Arborophila ardens) to climate change. We concluded that A. ardens survived through LGM climate changes, and its current distribution on Hainan Island was its in situ refuge. Range model results indicated that A. ardens once covered a much larger area than its current distribution. Demographic history described a relatively stable pattern during and following the LGM. In addition, weak population genetic structure suggests a role in promoting gene flow between populations with climate-induced elevation shifts. Human activities must be considered in conservation planning due to their impact on fragmented habitats. These first combined data for Hainan Partridge demonstrate the value of paired genetic and SDMs study. More related works that might deepen our understanding of the responses of the species in Southeast Asia to late Quaternary Climate are needed.Understanding the historical dynamics of animal species is critical for accurate prediction of their response to climate changes. During the late Quaternary period, Southeast Asia had a larger land area than today due to lower sea levels, and its terrestrial landscape was covered by extensive forests and savanna. To date, however, the distribution fluctuation of vegetation and its impacts on genetic structure and demographic history of local animals during the Last Glacial Maximum (LGM) are still disputed. In addition, the responses of animal species on Hainan Island, located in northern Southeast Asia, to climate changes during the LGM are poorly understood. Here, we combined phylogeographic analysis, paleoclimatic evidence, and species distribution models to examine the response of the flightless Hainan Partridge ( Pleistocene climatic changes left strong footprints on the diversity and structure of contemporary northern hemisphere wildlife by ice coverage and caused their distributions to shift Hainan Island has experienced connection-disconnection events with the Chinese mainland as well as Taiwan Island and Indochina due to sea level fluctuations throughout the late Pleistocene More recently, phylogeographic analysis has been integrated with species distribution models (SDMs) as a powerful multi-faceted approach to resolve the processes of how current distribution patterns of genes, populations, and species were shaped Arborophila ardens) and its conservation implications. The Hainan Partridge is mainly restricted in tropical evergreen forest, both broadleaved and mixed coniferous-broadleaved, usually between 600 m and 1,600 m on Hainan Island Here, we combined an analysis of population genetic structure and demographic history with SDMs to reveal the effect of historical climatic changes on the endemic Hainan Island species  genes and SDMs, the present study aimed to explore the responses of A. ardens are estimated to cover a total of 660 km2, of which 410 km2 is in natural reserves. Almost all of the extant populations are distributed in Bawangling, Jianfengling, Diaoluoshan, Limushan and Nanweiling Nature Reserves A. ardens on Hainan Island from a period of 2007–2009 . Collection permission was granted by the regional forestry departments, and the sample collection was carried out under the Guide for the Care and Use of Animals of the Institute of Zoology, Chinese Academy of Sciences with the approval of State Forestry Administration of China.Until now, suitable forest habitats of 5′- TCAACCACACTTCACACAGGC -3′), CytbH (5′-GGTTTACAAGACCAATGTTTTTCA -3′) (designed by the authors), and ND2L (5′- TATCGGGCCCATACCCCGAATAT-3′), ND2H (5′-CTTTGAAGGCCTTCGGTTTA -3′) EX Taq (TaKaRa). The PCR profile was 94°C for 5 min, followed by 35 cycles of 94°C for 40 s, 52°C for 40 s, 72°C for 1 min and 8 min at 72°C. Each round of PCR reactions also included one negative control to check for contamination. The PCR products were sequenced on a 3730 DNA sequencer (Applied BioSystems). Both strands of each PCR product were sequenced. Sequences were compared visually to the original chromatograms to avoid reading errors and were checked with published DNA data. To avoid amplifying mitochondrial DNA homologues from the nuclear genome (numts), we verified the sequences as follows: 1) designed specific primers, which included those amplifying longer segments of mtDNA (>1 kb); 2) checked the presence of PCR ghost bands, or extra bands; 3) detected the presence of sequence ambiguities, frame-shift mutation and stop codon; 4) used BLAST to do sequence similarity search and alignment with the published sequence databases of related species deposited in GenBank; and 5) detected whether unexpected phylogenetic placements existed in a phylogenetic analysis.Total DNA was extracted based on the TIANamp blood genomic DNA extraction kit . The whole mitochondrial CYTB and ND2 were amplified with primers CytbL . The number of segregating sites, haplotype diversity, and nucleotide diversity for each sampling location were estimated using DnaSP 5.0 A. ardens, a SDM based on current climate data was used. Assuming niche conservatism over time, this model was projected onto climate reconstructions for both the present-day and LGM A. ardens based on the largest-ever occurrence sites  throughout the Hainan Island based on our previous fieldwork www.worldclim.org) for constructing SDM. These climate layers for present-day are based on spatially interpolated values of temperature and precipitation gathered from weather stations around the world from 1950–2000 To obtain an independent perspective on the consequences of climate change for range fluctuations in b of mtDNA from 62 A. ardens individuals, we obtained ten haplotypes defined by twelve variable sites without stop codons and deletions/insertions (GenBank Accession number: JQ825228-JQ825239). The combined haplotype diversity (h) was 0.78 over samples, while it also exhibited an extremely low nucleotide diversity (π) ( = 0.062%) (as shown in With the combination of the complete 1041 bp ND2 and 1143 bp cyt A. ardens (Fst<0.001). Both Fu’s Fs and Tajima’s D statistics showed non-significantly negative value, and failed to reject the null hypothesis of neutral evolution and demographic equilibrium of the CYTB and ND2 (p = 0.3), which could be the indicator of rejection of the selective sweep. Moreover, the mismatch distributions were multimodal-shaped curves based on the combined sequences . Tests of sudden population growth based on mismatch distributions statistics could be rejected. In addition, by using the 2% Myr−1 molecular clock Minimum spanning network (MSN) parsimony showed a star-like topology, with a dominated haplotype shared between sampling locations and circled by all other rare haplotypes . The hap and ND2 . Likewisequences , with thA. ardens, we modeled extant potential climatic distribution areas  and mismatch distributions  were not consistent with each other well. Bayesian skyline plots indicate a relatively constant population size, with a slight trend of past population expansion and recent slight decrease. Although BSP confidence intervals were large, the stability of populations is supported by the SDMs. Therefore, relatively stable historical demographic history could be deduced from our molecular information of mtDNA. SDMs indicate that the amount of suitable habitat available was constant for A. ardens on Hainan Island, during both the present and LGM, and a dramatic increased distribution outside of Hainan Island during LGM. Postglacial expansion events from potential refugia by climate warming were not detected in this study. This pattern was similar to forest community results that species in Southeast Asia once have survived ice ages with relatively stable demographic history during the LGM A. ardens might have experienced local adaptation and coped with the glacial climate changes.For A. ardens without geographical isolation, but also the potential gene exchange caused by seasonal altitude shifts for foods and mating as described by The detected weak genetic differentiation of sampling regions could be attributed to not only the connected distributions of Arborophila, we did not detect more variable sites of HVR-1/CR than those of the cyt b and ND2 genes used in this study. Specifically, our previous work on the molecular diversity of Common Pheasant (Phasianus colchicus) in China also supported the lower diversity of HVR-1/CR than cyt bOur results and their interpretations are primarily dependent on the detected information of mtDNA for it can yield useful information about the historical processes behind intraspecific-level phylogeographic patterns, and its overall mutation rates tend to be higher than nuclear loci A. ardens on Hainan Island could be recognized as in situ refuge. That is, the total distribution of A. ardens before and during the LGM had once been located on the middle and southern areas of Hainan Island as the responses to the late Quaternary climate. Although the paleodistribution models of A. ardens supported the hypothesis of their in situ survival of the glaciations periods on Hainan Island, the extent of genetic diversity and demographic dynamics of A. ardens during the LGM did not coordinate well with the expansion of the modeled LGM distribution areas.We detected the pattern of more suitable distribution areas becoming available along the coast between Hainan Island and Taiwan Island with the retreat of the sea level, which implied the current distribution is partly representative of its past. Therefore, the current distribution of A. ardens has never been found in the savanna habitats; 2) because of the repeated sea-level changes. It seems conceivable that potential distribution on coastal areas and populations on those during the LGM might have vanished by gradually rising sea levels. A recent study showed there were >58 rapid sea-level rises of >40 m in the last 5 Myr in Southeast Asia A. ardens was more susceptible to cold temperature during the LGM. Field studies have demonstrated that this pheasant was sensitive to cold temperature. Some adults of A. ardens could freeze to death in winter There are four possible reasons for this: 1) because soils are particularly important in structuring tropical forest communities, the repeated submersion and exposure of soils on the shelf may have affected their structure and fertility significantly A. ardens on Hainan Island during glaciation cycles. They cannot exclude that extended land areas or more severe climate conditions might have influenced the divergence and demography of A. ardens populations earlier. Furthermore, such models do not take into account the potential effects of biotic exclusion, dispersal limitation, or historical contingency on species ranges. As such, it is important to recognize that these models reflect species potential ranges rather than their realized ranges. Transferable modeling involves projecting SDMs onto different geographic areas or different time periods and has been used for predicting species responses to global warming, reconstructing paleo-ranges of organisms, or modeling the spread of invasive species  only and not the entire ice age, which might have differed from the LGM at certain times during the Pleistocene. Therefore, our models just showed the possibility of the occurrence of ies e.g. . AlthougA. ardens might have formed local adaptations during those times and survived the late Quaternary glaciations. During the past decades, the fragmented habitat and small population size of extant A. ardens have been attributed to human-induced activities such as over-logging and poaching. Fortunately, the discovery of a number of new populations since 2002 and the subsequent protection of more forest where the species occurs suggests that this rate of decline may have slowed or even ceased in recent years A. ardens on Hainan Island. Moreover, due to the weak population genetic structure and current in situ refuge stage on Hainan Island, we suggest that extant populations of A. ardens on Hainan Island should be considered as one management unit.According to the relatively stable historical effective population size and stable distribution areas on Hainan Island through the LGM, Table S1Description of 19 bioclimatic variables used in ecological niche modeling. Variables are derived from the monthly temperature and rainfall values in order to generate more biologically meaningful variables. The bioclimatic variables represent annual trends , seasonality , and extreme or limiting environmental factors .(DOC)Click here for additional data file.


Nineteen male and 20 female source organs, and the remainder of the body, were studied to estimate time-integrated activity coefficients. The mean absorbed dose in each target organ and the effective dose were estimated from the time-integrated activity coefficients in the source organs. Biodistribution data from [18F] FACT in mice were also used to estimate absorbed doses and the effective dose in human subjects; this was compared with doses of [18F] FACT estimated from human PET data.Six healthy subjects  were enrolled in this study. An average of 160.8 MBq of [18F] FACT.The highest mean absorbed doses estimated using human PET data were observed in the gallbladder (333 ± 251 μGy/MBq), liver (77.5 ± 14.5 μGy/MBq), small intestine (33.6 ± 30.7 μGy/MBq), upper large intestine (29.8 ± 15.0 μGy/MBq) and lower large intestine (25.2 ± 12.6 μGy/MBq). The average effective dose estimated from human PET data was 18.6 ± 3.74 μSv/MBq. The highest mean absorbed dose value estimated from the mouse data was observed in the small intestine (38.5 μGy/MBq), liver (25.5 μGy/MBq) and urinary bladder wall (43.1 μGy/MBq). The effective dose estimated from the mouse data was 14.8 μSv/MBq for [18F] FACT PET study was acceptable for clinical purposes.The estimated effective dose from the human PET data indicated that the [ Therefore, several Aβ-binding probes dedicated for PET imaging have been developed PIB), has been extensively investigated with regard to its kinetics in the human body, and its effective radiation dose was found to be 4.74 μSv/MBq on average BF-227 [18F]FACT is derived from [11C]BF-227 by reducing its lipophilicity in order to reduce the nonspecific binding in the brain; AD patients showed significantly higher uptake of [18F]FACT in the neocortex region relative to controls [18F]FACT in humans has not yet been investigated.Fluorinated amyloid imaging compound , 76.3 ± 3.2 years). Subject characteristics are shown in Table This study was approved by the Ethics Committee on Clinical Investigations of Tohoku University School of Medicine and was performed in accordance with the Declaration of Helsinki. Written informed consent was obtained from all subjects after a complete description of the study had been made.18F-FACT. The radiochemical purity of the radioligand in the present clinical study ranged from 97.8% to 98.7% . The specific radioactivity ranged from 30.6 to 347.7 GBq/μmol at the time of injection .Figure Z-axis resolution of 6.5 mm FWHM at centre field of view FACT , a series of whole-body PET scans were performed. The schedule for the first and second transmission scans and the first, second, third and fourth emission scans was as follows: 6 positions × 4 min (24 min), 6 positions × 4 min (24 min), 6 positions × 3 min (18 min), 6 positions × 3 min (18 min), 6 positions × 3 min (18 min), and 6 positions × 4 min (24 min), respectively. The starting time of the second emission scan was different for each subject and was on average 55 min after the start of injection with a 5-min SD. The time gap between bed positions was 5 s. All emission data were reconstructed using OS-EM with iteration 16 and subset 2 after attenuation correction. Scatter correction was not performed because of the use of 2D mode data acquisition. The cross calibration factor of the scanner (Bq per ml/cps per voxel) was determined once per week using a cylindrical water phantom (25-cm length and 20-cm inner diameter) filled with 18F solutions and by measuring the sample activity of the 18F solutions at the well counter  FACT in the human subjects calculated from mouse data sets was compared with those of [18F]FACT from human whole-body PET scans. An average dose of 1.4 MBq of [18F]FACT was intravenously injected into ICR mice  without anaesthesia. In the [18F]FACT study, the mice were killed by cervical luxation at 2, 10, 30, 60 and 120 min ([18F]FACT) after administration (n = 4 at each time point). The masses of the blood, heart, lung, liver, spleen, small intestine, kidney, brain and urine samples were measured, and activity was also measured using a well counter. Thigh bone and muscle were also sampled. The average uptake of the radioligand into the male reference subject (70 kg) was extrapolated as follows [The experimental protocols were reviewed by the Committee on the Ethics of Animal Experiments at Tohoku University School of Medicine and performed in accordance with the Guidelines for Animal Experiments issued by the Tohoku University School of Medicine. Estimated radiation dose of FFACT in 18F]FACT.where the bodyweight of the mouse was assumed to be 30 for FACT in the human body. The highest accumulations of this radioligand were observed in the gallbladder, liver, intestine and urinary bladder. For subject no. 1, [18F]FACT contained in the bile was excreted from the liver to the duodenum through the biliary tract  in the gallbladder may be due to the quality and quantity of post-injection food intake FACT at different time points with an acquisition time of 3 min  and 4 min (fourth emission). There was no significant retention of [18F]FACT in the brain, and this may have been because the subject was normal.Figure 18F]FACT time-integrated activity coefficients in the source organs are shown in Table The [18F]FACT , liver (77.5 ± 14.5 μGy/MBq), small intestine (33.6 ± 30.7 μGy/MBq), upper large intestine (29.8 ± 15.0 μGy/MBq) and lower large intestine (25.2 ± 12.6 μGy/MBq). In mice, high absorbed doses were observed in the small intestine (38.5 μGy/MBq), liver (25.5 μGy/MBq) and urinary bladder wall (43.1 μGy/MBq) for [18F]FACT estimated from the clinical PET studies among other 18F-labelled PET amyloid radioligands were as follows: [18F]AV-45, 13 and 19.3 μSv/MBq [18F]GE067, 33.8 μSv/MBq [18F]BAY94-9172, 14.67 μSv/MBq [11C]PIB, Scheinin et al. normalised the %ID using the ratio of individual and reference subjects' body weights (Equation 4) FACT from the mouse experiments (14.8 μSv/MBq) was underestimated as compared with that from the human subject PET studies (18.6 μSv/MBq)  Table . This di18F amyloid ligand increased by a factor of three, this will only influence estimation of the effective dose within 1%; however, when the subject had taken medication that changed the function of the hepatic metabolism, the estimated effective dose will vary with a larger range FACT/year, and this injection dose limit allows two or three PET scans to be performed. Furthermore, amyloid imaging is mainly undertaken in elderly patients aged >50 years, even though for early detection of AD, patients aged <50 years will also have an amyloid PET scan. According to the guidance on medical exposures in medical and biomedical research by the European Commission FACT, was found to be acceptable for clinical study.The effective dose of the The authors declare that they have no competing interests.MS carried out the data analysis and interpretation and drafted the manuscript. TM, HT, YK and KY performed the study design and contributed to the intellectual discussion. NO, SF and RI performed the animal experiments and synthesis of PET probes. KF, SW, KH, MM and HA provided the clinical data. All authors read and approved the final manuscript.


Adherents to the Jewish faith have resided in numerous geographic locations over the course of three millennia. Progressively more detailed population genetic analysis carried out independently by multiple research groups over the past two decades has revealed a pattern for the population genetic architecture of contemporary Jews descendant from globally dispersed Diaspora communities. This pattern is consistent with a major, but variable component of shared Near East ancestry, together with variable degrees of admixture and introgression from the corresponding host Diaspora populations. By combining analysis of monoallelic markers with recent genome-wide variation analysis of simple tandem repeats, copy number variations, and single-nucleotide polymorphisms at high density, it has been possible to determine the relative contribution of sex-specific migration and introgression to map founder events and to suggest demographic histories corresponding to western and eastern Diaspora migrations, as well as subsequent microevolutionary events. These patterns have been congruous with the inferences of many, but not of all historians using more traditional tools such as archeology, archival records, linguistics, comparative analysis of religious narrative, liturgy and practices. Importantly, the population genetic architecture of Jews helps to explain the observed patterns of health and disease-relevant mutations and phenotypes which continue to be carefully studied and catalogued, and represent an important resource for human medical genetics research. The current review attempts to provide a succinct update of the more recent developments in a historical and human health context.The online version of this article (doi:10.1007/s00439-012-1235-6) contains supplementary material, which is available to authorized users. Since their emergence as a national and religious group in the Middle East over 2,000 years ago  level, but also at the levels of copy number variants (CNVs) and, where studied, identical-by-descent (IBD) segment sharing Fig. . The IBDThese studies also demonstrated that the history of the Jewish Diasporas could be observed in the genomes of Jewish people by patterns of admixture. A high degree of European admixture (30–60 %) was observed among Ashkenazi, Sephardic, Italian and Syrian Jews. The North African Jewish groups demonstrated North African and Middle Eastern admixture with varying European admixture. The proportion of European admixture among North African Jewish groups increased from east to west, with Moroccan Jews demonstrating the highest proportion. In contrast, the corresponding non-Jewish North African host populations demonstrated substantially higher inferred North African ancestry and less European ancestry.The closest genetic neighbors to most Jewish groups were the Palestinians, Israeli Bedouins, and Druze in addition to the Southern Europeans, including Cypriots. The genetic clusters formed by each of these non-Jewish Middle Eastern groups reflect their own histories of endogamy. Their proximity to one another and to European and Syrian Jews suggested a shared genetic history of related Semitic and non-Semitic Mediterranean ancestors who followed different religious and tribal affiliations. Earlier studies of Israeli Jewish, Palestinian and Druze populations made a similar observation by demonstrating the proximity of these two non-Jewish populations to Ashkenazi and Iraqi Jews  coordinated by Joel Zlotogora   Ostrer . The rea) Ostrer .The study of these conditions has been translated into public health genetics initiatives for Jews, and where the same conditions occur, non-Jews alike. These have included genetic screening programs for reproductive risks, presymptomatic identification of disease risk, and treatment of genetic disease. Screening for Tay-Sachs disease has been available for Ashkenazi Jews since the 1970s Kaback . This haThe second way in which genetic knowledge has impacted medically has been in the treatment of genetic disease. One of the dramatic examples is the treatment of Gaucher disease, a lysosomal storage disease that results in accumulation of glucocerebroside into the lysosomes of cells Charrow . People A third example relates to oncogenetics and in particular genetic testing for breast and ovarian cancer  for common diseases have been mounted for conditions that are either more prevalent among Ashkenazi Jews  or premised to have less genetic heterogeneity compared with European or European–American populations . Notably, these GWAS have identified disease-associated variants in the AJ population that were not identified in other populations  will provide an opportunity to identify the genetic factors that influence disease onset and progression and the therapeutic response.http://www.goldenhelix.org/server/israeli/ and supporting literature and online databases. (XLS 51 kb)Supplementary Table 1 Annotation of founder mutations in Jewish communities, based in part on information in the Israeli National Genetic Database Below is the link to the electronic supplementary material.


It was recently postulated that a few individual grey cattle still found in Estonia might be a relict of the old native cattle stock. Genotypes at 17 microsatellite loci from a total of 243 cattle from North European breeds and 11 grey cattle in Estonia were used in an attempt to clarify the genetic composition of the grey cattle.We characterize the genetic components of 11 examples of the grey cattle in Estonia at the population and individual levels. Our results show that the grey cattle in Estonia are most genetically similar to the Holstein-Friesian breed and secondarily to the Estonian Red cattle.Both Bayesian approaches gave similar results in terms of the identification of numbers of clusters and the estimation of proportions of genetic components. This study suggested that the Estonian grey cattle included in the analysis are a genetic composite resulting from cross-breeding of European dairy breeds. Conservation of farm animal genetic resources is of great value to the agricultural, economic, social and cultural sectors . This isca. 60 animals, which were postulated to be a relict of the old native cattle stock [Baltic cattle populations have been greatly affected by a few productive breeds such as Danish Red, Angeln, and Holstein-Friesian . As a rele stock ,5. TodaySo far the genetic composition of grey cattle relative to other existing breeds in Estonia is still unknown. In this study we use a panel of 17 microsatellite loci and Bayesian-based assignment techniques to evaluate the relationship of Estonian grey cattle to other breeds occurring in North Europe.n = 11, see Table n = 40, Estonian Red, n = 40, Finnish Holstein-Friesian, n = 43, Latvian Blue, n = 40, Latvian Danish Red, n = 40, Latvian Brown, n = 40) were included in the analysis. Seventeen  of 30 microsatellite loci recommended for genetic diversity studies in cattle http://www.projects.roslin.ac.uk/cdiv/markers.html were included in this investigation. The genotype data for the six parent populations were obtained from an earlier study [Genotypes of a total of 254 animals from seven cattle populations  using the Fragment Profiler program ver. 1.2 . International control samples were also genotyped in order to standardize the size of allele fragments. Blood sampling of the 11 Grey cattle in Estonia was taken by a veterinarian in a procedure according to the Estonian Veterinary and Food Board and satisfied all ethical concerns.Eleven Estonian grey cattle individuals from different stocks were blood-sampled. Particular efforts were made in all cases, using both the limited pedigree information (e.g. mostly only parent-offspring and full-sibling relationships) available and the knowledge of local herdsmen (e.g. the farm or village where the cattle originate from and the previous owners) via the interview questionnaire, to ensure that the animals were unrelated and had characteristics typical of the population . GenomicFST [Tests for genotypic linkage disequilibrium (LD) for each locus pair and tests for deviation from Hardy-Weinberg equilibrium (HWE) were analysed in GENEPOP version 3.4 . The gloFST  using FSFST . SignifiFST ).K value at 2 - 10 and ran the program assuming a model of admixture and correlated allele frequencies. We did not use any prior information about the population origin of the animals. A burn-in period of 200 000 generations and MCMC simulations of 500 000 iterations were used in all the above runs. The values of LnP(D) (the log probability of data) were estimated assigning a prior from 2 to 10 and the optimal K was chosen based on the delta K (ΔK) value. This criterion was originally described in Evanno et al. [Q) of the 11 grey cattle in Estonia to the K inferred clusters.A Bayesian clustering method was first employed to assess population structure using the program STRUCTURE version 2.2 . We perfo et al.  and was o et al. . We thenK almost always provided an overestimate of K, we used the number of clusters containing more than 3 individuals as a point estimate of K, as recommended by Tang et al. [K was correctly estimated, we calculated the average probability (q) of assignment to the 'correct' cluster ('correct' defined as q > 0.9 in the correct cluster). Individuals with a likelihood admixture ratio greater than 3.0 were considered to be significantly admixed.BAPS version 5.4  was run g et al. . For runFST analysis across breeds showed that 5.6% of the total genetic variation could be explained by the difference among populations. A low level of genetic differentiation was found between the grey cattle in Estonia and Finnish Holstein-Friesian  as well as between the grey cattle in Estonia and the Estonia Red cattle . Neither of the values was statistically significant at the 0.05 level (P > 0.05). No specific locus pairs showed a consistent deviation from LE that would have been in each, or even in most, of the populations. Deviations from HWE across the loci were present in the population of grey cattle in Estonia, which is most probably due to the small population size. However, no evidence for significant deviation from HWE was detected when a test was performed across all loci for all populations.The Q-values, the STRUCTURE program identified six clusters among the seven populations, but could not discern all seven populations  increased from K = 2 to K = 6, after which it began to decline, indicating the most likely value to be K = 6 (results not shown). When we used ΔK to infer the number of clusters, we found that K = 6 was clearly favoured (results not shown). At K = 6, all the grey cattle in Estonia were characterized, with the highest proportion of membership from the Finnish Holstein-Friesian cluster (QFiHF). Five grey cattle  showed high values of QFiHF > 0.9 and the remaining grey cattle are suggested to have large membership fractions in multiple clusters for the sampled populations. In particular, the grey cattle Le8 have similar values of Q for two distinct populations, Finnish Holstein-Friesian (QFiHF = 0.480) and the Estonian Red  of each grey cattle individual in each of the six identified clusters, while the corresponding values are presented in Table q values of 100% for the cluster of Holstein-Friesian and one (Le8) for Estonian Red. The remaining samples received proportions of membership from multiple clusters, while the higher average proportion of their membership was from Finnish Holstein-Friesian followed by Estonian Red  or Estonian Red, they can be valuable in the investigation of the genetics of the colour genes.The composite genetic components may explain their distinctive grey colour, which is a mixture of colours. This finding is also evidenced by the fact that a grey cow sometimes has grey and/or black-and-white calves in the same birth. Although the grey cattle are characterized as having most of their genetic components from the black-and-white dairy cattle , for exIn conclusion, given the low levels of genetic differentiation among the populations investigated, both Bayesian approaches gave similar results in terms of identification of the numbers of clusters and the estimation of proportions of genetic components. Our study shows that the Estonian grey cattle analysed were a genetically admixed population, most influenced by the Holstein-Friesian and Estonian Red cattle.The authors declare that they have no competing interests.MHL designed the study, performed the data analysis and wrote the manuscript. JK planned and coordinated the whole study, and contributed to the manuscript writing. AM collected information about Estonian grey cattle, contacted cattle owners and interviewed them. US participated in study design, sample collection of grey cattle in Estonia and the preliminary data analysis. All the authors read and approved the final manuscript.


Prunus africana, a tree species found across Afromontane regions, which is of great commercial interest internationally and of local value for rural communities, were defined with the aid of spatial analyses applied to a set of georeferenced molecular marker data (chloroplast and nuclear microsatellites) from 32 populations in 9 African countries. Two approaches for the selection of priority populations for conservation were used, differing in the way they optimize representation of intra-specific diversity of P. africana across a minimum number of populations. The first method (S1) was aimed at maximizing genetic diversity of the conservation units and their distinctiveness with regard to climatic conditions, the second method (S2) at optimizing representativeness of the genetic diversity found throughout the species’ range. Populations in East African countries  were found to be of great conservation value, as suggested by previous findings. These populations are complemented by those in Madagascar and Cameroon. The combination of the two methods for prioritization led to the identification of a set of 6 priority populations. The potential distribution of P. africana was then modeled based on a dataset of 1,500 georeferenced observations. This enabled an assessment of whether the priority populations identified are exposed to threats from agricultural expansion and climate change, and whether they are located within the boundaries of protected areas. The range of the species has been affected by past climate change and the modeled distribution of P. africana indicates that the species is likely to be negatively affected in future, with an expected decrease in distribution by 2050. Based on these insights, further research at the regional and national scale is recommended, in order to strengthen P. africana conservation efforts.Conservation priorities for  The identification of priority sites for conservation action remains a central issue in the implementation of conservation interventions, due to the fact that resources are usually limited and competition for land is high. Different approaches for making conservation choices when resources are scarce have been described previously Despite the recognized importance of evolutionary processes, they have often been excluded in conservation assessments and planning, which are more frequently based on species richness, rather than intra-specific diversity indicators Knowledge of the distribution of genetic diversity adds valuable information to support conservation efforts because the capacity of a species to adapt to changing environmental conditions depends on its heritable variation, which allows evolutionary processes to take place Some key questions continue to pose challenges. Determining the number of populations, and the number of individuals within a population, that are needed to capture useful genetic variation in a species, or within part of a species’ range, is not simple and there are many estimates in the published literature The concept of the evolutionarily significant unit, defined as “a population that merits separate management and has a high priority for conservation” Finally, conservation priorities have usually been defined based on a static snapshot of the current situation, but it is increasingly recognized that estimates of species vulnerability to global environmental changes should be incorporated in conservation planning decisions. This can be assessed in terms of predicted loss of climatically suitable areas for a particular species Prunus africana , with a broad but disjunct distribution across Afromontane regions, has been studied due to its economic importance: occurrence data from herbaria offer wide coverage and recently genetic data derived from a continent-wide collection have been made available. Over the past several decades, products from P. africana’s bark extracts have been the most widely exported of any African tree species for medicinal purposes, contributing to its overexploitation. Due to the threats to the species, mainly posed by overexploitation, but also by agricultural expansion and expected environmental changes, the need for a conservation strategy has been highlighted Among the African indigenous tree species, P. africana generated as part of two studies by Kadu et al. P. africana across geographical distance, due to its wide but disjunct distribution in Afromontane forest islands The current study builds on molecular marker data for P. africana, focusing primarily on the identification of priority populations for conservation. The formulation of a conservation strategy requires understanding a species’ spatial distribution. We generated a distribution map of P. africana based on predictions of the potential species’ distribution from records and environmental data from the same sites, using GIS (Geographic Information System) We address a series of issues related to the definition of a conservation strategy for P. africana. This is a very informative parameter, as the number of alleles per locus is dependent on effective population size, therefore a good indicator of past demographic changes that would have affected genes associated with adaptive traits as well as neutral markers We adopted the number of alleles  as a key measurement to determine priorities in the conservation of the genetic diversity in P. africana, we proposed an approach based on spatial analyses of genetic parameters and climatic variables. Geospatial methods can be applied to phylogeography in situ of species for which it is not possible to rely on ex situ conservation strategies. An example is that of important crop wild relatives, for which conservation can be achieved only by managing wild populations in situ, and which derive their properties from their adaptation to environmental conditions, favoured by evolutionary processes In order to define the location and number of populations needed to optimally conserve More specifically, we proposed the selection of a core set of priority populations based on a combination of two methods: one aimed at maximizing genetic diversity and distinctiveness of the conservation unit with regard to climatic conditions (S1), the other at optimizing representativeness of the genetic diversity found throughout the species’ range (S2). Both approaches attempt to maximize the species’ evolutionary potential through the identification and conservation of populations with the highest possible levels of genetic variation. The assumption is that this variation will allow evolutionary processes to take place and foster adaptation e.g., . These rvs. existing protected areas, and the expected threats posed by changing environmental conditions, using spatial analyses to build projected impacts of climate change on the distribution of the species Finally, we examined the conservation status of the populations selected, based on the location of our proposed sites for conservation P. africana is of great commercial interest due to the preparation of medicinal products from its bark, used to treat benign prostatic hyperplasia. The species also plays an important ecological role, providing food and home for pollinators and rare fauna, and supporting vascular and non-vascular canopy epiphytes P. africana, due to its close association with montane regions and low colonization potential P. africana, disproportionately affecting and ultimately causing the death of large, reproductively mature individuals P. africana has been reported as a pioneer The species has a broad but highly fragmented distribution, spanning the African continent from South Africa to Ethiopia and west to Cameroon, but is limited to montane regions where it can be locally common Typically, the species is found where the annual temperature range is 18–26°C, mean annual rainfall ranges from 890 to 2,600 mm, and at an elevation between 900 and 3,400 m, with increasing elevation range towards lower latitudes. Its distribution range is limited by high temperatures and by insufficient precipitation during the warmest months P. africana bark harvest has shifted from subsistence and local use to large-scale commercial use for international trade. Studies on the impacts of wild harvest on P. africana populations have shown that the practices adopted and the quantities extracted are not sustainable P. africana stands. However, enforcement issues and control problems persist, and there is considerable urgency to identify and implement sustainable management options, including conservation and appropriate domestication measures.Over the past 40 years, P. africana were used in this study: a) a set of genetic data from 32 populations collected in the near range-wide study by Kadu et al. Two main datasets on P. africana georeferenced chloroplast (cpSSRs) and nuclear microsatellites (nSSRs) consisted of: 7 chloroplast DNA loci from 582 individual trees; 6 nuclear loci from 484 individual trees. The DNA was isolated from leaf samples that were collected during 2007 and 2008 in natural stands  in 9 African countries. These are from 32 accurately georeferenced populations (a priori). Samples were collected by research partners from local institutions in different countries as indicated in Kadu et al. The ulations , spatialP. africana (mostly recorded between 1965 and 2010) that cover the range of the species and enabled us to define its ecological niche and to model its potential distribution. We assembled the dataset accessing the following sources: Mpumalanga Tourism & Parks Agency Lydenburg, South Africa (botanist Mervyn Lotter), University of Bangor , World Agroforestry Centre (ICRAF), GBIF We assembled a dataset of 1,500 georeferenced observations of P. africana’s potential distribution was modeled using the distribution modeling program Maxent. This program uses an algorithm of maximum entropy to calculate the ecological niche of a species and to define the areas of potential natural distribution P. africana were first filtered in DIVA-GIS (www.diva-gis.org) Two approaches to select priority populations for conservation were used. One method (S1) maximizes genetic diversity and distinctiveness of the conservation unit based on a combination of genetic and climatic criteria. A second method (S2) is based exclusively on genetic data and optimizes representativeness of the genetic diversity found throughout the species’ range. Each of the selected priority populations was further evaluated for urgency of conservation action on the basis of current protected status and the threat posed by predicted future climate conditions.We used the following steps to add populations to the list of priority sites: populations were clustered based on their genetic (chloroplast- and nuclear-based) and climatic similarity; in each genetic cluster, the population with highest ranking in allelic/haplotype richness and presence of locally common alleles was selected. The second step identified populations in different climatic clusters not represented in the selection above. In each climatic cluster, the population with the highest rank in both genetic parameters  was added to the priorities for conservation.1: Clustering of populations based on allelic composition: the degree of genetic similarity between populations was calculated in the R statistics environment version 2.14 2: Identification of populations with highest allelic/haplotype richness: the dataset analyzed in this study included 147 alleles at 6 nSSR loci and 19 alleles at 7 cpSSR loci. A total of 22 multilocus haplotypes were constructed by combining single cpSSR loci. An inherent difficulty with many diversity studies arises from the well-known property that diversity of a sample increases with sample size 3: Identification of populations with presence of locally common alleles: locally common alleles are those repeatedly observed but only in a small area relative to the species’ distribution; they are of high interest, especially if they are associated with adaptive traits P. africana observations were clustered based on climatic data, assuming that natural populations from different climate zones would show variable adaptive traits not captured by the analyses of SSR markers data. The 1,500 observations were clustered on the basis of 19 bioclimatic variables, extracted by point from the 2–5 minutes Wordclim dataset. The function “dist” in the R statistics environment 2.14.0 was used to calculate the Euclidian distance and hierarchical clustering was carried out using the UPGMA method. As for the clustering above, based on genetic similarity, we used the Kelley-Gardner-Sutcliffe penalty function, in order to derive an optimal number of clusters, and the cophenetic correlation coefficient to validate the clustering.4: Climate clustering: eserve selection’. It generates a selection of grid cells (30 minute cell size) that are complementary to each other in terms of diversity included in each cell, and that captures the maximum amount of diversity in the smallest number of cells possible  P. africana’s potential distribution that falls within the boundaries of protected areas of different types. WDPA includes detailed information on flora, fauna, and a wide range of climatic, environmental and socioeconomic data for 741 protected areas across 50 countries. More detailed results were presented for three countries  that host high genetic diversity for P. africana, in order to examine more closely specific threats, such as climate change and land conversion to croplands.The modeled potential distribution was combined with data on the location of protected areas. The portion of Loss of forest cover in Africa has been substantial over the past 20 years, with an average area of ca. 3.7 million ha/year converted to other land uses in the period 1990–2010 P. africana under the current climate, based on the species’ current distribution, with the potential distribution under future climatic conditions. Future climate projections were developed for 2050 under the A2 emission scenario (with constantly increasing emission rate) from the average of three different Global Circulation Models (GCMs) downloaded from the GCM Data Portal P. africana during the peak of the last glacial period  the GCM “CCSM: Last glacial maximum ” downloaded from WORLDCLIM (http://www.worldclim.org/past) was used [Source: Paleoclimate Modeling Intercomparison Project Phase II (PMIP2)].The potential threats from changes in climatic conditions at the regional scale were also assessed, by comparing the potential distribution of P. africana has been directly recorded in 22 African countries, while the modeled distribution extends to 34 countries  richness and allelic (nuclear SSRs) richness were determined after rarefaction  a,b. BotThe following populations had highest haplotype richness, in descending order : pop. NoThe set of priority populations that combined highest value of haplotype richness and highest presence of locally common alleles, in each of the 5 clusters identified, based on chloroplast markers, was the following in ascending cluster number : pop. NoAfter the selection above, additional populations were included in the priority list based on the analysis of climatic variables across the sites where the species is found. Those populations occurring in areas having unique climatic conditions, not selected based on the previous criteria, were added among the priorities for conservation. The rationale for this is that in the absence of quantitative genetic data, distinctive environmental conditions can be a proxy for useful adaptive variation.P. africana occurrence observations available , and one population each in Equatorial Guinea (pop. No. 4), Madagascar (pop. No. 15), Tanzania (pop. No. 22), and Zimbabwe (pop. No. 31). Of the 32 populations sampled for genetic analyses, 21 are included within official conservation areas, and 4 others are in sites proposed for special protection . An asseP. africana populations present the highest haplotype and allelic richness, revealed that a considerable portion of the area with suitable climate for P. africana is not covered by natural vegetation; large parts of it have been converted to cropland  where the sampled cropland . Naturalcropland . This meP. africana by year 2050 (red areas), as a result of changing climate, with large portions of modeled distribution disappearing from the map  and a very modest expansion of the species to new suitable habitats (1% of area expected to be occupied by the species in 2050), while the blue areas indicate continued suitability for the next 40 years.The predicted suitable habitat for the species, according to climate scenarios based on average values of three GCMs, is presented in P. africana in 2050 in Kenya, Uganda and Tanzania is predicted to be impacted by climate change  will be located at the margin of the modeled distribution in 2050. Of the 6 priority populations, 2 will be located at the margin of the modeled distribution: No. 13 in Kenya, and No. 22 in Tanzania.For two populations, No. 11 in Kenya and No. 29 in Uganda, the future climatic conditions are predicted to become unsuitable for P. africana, particularly in Kenya and Tanzania, which harbor a large portion of the genetic diversity found across the species’ range. At a country level, Kenya has unique opportunities to contribute to the conservation of the species, as discussed also in Muchugi et al. Both approaches presented (S1 and S2) to select priority populations indicate a high conservation priority for populations in the eastern part of the distribution of P. africanaThe patterns of genetic variation found in P. africana) are indicative of isolation and we might expect to find local adaptive variation on this basis.Obtaining genetic information specific to valuable traits requires considerable time and cost but, as genomic tools develop, their potential for describing useful variation of expressed genes will likely be a breakthrough for conservation genetics P. africana. In addition, the combination of the two methods described (S1 and S2) allows inclusion within priorities of those populations with highest genetic diversity across genetically separate clusters, but also of populations with lower diversity belonging to distinct climate clusters, potentially harbouring important adaptive properties. The clustering and ranking were obtained through a user friendly sequence of steps, with the support of freely available software, enabling conditions for a wide uptake.The approach proposed enables a reduction of the number of priorities to a minimum set of core sites optimally distributed across the range of P. africana often occurred within Afromontane forest habitats of global conservation significance including in protected areas The results reveal that although the species is not in danger of extinction, some important populations, with distinct characteristics, are threatened and their loss would reduce the livelihood potential for local people. Populations in Cameroon and Madagascar have been exposed to sustained high rates of exploitation P. africana populations of priority conservation value based on both approaches (S1 and S2) are found within the boundaries of protected areas. These populations constitute a starting point for a conservation strategy for P. africana at the continental scale, because conservation actions would be facilitated by the existing conservation measures in place. However, information on the respective IUCN conservation status is not available for all the identified priority sites for P. africana conservation; therefore it is not clear what level of protection is applied, and there are concerns about the effectiveness of protection despite the legal status.Only 4 Pinus, Eucalyptus and Acacia spp. has changed the landscape of the Afromontane region considerably, and increased fire susceptibility in the forest cover. These commercial species also encroach on protected areas and compete strongly with other tree species, including P. africana. Clearance for agriculture is a major threat affecting forested areas where P. africana occurs, as illustrated for the eastern part of the range of the species. It is not clear how climate change and further expansion of agriculture will interact with threats in different parts of the Afromontane regions, where P. africana would be pushed to higher elevations.Illegal logging and other types of encroachment into protected areas and fire frequently pose threats to the species. The situation might be worsened by a combination of different pressures on the forest cover. For example in Zimbabwe, Jimu P. africana, with the degree of reduction highly variable depending on the climate scenarios and approaches adopted.Potential threats from climate change in 2050 are envisaged for some highly diverse populations in Kenya, Tanzania and Uganda. The projections presented in this paper do not account for phenotypic plasticity, which affects the way a species responds to a change in environmental conditions. However, they are congruent with those generated by similar studies limited to East African countries Pinus oocarpa populations in Mexico suggest transferring seed at higher altitudes upwards, following a progressive change in climatic conditions It is not clear how populations in Cameroon, Equatorial Guinea, Nigeria, Angola and South Africa will respond to environmental challenges. They may be less well equipped as they have reduced levels of genetic diversity, probably attributable to their greater distance from the centre of origin of the species. On the other hand, trends are unclear; more peripheral lowland populations may contain more useful traits for climate change adaptation than other populations occurring at higher elevations. As an example, recommendations for A broad corridor was revealed by modeling the potential range of the species in the past; it connected Uganda with the Democratic Republic of the Congo and further with West African countries, like Cameroon and Nigeria. This may explain the current occurrence of the species and the possible migration route, considering the similarities found in the genetic profile of West African populations and those located on the western side of the Rift valley in eastern Africa P. africana identified in the present study include genetically unique and highly diverse P. africana populations. The areas selected are also representative of the main climatic conditions found across the species range and constitute a network of priority sites for conservation of P. africana across its range. Gap analyses at a finer scale should be carried out to identify areas with particular environmental conditions and elements of intraspecific diversity in P. africana not adequately represented inside the network of protected areas The priority areas for conservation of P. africana populations are available Encroachment and conversion of forest land to other uses threatens viability of isolated populations. Thus it is crucial to maintain a minimum population size in community forests or in a series of patches linked by pollinators in farmland. Preliminary results on gene flow in in situ conservation units for P. africana, spread across the range and representing the variety of conditions in which the species grows. Particular attention should be paid to those priority sites within protected areas but expected to be in marginal conditions by 2050, due to predicted climatic changes . In situ priority populations should be inventoried in terms of area and number of individuals, assessment of regeneration success and effective population sizes, phenological observations, biotic and abiotic factors that affect regeneration, and potential factors that could threaten individual populations.The present study highlights priority populations to be considered for inclusion in a core set of P. africana has been widely adopted by small-holder farmers in some countries such as Cameroon, where the intensive exploitation of the species started earlier, in the 1970s, and where only scattered remnants of natural populations can be found P. africana has been income, while the most important constraints are lack of good planting materials, destruction by animals and fire, low sale prices and lack of fertilizers as indicated in a recent survey on farmers’ decisions P. africana has been the international trade of its bark extract, increasingly monitored and subjected to regulation through application of quotas to quantities of bark extract exported from African countries. However, a growing interest for ethnoherbal remedies and a search for novel products from traditional medicinal plants have been documented in local markets; in Kenya, P. africana is among the most popular species traded in the local herbal industry Planting Attempts have been made to identify superior populations with regard to the chemical composition of the bark Ex situ conservation efforts should be coupled with in situ conservation, giving priority to threatened populations with known highest genetic diversity , or to the most isolated populations that consist of more than 500 mature individuals. Tests on seeds of P. africana have shown that seeds can tolerate desiccation under appropriate storage conditions


Bombyx mori L (Lepidoptera: Bombycidae) strains are currently available and these strains are maintained through continuous sibling mating. This repeated sibling mating makes the populations of each strain more homozygous, but leads to loss of unique and valuable genes through the process of inbreeding depression. Hence, it is essential to maintain a minimal degree of heterozygosity within the population of each silkworm strain, especially in the traditional geographic strains, to avoid such loss. As a result, accurate estimation of genetic diversity is becoming more important in silkworm genetic resources conservation. Application of molecular markers help estimate genetic diversity much more accurately than that of morphological traits. Since a minimal amount of heterozygosity in each silkworm strain is essential for better conservation by avoiding inbreeding depression, this article overviews both theoretical and practical importance of heterozygosity together with impacts of inbreeding depression and the merits and demerits of neutral molecular markers for measurements of both heterozygosity and inbreeding depression in the silkworm Bombyx mori.Silkworm genebanks assume paramount importance as the reservoirs of biodiversity and source of alleles that can be easily retrieved for genetic enhancement of popular breeds. More than 4000  Bombyx mori L. (Lepidoptera: Bombycidae) is a completely domesticated lepidopteran insect that has been used for silk production for more than 5000 years . Its production base is spread over 60 countries with Asian countries producing over 90% of mulberry silk production , thereby creating a wide genetic diversity among the strains; but this diversity becomes more homogenous within the strain. This repeated inbreeding in each traditional strain results in the loss of valuable genes through a process called inbreeding depression. Therefore, it is imperative to prevent this genetic erosion due to excessive inbreeding to conserve the genetic resources of geographical strains for their use in divergent breeding programmes. In this context it is worth bearing in mind that the process of conservation of the genetic resources of these strains does not only necessitate acquisition and physical possession of them, it is equally important to ensure that the original genetic characters are maintained and the rare and endemic strains are well protected from extinction. Therefore, all possible measures need to be taken topreserve the originality of the B. mori races as the gene banks are reservoirs of gene assemblies consisting of geographically isolated races, genetic stocks, and breeds collected from indigenous and exotic regions. Frankel  when compared to the wild strains (0.013). The rate of heterozygosity in domesticated strains was two times lower than that in wild varieties , which is most likely attributed to inbreeding experienced by domesticated lines. The study also shows clear genetic separation between domesticated and wild varieties and relatively less gene flow between them.Genetic diversity is the genetic variation within species, both among geographically separated populations and among individuals within a single population. Genetic diversity is an essential aspect in conservation biology because a fundamental concept of natural selection states that the rate of evolutionary change in a population is proportional to the amount of genetic diversity present in it . DecreasHeterozygosity is the state of having different alleles with regard to a given character. Allelic diversity, on the other hand, determines a population's ability to respond to long-term selection over many generations, and ultimately the survival of the population (perhaps even of the species). Individual heterozygosity is expected to be correlated between parents and their offspring. The expected correlation is maximal, approaching r = 0.50, when allelic frequencies are highly asymmetric, and it is zero when the allelic frequencies are equal to 0.50 . InbreedB. mori, it is speculated that the wild gene pool has undergone considerable selection in every generation during the course of its domestication, thougth it might be unintentional (http://unp.un.org/).Consequently the pool reached its current level of diversity of characteristics within the species. Genetic priciples and breeding techniques of modern genetic science were utilised for cross breeding and exploitation of the phenomenon of hybrid vigour in thesilkworm. A high level of heterosis/hybrid vigour for various quantitative characters was found to occur in the crosses of highly inbred lines and geographically divergent populations of the silkworm  where p and q are the allelic frequencies at a diallelic locus and F is the inbreeding coefficient. Individuals with the same expected inbreeding co-efficients vary in their degree of homozygosity hence there is opportunity for natural selection to modify the array of genotypic frequencies, particularly to favour the more heterozygous individuals. Inbreeding depression for a trait is a linear function of the inbreeding coefficient (ST), quantified by effective population size, is another parameter with which inbreeding depression within a population is measured. STF measures the decrease in heterozygosity due to the finite size of the population and the effective size. In fact STF gives a measure of the genetic drift among conceptual replicates of the population in an infinite nonrandom mating population, STF= 0 and ITF= IS,F rising toward a value which depends on the proportion of inbred matings. Thus, genetic drift ST)(F and decline in heterozygosity IT). The success and spread of silkworm rearing was mostly due to the use of F1 silkworm hybrids of exotic temperate silkworm male parental strains and native female tropical silkworm strains . Most estimates yield an order of magnitude of 0.01–0.05. However, sample size (N) must be considered as, even if the true correlation is zero, an average value of 1/(N-1) is expected by chance  is of the same magnitude as the r2 values observed. For multiple regressions (L loci as separate ‘heterozygosity’ variables) the null expectation for 2R is L/(N-1), a serious problem if L is not <N  historically appeared as an opportunity to develop marker-based genetics, an approach boosted by the discovery of allozyme polymorphism . Geneticy chance . As mosts not <N . Howeverosophila  and in mosophila , stronglosophila . HeterozB. mori, the few attempts which made recently have revealed that within strains genetic diversity is much less than it is among the strains. Staykova (Assessment of heterozygosity in each silkworm race is important for efficient management and conservation of genetic resources . GenotypStaykova  used isoStaykova showed tStaykova . Since tStaykova  they canStaykova . GeneralStaykova . The resStaykova , which iStaykova . Using iB. mori is a completely domesticated insect and has undergone intense artificial selection, (B. mori unable to survive in the wild. Therefore, natural selection induced genetic diversity is limited to few traits such as voltinism. Hence, it is essential to conserve and utilize the wild relatives of B. mori to broaden the genetic base of the domesticated races apart from utilizing the geographically divergent races, mutants, sex-limited races, evolved breeds, breeder's genetic stocks, and exotic resources. The wild relatives of Bombyx are ecologically very vulnerable and the vulnerability at different spatial and temporal scales is not known. The design of biodiversity network in sericulture involving the complementaries of wild relatives and domesticated B. mori is also not well established. Therefore, conservation of wild as well as domesticated seribiodiversity resources is very essential for sustainable development of sericulture since loss of genetic resources of domesticated and wild relatives of Bombyx species along with their unique genes may be disadvantage for future generations (lection, . Therefoerations .With the advent of co-dominant DNA markers that are superior to allozymes, the identification of heterozygosity among the populations of races, variety, or breeds of a species became easier. Use of SSR and SNP markers to identify the heterozygosity levels for conservation of genetic material for true-to-type genotype maintenance among germplasm stocks has become a regular practice. The use of DNA markers to relate genetic homozygosity and genetic distance of the parental strains to the manifestation of hybrid vigour, and understanding the geneticbasis of heterosis, will be quite useful to select suitable parental lines for hybridization programmes. Considering the large number of silkworm inbred lines that are maintained in the silkworm breeding stations, it is prudent to use molecular markers to identify parental lines that are most suitable for developing heterotic hybrids. Silkworm strain-specific markers could also be used for authentication of races, protection of breeders' rights, and periodical monitoring of the purity of varieties in the conservation centres. So far, evidence of effects of inbreeding on inbreeding coefficients in silkworm populations is limited. Hence understanding the effects of inbreeding for various traits can be very crucial points in the management of germplasm. Another important objective of silkworm genetic resource conservation is to protect the available seri-biodiversity from extinction. At present only few old Indian indigenous races survive, viz. Barapolu, Chotapolu, Nistari, Sarupat, and Moria; several indigenous races like Ichot, Nishmo, Itan, and the univoltine Kashmiri races are extinct . Therefo


Culex pipiens complex includes two widespread mosquito vector species, Cx. pipiens and Cx. quinquefasciatus. The distribution of these species varies in latitude, with the former being present in temperate regions and the latter in tropical and subtropical regions. However, their distribution range overlaps in certain areas and interspecific hybridization has been documented. Genetic introgression between these species may have epidemiological repercussions for West Nile virus (WNV) transmission. Bayesian clustering analysis based on multilocus genotypes of 12 microsatellites was used to determine levels of hybridization between these two species in Macaronesian islands, the only contact zone described in West Africa. The distribution of the two species reflects both the islands' biogeography and historical aspects of human colonization. Madeira Island displayed a homogenous population of Cx. pipiens, whereas Cape Verde showed a more intriguing scenario with extensive hybridization. In the islands of Brava and Santiago, only Cx. quinquefasciatus was found, while in Fogo and Maio high hybrid rates (∼40%) between the two species were detected. Within the admixed populations, second-generation hybrids (∼50%) were identified suggesting a lack of isolation mechanisms. The observed levels of hybridization may locally potentiate the transmission to humans of zoonotic arboviruses such as WNV.The  Calathus, Meladema, Pimelia, Tarphius), butterflies (Gonepteryx), and spiders  and Cx. quinquefasciatus are the most common and widespread species. The former is found primarily in temperate zones, whereas the latter occurs in tropical and subtropical zones. Cx. pipiens has a greater ecological range with populations found from the low subarctic of Siberia and Scandinavian countries to the semidesert regions of the Maghreb  prezygotic isolation may result from differences in species distribution and in mating behavior; and (2) intrinsic postzygotic may result from cytoplasmic incompatibility that creates unviable hybrids  to determine the degree of genetic differentiation among island populations; (2) to measure rates of hybridization and genetic introgression between the sibling species; and (3) to infer about the colonization process and their impacts in Macaronesian region.In this study, Bayesian model-based methods were applied to multilocus microsatellite genotypes to infer the genetic structure of the Cx. pipiens s.l. larvae were sampled in Cape Verde is shown in Indoor resting collections of adult mosquitoes using mechanical aspirators were carried out in four localities of Madeira Island in September 2006 and in June 2007 . Given tCx. pipiens, Cx. quinquefasciatus, and Cx. torrentium . For each locus, one of the primers was fluorescently labeled . Thermocycling conditions included an initial denaturation step of 5 min at 96ºC, followed by 30 cycles each of 96ºC for 30 sec, annealing at 52ºC-56ºC (locus-dependent) for 30 sec and 72ºC for 30 sec. After a final extension step of 5 min at 72ºC, reactions were stopped at 4ºC.Twelve microsatellites  at Yale DNA Analysis Facility (USA). Fragment sizes and genotypes were scored using the software GeneMarker 1.4. .FIS). Significance of FIS values was assessed by randomization tests. These analyses were performed using FSTAT v. 2.9.3.2. (Goudet AR), a measure of allele diversity adjusted for the lowest sample size, were obtained by the statistical rarefaction approach implemented in HP-RARE . The inference of the number of genetic clusters (K) in the Bayesian method implemented by STRUCTURE is not straightforward, and it is normally performed by ad hoc approaches: an estimation of ln[Pr(X|K)], described in the original publication  of 0.90. Individuals displaying 0.1 ≤ qi ≤ 0.90 were considered of admixed ancestry. The information from the outputs of each K (10 runs) was aligned by the Greedy method implemented in CLUMPP  and four hybrid . The approach of uniform priors was used because it reduces the influence of low-frequency alleles thus which may result from sampling and genotyping errors in closely related populations. Results were based on the average of five independent runs of 105 iterations. Following the suggestions of Anderson and Thompson (Tq) of 0.50.The Bayesian method implemented by NEWHYBRIDS 1.1.  tree based on Cavalli-Sforza and Edwards  chord diWhenever multiple testing was performed, the nominal significance level of rejection of the null hypothesis (α = 0.05) was corrected by the sequential Bonferroni procedure Holm .Cx. pipiens and were collected in Madeira (N = 190) and in Maio (N = 13). Culex quinquefasciatus was found in the four islands of Cape Verde (N = 115), and it was the only member of the complex present in the collections from Brava (N = 31) and Santiago (N = 54). Fifty-six mosquitoes displayed a heterozygous pattern for ACE-2 and were collected in Fogo (N = 14) and Maio (N = 42). The island of Maio was the only island where the two species and putative hybrids were found in sympatry.A total of 374 females  were analyzed by the molecular assay ACE-2 (Smith and Fonseca N = 374) and subsamples determined by ACE-2 identification and geographic location are shown in Genetic diversity estimates for the 12 microsatellites in whole sample (K = 1) and the intriguing scenario of Cape Verde with three possible subdivisions  was the most abundant in the archipelago with 91 specimens from three islands , all identified as Cx. quinquefasciatus by ACE-2 PCR. Cluster 2 (C2) was the smallest cluster with 25 specimens from Maio Island being classified as Cx. pipiens or hybrid by ACE-2 PCR. Cluster 3 (C3) includes individuals from Fogo and Maio Islands, and the majority (87.2%) of the specimens were identified as hybrids by ACE-2 PCR. Twenty-nine specimens, the majority of which from Maio and Fogo, were not assigned to any of the four clusters and were thus considered admixed.The combination of the Bayesian clustering results with the ACE-2 identification clarified the separation of sampled mosquitoes into four different clusters : ClusterCx. quinquefasciatus, while the majority of the individuals of C2 (96.0%) were classified as pure Cx. pipiens and one individual was classified as a backcross with Cx. pipiens (BxP). The majority of individuals of C3 (87.1%) were classified as hybrids. Of these, 10  were classified as F2 hybrids and nine individuals from Maio were backcrosses with Cx. pipiens . In Cape Verde, all the samples from C4 were classified as pure ens BxP; .Culex pipiens samples from Madeira (C1) and Maio (C2) displayed a high genetic distance but were still grouped in a common cluster separated from the remaining samples. Samples from cluster C3, composed mainly by hybrid individuals, displayed an intermediate position in the topology of the tree. Culex quinquefasciatus samples from cluster C4 shared the same cluster, but it was possible to observe significant divergence between the populations of the three islands .The Dc-based NJ tree was consistent with the presence of the four clusters identified in the analysis performed by STRUCTURE . Culex pCx. pipiens and Cx. quinquefasciatus were found to differ among islands of the Macaronesian region. Madeira showed a genetically homogenous Cx. pipiens population. In Cape Verde, it was possible to identify monospecific populations of Cx. quinquefasciatus in Brava and Santiago, while admixed populations between both species were observed in Maio and Fogo. The species diagnostic ACE-2 PCR was effective in the identification of each species in the allopatric populations of Madeira, Brava, and Santiago. However, in sympatric populations with interspecific admixture such as those of Maio and Fogo, repeated introgression and recombination lead to a disruption of the linkage between the diagnostic alleles and the respective genetic backgrounds of each species. Under these conditions, a more cautious interpretation of the results obtained by a single diagnostic marker such as the ACE-2 is needed for the correct identification of each species and hybrids . A possible explanation for this apparent contradiction could be insufficient sampling of the least abundant species. Factors that may have contributed for an insufficient sampling were the collection method used (immature captures) coupled with the low number of breeding sites positive for Cx. pipiens s.l. larvae. In Cape Verde, very low adult mosquito densities preclude the use of collection methods targeting adult mosquitoes for sampling sufficient numbers of individuals  are comparable with those recorded in the hybrid zone of North America  between infected males and uninfected females or females infected by incompatible strains  may explain the high levels of hybridization in Cape Verde. Molecular analysis of W. pipientis in these mosquito populations would help clarifying this hypothesis.High hybridization rates between Cx. pipiens indicate mating success of F1 individuals  suggesting a low effect of extrinsic postzygotic isolation mechanisms between the Cx. pipiens and Cx. quinquefasciatus in Cape Verde.Isolation between close species can be promoted by several mechanisms that may act in simultaneous. The lack or incomplete action of prezygotic (e.g. mating behavior) and intrinsic postzygotic (e.g. cytoplasmic incompatibility) mechanisms allows hybridization creating first generation (F1) hybrids. However, an extrinsic postzygotic mechanism such as hybrid sterility or hybrid low fitness can restrict gene flow to one generation avoiding introgression , avian pox virus, Usutu virus and WNV. The introduction of these pathogens may place the endemic bird populations in danger (Kilpatrick Cx. pipiens and Cx. quinquefasciatus may promote a more opportunistic feeding behavior increasing the chance for the accidental transmission of WNV to humans. A serologic survey in 1980s showed 40% positive cases of WNV in Cape Verdean children (Vieira Macaronesia is a passage route and breeding region for migratory birds (Garcia-del-Rey n Vieira . Howevern Vieira . Even son Vieira , or the n Vieira , highlig


Donax deltoides (Veneroida: Donacidae) is an intertidal, soft-sediment mollusc and an ideal surrogate for testing this relationship, given its lack of habitat constraints in this region, and its immense dispersal potential driven by year-long spawning and long-lived planktonic larvae. We assessed allele frequencies at 10 polymorphic microsatellite loci across 11 sample locations spanning the barrier region and identified genetic structure consistent with the major ocean currents of southeastern Australia. Analysis of mitochondrial DNA sequence data indicated no evidence of genetic structuring, but signatures of a species range expansion corresponding with historical inundations of the Bassian Isthmus. Our results indicate that ocean currents are likely to be the most influential factor affecting the genetic structure of D. deltoides and a likely physical barrier for passive dispersing marine fauna generally in southeastern Australia.Major disjunctions among marine communities in southeastern Australia have been well documented, although explanations for biogeographic structuring remain uncertain. Converging ocean currents, environmental gradients, and habitat discontinuities have been hypothesized as likely drivers of structuring in many species, although the extent to which species are affected appears largely dependent on specific life histories and ecologies. Understanding these relationships is critical to the management of native and invasive species, and the preservation of evolutionary processes that shape biodiversity in this region. In this study we test the direct influence of ocean currents on the genetic structure of a passive disperser across a major biogeographic barrier.  In many regions of the world discrete faunal boundaries have been identified in marine communities, including the Isthmus of Panama  markers. These markers have revealed deep phylogenetic splits in molluscs, echinoderms, arthropods, and cnidarians, indicating historical vicariant divergence likely associated with the Bassian Isthmus  directly influence contemporary structuring of D. deltoides, and compare these findings with those from species with various life histories to identify likely common drivers of gene flow in the region. We discuss these findings in context of biogeography, biodiversity conservation, and fisheries management of D. deltoides Australia.The present study describes broad patterns of genetic structure of Donax deltoides specimens were collected from 11 locations extending from Fraser Island in Queensland to Goolwa in South Australia  , and pairs of loci tested for linkage disequilibrium using a log-likelihood ratio test. In order to overcome potential limitations of FST calculations using multiallelic loci  and expected (HE) heterozygosity were determined using the Excel Microsatellite Toolkit , among sample sites within regions, and within sample sites. A factorial correspondence analysis (FCA), implemented in GENETIX version 4.05  ten independent simulations for K = 1–11 with 100,000 burn-in and 1,000,000 data iterations were run. Analyses were performed using the admixture model of population structure  and allele frequencies were set as independent among populations. The most likely K was estimated using Evanno's ΔK  value. A pilot analysis was performed initially to confirm that 50,000 sweeps with a 10,000 step burn-in stabilized the likelihood. K was then determined from five independent runs where the value was allowed to vary from 1 to11. After identifying the most likely K, 100 replicate analyses were performed using an admixture model and summarized using CLUMPP  at HWE to the heterozygosity expected (HEQ) at mutation-drift equilibrium for each sample locality. The significance of any observed excess was tested using a Wilcoxon's sign-rank test based on the stepwise mutation (SMM) and two-phase mutation (TPM) models  was amplified for 10 individuals from each sample site by PCR using primers HCO and LCO  of the mitochondrial cytochrome oxidase I gene  and pairwise measures of ΦST among sample locations. Significance was determined using 10,000 permutations and significance levels were corrected using the sequential Bonferroni procedure . Consensus sequences were imported into MEGA version 5.1  was estimated using an mtDNA COI mutation rate of 0.7–1.2% per million years  coalescent approach implemented in BEAST 1.7.3  at South Australian and western Victoria locations  compared with estimates from locations in the more easterly Victorian (Venus Bay and Paradise Beach) and New South Wales locations . Expected heterozygosities were moderate to high and ranged from 0.53 to 0.71 (mean HE = 0.65), with locations in South Australia and western Victoria also exhibiting the lowest genetic diversity  for 8 of the 11 locations, while only five were significant after corrections for multiple comparisons  indicating limited gene flow and genetic structuring amongst sampling locales. Pairwise population comparisons of FST indicate limited gene flow among three distinct regions  with significant values observed at all pairwise comparisons among these regions. Conversely, all pairwise comparisons of sites within regions did not differ significantly from zero indicating panmixia (P < 0.001), whereas variation between regions  explained 16% (P < 0.001) of the variation, and populations within regions explained 1.0% (P < 0.001). The high “between region” variation emphasizes the extent of genetic differentiation among D. deltoides populations from different Australian coastal areas.Global estimates of panmixia . An AMOVFST and the natural log of geographic distance . Regression showed this relationship to be positive and linear  indicating extensive gene flow within regions. The relationships between sample locales are best depicted by the two-dimensional FCA of the microsatellite variation  and those from South Australia and Victoria (west) with an area of strong overlap occurring around the Wilsons Promontory region . This pattern is somewhat consistent with an isolation-by-distance model, but in this case more likely to be reflecting an isolation-by-barrier scenario.Regression analyses and a Mantel test suggest moderate isolation by distance. There was a significant association between genetic distance and geographic distance with the Mantel test showing a moderate relationship between Slatkin's linearized ariation . When thK) within the data. STRUCTURE identified two populations corresponding exactly with those identified by TESS  indicating each site is at mutation-drift equilibrium. These findings indicate that effective population sizes have remained constant in the recent past.BOTTLENECK analysis found no evidence of recent bottlenecks across sample localities. Under each mutation model Wilcoxon's sign-rank test was nonsignificant  fragment of the mitochondrial COI gene was successfully amplified and sequenced for 118 individuals representing 11 populations of ustralia . Followiustralia .P < 0.001), whereas variation between sample sites explained only 6% (P < 0.001) of the total variation. The low “between site” variation indicates a lack of genetic differentiation among D. deltoides collection sites. Global ΦST was weak (ΦST = 0.06) yet significantly different from zero indicating potentially limited gene flow and genetic structuring between sample locations. However, all pairwise comparisons of ΦST were nonsignificant suggesting potential panmixia, or alternatively a lack of statistical power for resolving patterns of genetic structure with confidence.The majority of the variation in the mtDNA dataset was explained by variation within sample sites . Using the tau statistic (0.746) the time since expansion was estimated as 51,805–88,736 years. Similarly the Bayesian skyline plot indicates that the D. deltoides population had been stable prior to a range expansion event occurring ∼50,000 years ago . The potential influence of selection is more difficult to determine. Microsatellite loci are typically recognized as neutral genetic markers, however, it is possible that one or more loci are linked to genes or gene groups under selection  along the east coast of Australia to Eden  suggests extensive gene flow along the east coast of Australia, further complementing the previous findings of Murray-Jones and Ayre . The direction of larval movement is likely to be largely determined by the predominant current directions , however, bidirectional gene flow is likely within the respective populations given the nonsignificant results from our isolation-by-distance tests. Given the longevity of the larval phase and the year-long spawning cycle of D. deltoides it is possible the direction of larval movement is influenced by seasonal variation in current intensity and direction, surface currents driven by varying wind activities, and smaller scale eddie systems , providing opportunities for expansion events to occur. We propose the western population was likely colonized most recently, involving a relatively small number of founder individuals from the larger, more genetically diverse eastern population. Following this range expansion the western population has remained isolated due to contemporary physical oceanographic conditions. Although highly speculative potential vectors for historical movement across Bass Strait might include irregular ocean currents and climatic events, birds, and even Indigenous Australians who have a long history of harvesting the species as a food resource  have identified limited east-west gene flow and significant structuring in this region , however, we suggest that ocean currents are likely to be a common barrier to dispersal regardless of other ecological factors. This study contributes to our understanding of the physical drivers responsible for the biogeographic structuring of marine communities in southeastern Australia and the link between species life histories and dispersal across marine biogeographic barriers generally.We conclude that the major ocean currents of southeastern Australia influence the contemporary structure of