Salmo salar, L.), an economically, culturally, and ecologically significant species, that is also characterized by complex hierarchical population structure. Specifically, we focus on an in‐depth example of how our approach was applied to a previously data limited region of northern Canada that was defined by a single large DU. Application of our framework with newly available genetic and genomic data led to subdividing this DU into three new DUs. Although our approach was developed to meet criteria of COSEWIC, it is widely applicable given similarities in the definitions of a conservation unit.Conservation units represent important components of intraspecific diversity that can aid in prioritizing and protecting at‐risk populations, while also safeguarding unique diversity that can contribute to species resilience. In Canada, identification and assessments of conservation units is done by the Committee on the Status of Endangered Wildlife in Canada (COSEWIC). COSEWIC can recognize conservation units below the species level  if the unit has attributes that make it both discrete and evolutionarily significant. There are various ways in which a DU can meet criteria of discreteness and significance, and increasing access to “big data” is providing unprecedented information that can directly inform both criteria. Specifically, the incorporation of genomic data for an increasing number of non‐model species is informing more COSEWIC assessments; thus, a repeatable, robust framework is needed for integrating these data into DU characterization. Here, we develop a framework that uses a multifaceted, weight of evidence approach to incorporate multiple data types, including genetic and genomic data, to inform COSEWIC DUs. We apply this framework to delineate DUs of Atlantic salmon ( We focus our example on Labrador, as the original Labrador DU (DU‐02) defined by COSEWIC in 2010 encompassed an extensive area covering over 1300 km of coastline from the northern tip of Labrador, and south along the coast to the Napitipi River in Quebec. Given the large size of this geographic region, it was previously suggested that there was substantial potential for smaller DUs within this region, although data were lacking at that time  from almost 200 locations , our approach next focused on two types of genetic markers to inform discreteness, including microsatellite and SNP datasets. The primary datasets used to evaluate discreteness in Lehnert et al.  were chon = 1433; Table n = 1333; Table FST) or evaluating migration between units. However, we recommend using unsupervised analyses as a starting point, as this does not require prior knowledge of the species' population structure or for groups to be defined a priori. In addition, statistically significant FST values can be generated when gene flow between groups is high. Therefore, these metrics can be useful in conjunction with unsupervised analyses to further corroborate findings.In Labrador, there are 102 salmon‐bearing rivers within the boundaries of the original 2010 COSEWIC Labrador DU, and our genetic datasets include samples from 34 locations for the 101 microsatellite dataset , which ranged from 1 to 10 for each dataset. To determine support for the number of genetic clusters (K), the optimal number of K was determined based on the ∆K statistic  estimates to assess support for the number of genetic clusters , the criterion of multiple discrete genetic groups needed to recognize more than one DU would not be met Figure . If mult2.32.3.1Identifying higher level genetic groups); within the original Labrador DU, data were available for 19 locations from previously published  life‐history differences and (2) climate‐linked differences that are likely to give rise to local adaptation. Anadromous Atlantic salmon spend the first part of their life cycle in fresh water and later migrate to feeding grounds in the marine environment, before returning to fresh water to spawn  was used to infer evidence of local adaptation driving differences between the genetic groups, thus supporting criteria of “evolutionary significance.”As noted above, environmental differences can also be important for shaping life‐history variation and local adaptation  to provide a weight of evidence to fully support evolutionary significance of DUs Figure . InformaFinally, the last step after identifying one or multiple putative DU(s) within the higher level group was to evaluate evolutionarily significant differences between these putative DUs and any neighboring DUs in other higher level genetic groups. At this stage in the framework process, discreteness between the putative DUs was already established based on initial unsupervised analyses, and thus only criteria of evolutionary significance needed to be addressed following the same approach outlined above. In the case of Atlantic salmon, significance of DUs was compared against adjacent DUs, consistent with the biology of the species. Specifically, for Labrador, adjacent DUs to the south (Quebec Eastern North Shore DU) and north (Nunavik DU) were evaluated; however, their significance has been previously supported in prior evaluation of COSEWIC DUs for Atlantic salmon COSEWIC, , and we 33.13.1.1PCA of neutral genomic data  across Canadian Atlantic salmon populations revealed clear separation of Labrador populations from other regions in Canada. These populations corresponded to the previously recognized Labrador DU COSEWIC, , and we 3.1.22) with an extended estuary. Rivers north and south of this embayment represent the Northern Labrador and Southern Labrador genetic groups, respectively, with the rivers located within the embayment representing the Lake Melville genetic group  was 2 based on the ∆K statistic   Figure . For exae Figure . Based oe Figure . Genetice Figure , and in 3.23.2.1pcadapt, populations within the Lake Melville genetic cluster were clearly separated from coastal Labrador sites (Southern and Northern Labrador) along the first PC axis  and these loci were distributed across 27 of the 29 Atlantic salmon chromosomes  overrepresented based on the outlier dataset, with a large proportion of processes related to “fatty acid homeostasis”  with an adjusted R2 of 0.62. Both RDA axes were significant (p < 0.001), with RDA axis 1 separating Southern Labrador from other regions, and explaining 76.9% of the variance in the model. The highest loading variable on RDA axis 1 was precipitation of the coldest quarter (bio19)  Figure . ANOVA oSalvelinus alpinus), whereas the Lake Melville DU is mainly dominated by Atlantic salmon and sea‐run brook trout , and all three of these salmonid species are represented equally in Southern Labrador DU  within Labrador, which include (1) Northern Labrador, (2) Lake Melville, and (3) Southern Labrador, and full details supporting this determination are summarized in Table Finally, while we focus on Labrador DUs here, the evolutionary significance of these DUs must also be evaluated against any neighboring DUs , broad‐scale population structure across conservation units was evaluated from SNP data using PCA and by estimating individual ancestry coefficients  to evaluate discreteness among units using neutral genomic data , genomic data have been used to support evolutionarily significant differences among several DUs based on genetic–environment associations linked to ocean temperature  has revealed that current DU structure warrants changes, as data revealed evidence of parallel evolution of ecotypes, complex demographic histories, and patterns of introgression among DUs  and steelhead    has been identified that influences age‐at‐maturity in Atlantic salmon, accounting for almost 40% of variation in phenotype  salmon between two DUs, supporting evolutionarily significant differences. However, in our framework (and under COSEWIC), we would not consider interbreeding individuals within rivers with different genotypes (and thus different age‐at‐maturity) as different DUs because discreteness is not met in this scenario. This instead represents the diversity captured within the conservation units , such as RDA or random forest analysis (Forester et al., In our approach, we also capture additional sources of variation in the genome beyond SNPs, such as structural variants (Mérot et al., In conclusion, our work highlights how genetic, but more so genomic data, have the capacity to transform our ability to address criteria of conservation units, such as those defined under COSEWIC. Our framework has been applied to anadromous Atlantic salmon populations across eastern Canada, and in Labrador has resulted in the subdivision of a previously data limited single, large DU. These changes may improve conservation and management of Labrador populations and help protect unique biodiversity in the species, as currently the three DUs are experiencing different trends in abundance over time COSEWIC, . DiffereWe declare no conflict of interest.Figure S1.Click here for additional data file.


Lutjanus xanthopinnis, was recorded as a newly described species in the Indo-Pacific region in 2015. However, the knowledge of its biology, biogeography and ecology is scarcely understood, and, hence, its current conservation status is categorized as Data Deficient. The mitochondrial cytochrome c oxidase subunit I (COI) gene was examined to confirm species identification. We also examined the COI gene haplotypes of L. xanthopinnis in Brunei Darussalam and Malaysia together with other waters, i.e., Bangladesh, Indonesia, Japan, Singapore, Sri Lanka and Taiwan. Our molecular analyses found that Brunei Darussalam and eastern Peninsular Malaysia samples were genetically similar. However, the former showed higher genetic diversity than the latter. The samples from these two sites also showed signatures of population expansion. Furthermore, identical haplotypes could be found in different locations, suggesting the absence of spatial genetic structure. On the other hand, Lutjanus lutjanus showed a population structure associated with geographical locations, i.e., western Pacific Ocean, Indian Ocean and Maluku in Indonesia.The yellowfin snapper,  In addiLutjanus based on their external morphological features and meristic counts. However, due to their morphological similarity and overlapping diagnostic characteristics, their taxonomy is still ambiguous and confusing, and the phylogenetic relationships of Lutjanus species remain obscure [Several scientists have revised the taxonomy of  obscure . Accurat obscure .Lutjanus received recent attention from Ref. [Lutjanus xanthopinnis, was subsequently discovered. This is not surprising because L. xanthopinnis resembles Lutjanus madras; hence, it was previously mistaken as this species. In addition, L. madras also exhibits similar colour patterns with other members of the yellow-lined snapper complex including Lutjanus lutjanus and Lutjanus vitta. The discovery of L. xanthopinnis was made after the reexamination of the specimens collected in Indonesia, Japan, Sri Lanka and Taiwan, which was carried out using morphological characteristics, morphometric counts and DNA barcoding. The redescription of true L. madras also had to be made in the study [L. xanthopinnis is highly limited. Therefore, the current conservation status of L. xanthopinnis in the IUCN Red List is categorized as Data Deficient [L. xanthopinnis is still undetermined, hampering pertinent conservation measures for this species. This task is crucial and requires immediate attention since lutjanids represent some of the most beneficial fisheries resources, contributing to their substantial exploitation. Furthermore, snappers also constitute an essential food resource for communities that rely on artisanal fisheries [Indeed, the yellow-lined rom Ref.  and a nehe study . In sucheficient . Furtherisheries .L. lutjanus is distributed in the tropical waters of the Indo-West Pacific region, and its distribution range overlaps with that of L. xanthopinnis [L. lutjanus is a target species in fisheries and broadly consumed in this region, only a few research on its biology and ecology have been conducted [L. lutjanus were found between the western and eastern Peninsular Malaysia  through DNA barcoding [L. lutjanus within the distribution range is not well examined.The bigeye snapper hopinnis ,4. Althoonducted . In addiarcoding , the popc oxidase subunit I (COI) gene sequence of L. xanthopinnis in Brunei Darussalam and eastern Peninsular Malaysia. Mitochondrial DNA markers, particularly COI, have been proven to be a powerful tool for revealing phylogenetic patterns, species identification and genetic diversity of aquatic species [L. xanthopinnis from other sites , which would provide an insight into its population structure. We also similarly examined its closely related species, L. lutjanus.The aim of the present study was to analyse the mitochondrial cytochrome  species ,13. Furt species ,14. This22.1L. xanthopinnis from seven countries, i.e., Bangladesh, Indonesia, Japan, Malaysia, Singapore, Sri Lanka, and Taiwan were obtained from the GenBank database  and were approved by the animal ethics committee at UBD.Thirty-three and 20 specimens were collected in the coastal waters of Brunei Darussalam in January 2021 and in the coastal waters of Terengganu, eastern Peninsular Malaysia, Malaysia in January 2021, respectivelly, and 15 specimens/sequences of database , Table 1i et al. . After m2.2Taq PCR Master Mix according to the manufacturer's instructions. The PCR primers were fish universal primers [Genomic DNA was extracted using the QIAGEN DNeasy Blood & Tissue Kit according to the manufacturer's instructions. The partial mitochondrial COI gene was amplified by polymerase chain reaction (PCR) using the QIAGEN 2×  primers . The PCRST), sum of squared deviation (SSD), Harpending's raggedness index and neutrality tests. Network 10 (www.fluxus-engineering.com) was used to construct haplotype network using the median-joining method.MEGA X  was used3L. xanthopinnis (44 specimens) and a few L. vitta (9 specimens). With DNA barcoding, the BLAST results confirmed that 38 of 44 specimens were L. xanthopinnis with 99–100% identity. The other specimens were morphologically misidentified, in which 6 specimens were barcoded as L. xanthopinnis instead of L. vitta, and 9 specimens were barcoded as L. lutjanus instead of L. xanthopinnis or L. vitta, with 99–100% identity. Therefore, of 53 specimens, this study barcoded 44 specimens as L. xanthopinnis (GenBank accession no. OQ081715-OQ081758) and 9 specimens as L. lutjanus (OQ081759-OQ081767).This study collected and sequenced a total of 53 specimens form Brunei Darussalam and eastern Peninsular Malaysia. Initial morphological identification suggested that the specimens were mostly ST = −0.01, p > 0.05), suggesting that they were of one genetic population in the South China Sea.Based on the 573 bp of COI gene sequence, the samples from Brunei Darussalam showed a higher number of polymorphic sites (10 sites) than those from eastern Peninsular Malaysia (4 sites). Similarly, the haplotype diversity were 0.76 and 0.65, respectively, with the former having 8 haplotypes and the latter having 5 haplotypes. The within-group mean genetic distances  were 0.0025 and 0.0016, respectively. AMOVA resulted in a negative value for the within-group variation (−1.24%) and a value of 101.24% for the between-group variation. A pairwise comparison of the two collection sites showed that the samples were not significantly different from each other  were 0.83 and 0.0019, respectively, which was almost similar to the nucleotide diversity of L. xanthopinnis.Based on the 574 bp of COI gene sequence, 5 polymorphic sites and 6 haplotypes were identified in Haplotype analysis found 25 haplotypes (H1–H25 in L. lutjanus was slightly lower compared to the nucleotide diversity (π = 0.0027) of the Indian Ocean population . On the other hand, the haplotype diversity was higher in the western Pacific Ocean population  compared to the Indian Ocean population . The genetic diversity for the Maluku population was not determined due to the small sample size (2 samples). However, the mean genetic distance between the western Pacific and the Indian Ocean was 0.058, indicating that they were highly divergent. Compared with the Maluku population, the western Pacific Ocean population showed a higher between-group mean genetic distance (0.044) than the Indian Ocean population (0.016). AMOVA showed that the between-group variation was higher (95.76%) than the within-group variation (4.24%) for the three genetic populations. If the Maluku population was excluded, AMOVA similarly showed that the between-group variation was higher (95.98%) than the within-group variation (4.02%). The fixation indexes showed that the western Pacific Ocean population was significantly different compared to the Indian Ocean population . On the other hand, the Maluku population was more different compared to the western Pacific population  than to the Indian Ocean population .The nucleotide diversity (π = 0.0020) for the western Pacific population  of 4L. xanthopinnis in the waters of Brunei Darussalam and the east coast of Peninsular Malaysia. These two sites are located in the southern South China Sea (L. xanthopinnis, probably because the species was only recognized as distinct from L. madras and other Lutjanus species in 2015. Lutjanus xanthopinnis and L. madras together with other lutjanids occur sympatrically, and these fishes can be differentiated through several morphological features such as colour and scale pattern but their meristic numbers are overlapping among these lutjanids [L. lutjanus using DNA barcoding. Therefore, it is important to conduct species identification using DNA barcoding to validate snappers accurately including the yellow-lined snapper complex.The present study is the first to record hina Sea , and preL. xanthopinis showed that some geographically distant localities shared the same haplotypes (L. xanthopinnis might form a panmictic population. Life history and oceanographic characteristics are known to influence the population structure of marine species [L. xathopinnis may be responsible for connecting the species habitats within its distribution range.The study found that the specimens from the eastern Peninsular Malaysia had lower genetic diversity than Brunei Darussalam. This could probably be due to factors such as habitat differences or higher fishing pressure. Haplotype analysis of plotypes ; Fig. 3. species ,22, howeL. lutjnaus and found three genetic populations. Interestingly, these populations are linked to their geographical locations, i.e., the western Pacific Ocean, the Indian Ocean and Maluku in Indonesia. A previous study reported that the L. lutjanus specimens from the eastern and western coastlines of Peninsular Malaysia were genetically distinct. One group could potentially be considered a new species of L. lutjanus [L. lutjanus specimens from the western Pacific Ocean and Indian Ocean were highly divergent. In addition, the present study also found that the L. lutjanus specimens from Maluku in Indonesia might also be another distinct genetic population. This unique population structure could be brought about by limited migration between the geographical locations. Alternatively, this could reflect the historical population isolation caused by the lowering and rising of sea levels during the late Pleistocene, which is known to shape the biogeography in Southeast Asia [The present study also analysed lutjanus . The preast Asia . Thus, fL. xanthopinnis, some other snappers have already been categorized as Near Threatened or Vulnerable [L. xanthopinnis are among the most important fish resources and would be exploited for human consumption. The present study could contribute to further research on stock assessment and conservation measures in the future.Although the current conservation status of the IUCN Red List is Data Deficient for lnerable . This isConceptualization, T.A.; methodology, T.A. and H.T.; validation, T.A. and H.T.; formal analysis, H.T., N.A. and J.J.; investigation, N.A., J.J., S.A. and N.D.Z.; resources, T.A., J.J., S.A., T.N.A,M.J. and A.H.; writing—original draft preparation, T.A. and H.T.; writing—review and editing, T.A and H.T.; supervision, T.A. and H.T.; project administration, T.A.; funding acquisition, T.A. All authors have read and agreed to the published version of the manuscript.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


Genetic discrimination (GD) has not been discussed in East Asia as extensively as in Europe and North America. Influenced by UNESCO’s universal declaration in 1997, the Japanese government took a stringent approach toward GD by releasing the Basic Principles on Human Genome Research in 2000. However, Japanese society has mostly been ignoring the prevention of GD for decades, and the principle of prohibiting GD was never adhered to in any of the Japanese laws. We conducted anonymous surveys among the general adult population in 2017 and 2022 to explore their experiences of GD and attitudes toward laws carrying penalties to prevent GD in Japan. In both years, approximately 3% of the respondents had experienced some unfavorable treatment regarding their genetic information. They showed higher recognition of the benefits of using genetic information and lower recognition of concerns about using genetic information and GD in 2022 than in 2017. However, the awareness regarding the need for legislation with penalties on GD had increased over the five-year period. In 2022, the framework of a bill to promote genomic medicine and prevent GD without any relevant penalties was released by the Bipartisan Diet Members Caucus. Considering that the absence of regulations may be a barrier to obtaining genomic medicine, as the initial step toward making the prohibition of GD more effective, legislation that no form of GD will be tolerated may stimulate education and awareness regarding respect for the human genome and its diversity. Genetic discrimination (GD) is a classic example of ethical, legal, and social implications/issues in genetic research and medicine. UNESCO’s 1997 Universal Declaration on the Human Genome and Human Rights states that “no one shall be subjected to discrimination based on genetic characteristics that is intended to infringe or has the effect of infringing human rights, fundamental freedoms and human dignity , 3. CompInfluenced by UNESCO’s Declaration in 1997—which was one year after the Eugenic Protection Law of Japan was finally repealed—the Japanese government took a tough stance against GD by releasing the Basic Principles on Human Genome Research in 2000 . ArticleHowever, the principal nuance of the prohibition of GD disappeared from the Ethical Guidelines for Human Genome/Gene Analysis Research, which were developed jointly by the Ministry of Education, Culture, Sports, Science and Technology (MEXT), the Ministry of Health, Labour and Welfare (MHLW), and the Ministry of Economy, Trade and Industry (METI) in 2001 . The guiWith regard to guidelines on genetic medicine, 10 Japanese academic societies related to genetic medicine jointly issued a set of guidelines in 2003, which included the following: (1) even if the person tested provides consent, the test results must not be accessible to employers, insurance companies, or schools; (2) healthcare professionals engaged in genetic testing must always exercise caution and special care to ensure that genetic test results are not used in any discriminatory way; and (3) genetic test results must not be used to discriminate against a person who has been tested. As the people tested for the purpose of predicting the onset of a particular disease are generally healthy, strict protections of privacy and appropriate psychological support must be provided . In partThe Personal Data Protection Law was amended in 2015, and the terms “individual identification code” and “special-care-required personal information” were added. The description “base sequence constituting DNA taken from a cell” was included into the category of “individual identification code.” However, it was still unclear how these statutory definitions, especially “special-care-required personal information”, should be interpreted and operationalized in genomic medicine. The MHLW set up a task force to discuss how genetic test results, sequenced data, and relevant information fit into the new legal terms . ConsequThe report of the task force also acknowledged that “there are no legal regulations that directly prohibit discrimination based on genetic characteristics,” but the suggested response was vaguely expressed as “it is necessary to promote efforts to develop the social environment necessary for the promotion of genomic medicine.” Although several members of the task force mentioned the need for legislation on several occasions during the nine meetings , the MHLFor example, the MHLW’s recommendations to employers encouraged them to use only the aptitude and abilities necessary to perform a job as the criteria for employment selection. In the area of employment management, the MHLW has stipulated since 2004 that health information should not be handled beyond the scope necessary to ensure employees’ health. The Insurance Business Law stipulates that insurance companies must obtain approval from the Financial Services Agency (FSA) regarding their business methods, general insurance policy conditions, and methods of calculating premiums and policy reserves when selling new life insurance products. The FSA is responsible for approving insurance contracts to prevent unfair discrimination against people.The report of the MHLW task force concluded , “it is In the absence of a legal ban against GD, approximately 100 genetic tests are covered by universal health insurance in Japan. One such test, a tumor-profiling gene panel test, has been performed about 45,000 times since its inclusion in universal health insurance coverage in 2019.Although a new policy group on genomic medicine comprising bipartisan parliamentarians was formed in 2015, it took some time for them to decide whether to create any new legislation. While various needs were eagerly expressed by the long-neglected genomic medicine community, the lawmakers did not evince much interest in improving genomic medicine and prohibiting GD. However, the MHLW began its policy of promoting genomic medicine in earnest in 2017 without the government and the Diet taking any precautionary measures for GD, which resulted in great concerns among the patient community.With the support of Genetic Alliance in the US as a coalition of patient associations for hereditary diseases, Genetic Alliance Japan was established in 2017. The organization, headed by Ms. Makiko Dazai, submitted requests for a unified response to GD by relevant government ministries, academia, and industry, in addition to the establishment of legislation to prevent GD. The Japan Federation of Cancer Patient Groups, an umbrella organization of cancer patient associations headed by Mr. Shinsuke Amano, also submitted a request for legislation to the ruling parties in 2018 and held a rally at the Diet Members’ Building in December 2019 for the promotion of cancer genome medicine and the protection of patients and others from social disadvantages.In 2021, the Bipartisan Diet Members Caucus released the framework of a bill, the basic principles of which were the promotion of genomic medicine, the consideration of bioethics, and the prevention of GD. However, the group was unable to reach a consensus within the ruling Liberal Democratic Party, and the bill was not drafted.In April 2022, the presidents of the Federation of Medical Societies of Japan and the Japan Medical Association issued a joint statement urging the government and the Diet to regulate GD . In May In October 2022, the final framework of the bill was released by the Bipartisan Diet Members Caucus. Patient advocacy groups, industry, and academic societies jointly lobbied for the early passage of the bill, with approximately 250 groups supporting its framework. These requests led to the swift drafting of the bill, and the final version was released in December.The bill includes three basic principles. The first principle is to realize “world-class genomic medicine” in a wide range of medical fields and to make its benefits widely available to the public by promoting measures for research, development, and provision of genomic medicine in an organic, mutually coordinated manner. The second principle is “to ensure that appropriate consideration is given to bioethics at each stage of research, development, and provision of genomic medicine, given that some of these activities involve the manipulation of genes that can be passed on to progeny, which may have a significant impact on the preservation of human dignity”. This principle implies a cautious attitude toward germline editing in assisted reproductive technology; however, it fails to prohibit any activity. The third principle is that “genomic information obtained in the research and development and provision of genomic medicine should be sufficiently protected, and unjust discrimination should not be made based on the genomic information”. The bill requires the national and local governments to formulate a basic plan for “genomic medicine policies” based on these principles and does not define the relevant penalties.As of this writing , this bill has been passed by the House of Representatives and is awaiting debate in the House of Councillors. If this bill is enacted, it will be the legal responsibility of the Japanese government to create measures to address GD.More than 25 years have passed between the UNESCO Declaration and the completion of the bill containing the article preventing GD in Japan. Aside from the activities of patient communities, people’s views and experiences with GD remain unknown. We therefore conducted large-scale surveys to clarify people’s experiences of GD in Japan, their expectations and concerns about the use of genetic information, and their attitudes toward legal regulations to prevent GD and the inappropriate use of genetic information.The situation surrounding genomic medicine has changed since 2019, and the people of Japan are more familiar now with genomic medicine. As such, we also conducted comparisons between the survey results in 2017 and 2022.In February 2017 and April 2022, we distributed anonymous, cross-sectional online surveys to 44,360 and 45,488 adults aged 20–69 years from the general Japanese population. These samples were extracted based on national census data from the survey panel of INTAGE Inc.The survey questions covered three topics: genetics knowledge, attitudes toward the use of genetic information, and the need for regulations to prevent GD and the inappropriate use of genetic information. The inclusion of family medical history in the definitions of GD prevention laws varies from country to country, and it is not clear whether genetic information includes family medical history in Japan. To ascertain the impact of genetic information in the broadest sense, the surveys were administered by explaining to the respondents that family history was included in genetic information.We compared the characteristics and attitudes of the respondents toward the use of genetic information in 2017 and 2022 using chi-square tests and Wilcoxon rank-sum tests. Multivariate logistic regression analyses were performed using SAS version 9.4  to identify the factors associated with their attitudes toward legal regulations.n = 4982).The response rates in 2017 and 2022 were 24.5%  and 11.6% , respectively. We excluded 286 respondents who did not answer the questions on educational background in 2022 (Table p < 0.0001) and had less concerns about genetic information than those in 2017 (p < 0.0001).Those who believed genetic information would be useful for disease prevention (the sum of “agree” and “tend to agree”) comprised 65.4% of the respondents in 2017 and 68.9% in 2022 Table . NeverthIn both surveys, most of the respondents recognized the need for laws with penalties for inappropriate handling of genetic information and GD Table  and idenTable Our study revealed that in the period between the surveys, the Japanese public’s expectations of genomic medicine and desire for legal regulations, including penalties, had increased; however, their concerns about the use of genetic information and GD had decreased. A survey about attitudes toward the whole genome sequence conducted in 2021 showed that cancer patients and their families had more concerns about the use of genetic information and GD than the general public . Given tOur findings show that female and older respondents, those with higher genetic knowledge, and those with a higher degree of recognition of the benefits of using genetic information and concerns about GD were more likely to express the need for penalties. In previous research, it was found that female and older people had more privacy concerns in North America . Female It has been pointed out that more serious concerns exist with respect to GD in the context of marriage and pregnancy in Asia than in North America and Europe . The surOur surveys had some limitations. The respondents’ experiences of GD were based on self-recognition, which is difficult to verify objectively. Our study was also limited by a low response rate. This may indicate a lack of interest in genetic research and genetic testing. Nonetheless, our surveys were the first to examine public attitudes toward legal regulations regarding GD in Japan.In Japan, a principled bill will constitute a starting point from which to discuss definitions of GD and examples of GD that should be eliminated from society. As noted by Joly et al. 2022), creating a law prohibiting GD does not necessarily prevent all instances of GD [22, creatThe government is responsible for breaking the chains of GD, which can span generations. It is expected that the future definitions and monitoring of unfair and discriminatory treatment will become more complex due to genome editing interventions on germ cells and embryos and advances in behavioral genetics and social science genomics. The government must not be content with merely enacting the current bill; instead, it must constantly strive to collect GD cases, review definitions, and raise awareness.


Leptotila wellsi is threatened by accelerated loss of genetic diversity resulting from ongoing habitat fragmentation. Small, threatened populations are difficult to sample directly but advances in molecular methods mean that non‐invasive samples can be used. We performed the first assessment of genetic diversity of populations of Grenada Dove by (a) assessing mtDNA genetic diversity in the only two areas of occupancy on Grenada, (b) defining the number of haplotypes present at each site and (c) evaluating evidence of isolation between sites. We used non‐invasively collected samples from two locations: Mt Hartman (n = 18) and Perseverance (n = 12). DNA extraction and PCR were used to amplify 1751 bps of mtDNA from two mitochondrial markers: NADH dehydrogenase 2 (ND2) and Cytochrome b (Cyt b). Haplotype diversity (h) of 0.4, a nucleotide diversity (π) of 0.00023 and two unique haplotypes were identified within the ND2 sequences; a single haplotype was identified within the Cyt b sequences. Of the two haplotypes identified, the most common haplotype (haplotype A = 73.9%) was observed at both sites and the other (haplotype B = 26.1%) was unique to Perseverance. Our results show low mitochondrial genetic diversity and clear evidence for genetically isolated populations. The Grenada Dove needs urgent conservation action, including habitat protection and potentially augmentation of gene flow by translocation in order to increase genetic resilience and diversity with the ultimate aim of securing the long‐term survival of this critically endangered species.As an island endemic with a decreasing population, the critically endangered Grenada Dove  We performed the first assessment of genetic diversity of populations of Grenada Dove by (a) assessing mtDNA genetic diversity in the only two areas of occupancy on Grenada, (b) defining the number of haplotypes present at each site and (c) evaluating evidence of isolation between sites. Our results show low mitochondrial genetic diversity, a non‐expanding population and clear evidence for genetically isolated populations. The Grenada Dove needs urgent conservation action, including habitat protection and potentially augmentation of gene flow by translocation in order to increase genetic resilience and diversity with the ultimate aim of securing the long‐term survival of this critically endangered species. We used this information to infer whether populations were isolated based on site‐specific haplotypes and to assess the level of genetic diversity both within and between these populations.22.1n = 194) and eggshell samples (n = 3) were stored at 4°C and imported to the United Kingdom using CITES import  and export licences  in September 2016 and June 2018 where they were cleaned with 70% ethanol and stored at −20°C.Moulted feathers and eggshell samples from Grenada Dove were collected opportunistically in the months of May and July between 2014 and 2018 by members of the Forest and National Parks Department of the Government of Grenada from two known dove habitats: Mt Hartman Estate and Perseverance, Grenada. We used non‐invasive sampling methods to ensure minimal disturbance to this critically endangered species. Information such as site and date were recorded, and where possible the location using a handheld GPS, to accompany each sample. Feather samples . Samples were visualised on a 1% agarose gel  using a BioRad Gel Doc™ EZ Imager and quantified using Image lab 4.0 software (Bio‐Rad Laboratories 2017). Samples were prepared and submitted to Eurofins Genomics following the Mix2Seq kit  instructions for sequencing using Sanger sequencing methods. Sequence assembly was performed using Sequencher 5.4.6  using the optimised protocol outlined in Peters et al. . Positivs et al. . Sampless et al.  prior to2.3Samples were sexed using the universal avian sexing primers P8 and P2 ; eggshells [n = 2]) and Perseverance (n = 12: feathers (n = 12); Table h) and nucleotide (π) diversity were calculated. Sequences with low average quality score ND2 (n = 7) and Cyt b (n = 1) were removed from mtDNA genetic analyses.The final sample set used for each site was as follows: Mt Hartman (33.1n = 23) were amplified which is comprised of 965 bps of ND2 and 786 bps of Cyt b. Two unique haplotypes were identified using the 1751 bps combined gene regions with a haplotype diversity (h) of 0.4 and a nucleotide diversity (π) of 0.00024. The most common haplotype (A = 73.9%) was found at both sites, and the second haplotype (B = 26.1%) was found only in Perseverance  included one single transitional polymorphism at the 882 bp position, and as such, two haplotypes were present (Table Cyt b gene (n = 29).Variation within the 965 bp of 3.2n = 23) was higher at Perseverance  in comparison to Mt Hartman which showed no variation in any of the samples from that site (Table n = 15) was comprised solely of haplotype A: 15 (100%) whereas Perseverance (n = 8) comprised of haplotype A: two (25%) and haplotype B: six (75%)  with the sex unknown for the remaining samples due to amplification failure for this marker.4This study is the first to evaluate genetic diversity in populations of the critically endangered Grenada Dove and has revealed extremely low haplotype diversity based on the examination of two mitochondrial genes obtained from non‐invasively collected samples. The two extant sites that constitute the entire range of the species are likely isolated from each other, as shown by the unique haplotype found in the West but not the Southwest population. Molecular sexing revealed a male sex bias in the samples used in this study. Given the estimated population size of around 160 individuals  was much higher than for the Grenada Dove  and nucleotide polymorphisms (1) obtained in this investigation are low in comparison to similar mtDNA studies carried out on other 4.2n = 8) compared to Mt Hartman (n = 15), which had no genetic variation despite a larger sample size and larger population  was shared between both locations, whereas the other haplotype identified  was unique to Perseverance. Higher haplotype and nucleotide diversities were observed within the samples at Perseverance ; data curation (lead); formal analysis (lead); funding acquisition ; investigation (lead); methodology (lead); project administration (lead); validation (lead); visualization (lead); writing – original draft (lead); writing – review and editing (lead). Matthew Geary: Conceptualization ; formal analysis (supporting); investigation (supporting); methodology (supporting); project administration (supporting); supervision (lead); visualization (supporting); writing – original draft (supporting); writing – review and editing (supporting). Charlotte Hosie: Project administration (supporting); supervision (supporting); writing – review and editing (supporting). Howard Nelson: Conceptualization (supporting); funding acquisition (supporting); project administration (supporting); supervision (supporting); writing – review and editing (supporting). Bonnie Rusk: Conceptualization (supporting); writing – review and editing (supporting). Anna Muir: Conceptualization (supporting); formal analysis (supporting); investigation (supporting); methodology (supporting); project administration (supporting); supervision (supporting); validation (supporting); visualization (supporting); writing – original draft (supporting); writing – review and editing (supporting).This work was supported by the University of Chester Biological Sciences department.The authors have no relevant financial or non‐financial interests to disclose.Appendix S1.Click here for additional data file.Appendix S2.Click here for additional data file.


Mycobacterium tuberculosis encounters diverse microenvironments, including oxidative assault (ROS and RNS), when it attempts to establish itself within its human host. Therefore, redox sensory and regulation processes are assumed significant importance, as these are essential processes for M. tuberculosis to survive under these hostile conditions. M. tuberculosis contains thioredoxin system to maintain redox homeostasis, which establish a balance between the thiol/dithiol couple.M. tuberculosis thioredoxin proteins  and a thioredoxin-like protein, NrdH, under aerobic and hypoxic conditions by performing thioredoxin trapping chromatography followed by mass spectrometry. We found that TrxC captured the maximum number of targets in both the physiological conditions and most of the targets of TrxB and NrdH showing overlap with targets of TrxC, indicating that TrxC acts as main thioredoxin. Further the PANTHER classification system provides involvement of targets in various metabolic processes and Gene Ontology analysis suggests that glutamine biosynthetic process and Fe–S cluster biosynthesis are the most enriched processes in the target list of TrxC and TrxB respectively. Also, we suggest that the thioredoxin system might play an important role under hypoxia by targeting those proteins which are responsible to sense and maintain hypoxic conditions. Furthermore, our studies establish a link between TrxB and iron-sulfur cluster biogenesis in M. tuberculosis. Ultimately, these findings open a new direction to target the thioredoxin system for screening new anti-mycobacterial drug targets.Still very less is known about it. In the present study, we attempted to capture the targets of all the   •Thioredoxin (Trx) system performs a major role to survive under redox stress, generated by the host.•TrxC captured the highest number of targets under aerobic and hypoxic conditions.•TrxC acts as a major reducing agent and TrxB is dedicated to perform special functions.•TrxB and TrxC interact with proteins responsible for survival under hypoxic conditions.•TrxB performs a specialized function in the Fe–S cluster biogenesis by interacting with Rv1465. Vectors2.73 as an iron source and Li2S as a source for sulfur were used to incorporate the Fe–S cluster on TrxB. A total of 1 mM TrxB was reduced with 5 mM DTT at room temperature for 1 h in anaerobic chamber. For the subsequent Fe–S cluster reconstitution, TrxB was diluted to the final concentration of 150 μM with a buffer containing 50 mM Tris-Cl pH 8, 150 mM NaCl and 2 mM DTT. FeCl3 was added to the final concentration of 600 μM and incubated for 5 min at room temperature. Li2S was added at the same molar concentration as FeCl3 and incubated for 10 min at room temperature under anaerobic chamber. Further to remove unbound reaction components, the reconstituted reaction was allowed to bind with 100 μl of Ni-NTA resin, pre-equilibrated with 50 mM Tris-Cl pH 8, 150 mM NaCl for 10 min. Washing was done for 10 column volumes with a buffer containing 50 mM Tris-Cl pH 8, 150 mM NaCl. Reconstituted TrxB was eluted with a buffer containing 50 mM Tris-Cl pH 8, 150 mM NaCl, 250 mM imidazole. To confirm the Fe–S cluster reconstitution, absorbance at 420 nm was observed.Chemical mode of the Fe–S cluster reconstitution was applied to incorporate Fe–S cluster on TrxB, as described by Freibert et al. with minor modifications . All sto3Thioredoxins being the major buffer against redox fluctuations in cells, it is important to understand the mechanism of how they regulate response to redox changes. Typically, thioredoxins participate in these processes by thiol disulfide exchange reactions and by partnering with different proteins in the cells, protecting them from such perturbations. Thioredoxins have the ability to attack on the substrates having S–S bridge and reduce them. Thioredoxins contain active site CXXC motif. One Cys with lower pKa value (attacking Cysteine) acts as a nucleophile (thiolate ion). The thiolate ion attacks on the disulfide bond of the target and forms a mixed disulfide bond between target and enzyme. At this stage another Cys residue (resolving Cysteine) attacks on the mixed disulfide bond, leaving the target in reduced form.M. tuberculosis has been shown to possess three thioredoxins, out of which two are known to be active, and some glutaredoxin-like proteins, such as NrdH with thioredoxin-like activity /pUAB200-GCN4[F3], provides the viable phenotype on the 7H11 medium containing trimethoprim and acts as positive control for the study. Similarly, the interaction of TrxB and Rv1465 allows the functional reconstitution of both the fragments of mDHFR and displayed viable phenotype of co-transformants containing, MtbtrxB in pUAB300 and MtbRv1465(F3) in pUAB400 on media containing 20 μg/ml trimethoprim . Interestingly the coloration property was only observed on TrxB, but not on TrxC. This observation encouraged us to perform in vitro Fe–S cluster reconstitution experiments.Apart from the interaction of TrxB with cluster biosynthesis proteins we also observed the unexpected brown-red color on theNi-NTA affinity purified recombinant fraction of TrxB expressed in in vitro reconstitution of Fe–S cluster was done. As a result, reconstituted reaction showed the presence of characteristic peak at 420 nm indicates the successful reconstitution of Fe–S clusters on TrxB  with thioredoxin and presence of Fe–S cluster on thioredoxin [B. subtilis by providing the in vivo data of interaction between TrxB and Rv1465 using M-PFC assay. Moreover, result of in-vitro Fe–S cluster reconstitution experiment confirms the binding of Fe–S cluster on TrxB by providing a peak at 420 nm. The binding of the Fe–S cluster to TrxB is an interesting and unexpected finding of our study.A similar observation to our present study was made in oredoxin . FurtherOn the basis of evidences of interaction between TrxB and Rv1465 and previously available literature, we hypothesized the role of TrxB in Fe–S cluster biogenesis. The function of Rv1465 is to perform the transfer of terminal sulfide from Rv1464 to scaffold proteins for the synthesis of Fe–S cluster. We hypothesized that this step of terminal sulfide transfer requires a reductant and possibly that reductant is TrxB. Interestingly, we found that Rv1465 was captured exclusively by TrxB Whereas, TrxC captured Rv1461, Rv1462 and Rv1463, which together serve as scaffold for synthesis of Fe–S cluster. Here both the thioredoxins displayed the specificity for their targets despite having similar modes of action. The present study suggests the specific interaction between TrxB with Rv1465 while TrxC acts as a main reductase and interacts with multiple SUF proteins to maintain the redox balance.M. tuberculosis  The conception and design of the study: SCM and SS. Acquisition of data: SS, VR, KT, SJ. Analysis and interpretation of data: SS, SCM, SR, VN. (2) Drafting the article and revising it critically for important intellectual content: SS, SCM. (3) Final approval of the version to be submitted: All authors.10.13039/501100001407Department of Biotechnology, Government of India (BT/PR10855/BRB/10/1330/2014) for funding Orbitrap mass spectrometer to NCCS. We also thank the 10.13039/501100001407Department of Biotechnology, Government of India for support through Centre of Excellence grant (BT/PR15450/COE/34/46/2016) to SCM.The authors acknowledge the The authors report no competing interests.


PRCD variant  and the collie eye anomaly (CEA)-associated NHEJ1 deletion  provided by the commercial genetic testing company . These data were analyzed using the chi-square goodness-of-fit test, time-trend graphical analysis, and regression modeling in order to evaluate how test results changed over time. The results span fifteen years, representing 82 countries and 67 breeds/breed mixes. Both diseases exhibited significant differences in genotype frequencies (p = 2.7 × 10−152 for prcd-PRA and 0.023 for CEA) with opposing graphical trends. Regression modeling showed time progression to significantly affect the odds of a dog being homozygous or heterozygous for either disease, as do variables including breed and breed popularity. This study shows that genetic testing informed breeding decisions to produce fewer affected dogs. However, the presence of dogs homozygous for the disease variant, especially for prcd-PRA, was still observed fourteen years after test availability, potentially due to crosses of unknown carriers. This suggests that genetic testing of dog populations should continue.Hundreds of genetic variants associated with canine traits and disorders have been identified, with commercial tests offered. However, the geographic distributions and changes in allele and genotype frequencies over prolonged, continuous periods of time are lacking. This study utilized a large set of genotypes from dogs tested for the progressive rod-cone degeneration–progressive retinal atrophy (prcd-PRA) G>A missense  With the advent of genetic testing for dogs, canine health care could begin to embrace preventative genetic screening; when tests became available for fully penetrant autosomal recessive diseases, breeders now had accurate tools to predict outcomes and carefully plan matings to avoid producing affected puppies. To date, over 320 Mendelian traits and conditions have had a likely causal variant(s) described ,2 and soIn order to obtain a larger and more complete perspective, including the changing genotype and allele frequencies for numerous breeds over time and with a global context, data are best harnessed from commercial canine genetic testing providers. In the present study, we utilize current data from a large provider merged with historical testing data from a single genetic testing provider (OptiGen) that offered the examined tests nearly exclusively for the first ten years of their lifespan. The latter means that the captured picture (from 2004 or 2005 to 2013) essentially represents the global population of dogs being tested. Two ophthalmic diseases that fit this profile were selected for the present study: progressive rod-cone degeneration–progressive retinal atrophy (prcd-PRA) (OMIA 001298-9615) and collie eye anomaly (CEA) (OMIA 000218-9615).PRCD ; the most recent year with publicly available data for number of new registrations per breed) and the sample number of that breed tested in our dataset  reveal that the frequency of coloboma has increased within the Collie population alongside the less-severe chorioretinal hypoplasia. Another potential factor responsible for the observed trends is breeders subscribing to a long-held belief within the Collie breeder community; this belief is that the NHEJ1 mutation is strongly associated with the desirable “almond eye” shape, explaining the high prevalence of CEA among show-quality Collies . To date, there is no scientific literature supporting this proposed relationship. However, since CEA was typically considered a “mild” phenotype (not counting colobomas), and the breeders placed higher priorities elsewhere, CEA genotypes may not have weighed as heavily in mating decisions .Collies showed a significant departure from other breed trends in the CEA data. Collies not only had the greatest relative probability of affected genotype or carrier genotype for the CEA-associated variant (among breeds with e points ) but alse points A–C. In te points D–F. One The variable of country was owner-reported and represented the year in which the tests were submitted. As such, the countries analyzed in this study may not accurately reflect the current geopolitical relations and borders. Furthermore, a caveat to country (and breed) information is that it was owner-entered and, therefore, subject to possible entry error. This is demonstrated by looking into the single sample from Bouvet Island, which upon further investigation, likely originated from South Carolina, USA, although there is no way to definitively verify its origin. Such errors likely do not have a large overall effect, and in most cases, are nearly impossible to detect. Logistic and multinomial regression models for both diseases indicated that no country had a statistically significant effect on log odds of affected or carrier genotype status . Althougp < 2 × 10−16 for both diseases)  . Given tiseases) . RecogniPRCD or NHEJ1 positioned on a chromosome close to a breed-linked behavior  locus with strong influence, selection for those behaviors might affect the prevalence of prcd-PRA and CEA-associated variants due to linkage disequilibrium. However, the genomic locations of seven herding-related genes  [LRRTM4, JAK2, and MEIS1) [CNIH1) [PRCD and NHEJ1 according to CanFam3.1, and only ASIC2 was on the same chromosome (chromosome 9) as PRCD. None shared a chromosome with NHEJ1. On CFA9, ASIC2 and PRCD are separated by approximately 34 million base pairs. Using CanFam3.1 positions and an estimated sex average rate of recombination of 0.91 cM/Mb on CFA9 [Alternatively, breed genotype and allele frequency trends may relate more to a dog’s function, i.e., what a dog was bred to do, as captured by AKC and FCI groupings. While a large number of represented groups under the AKC and FCI were significant for prcd-PRA , only thnd CHL1) ,55, thred MEIS1) , and one on CFA9 , the twoFurther investigation of genetic relationships and how they may impact prcd-PRA and CEA breed trends was undertaken using a model to investigate the impact of genetic clade ,49. WhilOverall, this study demonstrates that commercial genetic testing for prcd-PRA and CEA is being used to inform breeding decisions to improve canine health, as evidenced by the progression of time being associated with significantly decreasing log odds of affected and carrier genotypes . While bThis study is unique in terms of the large sample size and lengthy continuous time period; however, there are limitations to its scope and conclusions. First, while these data include samples from all seven continents, the majority represent North America and Europe. Therefore, in countries and continents with smaller sample sizes, these analyses are unlikely to accurately reflect the full extent of genotype and allele frequency trends. Moreover, in this dataset, as mentioned above, country and dog breed or breed mix were owner-reported and, therefore, subject to human data entry error. Also, while likely to reflect the location an owner was living at the time of test submission, owner-reported data do not guarantee the country where the dog was born or where the dog’s bloodlines originated. And, like all large datasets, the size of this study also potentially allows for some insubstantial differences to be statistically significant, so some caution may need to be considered.NHEJ1 deletion. In addition, some level of sampling bias likely affects these data, since a dog’s clinical disease status can influence genetic testing decisions. For example, dogs diagnosed with either disease may then undergo genetic testing as confirmation. Sampling bias also results when dogs designated as “parentage clear” (due to both parents testing homozygous clear) do not undergo testing. Therefore, affected genotype, carrier genotype, and variant allele frequencies in this study may be greater than the true frequencies. This could be rectified via a combined genetic and hereditary status approach [An additional limitation of this study is the lack of clinical information. The present study exclusively presents genetic testing results, and no information is available on any dog’s diagnosis, either before or after test submission. Although there is variation in the rate of retinal degeneration, dogs of any breed with two copies of the prcd-PRA variant almost invariably go blind as they age. Conversely, because CEA has a wide array of phenotype severity, including normal vision, no firm conclusions about disease status or clinical signs can be drawn about dogs homozygous for the approach , which wFinally, it must be pointed out that while these trends are likely the outcome of direct action by breeders, it is also possible that they are the result of genetic drift and, therefore, simply artifacts of breeding decisions made with respect to other prioritized traits . Such decision paradigms are undoubtedly influenced by the severity of the disease, with PRA likely motivating more direct action compared with CEA, but are also influenced by each breed community’s approaches and rules regarding genetic testing. While the trends identified in this study are highly encouraging, the remaining presence of homozygous-affected genotypes for both diseases in the final year of study (and supported by other work ) shows tThis study is a first-of-its-kind examination of how commercial genetic testing is used, and to what extent, to influence disease-associated genotype frequencies for two congenital ocular diseases, utilizing a global dataset with over 120,000 individual genetic test results. This study presents a continuous fifteen-year cohort of 67 breeds and breed mixes and 82 countries globally using commercial genetic testing results to reduce the frequency of affected and carrier dogs, as well as the overall disease-associated  allele frequency. Generally, the geographic distribution of tests did not have a significant impact on genotype probability, although breed, AKC group, FCI group, and breed popularity did. Prcd-PRA, the more severe of the two diseases, exhibited a more significantly decreasing frequency trend compared with CEA, which may indicate a stronger priority amongst breeders to remove prcd-PRA from the population. Considering that affected dogs were observed for both CEA and prcd-PRA in the 2019 data, it is important that genetic testing continues, with careful test-and-replace strategies in order to maintain genetic diversity because while progress has been made, there is still more work that needs to be carried out. Future work should continue this line of study, perhaps by pooling genetic testing data from many genetic testing providers over the same time period in order to gain a truly globally distributed dataset with a larger cohort of dogs for the two present and other different diseases; this would provide highly accurate global trends. Additionally, updated analyses including data from 2020 to the present would allow scrutiny of the most current trends. To aid in this strategy, lay public-accessible summary tables are available in the


Genetic counseling is a fast-growing profession worldwide, with genetic counselors taking on increasingly comprehensive and autonomous roles in the healthcare sector. However, the absence of appropriate legal frameworks could potentially create risks of harm to the public. Legal recognition serves to protect the public from risk of harm by regulating the safe and competent practice of healthcare professionals. Genetic counseling is not legally recognized in most world jurisdictions. Examination of the legal status of genetic counseling in different jurisdictions and whether existing legal mechanisms are adequate to address potential risks of harm is therefore timely. This paper examines the different roles of genetic counselors in the Canadian province of Quebec and the state of Qatar, the authors’ respective jurisdictions. It considers the types of harms that may be created where appropriate legal mechanisms are lacking, considering the socio-political and legal differences between the two jurisdictions. Moreover, it examines the legal status of genetic counseling in Quebec and Qatar to determine whether these statuses appropriately address the identified risks of harm. The authors argue that existing legal frameworks are inadequate to address these risks and recommend that additional regulatory mechanisms be implemented to properly protect the public from risks of harm. Genetic counselors are healthcare practitioners with specialized education and training in medical genetics who counsel patients and their families on the medical, psychological and familial implications of genetic diseases and genetic testing options . While tIndeed, genetic counselors are taking on increasingly comprehensive and autonomous roles in the healthcare sector. For instance, a growing number of genetic counselors are working in non-genetic clinical settings with non-geneticist physicians  . The ratGenetic counseling is usually introduced as a non-directive service, with the goal of providing neutral information, predictable outcomes and statistical probabilities without attempting to influence the decisions of the concerned parties. However, the implementation of the mandatory premarital screening program and the corresponding need for genetic counseling raises several potential risks of harm to the public. The scope of these risks can be quite broad, encompassing religious, ethical and socio-cultural considerations. Risks associated with the disclosure of confidential information and potential violations of privacy have been particularly emphasized by the literature . Such coIndeed, one of the unique aspects of the genetic counseling process for discussing premarital screening results is the involvement of two parties, the prospective spouses. Both parties’ genetic information must be examined to determine whether it will be beneficial or harmful, from a genetic perspective, to proceed with the planned marriage . Revealing information about one’s own genetic profile could also entail information about one’s genetically related relatives. Keeping in mind the social significance of the family institution in a Muslim-majority country like Qatar , disclosIt is therefore clear that in both Quebec and Qatar, genetic counseling may pose risks of harm to the health and safety of the public if not properly regulated. These harms may include physical, psychological, and social harms. In Quebec, physical and psychological harms have been particularly emphasized, whereas social harms have been identified as significant in Qatar, given its socio-cultural and religious context. In light of these risks, we will now examine the legal status of genetic counseling in Quebec and Qatar to determine whether existing regulatory frameworks adequately address these risks and protect the public from risk of harm.Canada is a federation in which legislative powers are shared between the federal and provincial governments, the latter of which is responsible for the regulation of professions . Quebec,In Quebec, the Professional Code governs the provincial professional system, which comprises those professions which have been legally recognized by the provincial government through the implementation of specific public protection measures. These include regulating access to specific professions, protecting professional titles, defining professional scopes of practice, and implementing public accountability measures . GeneticThree models of legal recognition are available under the Professional Code: 1) constitution of a professional order, 2) inclusion in an existing professional order, and 3) delegation . Under tDelegation provides a more limited scope of public protection, specifically regulating the performance of specific acts, such as the communication of a diagnosis or the prescription of clinical tests or medication. It does, however, ensure some degree of public protection through the oversight of the performance of these acts . In ManiNone of these models apply to genetic counselors in Quebec, which limits legal recourse for patients in the event of harm. Under Quebec law, genetic counselors may, however, be considered service providers . ConsequQatar is a unitary state bound not only by legal norms, but also by religious edicts rooted in the Islamic tradition. Indeed, Sharia, which refers to the body of Islamic religious law, is a principal source of legislation. The Ministry of Health recently implemented national registration requirements for genetic counselors in Qatar, thereby legally recognizing the genetic counseling profession. Under this framework, genetic counselors must apply for licensure to the Department of Healthcare Professions (DHP) . The minprima facie principle is that disclosing confidential information is prohibited and disclosing such information without a compelling reason may lead to legal penalties, unless the disclosure is permitted on exceptional grounds.The guidelines are silent, however, on how to deal with the risks of harm that may result from the genetic counseling process, including potential breaches of confidentiality. They state that genetic counselors must always “practice in accordance with relevant legislative, regulatory and policy guidelines”, but do not address specific public protection measures or legal recourses in the event of patient harm , s. 1.3.In the case of the above-mentioned mandatory premarital screening program, it seems particularly challenging to provide genetic counseling where the potential risk of harm can be mitigated or minimized. For practicing Muslim couples, the marital contract entails religious commitments with mutual rights and obligations. The legal enforcement of genetic testing raises concerns about the religiously protected right of autonomous individuals to decide what to do with their body and to choose their marriage partner. To mitigate this harm, genetic counselors in Qatar should make it clear to to-be-married couples that the mandatory nature of the testing is not meant to influence the religious validity of the marital contract. Furthermore, priority should be given to increasing public awareness of the significance of these genetic tests through public communications such as newsletters. Any laws and related executive orders associated with genetic testing should also be reviewed on a regular basis to determine if they are effective, if certain modifications are needed, or if they are still required. To avoid harms resulting from inadequate knowledge of the various Islamic perspectives on these questions, special training programs should be developed for genetic counselors to improve their religio-cultural competence. To meet Qatar’s need for genetic counselors and to have competent and religio-culturally sensitive counselors, Qatar University, the main national university in Qatar, has established a 2-year Master Program in Genetic Counselling in 2018 . GraduatThe roles and responsibilities of genetic counselors in Quebec and Qatar are overall similar, despite the socio-cultural and legal differences between these two jurisdictions. This is consistent with research on the roles and responsibilities of genetic counselors globally, which finds similarities in the practice of genetic counseling across countries . HoweverThese differences are not only limited to the roles of genetic counselors and the risks of patient harm in Quebec and Qatar, but also to the manners in which their respective legal systems respond to these risks. Whereas professions that pose a risk of harm to the public may become legally recognized in Quebec under its professional legislative scheme, this process has yet to be extended to genetic counselors. Consequently, there are few public protection mechanisms in place to mitigate potential risks of harm. Instead, patients who suffer injury from the genetic counseling process must use the legal recourses available under Quebec’s general civil liability rules. Though they afford some form of protection, these rules offer more limited protections and are more reactive than proactive.Unlike Quebec, Qatar’s Ministry of Public Health has recently legally recognized the genetic counseling profession through the adoption of national guidelines. While these guidelines provide licensure requirements and create obligations for genetic counselors, including the obligation to maintain patient confidentiality, they do not provide information on how to deal with potential risks of harm.Genetic counseling is a rapidly growing and evolving profession. Genetic counselors are becoming increasingly autonomous and taking on more comprehensive roles in healthcare systems across the world. This professional expansion, without appropriate regulatory mechanisms to protect the public, may create risks of harm. This is true of the Canadian province of Quebec, where the lack of legal recognition limits the measures that can help protect the public from risk of harm. In Qatar, genetic counseling has recently been legally recognized by the Ministry of Public Health.status quo in Quebec, whereby instances of patient harm are governed by ordinary civil liability rules, do not sufficiently protect the public from harm. Legal recognition is therefore required to ensure the safe, competent, and ethical provision of genetic counseling services in the province.While the types of harm may differ between these two jurisdictions, the potential for risk of harm warrants the implementation of public protection measures. In Quebec, we recommend that genetic counseling be legally recognized and subject to the public protection mechanisms set out in the Professional Code. The potential risks of harm engendered by the unregulated practice of genetic counseling are sufficiently compelling to warrant legal recognition. The legal While genetic counseling is now a legally recognized profession in Qatar, the current regulatory framework does not provide for mechanisms to adequately address potential risks of harm. We recommended that the ministerial guidelines provide for mechanisms to ensure a wider ambit of protection for the public. We also recommend tailoring training programs to improve genetic counselors’ knowledge of the Islamic perspectives on risks of harms, thereby avoiding harms which could result from communicating incorrect or incomplete information during the genetic counseling process.In summary, while the types of risks of harm may differ due to legal and socio-cultural differences, the case for the legal recognition of genetic counseling and for the implementation of public protection measures in both Quebec and Qatar is compelling. Whether they are implemented through legislation or through the application of religious and cultural norms, these measures are necessary to ensure safer and higher quality genetic counseling services for patients.


Wolves were the first animal species to become domesticated by humans, approximately 30,000–50,000 years ago. Human-directed dog breeding over thousands of generations has generated more than 350 recognized breeds displaying surprisingly different phenotypes with respect to morphology, behavior and disease predispositions. The domestication of wolves and the subsequent breeding of dogs can be viewed as one of humankind’s oldest and largest genetic experiments and provides us with unique opportunities for research. Dogs have not only become human’s best friend but were also described as geneticists’ best friend in a past issue of Science. In recognition of the importance of canine genetics, this Special Issue, entitled “Canine Genetics 2”, was compiled. It represents a sequel to the former Special Issue “Canine Genetics”, which was published in 2019. During the last 15 years, the canine community has heavily relied on a reference genome derived from the female Boxer Tasha. “Canine Genetics 2” includes an article describing a greatly improved version of this important community resource. This Special Issue further contains several reports related to monogenic or complex inherited diseases in dogs. Finally, important aspects of wild canid research, genetic diversity in different populations and canine morphology were investigated. Canis lupus familiaris) represent the species with the greatest phenotypic variability among mammals [Canis lupus); feral populations of descendants from early domesticated dogs, such as the Australian Dingo; so-called village dogs; various kinds of random-bred and mixed-breed dogs; and finally purebred dogs, which are bred within strictly isolated populations. All purebred dog populations underwent very tight bottlenecks at breed formation, which took place between the 18th century and today [Dogs  is a highly endangered wild canid species. Miller-Butterworth et al. investigated the genetic diversity in the captive population of North American zoos to provide a basis for the future breeding management and conservation of this iconic species [The Norwegian Lundehund breed is characterized by extreme inbreeding and several disease predispositions that are hypothesized to result from inbreeding depression. Melis et al. report on a genetic rescue experiment that uses introgression of the phenotypically similar Norwegian Buhund in an attempt to increase genetic diversity in the Norwegian Lundehund . The Afr species .Ancient DNACuon alpinus) whose fossils were found in Europe. These data will help to better understand the migration and decline of the once wide-spread species that is now restricted to South and South-East Asia [Finally, the Special Issue “Canine Genetics 2” contains a study on the mitochondrial DNA diversity of six 25,000–45,000-year-old Asiatic wild dogs or dholes (ast Asia .ConclusionsThe 21 articles compiled in this Special Issue nicely represent the breadth of canine genetic research. These research findings advance our basic understanding of gene functions, genetic diversity and evolution. Many studies enable direct applications, such as the introduction of genetic testing or improved breeding management, that will foster the health and wellbeing of dogs and wild canids.


Threatened animal species live in small and isolated populations, often with a reduced number of individuals and reduced genetic diversity. In this review, we summarise studies on amphibian species that examined the effect of genetic diversity on the fitness of individuals or populations. We found evidence for the positive association between genetic diversity and different estimates of fitness where low genetic diversity is associated with lower fitness. Published studies are however scarce, concentrated on a handful species and with a high proportion of non-significant results reported. For the maintenance of amphibian and other animal populations, it is important to increase the research effort in this topic and, for imperilled populations, enhance genetic diversity through different types of conservation measures.Ranavirus. By employing experiments, researchers investigate whether changes in genetic diversity within or among isolated populations affect amphibian fitness. While genetic diversity estimates are based on molecular markers, typically microsatellites, fitness is mostly measured as tadpole performance in rearing experiments often under varying environmental conditions. Tadpole performances  have been found to be negatively affected by low genetic diversity, as several studies have found a positive association between genetic diversity and these fitness traits. Moreover, infection with pathogens also seems to be more likely in individuals or populations with lower genetic diversity. Overall, these genetic–fitness correlations seem to be more pronounced or detectable in smaller, declining populations but not in larger populations. Genomic studies, which sample a larger fraction of the genome, are still scarce in the conservation genetic literature on amphibians. These are likely to increase in upcoming years and may reveal adaptive variants that protect against dangerous pathogens or environmental changes. Altogether, genetic–fitness correlation studies should be a priority in order to develop effective management plans for the genetic rescue of isolated, imperilled amphibian populations.Endangered animals suffer from isolation of their habitats. Isolation leads to a reduction in population size as well as a decrease in genetic diversity and a concomitant increase in the risk of extinction. Amphibians are the most endangered vertebrate class. Besides habitat loss, fragmentation and isolation, amphibians are threatened by emerging diseases e.g., chytrid fungus or  Genetic diversity is highly important to maintain healthy animal and plant populations and assure their long-term persistence ,2. If poGD) in vertebrates, invertebrates and plants correlates significantly with population fitness and decreases when population sizes decline. For example, in deer mice from nine populations of Michigan, parasite load (nematode Capillaria hepatica) is a proxy for fitness that negatively correlates with GD  [GD for the species and their conservation are not always clear. Populations of some species seem to persist or even grow although their GD is low  . Howevercalamita , or Alcees alces ; reviewees alces ), while es alces ,12,13).GD as well as certain genetic factors is important for the persistence of populations in many taxa [GD and reduce their chance of extinction resulting in a genetic rescue [GD has been traditionally measured with neutral molecular markers such as microsatellites [Despite some discrepancies in the research results, the role of any taxa ,15. The c rescue ,17. GD htellites ; howevertellites .ST values) indicating population fragmentation have been found for amphibians  than for other taxa  in most toad localities [GD are the Oregon spotted frog Rana pretiosa from the Pacific Northwest [Salamandra inframaculata from northern Israel [Amphibians are particularly vulnerable to anthropogenic activities that often result in declines in population sizes. Compared to other vertebrates, a higher proportion of amphibian species  are endangered ,21. Amphcalities ,25. Othen Israel . GD, population bottlenecks and inbreeding. Together these challenges result in fitness reductions, especially under stressful conditions . TheGFC) or heterozygosity and fitness (HFC). Therefore, we used the global citation databases Web of Science and PubMed in February 2023. We used the following key words for the search: amphib* OR frog* OR toad* OR newt* OR salamander* OR caecilian* OR anura* OR caudata* OR gymnophiona* AND fitness* OR infection* AND genetic diversity* OR heterozygosity fitness correlation* OR genetic fitness correlation* OR inbreeding*. From all articles displayed (Web of Science: N = 379 and PubMed: N = 262), we read the abstract and methods to decide whether the article contained the desired information for the topic. In total, we found 35 articles that met our criteria. The criteria were as follows: estimates of genetic diversity (GD) and a statistical test of association with fitness traits of an amphibian species were reported , the proxies for fitness , the applied statistical method, the presence or absence of correlations or associations between GD or certain genetic elements  with the fitness proxies, and finally a short summary or conclusion from the study. We additionally report—only in cases where this information was available in the publication—the population size of the focal species at the sample site and values (or ranges) of genetic diversity estimates. In cases where the genetic population structure has been evaluated, we report the number of genetic clusters found (K) via Bayesian Assignment algorithms  and FST values. In experiments or when differences in the environment in the field have been measured, we shortly outline the treatments or ecological factors. Since the studies differed in the genetic markers used and analytical procedures, the results  among studies cannot be directly compared. In most studies the GD measures are reported for populations (Ho or He), in some studies however GD and fitness were measured on the individual level . This is then indicated as “indiv.” genetic variation. If available, we transferred values of correlations (r or R2) and probabilities (P) from the publication into the table. However, since the statistical models applied are highly diverse among studies this information could not be standardised. Since the information content in the table is complex we simplified the content by dividing it into three tables. The first table covers the genetic markers used to measure GD and how different fitness parameters are affected by GD and certain adaptive loci. The results of the experiments which analysed the effects of ecological conditions on fitness parameters and how these interact with GD and those that analyse the effect of small population size and/or population isolation on amphibian fitness were also tabulated.For this review, we searched for literature on amphibian species that report correlations between genetic variation and fitness  and the amphibian species studied in each case. To summarise the phylogenetic diversity of the studied species, we used a comprehensive amphibian phylogeny [phytools package [To graphically summarise the compiled data we aggregated the diversity of the phenotypic variables measured in the original articles into eight main fitness trait categories as follows: development , growth rate (tadpole growth rate and juvenile growth rate), fertility (adult reproductive investment and clutch size), health , home range, size , and survival . We then registered the reported statistical effect of the hylogeny  pruned t package .GD and at least one fitness-related trait for one up to 38 study sites (SS) in different amphibian species , observed HO and expected heterozygosity HE for populations as well as multilocus heterozygosity (MHL) for individuals. Allele frequencies and heterozygosity, gene diversity (π) and Tajimas D (TD) were also reported for MHC results, and the three CytB/mtDNA studies reported gene diversity (π) and/or Tajimas D  were the obvious preferred method in the lab; however, allozymes (N = 2), minisatellites (N = 1), RAPD (N = 1), AFLPs (N = 1), Cytb (N = 2) and unspecified mtDNA (N = 1) also served as measures of neutral genetic variation. Both Cytb studies were performed in combination with microsatellites. Eight studies used an adaptive immunological locus, the MHC IA or II β (exon 1 or 2) loci, four of those also used microsatellites, one combined with mtDNA, and two also used a genomic approach. Furthermore, five studies used genomic approaches: four studies sequenced SNPs (single nucleotide polymorphisms), while one study applied transcriptome sequencing . The meaajimas D . Several studies ,37,38, w studies ,40,41,42ulations . Accordi habitat  or NGSadNGSadmix . The nums boreas  to K = 1rreauxii ; thus, aGD have been conducted in all but one study , but th studies . The stuD . Six sts . Only orrelated . Two stuulations ,51, whilulations  found thGD or negatively affected by particularly low GD, while 34 fitness traits showed no relationship with GD and four fitness traits  were associated with lower GD. Since some studies involved two or more molecular markers, it is possible that fitness traits were correlated with the GD measured at one marker but no correlation was found for GD measured with another marker . For. ForGD oiversity .GD and fitness traits in 18 species. Four species  were the most studied and accounted for 57% of all the statistical tests conducted. Overall, the majority of the reported GD effects on fitness traits were positive (>48% positive vs. 6% negative), although the number of non-significant effects reported was high overall (>45%)  .animalsGD and treatments: four studies found evidence of fitness costs associated with low genetic variation under severe conditions [Rana sylvatica mortality was higher in tadpoles of low GD when exposed to UV light [B. calamita a higher GD improved survival of tadpoles in water of cold temperatures, where survival was highest, but not in water of variable temperatures, a situation which was supposed to be stressful for the toads [GD improves amphibian fitness under certain ecological conditions, while a low GD was never associated with better fitness. Tadpoles were commonly used for fitness experiments in which tadpole development or survival was measured under different ecological conditions  found that Bd prevalence and intensity was highest in two isolated and inbred populations [Litoria verrauxii from Australia Bd infection was absent in an isolated population [Bd infection was more intense in fragmented populations as compared to non-fragmented populations [In amphibians as well as in other animals, isolation and fragmentation of populations is associated with small population sizes and therefore lower s review , thirtees review . Egg and arborea ; R. latalatastei ; and R. mporaria ) one Nor sevosus  and two iventris ). With rulations , while ipulation . Across ulations .GD and fitness traits, in some cases together with population size and fragmentation, in amphibian populations. Most studies did not contain information on all of these items, and the number of species investigated is extremely small (N = 20 species) when considering the 8689 known amphibian species [GD, with a concomitant negative effect on fitness. This is in accordance with earlier reviews on GFC in amphibians, pointing out the relationship between small population size, genetic diversity depletion or inbreeding, and the unfavourable consequences on survival [Our review analysed the relationships among  species . Even wisurvival ,71. The survival ,73, and survival ,75. AddiGD, mostly measured at microsatellite loci, is often positively correlated with fitness traits. Fitness traits were mostly measured as traits that are related to the development of eggs and tadpoles or in some cases as Bd infection. The fitness traits of eggs and tadpoles  are generally positively correlated with increasing GD meaning that individuals in populations with a higher GD are more likely to survive and reach adulthood. In this report, it became apparent that Bd. We considered Bd infection an adverse fitness trait since Bd infection has led to disease outbreaks, mass mortality and population declines. Even though the fungus is widely distributed—it has been detected in 86 of 119 countries (72%) and in 1062 of 1966 tested species —the moses . The host specificity of this fungal parasite is largely restricted to Caudata [GD and Bsal infection appear to have not been studied or published to date.Interestingly we found three studies that documented  with GD ,51,52. Oathogens . By the rreauxii , in whicariegata . Those s Caudata , and assGD and fitness proxies, while only a few studies showed negative correlations, nearly 46% of the calculated statistical tests were not significant  because only outbred, genetically more diverse individuals survive the tadpole stage. This case emphasizes the need to measure amphibian inbreeding at different developmental stages [In most studies reviewed for this report, inbreeding coefficients and estimates for population sizes are not calculated or presented. Nevertheless, we found substantial evidence for the positive association between connectivity and population size with ulations . Notablyl stages . Bd infection  together with l design . Four ofs higher ,55,57, wer study . Even thGD on tadpole fitness, and GD was mostly evaluated with microsatellites, most of them varied in the fitness proxies that were measured (Pelobates fuscus fuscus) was weakly affected by variations in size and timing at metamorphosis [GD as population HO, HE and individual MLH, and providing at least a rough estimate of population size or effective population size (NE). Currently, the use of microsatellites is declining and is increasingly being replaced by more informative methods like SNPs, where the number of molecular markers is increased a thousandfold . The efd  of more than a thousand individuals [E are often unknown and hard to infer from the population size, N, in amphibians, but fortunately they can be estimated from genetic markers [GD facing extinction risk should be reconnected to larger, geographically close meta-populations via steppingstones and artificial crossing aids such as ditches, tunnels or green bridges. If this is not possible, genetic rescue by the introduction of individuals from larger populations that are genetically diverse or by mixing of individuals from several source populations for translocation [GD. In the latter case, the risk of outbreeding depression can be avoided by applying some simple rules developed by experts [Multiple studies have shown the importance of keeping ulations ,87. Low ividuals . One pro markers . In amphlocation ,90 could experts  and by l experts  on genetAnaxyrus canorus, 24 candidate genes have been identified that seem to be under climate-related selection [GD within and among populations of all species  by applying genomic methods. Once those genes have been identified, beneficial variants should be introduced into the affected threatened populations. For example, in the Yosemite toad, election . Identifrk, GBF, ).