, as males disperse over significantly longer distances than females12. When the target species have gender differences in dispersion distance, only males are distributed in the foremost part of the distribution expansion area, and a low-density region “lag-phase” is formed, where the Allee-effect13 retards population growth. Thereafter, the female spreads its distribution similar to “petals on a rose”14, and an “increase-phase” is formed, where the number of animals increases rapidly. Although it is difficult to achieve eradication, maintaining lag-phase by applying capture pressure is possible. We think that this lag-phase management can effectively prevent the expansion of the distribution area. For that purpose, it is important to understand the current genetic structure and set an appropriate management unit. This enables adaptive management by monitoring whether it occurs to exchange with other regions and whether countermeasures are successful.Implementing biological control for invasive species often requires both systematic measures and adaptive management approachesProcyon lotor) is a carnivore originally native to North and Central America16. Due to its invasiveness in terms of wide dietary niche, habitat generalist, high density, and rapid population growth, feral raccoons are a serious threat to local ecosystems and eliminate native species through predation, competition, and pathogen transmission2. Despite the urgency and need for raccoon population management, it has not been properly managed in Europe10 and Japan17, because of increasing population trends, unlimited range expansion, and an inefficient management strategy18.Raccoon . Since the relationship between raccoon expansion process and genetic exchange in Japan might be extremely complex19, it is difficult to estimate the details of range expansion only from sighting records. However, if we focus on a geographically limited area, we might be able to examine the details of range expansion in association with landscape and genetic structures within a particular region.To take effective management, we tried to understand feasible management units for retarding population growth. We explored molecular genetic methods for understanding the process of range expansion and degree of genetic exchange in various landscapes21. We have discussed the recognition of management units for feral raccoons in a semi-confined landscape of the Boso Peninsula as a model system.In this study, we analyzed the mitochondrial D-loop and microsatellite loci of raccoons in the Chiba Prefecture to detect the area of introduced populations and expansion process, supplemented with information on past distribution2), protruding into the Pacific Ocean from the Kanto Plain , and preserved in medium with 70% concentration of ethanol until DNA extraction.Tissue samples were collected from 179 carcasses of feral racoons that were euthanized for pest control in Chiba Prefecture from November 2014 to August 2019. Mitochondrial DNA (mtDNA) haplotype and nuclear microsatellite genotypes were determined for the tail tissue preserved in 99.5% ethanol or buccal cells collected with a cotton swab, rinsed in phosphate-buffered saline (PBS) solution . The tissue samples were cut into small pieces and DNA was extracted according to the manufacturer’s instructions. The buccal cell sample was rinsed three times in PBS solution and DNA was extracted. The DNA was eluted using 200 µL AE buffer.26 and PLO-CRL1 (5′-CGCTTAAACTTATGTCCTGTAACC-3′)27 primers. The PCR amplifications were performed in 15 µL total volume containing 1 µl template, 0.5 U Expand High Fidelity Enzyme Mix , 150 µM of each dNTP, and 300 nM of each primer. After an initial incubation at 95 °C for 5 min, PCR was performed for 35–40 cycles of denaturation at 94 °C for 30 s, annealing at 56 °C for 30 s, and extension at 72 °C for 30 s, followed by a final extension at 72 °C for 30 min. The PCR products were purified using EXoSAP-IT  and sequenced using a Big Dye Terminator v3.1 Cycle Sequencing Kit  using the FASMAC sequencing service . The sequences were aligned using MEGA 10 software28.A part of the mitochondrial D-loop region was amplified using PLO-L15997 (5′-CCATCAGCACCCAAAGCT-3′)31 were analyzed via multiplex PCR  using Multiplex Manager v1.236 , was constructed using Network 5 (http://www.fluxus-engineering.com)38. We followed the categorization of lineages in the network described by Cullingham et al.27. Thereafter, the genetic diversity (He) of microsatellite loci was compared among the feral raccoons from Chiba, Hokkaido19, those from Spain, Germany, Czech Republic, and Poland42, and the native raccoons from North America, including Ontario, Indiana, Columbia, Missouri and Illinois43 with a sign test using R version 3.6.2. Thus, we downloaded the Excel file with geographical coordinates of each animal and the allelic fragment length at every gene locus from the Dryad database (10.5061/dryad.412h3)40. Information regarding the genetic diversity of raccoons of Germany40 was calculated using GenAlEx 6.502.For the mtDNA control region, 406 bp sequences that overlapped with the same region used in a previous studyNa) in each locus, the effective number of alleles (Ne), observed heterozygosity (Ho), expected heterozygosity (He), private alleles (pA), and allelic richness using GenAlEx 6.50245 and FSTAT 2.9.3.246. We regarded each city as a raccoon district group, and the adjacent districts of Kimitsu and Futtsu were combined due to the small sample size. Narita, Chiba, and Sosa were excluded from the analysis of the genetic differentiation due to the low sample size. The He among the districts were compared through a pairwise t-test using R version 3.6.2 (R Core Team 2019) and corrected via the Bonferroni method.Genetic diversity and differentiation among district groups of the raccoons were described initially by genotyping 179 raccoons sampled from the various localities of Chiba Prefecture, and by calculating the number of alleles . The mean Q was calculated from 10 trials in the animal using CLUMPP 1.1.249 and the Q value of each animal was illustrated using distruct 1.150. We set a criterion to include or exclude individual raccoons in particular clusters by considering the Q value of individual raccoons. We considered individual raccoons were to be from a single cluster when the highest Q value was > 0.7. Raccoons with the highest Q value of < 0.7 were grouped as a mixture that did not belong to a particular cluster. We then calculated the district groupwise cluster frequency.Individual-based genetic clustering was performed to determine the number (K) of genetically different groups  using STRUCTURE 2.3Five haplotypes  were identified in the Chiba Prefecture. Among them, three haplotypes  were first found in Chiba and the sequences have been registered in the DNA Data Bank of Japan nucleotide sequence database . Only one single nucleotide polymorphism (SNP) was detected between haplotypes A and E and between haplotypes C and D. The median-joining network, including published haplotypes, is shown in Fig. pA was highest in Noda. The values of others were similar among district groups. No significant difference was found in He between district groups .Table He in Chiba Prefecture and that reported in previous studies. The number of loci used for comparison was different because the markers used in different studies were different. There is no significant difference between the mean He between Chiba Prefecture, native areas, central Europe, Germany, and Hokkaido. Although the mean He in Spain was not significantly different from that in the Chiba Prefecture, the genetic diversity tended to be lower than that in the Chiba Prefecture (P = 0.07).Table Based on the ΔK value, the most probable structure clustering was K = 3. Figure 19. The Invasive Alien Species Act of Japan enacted in 2005 has banned the pet trade of raccoon, and strictly regulated the transportation of living animals within Japan since 2005. Thus, the newly released or escaped captive raccoons or animals dispersed from adjacent areas should account for the origin of raccoon populations in Boso Peninsula. Boso Peninsula is almost isolated from the adjacent prefecture by the sea and the large rivers of Tonegawa River and Edogawa River  were detected in the nuclear DNA of raccoons of the Chiba Prefecture using STRUCTURE analysis, and these three nuclear genetic groups corresponded to mtDNA haplotypes E, A, and B, respectively. Therefore, these three groups, two in the east  and west  of southern Boso Peninsula, and one in the northwest corner  of the Chiba Prefecture Fig. , are comEven though there are significant gaps in the abundance and genetic compositions among the northwestern and southern groups Fig. , the geo51, genetic admixture extent differs between the sexes. Variation in mtDNA haplotypes is transmitted maternally; hence, an admixture of mtDNA haplotypes would be much slower than the nuclear genetic variations. In the case of the genetic admixture of mtDNA haplotypes between Isumi and Kimitsu-Futtsu in the Tateyama-Minamiboso area, the straight-line distance from Isumi to Tateyama is ca. 50 km, and the female range expansion proceeds with an average speed of ca. 2.9 km per year from 1998 to 2015, being much faster than the estimated speed of 0.6 km in the native area12.Due to the male-biased long-distance dispersal in raccoons52. By analysis of nuclear DNA, it was shown that the raccoon population in Chiba Prefecture is currently divided into a northern population, a southwestern population, and a southeastern population. A wide range of gene flow has already occurred in the southern part of Chiba Prefecture. On the other hand, there is no genetic exchange yet between the northern and southern parts. Accordingly, in Chiba Prefecture, we think that it is possible to further suppress the rate of distribution expansion by setting the northern population and the southern population as separate management units and focusing on capture measures from the breaks in the distribution of each population.To exterminate and manage alien species, it is necessary to set up a management unit for the target population in consideration of the movement of organisms to prevent re-invasionWhen taking management, we think that is important to monitor DNA at the same time as capture measures. To reduce the distribution area of the target population and reduce the density, there must be no invasion from the outside, especially female invasion that has a huge influence on the increase in the population. Haplotypes of mtDNA between the northern populations and the southern population in Chiba Prefecture are not mixed. Therefore, by monitoring mtDNA derived from females, it is possible to determine whether or not females have invaded from another population.Because the expansion speed of the feral raccoons is very fast, it is essential to adjust the measures in large administrative units such as Chiba Prefecture, instead of the measures in narrow administrative units where the methods and efforts differed for each municipality as current. In this study, we showed Chiba Prefecture for the model area, we believe that these ideas are not limited to raccoons but can be applied to population management measures for many alien species.


In recent years, genetic counseling has started playing a major role in the field of genomic medicine. There are currently about 7,000 genetic counselors in more than 28 countries, with 267 certified genetic counselors in Japan alone . While the rapid advancement of genomic medicine has expanded this field, the challenges genetic counselors face are also evolving. This article aims to provide an overview of the institutional and social issues surrounding genetic counselors in Japan and discuss implications for the global community. In Japan, with the rapid changes in genomic medicine and the establishment of a delivery mechanism within the healthcare system, several issues need to be discussed. First, many genetic testing, counseling, and preventive healthcare programs are not covered by public health insurance. Second, reducing human resource shortages for genetic counseling is an urgent issue. Third, the lack of a national qualification in the profession is critically important issue in the field. Fourth, research on the role and value of genetic counselors is still limited. To address these issues, discussions among relevant stakeholders, including genetic counselors, professionals in genomic medicine, and lawmakers, are necessary. Additionally, we believe that research by genetic counselors to evaluate and improve their practice and examine institutional and social issues is crucial for developing their profession’s activities and delivering high-quality healthcare to many people. To establish the position and role of the relatively new profession of genetic counselor, sharing information and collaborating on institutional and social challenges faced by genetic counselors globally will be beneficial. In recent years, genomics as a specialization in medicine has progressively expanded and attracted attention. Genetic counselors play a major role in genomic medicine. The first genetic counseling training program was established at the Sarah Lawrence College in New York, United States, in 1969. There are about 7,000 genetic counselors in more than 28 countries, as of 2018 .In Japan, there is a certification program called “Certified Genetic Counselor” (CGC), which is a joint qualification obtained under the Japan Society of Human Genetics and the Japanese Society of Genetic Counseling . To becoGlobally, genetic counselors face various institutional and social challenges in their work, and the situation varies across countries . AlthougJapan is an island country covering 377,974 square kilometers. It is made up of 47 prefectures and has a population of about 126 million , with moMost medical professions are regulated by qualification laws that define the permitted scope of work. To obtain a license, individuals must complete a required curriculum at an educational institution established in accordance with the law, and pass a national examination. In Japan, these licenses for healthcare workers are granted by Ministry of Health, Labour, and Welfare. In this article, when referring to these systems, we use the term “national qualification.”There are several government organizations involved in healthcare in Japan; the Ministry of Health, Labour, and Welfare is mainly responsible for administration related to medical care and welfare . In addiIn Japan, as in other countries, there have been rapid advancements in genomic medicine, in line with advancements in genome-wide analysis technology. These changes have been particularly significant during the past few years, with the implementation of various large-scale clinical studies and the launch of new healthcare services related to genomic medicine.In 2013, American actress Angelina Jolie announced that she had hereditary breast and ovarian cancer syndrome (HBOC) . This caThere have been some major changes in healthcare in Japan during the last few years. In 2018, one of the national healthcare and research projects—known as “cancer genomic medicine,” which utilizes cancer gene panel tests—began to develop a healthcare delivery system for tumor profiling testing, which was covered by public health insurance . This, iGenomic medicine requires the provision of individualized prevention and treatment based on appropriate diagnoses for individuals at risk, not merely those with symptoms. Improving healthcare delivery and securing human resources are an important factor in ensuring that such services are available to those who need it.BRCA1/2 genetic test was already covered as a companion diagnosis for the PARP inhibitor olaparib in 2018. As a result of the 2020 reforms in the medical reimbursement system, the government committee for national health insurance decided to extend the application of genetic testing to individuals with a family history of HBOC and breast and ovarian cancer patients. Further, reduced-risk mastectomy or risk-reducing salpingo-oophorectomy for cancer patients with BRCA1/2 associated HBOC are also covered by insurance following these reforms. In the 2020 reforms in the medical reimbursement system, the number of diseases diagnosed by genetic testing covered by public insurance increased by 65—that is, from 75 to 140 intractable diseases—including genetic diseases occurring in children and intractable diseases that are covered by the national subsidy system for healthcare expenses   and 267 Training genetic counseling professionals has not been able to keep up with the rapid growth in demand. Owing to the practical difficulties of training numerous people at once, each graduate school can only train a few people per year, which results in a shortage of genetic counselors. Further collaboration between stakeholders is necessary, as there is a need to improve the system to provide high-quality genetic counseling promptly to those who need it.A critical challenge faced by the genetic counseling industry in Japan is the professional qualification of genetic counselors. While other medical professions specializing in genomic medicine—including physicians and nurses—have a national qualification to pursue, CGC is not yet a national qualification but a certification accredited by academic societies. This means that the scope of genetic counselors in the context of the Japanese healthcare system is unclear, whereas the scope of practice of other nationally qualified professionals is clearly defined by law. When the current CGC certification system was established in Japan, it had considerably little recognition in the medical field and offered few opportunities for employment . HoweverGlobally, the state of genetic counselor certification, legislation, and licensing systems vary. Countries with national and state regulations are limited, such as South Africa, Malaysia, and parts of the United States; Australia is currently lobbying for the regulation of the profession . In JapaWith the abovementioned advancements in genomic medicine, the role of the genetic counselor is expanding, as opportunities to handle genome-wide information and areas of engagement are also expanding. The nature of the counselor’s work is changing from genetic to genomic counseling . There aAnother challenge for genetic counselors is increasing their capacity to carry out research, as many genetic counselors work in clinical settings. In the United States, there has been an increase in the proportion of genetic counselors working in non-clinical settings such as testing companies, training programs, and research institutions . In JapaHowever, to develop activities as part of the genetic counseling profession, it is necessary to evaluate and improve practice. Simultaneously, institutional and social issues—such as the position of the profession in collaboration with multiple professions and the regulation of qualifications—need to be considered. To implement these things, we believe that research is a powerful tool that can capture and evaluate phenomena from an objective standpoint and consider issues and identify solutions.The importance of genetic counselors conducting research has already been highlighted . NationaThe Japanese Association of Certified Genetic Counselors clearly states in its Code of Ethics that research is one of its duties “to contribute to the development of professional knowledge and skills as a CGC through efforts in practice, research and education” . AlthougHowever, few studies in Japan have examined the institutional and social issues surrounding genetic counselors, such as workforce issues, evaluation of genetic counseling by the public, and its cultural impact. There is limited information, such as published statistical data, on these issues. Many of them are also written in Japanese. There is limited information from other countries, especially those with a short history of genetic counselors. We believe that expanding these areas of studies will be useful in promoting an understanding of the genetic counseling profession among stakeholders such as healthcare policymakers. Future developments in this area are desirable, including collaboration with researchers in other countries, as a certain number of samples and experience will be necessary to conduct research in these areas.As more and more healthcare professionals begin to get involved in genomic medicine, genetic counselors with specialized training programs in genetic counseling will become increasingly important in the future. As this occupation has a short history, in Japan and many other countries, it will be necessary to consider various aspects to make improvements in the future. We hope that genetic counselors will take various research initiatives and report their findings both domestically and internationally. This will stimulate discussions on various issues and lead to the development of the field.In Japan, genetic counseling is new, compared with many other healthcare professions. This means that there are also many institutional and social challenges surrounding the profession and the necessity for it to respond to a rapidly growing need among patients. During the past 15 years, the foundation of the profession has been laid based on precedents from both Japan and abroad, and it appears that genetic counselors are moving to the next step in their development as a profession, which will support the future of medical care and deliver high-quality genetic counseling to those who need it. Although there are differences in the medical systems and cultural backgrounds of all countries, we can learn a lot from the variety of efforts in other countries, such as how to tackle workforce issues and improve the quality of genetic counseling. Therefore, it will be important for Japanese genetic counselors to take the initiative in promoting discussion and creating a professional role in genomic medicine, while drawing inspiration from their global colleagues.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.YA and KK discussed institutional and social issues surrounding genetic counselors in Japan and came up with the idea for the work. YA investigated the relevant literature and wrote the first draft. All authors discussed and refined the manuscript and completed it.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


N = 33,375). Moderate genetic diversity and relatedness were observed across the foxes sampled. Low to moderate levels of inbreeding, high-levels of identity-by-state values, as well as high identity-by-descent values were also found. There was limited evidence for population genetic structure among the foxes across the landscape sampled, supporting the presence of a single population across the study area. This indicates that there may be no barriers hindering fox dispersal across the landscape.The use of genetic information in conservation biology has become more widespread with genetic information more readily available for non-model organisms. It has also been recognized that genetic information from invasive species can inform their management and control. The red fox poses a significant threat to Australian native fauna and the agricultural industry. Despite this, there are few recently published studies investigating the population genetics of foxes in Australia. This study investigated the population genetics of 94 foxes across the Illawarra and Shoalhaven regions of New South Wales, Australia. Diversity Array sequencing technology was used to genotype a large number of single nucleotide polymorphisms ( Vulpes vulpes, has been recognised as an invasive species and key threat to Australian fauna, following the deliberate introduction and establishment of the species in southern Victoria about 140 years ago [Dasyurus maculatus), with a conservation status of near-threatened, where there is extensive overlap in habitat and resource use [The European red fox, ears ago . Foxes oears ago . It is wears ago . Their mears ago ,7,8. Reiears ago ,10,11,12urce use . Field sThe most widely used method to control foxes remains sodium fluoroacetate (compound 1080) baiting . HoweverThus far, markers used for tracking genetic diversity of wildlife had focused on the use of highly informative microsatellite markers. Based on known dog breeds’ microsatellite loci, Atterby et al.  used 14 Here, we report on the genetic diversity and genetic relatedness of red foxes across an agricultural landscape in south-eastern Australia, using SNPs. The aim of this study was to improve the understanding of red fox genetics in south-eastern Australia. Investigating the population genetics of this key invasive species is a fundamental step in improving fox control, and therefore reducing their negative impact in Australia.2 , following the recommended manufacturer’s protocol. Extracted DNA was subjected to quality control measures, including visualization by agarose gel electrophoresis and spectrophotometric quantification at 260 nm using a NanodropTM 2000 spectrophotometer .Approximately 0.5 cmGenotyping of the 94 foxes was undertaken by Diversity Arrays Technology Canberra (DArT) Pty Ltd.  using DArTseq technology. The method uses complexity reduction through the use of restriction enzymes, as well as preferentially targeting low-copy genomic regions over repetitive DNA fragments, increasing assay sensitivity, and allowing for detection of a high number of informative SNPs across the genome ,30. The p < 0.05, and those with >1% minor allele frequencies, using the package dartR in R v4.0.0 [The DArT SNP data and associated basic data provided were converted into a genlight object using the package adegenet  to assisR v4.0.0 .OH) heterozygosity and expected (EH) heterozygosity using dartR and HIERFSTAT v0.04-22 [STF) [ISF) was calculated using PLINK after having randomly removed one individual from potential first-degree relative pairs. The program STRUCTURE v2.3.4 [K = 1 to 9 for 10 iterations. Structure Harvester v0.6.94 [K to explore the number of possible genetically distinct clusters. In addition, the Principal Coordinate Analysis (PCoA), as available in the dartR program package, was used to allocate single foxes into potential genetic clusters. Finally, PLINK v1.07 was used to explore any study-wide significant allele frequencies (p < 0.0001) between the two postulated genetic clusters.The filtered dataset was used to estimate observed (v0.04-22 . Nei’s p22 [FST)  was calc22 [FST) . The inbE v2.3.4  was used v0.6.94  was usedp < 0.05.Sex-biased dispersal was tested for using the R package HIERFSTAT. The test was run using the mean allelic index count model based on the method presented by Goudet et al.  for 10,0Pairwise identity-by-state (IBS) was calculated using PLINK. Identity-by-descent values were calculated using PLINK to determine the potential kinship within the sampled foxes. The type of relationship was estimated in consultation with the demographic data available for the foxes see . This inThe DNA of 93 foxes was successfully genotyped. One sample did not return genotypes and was therefore excluded from further analysis. In total, 33,735 SNPs were reported. Filtering for a 90% call rate removed 7092 SNPs; filtering for Hardy–Weinberg Equilibrium at a nominal α value of α = 0.05 (without Bonferroni correction for multiple testing) removed a further 8202 SNPs. A final minor allele frequency filter for SNPs with frequencies <1% excluded a further 543 SNPs. Minor allele frequencies for the remaining 17,898 SNPs ranged from 0.012 to 0.5. The large majority (49.7%) of SNPs had minor allele frequencies >0.15 and <0.4, indicative of having a high information content for genetic analysis. Complete genotyping was observed for 11,307 or 63% of the SNPs. Only four foxes had more than 5% (but less than 8%) of their genotype missing.K = 2.The ΔK plot  indicateK = 1, we decided to model two different scenarios. The first was a model for a single genetic cluster in which all foxes (N = 93) were grouped. Assuming this model, a moderate heterozygosity (OH) was observed (OH = 0.280). Moderate genetic diversity, estimated by expected heterozygosity (EH), was observed across all foxes (EH = 0.302). Values of inbreeding were estimated (ISF = 0.057 (−0.277–0.343)), with some larger values of inbreeding observed for a small portion of foxes .Taking into account that Structure Harvester cannot accurately estimate the number of genetic clusters if K = 2 best-fit genetic cluster model, both from the log probability (LnP) and highest ΔK (ΔK = 50.8)  .Cluster 1 was 42  and to Cluster 2 was 51 individuals , respectively. Foxes belonging to Cluster 2 were mainly sourced from the area of the Kangaroo Valley, whereas foxes from Cluster 1 were more identified either north or south of the Kangaroo Valley  in allele frequencies   . The oveWe used PLINK to calculate the IBD and IBS values to construct potential lineages within the foxes. The following key parameters were observed: A Z0 value referring to the proportion of no alleles shared between two individuals. A Z1 value referring to the proportion for which one allele is shared between the two individuals. Finally, a Z2 value referring to the proportion for which two (both) alleles are shared between the two individuals. A he foxes .These family kinships were found to be sampled less than 1.6 km of their respective kinship groups. All of these assumed pedigrees were dyads, i.e., one parent and one or two offspring. We tested these kinships by constructing ‘family trees’ and tested the Mendel error rates . Comparep < 0.05. Sex-biased dispersal was found to be not significant, neither for the one-cluster model (p = 0.838) nor the two-cluster model (p = 0.337).Sex was not provided for 14 foxes; therefore, these foxes were excluded from this analysis. The HIERFSTAT test was performed assuming a one-cluster model and a two-cluster model, independently. Significance was considered at a nominal value of 2 landscape) of south-eastern Australia using SNPs. We were able to collect DNA from the ear tissue of 94 red foxes, which were donated to us by hunters from a regional fox control program. We employed a rigorous genotyping data quality control process before analysing the data, making sure that we did not introduce any unintentional bias. We demonstrated that the foxes sampled most likely belonged to one panmictic population, with moderate evidence for reduced genetic diversity and moderate evidence for genetic relatedness within this sample.The aim of our study was to explore the population genetics of the red fox in a confined region  in New Zealand [Petaurus breviceps) in Tasmania [Camelus dromedarius) in central Australia [Genetic diversity was estimated using HIERFSTAT. Our data indicated some limited genetic diversity of the studied fox population. A more recent study by Walton et al. , using fn et al.  is close to 0.75 ,44. Therustralia . It is tustralia . The net Zealand , sugar gTasmania , and droustralia .STF estimated values and a lack of a difference in heterozygosity values supported one panmictic population across the 1170 km2 area used in this study. This interpretation was supported by a limited number of loci, which revealed study-wide significant differences in minor allele frequencies between an experimentally constructed two-cluster model. Only 19 of the 17,898 loci used in this study showed significant differences in minor allele frequencies between the two postulated genetic clusters, as determined by association analysis in PLINK. We would like to point out that all SNPs in our study were subjected to strict quality control measures. Specifically, we used stringent inclusion criteria for genotype distribution expectations according to Hardy–Weinberg equilibrium. While the number of male and female foxes is significantly different for the two hypothetically constructed genetic clusters , the genotype distribution for these 19 loci does not seem to be related to sex chromosomal location. Therefore, our study is in alignment with what is expected, as the observed genetic clusters are in close geographic proximity. Given the very low number of these SNPs with such dramatic differences suggests a lack of landscape barriers that otherwise could limit gene flow and dispersal across the region investigated, enabling unmitigated gene flow across the landscape.When exploring various population models using STRUCTURE, our study did not find strong evidence for a two-cluster model. Specifically, the Cluster 2 with a limited number of samples from Cluster 1, whereas the samples collected close to the town of Nowra, and the samples collected close to the town of Bowral, comprised exclusively of foxes that were allocated to Cluster 1. Therefore, foxes may be using the main road that connects these towns as a movement corridor. This information is consistent with findings from previous studies that show fox preferences for manmade roads and tracks [The study site is geographically and ecologically characterized by a relatively narrow coastal plain and an escarpment, which rises in elevation up to about 450 m above sea level, opening towards the Southern Highlands. The escarpment, with pronounced valleys, provide eastward drainage for rivers or creeks draining through wet/dry sclerophyll vegetation types. The coastal plain and the highlands are mostly cleared agricultural land with highly fragmented remnant native vegetation. While it may be possible that the escarpment acts as a landscape-scale barrier to the foxes east–west dispersal, it should be noted that this is mitigated by the numerous roads and highways within this location. For example, when main roads are taken into consideration, it is important to note the clustering of foxes near a single location in the Kangaroo Valley. The Kangaroo Valley cluster comprised mainly samples from d tracks . Howeverd tracks .When exploring the possible sex-biased dispersal of foxes, no supportive evidence was found. Although it is stated otherwise, in that male foxes will disperse greater distances from their birth den, the results of this study are not consistent with the published literature ,54. GiveWe have shown that across a landscape-scale study in south-eastern Australia, there are insufficient geographic barriers to mitigate geneflow of the foxes sampled. Long-term fox control should focus on identifying key local geographic source populations and concentrate control efforts in these locations. A push toward reduction of gene flow, the structuring of populations across the landscape, and allowing genetic drift to assist in control may assist in fox control . SimilarThe goal of control in the region should be to reduce gene flow across the region and to strive for the genetic isolation of foxes. The use of natural genetic control to promote genetic drift and allowing selective pressures through the reduction of advantageous alleles may allow for the reduction of fox population densities and a shift to genetically structured populations.