Modeling Problems in Conservation Genetics Using Captive Drosophila Populations: Consequences of Equalization of Family Sizes

Equalization of family sizes is recommended for use in captive breeding programs, as it is predicted to double effective population sizes, reduce inbreeding, and slow the loss of genetic variation. The effects of maintaining small captive populations with equalization of family sizes versus random choice of parents on levels of inbreeding genetic variation, reproductive fitness, and effective population sizes ( N _e) were evaluated in 10 lines of each treatment maintained with four pairs of parents per generation. The mean inbreeding coefficient ( F ) increased at a significantly slower rate with equalization than with random choice (means of 0.35 and 0.44 at generation 10). Average heterozygosities at generation 10, based on six polymorphic enzyme loci, were significantly higher with equalization (0.149) than with random choice (0.085), compared to the generation 0 level of 0.188. The competitive index measure of reproductive fitness at generation 11 was more than twice as high with equalization as with random choice, both being much lower than in the outbred base population. There was considerable variation among replicate lines within treatments in all the above measures and considerable overlap between lines from the two treatments. Estimates of N _e for equalization were greater than those for random choice, whether estimated from changes in average heterozygosities or from changes in F. Equalization of family sizes can be unequivocally recommended for use in the genetic management of captive populations.     

Founder Effects,Inbreeding, and Loss of Genetic Diversity in Four Avian Reintroduction Programs

Abstract: The number of individuals translocated and released as part of a reintroduction is often small, as is the final established population, because the reintroduction site is typically small. Small founder and small resulting populations can result in population bottlenecks, which are associated with increased rates of inbreeding and loss of genetic diversity, both of which can affect the long‐term viability of reintroduced populations. I used information derived from pedigrees of four monogamous bird species reintroduced onto two different islands (220 and 259 ha) in New Zealand to compare the pattern of inbreeding and loss of genetic diversity among the reintroduced populations. Although reintroduced populations founded with few individuals had higher levels of inbreeding, as predicted, other factors, including biased sex ratio and skewed breeding success, contributed to high levels of inbreeding and loss of genetic diversity. Of the 10–58 individuals released, 4–25 genetic founders contributed at least one living descendent and yielded approximately 3–11 founder–genome equivalents (number of genetic founders assuming an equal contribution of offspring and no random loss of alleles across generations) after seven breeding seasons. This range is much lower than the 20 founder–genome equivalents recommended for captive‐bred populations. Although the level of inbreeding in one reintroduced population initially reached three times that of a closely related species, the long‐term estimated rate of inbreeding of this one population was approximately one‐third that of the other species due to differences in carrying capacities of the respective reintroduction sites. The increasing number of reintroductions to suitable areas that are smaller than those I examined here suggests that it might be useful to develop long‐term strategies and guidelines for reintroduction programs, which would minimize inbreeding and maintain genetic diversity.     

Application of the One-Migrant-per-Generation Rule to Conservation and Management

Abstract:  Endangered species are commonly found in several (partially) isolated populations dispersed on different fragments of a habitat, natural reserve, or zoo. A certain level of connectivity among such populations is essential for maintaining genetic variation within and between populations to allow local and global adaptation and for preventing inbreeding depression. A rule of thumb widely accepted by the conservation community is that one migrant per generation (OMPG) into a population is the appropriate level of gene flow. This rule is based on Wright's study of his island model under a long list of simplifying assumptions. I examined the robustness of the OMPG rule to the violation of each of the many assumptions, quantifying the effect with population genetics theory. I showed that, when interpreted as one effective migrant per generation, OMPG is generally valid for real populations departing from the ideal model in the discrepancies of actual (  N ) and effective (  N_e  ) population sizes and actual ( m ) and effective ( m_e  ) migration rates. I also addressed the issue of converting the effective number of migrants (  M_e= N_em_e  ) into the actual number of migrants ( M = Nm  ) of a certain age and sex. In particular, N_e < N , a case common for natural populations, did not necessarily require M > M_e to maintain a certain level of differentiation among populations. Rather, translating the elusive M_e into the manageable M depends on the specific causes (e.g., biased sex ratio, reproductive skew) that lead to N_e < N .     

Genetic Diversity, Population Size, and Fitness in Central and Peripheral Populations of a Rare Plant Lychnis viscaria

Abstract: Genetic diversity is expected to decrease in small and isolated populations as a consequence of bottlenecks, founder effects, inbreeding, and genetic drift. The genetics and ecology of the rare perennial plant Lychnis viscaria (Caryophyllaceae) were studied in both peripheral and central populations within its distribution area. We aimed to investigate the overall level of genetic diversity, its spatial distribution, and possible differences between peripheral and central populations by examining several populations with electrophoresis. Our results showed that the level of genetic diversity varied substantially among populations (  H _exp = 0.000–0.116) and that the total level of genetic diversity (mean H _exp = 0.056) was low compared to that of other species with similar life-history attributes. The peripheral populations of L. viscaria had less genetic variation (mean H _exp = 0.034) than the central ones (0.114). Analysis of genetic structure suggested limited gene flow (mean F _ST = 0.430) and high differentiation among populations, emphasizing the role of genetic drift (  N _e m = 0.33). Isolation was even higher than expected based on the physical distance among populations. We also focused on the association between population size and genetic diversity and possible effects on fitness of these factors. Population size was positively correlated with genetic diversity. Population size and genetic diversity, however, were not associated with fitness components such as germination rate, seedling mass, or seed yield. There were no differences in the measured fitness components between peripheral and central populations. Even though small and peripheral populations had lower levels of genetic variation, they were as viable as larger populations, which emphasizes their potential value for conservation.     

Effective Population Size and Maintenance of Genetic Diversity in Captive-Bred Populations of a Lake Victoria Cichlid

Abstract: We used microsatellite DNA markers to investigate the maintenance of genetic diversity within and between samples of subpopulations (spanning five captive-bred generations) of the haplochromine cichlid Prognathochromis perrieri . The subpopulations are maintained as part of the Lake Victoria Cichlid species survival plan. Changes in the frequencies of 24 alleles, over four polymorphic loci, were used to estimate effective population size (   N _e   ). Point estimates of N _e ranged from 2.5 to 7.7 individuals and were significantly smaller than the actual census size (   N _obs  ) for all subpopulations (32–243 individuals per generation), with the corresponding conservative N _e   /  N _obs ratios ranging from 0.01 to 0.12. Approximately 19% of the initial alleles were lost within the first four generations of captive breeding. Between-generation comparisons of expected heterozygosity showed significant losses ranging from 6% to 12% per generation. Seven private alleles were observed in the last sampled generation of four subpopulations, and analysis of population structure by F _ST indicated that approximately 33% of the total genetic diversity is maintained between the subpopulations from different institutions. To reduce the loss of genetic variation, we recommend that offspring production be equalized by periodically removing dominant males, which will encourage reproduction by additional males. Consideration should also be given to encouraging more institutions to maintain populations, because a significant fraction of the genetic variation exists as among-population differences resulting from random differentiation among subpopulations.     

Longitudinal monitoring of neutral and adaptive genomic diversity in a reintroduction

Restoration programs in the form of ex-situ breeding combined with reintroductions are becoming critical to counteract demographic declines and species losses. Such programs are increasingly using genetic management to improve conservation outcomes. However, the lack of long-term monitoring of genetic indicators following reintroduction prevents assessments of the trajectory and persistence of reintroduced populations. We carried out an extensive monitoring program in the wild for a threatened small-bodied fish (southern pygmy perch, Nannoperca australis) to assess the long-term genomic effects of its captive breeding and reintroduction. The species was rescued prior to its extirpation from the terminal lakes of Australia's Murray-Darling Basin, and then used for genetically informed captive breeding and reintroductions. Subsequent annual or biannual monitoring of abundance, fitness, and occupancy over a period of 11 years, combined with postreintroduction genetic sampling, revealed survival and recruitment of reintroduced fish. Genomic analyses based on data from the original wild rescued, captive born, and reintroduced cohorts revealed low inbreeding and strong maintenance of neutral and candidate adaptive genomic diversity across multiple generations. An increasing trend in the effective population size of the reintroduced population was consistent with field monitoring data in demonstrating successful re-establishment of the species. This provides a rare empirical example that the adaptive potential of a locally extinct population can be maintained during genetically informed ex-situ conservation breeding and reintroduction into the wild. Strategies to improve biodiversity restoration via ex-situ conservation should include genetic-based captive breeding and longitudinal monitoring of standing genomic variation in reintroduced populations.     

Unprecedented Low Levels of Genetic Variation and Inbreeding Depression in an Island Population of the Black-Footed Rock-Wallaby

Abstract: It has been argued that demographic and environmental factors will cause small, isolated populations to become extinct before genetic factors have a significant negative impact. Islands provide an ideal opportunity to test this hypothesis because they often support small, isolated populations that are highly vulnerable to extinction. To assess the potential negative impact of isolation and small population size, we compared levels of genetic variation and fitness in island and mainland populations of the black-footed rock-wallaby ( Petrogale lateralis [Marsupialia: Macropodidae]). Our results indicate that the Barrow Island population of P. lateralis has unprecedented low levels of genetic variation (  H _e = 0.053, from 10 microsatellite loci) and suffers from inbreeding depression (reduced female fecundity, skewed sex ratio, increased levels of fluctuating asymmetry). Despite a long period of isolation ( ∼ 1600 generations) and small effective population size (  N _e ∼ 15), demographic and environmental factors have not yet driven this population to extinction. Nevertheless, it has been affected significantly by genetic factors. It has lost most of its genetic variation and become highly inbred (  F _e = 0.91), and it exhibits reduced fitness. Because several other island populations of P. lateralis also exhibit exceptionally low levels of genetic variation, this phenomenon may be widespread. Inbreeding in these populations is at a level associated with high rates of extinction in populations of domestic and laboratory species. Genetic factors cannot then be excluded as contributing to the extinction proneness of small, isolated populations.     

Relative Contribution of Inbreeding Depression and Eroded Adaptive Diversity to Extinction Risk in Small Populations of Shore Campion

Abstract:  To study the relative importance of inbreeding depression and the loss of adaptive diversity in determining the extinction risk of small populations, we carried out an experiment in which we crossed and self-fertilized founder plants from a single, large population of shore campion ( Silene littorea Brot.). We used the seeds these plants produced to colonize 18 new locations within the distribution area of the species. The reintroduced populations were of three kinds: inbred and genetically homogeneous, each made up of selfed seed from a single plant; inbred and mixed, made up of a mixture of selfed seeds from all founder plants; and outbred and mixed, made up of a mixture of seeds obtained in outcrosses between the founders. We compared the inbred homogeneous populations with the inbred mixed to measure the effect of genetic diversity among individuals and the inbred mixed with the outbred mixed to measure the effect of inbreeding. Reintroduction success was seriously limited by inbreeding, whereas it was not affected by genetic diversity. This observation and the nonsignificant interaction between family and reintroduction location for individual plant characters suggest that the fixation of overall deleterious genes causing inbreeding depression posed a more serious threat to the short-term survival of the populations than the loss of genes involved in genotype and environment interactions. Thus, reintroduction success was related to adaptive diversity. Preventing such fixation might be the most important consideration in the genetic management and conservation of shore campion populations.     

Genomic erosion in a demographically recovered bird species during conservation rescue

The pink pigeon (Nesoenas mayeri) is an endemic species of Mauritius that has made a remarkable recovery after a severe population bottleneck in the 1970s to early 1990s. Prior to this bottleneck, an ex situ population was established from which captive-bred individuals were released into free-living subpopulations to increase population size and genetic variation. This conservation rescue led to rapid population recovery to 400–480 individuals, and the species was twice downlisted on the International Union for the Conservation of Nature (IUCN) Red List. We analyzed the impacts of the bottleneck and genetic rescue on neutral genetic variation during and after population recovery (1993–2008) with restriction site-associated sequencing, microsatellite analyses, and quantitative genetic analysis of studbook data of 1112 birds from zoos in Europe and the United States. We used computer simulations to study the predicted changes in genetic variation and population viability from the past into the future. Genetic variation declined rapidly, despite the population rebound, and the effective population size was approximately an order of magnitude smaller than census size. The species carried a high genetic load of circa 15 lethal equivalents for longevity. Our computer simulations predicted continued inbreeding will likely result in increased expression of deleterious mutations (i.e., a high realized load) and severe inbreeding depression. Without continued conservation actions, it is likely that the pink pigeon will go extinct in the wild within 100 years. Conservation rescue of the pink pigeon has been instrumental in the recovery of the free-living population. However, further genetic rescue with captive-bred birds from zoos is required to recover lost variation, reduce expression of harmful deleterious variation, and prevent extinction. The use of genomics and modeling data can inform IUCN assessments of the viability and extinction risk of species, and it helps in assessments of the conservation dependency of populations.     

Low Genetic Variability in the Hawaiian Monk Seal

The Hawaiian monk seal (   Monachus schauinslandi) is a critically endangered species that has failed to recover from human exploitation despite decades of protection and ongoing management efforts designed to increase population growth. The seals breed at five principal locations in the northwestern Hawaiian islands, and inter-island migration is limited. Genetic variation in this species is expected to be low due to a recent population bottleneck and probable inbreeding within small subpopulations. To test the hypothesis that small population size and strong site fidelity has led to low within-island genetic variability and significant between-island differentiation, we used two independent approaches to quantify genetic variation both within and among the principal subpopulations. Mitochondrial control region and tRNA gene sequences (359 base pairs) were obtained from 50 seals and revealed very low genetic diversity (0.6% variable sites), with no evidence of subpopulation differentiation. Multilocus DNA fingerprints from 22 individuals also indicated low genetic variation in at least some subpopulations (band-sharing values for "unrelated" seals from the same island ranged from 49 to 73%). This method also provided preliminary evidence of population subdivision (  F'_st estimates of 0.20 and 0.13 for two adjacent island pairs). Translocations of seals among islands may therefore have the potential to relieve local inbreeding and possibly to reduce the total amount of variation preserved in the population. Genetic variation is only one of many factors that determine the ability of an endangered species to recover. Maintenance of existing genetic diversity, however, remains an important priority for conservation programs because of the possibility of increased disease resistance in more variable populations and the chance that inbreeding depression may only be manifest under adverse environmental conditions.     

Genetic Diversity in the Endangered Lily Harperocallis flava and a Close Relative, Tofieldia racemosa

We examined genetic diversity in 464 individuals of the monotypic lily Harperocallis flava in its two habitats (seepage bogs and a roadside right-of-way) and five populations of a co-occurring related lily, Tofieldia racemosa. The endangered H. flava, endemic to the Apalachicola lowlands of the Florida panhandle, was monomorphic for the 22 loci scored. In contrast, T. racemosa had a high proportion of polymorphic loci ( P_s = 68.2%; P_p = 47.7%) with moderate genetic diversity (   H_es = 0.134; H_ep = 0.114). Estimated gene flow was moderately high ( Nm = 2.07) for T. racemosa, with most (93%) of the total genetic diversity found within populations. Despite the low level of genetic divergence, some isolation by distance was detected among T. racemosa populations. Harperocallis flava and other species without discernable genetic variation pose special problems for conservation biologists because genetic criteria are not available for the development of ex situ and in situ conservation and management strategies.     

Rapid Loss of Genetic Variation in Large Captive Populations of Drosophila Flies: Implications for the Genetic Management of Captive Populations

Levels of variation in eight large captive populations of D. melanogaster (census sizes ∼ 5000) that had been in captivity for periods from 6 months to 23 years (8 to 365 generations) were estimated from allozyme heterozygosities, lethal frequencies, and inversion heterozygosities and phenotypic variances, additive genetic variances ( V _A), and heritabilities ( h ²) for sternopleural bristle numbers. Correlations between all measures of variation except lethal frequencies were high and significant. All measures of genetic variation declined with time in captivity, with those for average heterozygosities, V _A, and h ² being significant. The effective population size ( N _e) was estimated to be 185–253 in these populations, only 0.037–0.051 of census size (N). Levels of allozyme heterozygosities declined rapidly in two large captive populations founded from another wild stock, being reduced by 86% and 62% within 2.5 years in spite of being maintained at sizes of approximately 1000 and 3500. Estimates of N _e/ N for these populations were only 0.016 and 0.004. Two estimates of N _e/ N for captive populations of D. pseudoobscura from data in the literature were also low at 0.036 and 0.012. Consequently, the rate of loss of genetic variation in captive populations and endangered species may be more rapid than hitherto recognized. Merely maintaining captive populations at large census sizes may not be sufficient to maintain essential genetic variation.     

Endangered Greater One-horned Rhinoceros Carry High Levels of Genetic Variation

Abstract: The population of Rhinoceros unicornis in the Chitwan Valley, Nepal, was reduced to an estimated effective population size (N_e of 21–28 individuals (60–80 total animals) in 1962. Protein electrophoresis shows that heterozygosity remains very high in this population (H_o= 9.9%) despite its near extinction. We attribute this high heterozygosity to large N_e's prior to the population bottleneck, the recent occurrence of the bottleneck, and long generation time. These results illustrate the importance of considering historical demography and life history parameters when evaluating the possible genetic effects of bottlenecks in wild populations. They also offer support to recent arguments that the erosion of genetic diversity attributed to bottlenecks may be overemphasized.     

Evaluation of the Contribution of Genetic Research to the Management of the Endangered Plant Zieria prostrata

Abstract: Zieria prostrata (Rutaceae) is known from only four headlands within a 3-km stretch of coastline in New South Wales, Australia. The species was presumed to have occurred at a headland 24 km south of its present range. We used random amplified polymorphic DNA analysis to assess patterns of genetic variation within and among the extant populations. The analysis also included an individual reputedly rescued from the now extinct population. A high level of population divergence was revealed by principal coordinate analysis and an analysis of molecular variance (AMOVA; 37% among populations). Our genetic findings provide implications for the conservation management of the species. First, the loss of any one population would lead to a severe loss of genetic variation. Second, an adequate ex situ collection must sample the full range of genetic diversity from all populations. Third, the consequences of mixing populations may be an important conservation consideration if further translocations proceed. Fourth, the individual apparently sampled prior to its population extinction is genetically similar to individuals from one of the extant sites. This degree of similarity was unexpected and, after further investigation, led to the conclusion that prior existence of the species at the site is doubtful. Subsequently, a planned reintroduction program was abandoned. So far, of these four management implications, only the last has had a direct management outcome. Those implications that failed to lead to practical management outcomes did so because the same management recommendations could be obtained without genetic research. Clearly, the challenge for more effective conservation is to identify those cases in which genetic studies are likely to produce practical outcomes for conservation managers. This may be best achieved by assessing the outcomes of genetic studies already conducted.     

Genetic Diversity of Lepilemur mustelinus ruficaudatus, a Nocturnal Lemur of Madagascar

The genetic polymorphism of natural populations of Lepilemur mustelinus ruficaudatus was studied by protein electrophoresis. We sampled blood from 72 individuals from four populations separated by geographic or anthropogenic barriers from southwestern Madagascar. Six out of 22 enzyme loci showed genetic variation with a degree of polymorphism of 0.273. The expected and observed degree of genetic heterozygosity over all loci is similar to that of other primates (H_e = 0.058, H_o = 0.036). The F-statistics revealed that the four subpopulations were similar with respect to gene structure (F_ST = 0.065, p = 0.016), but the genotypic structures within subpopulations were inconsistent with random mating. For the total of the four subpopulations the proportion of heterozygous individuals was significantly smaller than expected under random mating (F_IS = 0.373, F_IT = 0.414, p < 0.01). These results correspond closely to what is expected considering the low migration ability of individuals of L. m ruficaudatus leading to small and rather isolated inbred populations.     

Developing a Sampling Strategy for Baptisia arachnifera Based on Allozyme Diversity

We analyzed the amount and distribution of genetic variation in Baptisia arachnifera Duncan to develop a sampling strategy for ex situ research. Baptisia arachnifera is an endangered plant species endemic to the coastal plain of Georgia (U.S.) where all populations are within 16 km of each other. A reduction in numbers of individuals has been observed during the last 50 years. Baptisia arachnifera was polymorphic at 24% of the 37 loci examined with an average of 1.32 alleles per locus. The genetic diversity index was relatively low ( H_e = 0.097) as expected for endemic species. Populations were in Hardy-Weinberg equilibrium, suggesting that the species is outcrossing. Consistent with this conclusion is the observation that the majority (approximately 90%) of the genetic variation present in the species is found within individual populations. Indirect evidence of gene flow between populations was detected (   Nm = 2.35). The close proximity of the populations and the recent reduction in population sizes suggest that the populations surveyed may be fragments of a once more continuous gene pool. Based on the observed distribution of genetic diversity among populations (G_ST = 0.096), sampling two populations would capture 99% of the allozyme diversity surveyed. Allozyme data were used to determine which 2 of the 10 populations surveyed should be sampled to maximize the ex situ conservation of genetic diversity. Although the paper-producing companies that own most of the land where Baptisia arachnifera occurs are modifying their harvesting techniques, the species could become extinct without more effective management and preservation efforts.     

Genetic Effects of Multiple Generations of Supportive Breeding

Abstract: The practice of supporting weak wild populations by capturing a fraction of the wild individuals, bringing them into captivity for reproduction, and releasing their offspring into the natural habitat to mix with wild ones is called supportive breeding and has been widely applied in the fields of conservation biology and fish and wildlife management. This procedure is intended to increase population size without introducing exogenous genes into the managed population. Previous work examining the genetic effects of a single generation of supportive breeding has shown that although a successful program increases the census population size, it may reduce the genetically effective population size and thereby induce excessive inbreeding and loss of genetic variation. We expand and generalize previous analyses of supportive breeding and consider the effects of multiple generations of supportive breeding on rates of inbreeding and genetic drift. We derived recurrence equations for the inbreeding coefficient and coancestry, and thereby equations for inbreeding and variance effective sizes, under three models for selecting captive breeders: at random, preferentially among those born in captivity, and preferentially among those born in the wild. Numerical examples indicate that supportive breeding, when carried out successfully over multiple generations, may increase not only the census but also the effective size of the supported population as a whole. If supportive breeding does not result in a substantial and continuous increase of the census size of the breeding population, however, it might be genetically harmful because of elevated rates of inbreeding and genetic drift.     

Predicted Genetic Consequences of Strong Fertility Selection Due to Pollinator Loss in an Isolated Population of Primula sieboldii

Previous studies demonstrated strong fertility selection for a self-fertile, homostyle morph due to pollinator loss in an isolated population of Primula sieboldii , an endangered heterostylous species. To predict genetic consequences of the selection we developed a deterministic genetic model based on a classical "supergene" model, and we studied the effects of pollinator availability and inbreeding depression on temporal changes of morph frequencies through model simulation. Because of the severe pollinator limitation experienced by the population, fast, irreversible loss of the thrum morph from the population was predicted, even if high inbreeding depression was assumed. To prevent the breakdown of the normal breeding system of the species, morph frequency monitoring for timely active management should be implemented. Active management should include hand pollinations and pollinator therapy—reintroduction and reestablishment of suitable pollinator populations. The method we adopted in this study to parametrize pollinator availability can be used widely in conservation modeling for a range of plant species that have multiple mating types with different degrees of self-incompatibility.     

Genetic Effects of Habitat Fragmentation on Common Species of Swiss Fen Meadows

Abstract:  The area of Caricion davallianae alliance in Switzerland has been considerably reduced and fragmented during the last 150 years. We assessed the genetic variability, inbreeding level, and among-population differentiation of two common habitat-specific plant species, Carex davalliana SM. and Succisa pratensis Moench, in 18 Caricion davallianae fen meadows subjected to fragmentation. We used a spatial field design of fen systems (six systems total), each consisting of one large habitat island and two small habitat islands. We used allozyme electrophoresis to derive standard genetic parameters ( A, P, H_O, H_E, F_IS, F_ST ). In Carex we identified a consistently lower A in isolated habitat islands; furthermore, H_E was lower in small habitat islands than in large habitat islands. In Succisa we identified a lower H_O in small habitat islands than in larger ones. Small habitat islands were marginally significantly differentiated (  F_ST ) from large islands for Succisa . For both species, no effects were evident for F_IS ; therefore, we argue that genetic drift rather than inbreeding is the main cause of the observed differences. The genetic structure of Carex and Succisa in small habitat islands differed from that in large habitat islands, but differences were small. It appears that the observed differences in genetic variability among fen meadows correspond to observed differences in fitness and demographic traits. We show that habitat fragmentation affects not only the rare species in an ecosystem but also reduces the survival probabilities of common species. One of the main goals of conservation should be to mitigate fragmentation of natural habitats in order to increase population sizes and connectivity.