An investigation of inbreeding depression and purging in captive pedigreed populations

We use regression models to investigate the effects of inbreeding in 119 zoo populations, encompassing 88 species of mammals, birds, reptiles and amphibians. Meta-analyses show that inbreeding depression for neonatal survival was significant across the 119 populations although the severity of inbreeding depression appears to vary among taxa. However, few predictors of a population's response to inbreeding are found reliable. The models are most likely to detect inbreeding depression in large populations, that is, in populations in which their statistical power is maximised. Purging was found to be significant in 14 populations and a significant trend of purging was found across populations. The change in inbreeding depression due to purging averaged across the 119 populations is <1%, however, suggesting that the fitness benefits of purging are rarely appreciable. The study re-emphasises the necessity to avoid inbreeding in captive breeding programmes and shows that purging cannot be relied upon to remove deleterious alleles from zoo populations.     

Evaluation of ancestral inbreeding coefficients: Ballou’s formula versus gene dropping

Estimation of the purging of detrimental effects through inbreeding and selection is an important issue in conservation genetics opening new perspectives for the management of small populations. In 1997 Ballou proposed the ancestral inbreeding coefficient, which is calculated recursively via pedigree inbreeding coefficients, as a tool for evaluating the purging of deleterious alleles in zoo populations. The formula of Ballou assumes independence of inbreeding and ancestral inbreeding coefficients at any stage of the recursion. This study investigates the consequences of this inaccuracy on the estimation of true ancestral inbreeding, i.e. the proportion of alleles within a genome that has undergone inbreeding in the past. As an alternative we propose the estimation of ancestral inbreeding by the method of gene dropping. The methods are compared by stochastic simulation for various models with respect to mode of inheritance (neutral, detrimental and lethal alleles) and different settings for population size and initial allele frequencies. In all scenarios the proportion of alleles within a genome that has undergone inbreeding in the past was overestimated by Ballou’s formula. The overestimation was more pronounced in smaller populations but was not affected by genetic model or initial allele frequency. In contrast, the ancestral inbreeding coefficient calculated by gene dropping provided a robust estimate of ancestral inbreeding in most models and settings. A marginal overestimation was observed only in models with lethal alleles. Therefore, we recommend applying the gene dropping approach to estimate ancestral inbreeding coefficients.     

SEVERE INBREEDING DEPRESSION AND NO EVIDENCE OF PURGING IN AN EXTREMELY INBRED WILD SPECIES—THE CHATHAM ISLAND BLACK ROBIN

Although evidence of inbreeding depression in wild populations is well established, the impact of genetic purging in the wild remains controversial. The contrasting effects of inbreeding depression, fixation of deleterious alleles by genetic drift, and the purging of deleterious alleles via natural selection mean that predicting fitness outcomes in populations subjected to prolonged bottlenecks is not straightforward. We report results from a long‐term pedigree study of arguably the world's most inbred wild species of bird: the Chatham Island black robin Petroica traversi, in which conditions were ideal for purging to occur. Contrary to expectations, black robins showed a strong, negative relationship between inbreeding and juvenile survival, yielding lethal equivalents (2B) of 6.85. We also determined that the negative relationship between inbreeding and survival did not appear to be mediated by levels of ancestral inbreeding and may be attributed in part to unpurged lethal recessives. Although the black robin demographic history provided ideal conditions for genetic purging, our results show no clear evidence of purging in the major life‐history trait of juvenile survival. Our results also show no evidence of fixation of deleterious alleles in juvenile survival, but do confirm that continued high levels of contemporary inbreeding in a historically inbred population could lead to additional severe inbreeding depression.     

Role of inbreeding depression and purging in captive breeding and restoration programmes

Inbreeding depression is a major force affecting the evolution and viability of small populations in captive breeding and restoration programmes. Populations that experience small sizes may be less susceptible to future inbreeding depression because they have been purged of deleterious recessive alleles. We review issues related to purging, as they apply to the management of small populations, and discuss an experiment we conducted examining purging in populations of mosquitofish (Gambusia affinis). Purging is an important process in many small populations, but the literature contains a diversity of responses to purging both within and among studies. With the exception that slow inbreeding results in more purging and less threat to population viability, there seem to be few consistent trends that aid in prediction of how a purging event will affect a population. In our examination of purging on population viability in mosquitofish, single or multiple bottlenecks do not appear to have resulted in any purging of the influence of genetic load on population growth. Rather, serial bottlenecks resulted in a marked decline in population growth and an increase in extinction. Our results, taken together with those of reviewed studies, suggest that in small populations there is great uncertainty regarding the success of any single purging event in eliminating inbreeding depression, together with the high likelihood that purging will depress population viability through the fixation of deleterious alleles. In management of captive breeding and restoration programmes, the common practice of avoiding inbreeding and small population sizes should be followed whenever possible.     

Population type can influence the magnitude of inbreeding depression in Clarkia concinna (Onagraceae)

Fragmented populations may face high risk of extinction due to the deleterious consequences of increased inbreeding or of genetic drift in small and isolated populations. Theories on the mechanisms of inbreeding depression predict that the severity of inbreeding depression can eventually decrease in populations that persistently inbreed, and hence populations that are isolated through habitat fragmentation might experience a decrease in inbreeding depression over time. In this study, we tested this hypothesis using the patchily distributed, outcrossing annual plant, Clarkia concinna concinna (Onagraceae), which naturally experiences many fragmentation effects. We collected seeds from isolated and central subpopulations and created artificially inbred and outcrossed lines. Progeny from these crosses were planted into the field and greenhouse and assayed for fitness traits over the course of a growing season. Overall, inbreeding depression was substantial, ranging as high as 0.76 (for cumulative fitness in the field), and significant for plant height, fecundity, and above-ground biomass in all experiments. No inbreeding depression was detected for germination or survival rates in the greenhouse experiments, but in the field, survival was significantly depressed for inbred progeny. There was no evidence to support our hypothesis that increased inbreeding in isolated populations would lead to the purging of deleterious alleles and a decrease in the severity inbreeding depression. The most likely hypothesis to explain our results is that purging is not occurring more strongly in the isolated populations due to details of a number of genetic factors (e.g., selection against deleterious alleles is inconsistent or insufficient, or drift has caused fixation of deleterious alleles in these populations). This study supports the view that even when inbreeding depression is predicted to be less problematic, it may still be an important force influencing the fitness of populations. This revised version was published online in July 2006 with corrections to the Cover Date.     

Ancestral inbreeding reduces the magnitude of inbreeding depression in Drosophila melanogaster

The influence of natural selection on the magnitude of inbreeding depression is an important issue in conservation biology and the study of evolution. It is generally expected that the magnitude of inbreeding depression in small populations will depend upon the average homozygosity of individuals, as measured by the coefficient of inbreeding (F). However, if deleterious recessive alleles are selectively purged from populations during inbreeding, then inbreeding depression may differ among populations in which individuals have the same inbreeding coefficient. In such cases, the magnitude of inbreeding depression will partly depend on the ancestral inbreeding coefficient (fa), which measures the cumulative proportion of loci that have historically been homozygous and therefore exposed to natural selection. We examined the inbreeding depression that occurred in lineages of Drosophila melanogaster maintained under pedigrees that led to the same inbreeding coefficient (F = 0.375) but different levels of ancestral inbreeding (fa = 0.250 or 0.531). Although inbreeding depression varied substantially among individual lineages, we observed a significant 40% decrease in the median level of inbreeding depression in the treatment with higher ancestral inbreeding. Our results demonstrate that high levels of ancestral inbreeding are associated with greater purging effects, which reduces the inbreeding depression that occurs in isolated populations of small size.     

Experimental evolution of the genetic load and its implications for the genetic basis of inbreeding depression

The degree to which, and rapidity with which, inbreeding depression can be purged from a population has important implications for conservation biology, captive breeding practices, and invasive species biology. The degree and rate of purging also informs us regarding the genetic mechanisms underlying inbreeding depression. We examine the evolution of mean survival and inbreeding depression in survival following serial inbreeding in a seed-feeding beetle, Stator limbatus, which shows substantial inbreeding depression at all stages of development. We created two replicate serially inbred populations perpetuated by full-sib matings and paired with outbred controls. The genetic load for the probability that an egg produces an adult was purged at approximately 0.45-0.50 lethal equivalents/generation, a reduction of more than half after only three generations of sib-mating. After serial inbreeding we outcrossed all beetles then measured (1) larval survival of outcrossed beetles and (2) inbreeding depression. Survival of outcrossed beetles evolved to be higher in the serially inbred populations for all periods of development. Inbreeding depression and the genetic load were significantly lower in the serially inbred than control populations. Inbreeding depression affecting larval survival of S. limbatus is largely due to recessive deleterious alleles of large effect that can be rapidly purged from a population by serial sib-mating. However, the effectiveness of purging varied among the periods of egg/larval survival and likely varies among other unstudied fitness components. This study presents novel results showing rapid and extensive purging of the genetic load, specifically a reduction of as much as 72% in only three generations of sib-mating. However, the high rate of extinction of inbred lines, despite the lines being reared in a benign laboratory environment, indicates that intentional purging of the genetic load of captive endangered species will not be practical due to high rates of subpopulation extinction.     

Reduced population size can induce quick evolution of inbreeding depression in the invasive ladybird <Emphasis Type="Italic">Harmonia axyridis</Emphasis>

Understanding biological invasion is currently one of the main scientific challenges for ecologists. The introduction process is crucial for the success of an invasion, especially when it involves a demographic bottleneck. A small introduced population is expected to face a higher risk of extinction before the first stage of invasion is complete if inbreeding depression, caused by the expression of deleterious alleles, is important. Changes in mating regimes or in population size can induce the evolution of deleterious allele frequencies, either by selection or by drift, possibly resulting in the purging or the fixation of such alleles within the population. The harlequin ladybird Harmonia axyridis became invasive on several continents following a scenario including at least one event of demographic bottleneck. Although native populations suffered from severe inbreeding depression, it was greatly reduced in invasive ones suggesting that deleterious alleles were purged during the invasion process. In this study, we performed an experiment designed to manipulate the effective population size of H. axyridis across successive generations to mimic contrasting introduction events. We used the measurement of two fitness-related phenotypic traits in order to test (1) if inbreeding depression can evolve at the time-scale of an invasion; and (2) if the changes in inbreeding depression following a bottleneck in laboratory conditions are compatible with the purging of deleterious alleles observed in this species. We found that two generations of very low population size are enough to induce a substantial change in inbreeding depression. Although the genetic changes mostly consisted in fixation of deleterious alleles, purging did also occur, sometimes simultaneously with fixation.     

Temporal change in inbreeding depression in life‐history traits in captive populations of guppy (Poecilia reticulata): evidence for purging?

Inbreeding depression, which generally affects the fitness of small populations, may be diminished by purging recessive deleterious alleles when inbreeding persists over several generations. Evidence of purging remains rare, especially because of the difficulties of separating the effects of various factors affecting fitness in small populations. We compared the expression of life-history traits in inbred populations of guppy (Poecilia reticulata) with contemporary control populations over 10 generations in captivity. We estimated inbreeding depression as the difference between the two types of populations at each generation. After 10 generations, the inbreeding coefficient reached a maximum value of 0.56 and 0.16 in the inbred and control populations, respectively. Analysing changes in the life-history traits across generations showed that inbreeding depression in clutch size and offspring survival increased during the first four to six generations in the populations from the inbred treatment and subsequently decreased as expected if purging occurred. Inbreeding depression in two other traits was weaker but showed similar changes across generations. The loss of six populations in the inbred treatment indicates that removal of deleterious alleles also occurred by extinction of populations that presumably harboured high genetic load.     

Long-term exhaustion of the inbreeding load in Drosophila melanogaster

Inbreeding depression, the decline in fitness of inbred individuals, is a ubiquitous phenomenon of great relevance in evolutionary biology and in the fields of animal and plant breeding and conservation. Inbreeding depression is due to the expression of recessive deleterious alleles that are concealed in heterozygous state in noninbred individuals, the so-called inbreeding load. Genetic purging reduces inbreeding depression by removing these alleles when expressed in homozygosis due to inbreeding. It is generally thought that fast inbreeding (such as that generated by full-sib mating lines) removes only highly deleterious recessive alleles, while slow inbreeding can also remove mildly deleterious ones. However, a question remains regarding which proportion of the inbreeding load can be removed by purging under slow inbreeding in moderately large populations. We report results of two long-term slow inbreeding Drosophila experiments (125–234 generations), each using a large population and a number of derived lines with effective sizes about 1000 and 50, respectively. The inbreeding load was virtually exhausted after more than one hundred generations in large populations and between a few tens and over one hundred generations in the lines. This result is not expected from genetic drift alone, and is in agreement with the theoretical purging predictions. Computer simulations suggest that these results are consistent with a model of relatively few deleterious mutations of large homozygous effects and partially recessive gene action.Subject terms: Quantitative trait, Inbreeding     

Inbreeding rate modifies the dynamics of genetic load in small populations

The negative fitness consequences of close inbreeding are widely recognized, but predicting the long-term effects of inbreeding and genetic drift due to limited population size is not straightforward. As the frequency and homozygosity of recessive deleterious alleles increase, selection can remove (purge) them from a population, reducing the genetic load. At the same time, small population size relaxes selection against mildly harmful mutations, which may lead to accumulation of genetic load. The efficiency of purging and the accumulation of mutations both depend on the rate of inbreeding (i.e., population size) and on the nature of mutations. We studied how increasing levels of inbreeding affect offspring production and extinction in experimental Drosophila littoralis populations replicated in two sizes, N = 10 and N = 40. Offspring production and extinction were measured over 25 generations concurrently with a large control population. In the N = 10 populations, offspring production decreased strongly at low levels of inbreeding, then recovered only to show a consistent subsequent decline, suggesting early expression and purging of recessive highly deleterious alleles and subsequent accumulation of mildly harmful mutations. In the N = 40 populations, offspring production declined only after inbreeding reached higher levels, suggesting that inbreeding and genetic drift pose a smaller threat to population fitness when inbreeding is slow. Our results suggest that highly deleterious alleles can be purged in small populations already at low levels of inbreeding, but that purging does not protect the small populations from eventual genetic deterioration and extinction.     

Inbreeding load, average dominance and the mutation rate for mildly deleterious alleles in Mimulus guttatus

The goal of this study is to provide information on the genetics of inbreeding depression in a primarily outcrossing population of Mimulus guttatus. Previous studies of this population indicate that there is tremendous inbreeding depression for nearly every fitness component and that almost all of this inbreeding depression is due to mildly deleterious alleles rather than recessive lethals or steriles. In this article I assayed the homozygous and heterozygous fitnesses of 184 highly inbred lines extracted from a natural population. Natural selection during the five generations of selfing involved in line formation essentially eliminated major deleterious alleles but was ineffective in purging alleles with minor fitness effects and did not appreciably diminish overall levels of inbreeding depression. Estimates of the average degree of dominance of these mildly deleterious alleles, obtained from the regression of heterozygous fitness on the sum of parental homozygous fitness, indicate that the detrimental alleles are partially recessive for most fitness traits, with h approximately 0.15 for cumulative measures of fitness. The inbreeding load, B, for total fitness is approximately 1.0 in this experiment. These results are consistent with the hypothesis that spontaneous mildly deleterious mutations occur at a rate >0.1 mutation per genome per generation.     

SEXUAL SELECTION COUNTERACTS EXTINCTION OF SMALL POPULATIONS OF THE BULB MITES

Genetic drift in small populations can increase frequency of deleterious recessives and consequently lead to inbreeding depression and population extinction. On the other hand, as homozygosity at deleterious recessives increases, they should be purged from populations more effectively by selection. Sexual selection has been postulated to strengthen selection against deleterious mutations, and should thus decrease extinction rate and intensify purging of inbreeding depression. We tested these predictions in the bulb mite Rhizoglyphus robini. We created 100 replicate lines of small populations (five males and five females) and in half of them experimentally removed sexual selection by enforcing monogamy. The lines were propagated for eight generations and then assayed for purging of inbreeding depression. We found that proportion of lines which went extinct was lower with sexual selection than without. We also found evidence for purging of inbreeding depression in the lines with sexual selection, but not in lines without sexual selection. Our results suggest that purging of inbreeding depression was more effective against mutations with relatively large deleterious effects. Thus, although our data clearly indicate a positive impact of sexual selection on short‐term survival of bottlenecked populations, long‐term consequences are less clear as they may be negatively impacted by accumulation of deleterious mutations of small effect.     

A simple method to account for natural selection when predicting inbreeding depression

It has been widely appreciated that natural selection opposes the progress of inbreeding in small populations, thus limiting the actual inbreeding depression for fitness traits. However, no method to account for the consequences of this process has been given so far. I give a simple and intuitive method to predict inbreeding depression, taking into account the increase in selection efficiency against recessive alleles during inbreeding. It is based on the use of a “purged inbreeding coefficient” g_t that accounts for the reduction of the probability of the deleterious homozygotes caused by the excess d of detrimental effect for deleterious alleles in the homozygous condition over its additive expectation. It is shown that the effect of purging can be important even for relatively small populations. For between-loci variable deleterious effects, accurate predictions can be obtained using the effective homozygous deleterious excess d_e, which can be estimated experimentally and is robust against variation of the ancestral effective population size. The method can be extended to any trait and it is used to predict the evolution of the mean viability or fecundity in a conservation program with equal or random family contributions.     

Royal dynasties as human inbreeding laboratories: the Habsburgs

The European royal dynasties of the Early Modern Age provide a useful framework for human inbreeding research. In this article, consanguineous marriage, inbreeding depression and the purging of deleterious alleles within a consanguineous population are investigated in the Habsburgs, a royal dynasty with a long history of consanguinity over generations. Genealogical information from a number of historical sources was used to compute kinship and inbreeding coefficients for the Habsburgs. The marriages contracted by the Habsburgs from 1450 to 1750 presented an extremely high mean kinship (0.0628±0.009), which was the result of the matrimonial policy conducted by the dynasty to establish political alliances through marriage. A strong inbreeding depression for both infant and child survival was detected in the progeny of 71 Habsburg marriages in the period 1450–1800. The inbreeding load for child survival experienced a pronounced decrease from 3.98±0.87 in the period 1450–1600 to 0.93±0.62 in the period 1600–1800, but temporal changes in the inbreeding depression for infant survival were not detected. Such a reduction of inbreeding depression for child survival in a relatively small number of generations could be caused by elimination of deleterious alleles of a large effect according with predictions from purging models. The differential purging of the infant and child inbreeding loads suggest that the genetic basis of inbreeding depression was probably very different for infant and child survival in the Habsburg lineage. Our findings provide empirical support that human inbreeding depression for some fitness components might be purged by selection within consanguineous populations.     

Dynamics of inbreeding depression due to deleterious mutations in small populations: mutation parameters and inbreeding rate

A multilocus stochastic model is developed to simulate the dynamics of mutational load in small populations of various sizes. Old mutations sampled from a large ancestral population at mutation-selection balance and new mutations arising each generation are considered jointly, using biologically plausible lethal and deleterious mutation parameters. The results show that inbreeding depression and the number of lethal equivalents due to partially recessive mutations can be partly purged from the population by inbreeding, and that this purging mainly involves lethals or detrimentals of large effect. However, fitness decreases continuously with inbreeding, due to increased fixation and homozygosity of mildly deleterious mutants, resulting in extinctions of very small populations with low reproductive rates. No optimum inbreeding rate or population size exists for purging with respect to fitness (viability) changes, but there is an optimum inbreeding rate at a given final level of inbreeding for reducing inbreeding depression or the number of lethal equivalents. The interaction between selection against partially recessive mutations and genetic drift in small populations also influences the rate of decay of neutral variation. Weak selection against mutants relative to genetic drift results in apparent overdominance and thus an increase in effective size (Ne) at neutral loci, and strong selection relative to drift leads to a decrease in Ne due to the increased variance in family size. The simulation results and their implications are discussed in the context of biological conservation and tests for purging.     

THE ROLE OF GENES OF LARGE EFFECT ON INBREEDING DEPRESSION IN MIMULUS GUTTATUS

Severe inbreeding depression is routinely observed in outcrossing species. If inbreeding load is due largely to deleterious alleles of large effect, such as recessive lethals or steriles, then most of it is expected to be purged during brief periods of inbreeding. In contrast, if inbreeding depression is due to the cumulative effects of many deleterious alleles of small effect, then it will be maintained in the face of periodic inbreeding. Whether or not inbreeding depression can be purged with inbreeding in the short term has important implications for the evolution of mating systems and the probability that a small population will go extinct. In this paper I evaluate the extent to which the tremendous inbreeding load in a primarily outcrossing population of the wildflower, Mimulus guttatus, is due to alleles of large effect. To do this, I first constructed a large outbred “ancestral” population by randomly mating plants collected as seeds from a natural population. From this population I formed 1200 lines that were maintained by self-fertilization and single seedling descent: after five generations of selling, 335 lines had survived the inbreeding process. Selection during the line formation is expected to have largely purged alleles of large effect from the collection of highly inbred lines. Because alleles with minor effects on fitness should have been effectively neutral, the inbreeding depression due to this class of genes should have been unchanged. The inbred lines were intercrossed to form a large, outcrossed “purged” population. Finally, I estimated the fitness of outbred and selfed progeny from the ancestral and purged populations to determine the contribution of major deleterious alleles on inbreeding depression. I found that although the average fitness of the outcrossed progeny nearly doubled following purging, the limited decline in inbreeding depression and limited increase in inbred fitness indicates that alleles of large effect are not the principle cause of inbreeding depression in this population. In aggregate, the data suggest that lethals and steriles make a minority contribution to inbreeding depression and that the increased outbred fitness is due primarily to adaptation to greenhouse conditions.     

Levels of inbreeding depression over seven generations of selfing in the androdioecious clam shrimp, Eulimnadia texana

Androdioecy (mixtures of males and hermaphrodites) is a rare mating system in both plants and animals. Theory suggests that high levels of inbreeding depression can maintain males in androdioecious populations if hermaphrodites commonly self-fertilize. However, if inbreeding depression (delta) can be 'purged' from selfing populations, maintaining males is more difficult. In the androdioecious clam shrimp, Eulimnadia texana, delta is estimated to be as high as 0.7. Previous work suggests that this high level is maintained in the face of high levels of inbreeding due to an associative overdominance of fitness-related loci with the sex-determining locus. Such associative overdominance would make purging of inbreeding depression difficult to impossible. The current experiment was designed to determine if delta can be purged in these shrimp by tracking fitness across seven generations in selfing and outcrossing treatments. Evidence of purging was found in one of four populations, but the remaining populations demonstrated a consistent pattern of delta across generations. Although the experimental design allowed ample opportunity for purging, the majority of populations were unable to purge their genetic load. Therefore, delta in this species is likely due to associative overdominance caused by deleterious recessive alleles linked to the sex determining locus.     

Trait specific consequences of fast and slow inbreeding: lessons from captive populations of <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

The increased homozygosity due to inbreeding leads to expression of deleterious recessive alleles, which may cause inbreeding depression in small populations. The severity of inbreeding depression has been suggested to depend on the rate of inbreeding, with slower inbreeding being more effective in purging deleterious alleles of smaller effect. The effectiveness of purging is however dependent on various factors such as the effect of the deleterious, recessive alleles, the genetic background of inbreeding depression and the environment in which purging occurs. Investigations have shown inconclusive results as to whether purging efficiently diminish inbreeding depression. Here we used an ecologically relevant inbreeding coefficient (f ≈ 0.25) and generated ten slow and ten fast inbred lines of Drosophila melanogaster by keeping the effective population size constant at respectively 32 and 2 for 19 or 2 generations. These inbred lines were contrasted to non-inbred control lines. We investigated the effect of inbreeding and inbreeding rate in traits associated with fitness including heat, cold and desiccation stress resistance, egg-to-adult viability, development time, productivity, metabolic rate and wet weight under laboratory conditions. The results showed highly trait specific consequences of inbreeding and generally no support for the hypothesis that slow inbreeding is less deleterious than fast inbreeding. Egg-to-adult viability and development time were investigated under both benign and heat stress conditions. Reduced viability and increased developmental time were observed at stressful temperatures and inbreeding depression was on average more severe at stressful compared to benign temperatures.     

Perspective: purging the genetic load: a review of the experimental evidence

Inbreeding depression, the reduction in fitness that accompanies inbreeding, is one of the most important topics of research in evolutionary and conservation genetics. In the recent literature, much attention has been paid to the possibility of purging the genetic load. If inbreeding depression is due to deleterious alleles, whose effect on fitness are negative when in a homozygous state, then successive generations of inbreeding may result in a rebound in fitness due to the selective decrease in frequency of deleterious alleles. Here we examine the experimental evidence for purging of the genetic load by collating empirical tests of rebounds in fitness-related traits with inbreeding in animals and plants. We gathered data from 28 studies including five mammal, three insect, one mollusc, and 13 plant species. We tested for purging by examining three measures of fitness-component variation with serial generations of inbreeding: (1) changes in inbreeding depression, (2) changes in fitness components of inbred lines relative to the original outbred line, and (3) purged population (outcrossed inbred lines) trait means as a function of ancestral outbred trait means. Frequent and substantial purging was found using all three measures, but was particularly pronounced when tracking changes in inbreeding depression. Despite this, we found little correspondence between the three measures of purging within individual studies, indicating that the manner in which a researcher chooses to estimate purging will affect interpretation of the results obtained. The discrepancy suggests an alternative hypothesis: rebounds in fitness with inbreeding may have resulted from adaptation to laboratory conditions and not to purging when using outcrossed inbred lines. However, the pronounced reduction in inbreeding depression for a number of studies provides evidence for purging, as the measure is likely less affected by selection for laboratory conditions. Unlike other taxon-specific reviews on this topic, our results provide support for the purging hypothesis, but firm predictions about the situations in which purging is likely or the magnitude of fitness rebound possible when populations are inbred remain difficult. Further research is required to resolve the discrepancy between the results obtained using different experimental approaches.