The importance of mating system in translocation programs: reproductive success of released male bridled nailtail wallabies

Translocation is an important tool for the conservation of species that have suffered severe range reductions. The success of a translocation should be measured not only by the survival of released animals, but by the reproductive output of individuals and hence the establishment of a self-sustaining population. The bridled nailtail wallaby is an endangered Australian macropod that suffered an extensive range contraction to a single remaining wild population. A translocated population was established and subsequently monitored over a four year period. The aim of this study was to measure the reproductive success of released males using genetic tools and to determine the factors that predicted reproductive success. Captive-bred and wild-caught animals were released and we found significant variation in male reproductive success among release groups. Variation in reproductive success was best explained by individual male weight, survival and release location rather than origin. Only 26% of candidate males were observed to sire an offspring during the study. The bridled nailtail wallaby is a sexually dimorphic, polygynous macropod and reproductive success is skewed toward large males. Males over 5800 g were six times more likely to sire an offspring than males below this weight. This study highlights the importance of considering mating system when choosing animals for translocation. Translocation programs for polygynous species should release a greater proportion of females, and only release males of high breeding potential. By maximizing the reproductive output of released animals, conservation managers will reduce the costs of translocation and increase the chance of successfully establishing a self-sustaining population.     

Effects of supplementary feeding on the offspring sex ratio of kakapo: a dilemma for the conservation of a polygynous parrot

The kakapo (Strigops habroptilus) is a large, flightless, nocturnal parrot, endemic to New Zealand. It is critically endangered, with a world population of ca. 62 individuals and a male-biased adult sex ratio. The species has a polygynous “lek” mating system and adult males typically weigh 30-40% more than females. The kakapo is subject to intensive conservation efforts, including the provision of supplementary food to wild birds to encourage successful nesting. There is mounting evidence that, in polygynous species with large variance of male reproductive success, females in better condition may maximise their fitness by producing more offspring of the larger, more costly sex to be reared. We used data on the sex ratio of progeny of female kakapo that had or had not received supplementary food, to test the hypothesis that supplementary feeding might cause a male-biased offspring sex ratio. There was a significant excess of males in the clutches of females provided with supplementary food, suggesting that changes need to be made to the feeding regime to increase recruitment of females. This is an example of applying evolutionary theory to a practical conservation problem.     

Sperm storage and low incidence of multiple paternity in the European pond turtle, Emys orbicularis: A secure but costly strategy?

The freshwater pond turtle, Emys orbicularis, has recently suffered from population declines throughout its range, mainly due to habitat destruction. The mating strategies of this species were studied using genetic data from successive clutches within and between years. To test for the occurrence and frequency of multiple paternity and sperm storage, genetic paternity at six microsatellite markers was assessed in 114 embryos and hatchlings from single and subsequent clutches of 11 females (including clutches from the same or consecutive years). Multiple paternity was rare and only found in two out of 20 clutches from 11 females. All annual successive clutches and 58% of the clutches in the next year, were fertilized with sperm from the same male. The use of stored sperm is thus a frequent strategy in E. orbicularis. However, hatching rate, hatchling mass, and hatchling length decreased in clutches fertilized by stored sperm, suggesting sperm depletion or deterioration through time. The occurrence of stored sperm despite an associated reduced reproductive output indicated that mating and/or the fertilization process is costly to females. The low incidence of multiple paternity may simply be the residual consequence of the capacity to store viable sperm. These results provide important and innovative insights for the conservation of E. orbicularis. In threatened populations, management strategies may aim to enhance effective copulations in order to increase the reproductive output of females.     

Stochastic variation in reproductive success of a rare frog, Rana sevosa: implications for conservation and for monitoring amphibian populations

Although amphibian populations are thought to be declining in many parts of the world, detailed information on populations in decline are often not available. From 1988 to 2001, we studied temporal variation in the reproductive biology of the only known population of dusky gopher frogs, Rana sevosa Goin and Netting. We found high annual variation in reproductive effort, mortality at the egg and larval stages, and hydroperiod length. No overall trends were apparent in terms of either number of egg masses deposited or in reproductive success, as we found extensive variation among years in the number of egg masses deposited, a high rate of reproductive failure, and no consistent relationship between the number of females present, the number of eggs deposited, and the number of metamorphs emerging. Given the complete isolation of this population from other gopher frogs and the high rate of reproductive failure, the probability of extinction of this population appears to be quite high (0.125-0.316).     

Long-term apparent survival of translocated gopher tortoises: A comparison of newly released and previously established animals

Most turtle species require high adult survivorship to maintain stable populations. Translocations are often implemented to conserve turtle populations but may cause demographic disturbance as a result of increased mortality or dispersal of released animals. The gopher tortoise (Gopherus polyphemus) is one of the most frequently translocated turtle species. Short-term monitoring indicates that dispersal by released tortoises is common, but few long-term data are available to determine if losses of translocated animals continue for multiple years. We used 12 years of mark-recapture data to investigate long-term apparent survival of two groups of gopher tortoises translocated during separate periods to St. Catherines Island, Georgia, USA. We analyzed capture histories in program MARK to compare apparent survival of newly released tortoises and previously established translocated tortoises and also to determine whether apparent survival varied with sex or maturity. Apparent annual survival did not vary between adult males and females (0.98 ± 0.01), but was lower in sexually immature tortoises (0.84 ± 0.05). We documented a temporary reduction in apparent survival of newly released adult (0.75 ± 0.06) and immature tortoises (0.45 ± 0.26) during the first year after release that may be attributed in part to permanent dispersal. However, for both maturity classes, apparent survival of newly released tortoises was consistently high and matched that of previously established animals during the remainder of the study. Additional long-term studies of both translocated and naturally-occurring populations are needed to improve management of remaining tortoise populations.     

Conservation of the red-winged grasshopper, Oedipoda germanica (Latr.): the influence of reproductive behaviour

Reproductive isolation can function as a mechanism to maintain locally adapted gene complexes while decreasing the heterozygosity in distinct populations. As a result, reproductive behaviour should be considered a fundamental factor influencing reproductive isolation. This is of interest to conservation biology when one desires to regulate gene flow between two populations either by creating opportunities for increased dispersal, by relocation of individuals, or by re-introduction of a species to its natural habitat. Reproductive behaviour can also influence the effective population size and the actual population size through the Allee effect. We investigated the reproductive behaviour of individuals from two isolated populations of the red winged grasshopper, Oedipoda germanica, an endangered species in Central Europe. We detail several methods to show how several aspects of the reproductive behaviour of this species interact with the conservation of this species. Foreign males were not disfavoured in mate choice and male body size was also unimportant in mating success. Heterogamic matings were as productive as homogamic matings in terms of total number of eggs per female, egg hatching rate, or nymph survival. Therefore, we suggest that cross-matings of individuals from different populations do not positively influence population size by heterosis effects nor act they negatively in the form of outbreeding depression. We found that female O. germanica were able to store viable sperm for extended periods but egg pods showed a decrease in hatching rate when these females were deprived of additional mating opportunities. Multiple mated females laid more eggs than once-mated females. Hence, females are capable of founding new populations even after only one mating but their reproductive output may be reduced. Present-day gene flow between populations of O. germanica probably does not occur. Consequently, there is an urgent need to pursue habitat management and release programmes that maintain current population sizes of this species. With regard to the reproductive behaviour of the red-winged grasshopper, we suggest that relocation programmes release males and female together and at an early adult stage. Also, because female reproductive output increases with mating activity, the initial release should involve excess females. Because males have a shorter lifespan than females a subsequent release of males at a later time may enhance the overall success of local conservation efforts by resulting in more offspring per female.     

Return to the wild: Translocation as a tool in conservation of the Desert Tortoise (Gopherus agassizii)

Translocation could be used as a tool in conservation of the threatened Mojave Desert Tortoise (Gopherus agassizii) by moving individuals from harm’s way and into areas where they could contribute to conservation of the species. Numerous factors may affect the success of translocations, including the conditions experienced by tortoises in holding facilities while awaiting translocation. The tortoises available for our translocation study had been provided supplemental water during their years spent in a captive holding facility, potentially inducing carelessness in water conservation. In addition to generally investigating the efficacy of translocation, we compared the effects of continuing with the effects of ceasing the holding facility’s water supplementation regimen. After exposure to one of the two water regimens, all tortoises were given the opportunity to hydrate immediately prior to release. We examined behavior, body mass, carapace length, movement, and mortality of tortoises for two activity seasons following release to the wild. Water supplementation was correlated with high rates of carapace growth and distant movements by males after release. Lengthy movements following translocation may be problematic for conservation planning, but this should be evaluated in light of the goals and circumstances of each translocation project. Although the mortality rate was 21.4% in 1997, data suggest that drought conditions at the site rather than the translocation itself negatively affected the tortoises. None of the tortoises died during their second season at the site. Our results indicate that translocation should be considered a useful tool in conservation of the Desert Tortoise.     

Sampling on private property to evaluate population status and effects of land use practices on the gopher tortoise, Gopherus polyphemus

Although private properties are predicted to play an increasingly significant role in conservation, surveys of species of special concern are rare on these lands. We created a template for a multi-county survey of randomly selected sites and sampled for burrows of the gopher tortoise (Gopherus polyphemus) in south-central Georgia, USA. Current land use was strongly correlated with tortoise population condition. The highest densities of tortoise burrows were found on lands with open-canopied pine stands that were managed with prescribed fire, a practice associated with types of selection forestry and/or wildlife management. Agricultural sites and unburned areas provided poor habitat and pine plantations were only slightly better. Our estimates of tortoise population densities indicated that the current landscape supports less than 20% of the animals present before implementation of modern land use practices. In addition, our estimate for density of active burrows was approximately one third of that projected for the entire state range 20 years ago by Auffenberg and Franz [Auffenberg, W., Franz, R., 1982. The status and distribution of the gopher tortoise (Gopherus polyphemus). In: Bury, R.B. (Ed.), North American Tortoises: Conservation and Ecology (US Fish and Wildlife Service Wildlife Research Report 12). pp. 95-126]. However, some good sites for gopher tortoises remain in south Georgia and our data also suggested that extraordinary conservation actions may not be required if ways can be developed to retain traditional land management practices on private property.     

Life history traits and food supplementation affect productivity in a translocated population of the endangered Hihi (Stitchbird, Notiomystis cincta)

We studied the effect of food supplementation, female age and clutch order on productivity in a translocated species. Food supplementation increased clutch size from 3.9 to 4.4 (average) eggs per nesting attempt, and more than doubled fledging and recruitment success. Supplemented females started a second clutch 9 days sooner after fledging first clutch chicks than unfed females. During second clutches, supplemented females incubated the eggs for a shorter period of time (15.2 days vs. 16.8). Older females laid larger clutches (4.6 vs. 3.7 eggs) than yearlings and incubated second clutch eggs for a shorter period (15.4 vs. 16.6 days). Females laid more eggs in first clutches (4.2 vs. 3.8 eggs), and those eggs took approximately 30% longer to lay than eggs in second clutches. The successful maintenance of hihi populations on the available islands may be dependent on the permanent provision of supplemental food at nest sites.     

Florida panther dispersal and conservation

We studied dispersal in 27 radio-collared Florida panthers Puma concolor coryi in southern Florida from 1986 to 2000. Male dispersal was longer (mean=68.4 km) than that of females (mean=20.3 km), tended to be circular, frustrated, and of insufficient length to ameliorate inbreeding. Females were philopatric and established home ranges that were less than one home range width away from their natal ranges. All females were successful in establishing territories, whereas males were successful 63% of the time. Dispersing panthers avoided moving toward the southeast and into an area of limited forest cover. Independence and the initiation of dispersal occurred at about 14 months of age and lasted for an average of 7.0-9.6 months for females and males, respectively. On average, Florida panthers disperse shorter distances than are typical for western populations of Puma concolor. A recent increase in long distance male dispersal events may be related to an increase in reproduction and population density resulting from the introduction of female cougars P. c. stanleyana into south Florida. Although the population exhibits the behavioral ability to colonize nearby vacant range, females have yet to do so. Successful dispersal to these areas could be facilitated by habitat restoration and translocation of females.     

Hybridization Between Wild and Domesticated Types of Phaseolus vulgaris L. (Fabaceae) in Argentina

A polymorphic population of Phaseolus vulgaris L. var. aborigineus (Burkart) Baudet growing in northwestern Argentina was studied. In order to know the origin of this polymorphism, plants belonging to the var. aborigineus, others showing floral dimorphism and individuals with weedy type characters were collected. Their seeds, obtained after fieldwork treatments of autogamy and free pollination in order to know their reproductive system, were sown in a greenhouse. Information of their growth was recorded and several numerical analyses were performed. With the results obtained, we concluded that those individuals that showed floral dimorphism are probably a result of hybridization and/or introgression between the var. aborigineus and cultivated forms that grow in the area. This hypothesis is supported by the presence of divergent segregation in the offspring. Also, the offspring of the F1 was followed and allowed us to conclude the possible existence of degeneration of the hybrid progeny. This degeneration indicates that an unwanted gene flow in the area could lead to a decline in the wild bean population. The vigour and high reproductive success of the offspring belonging to the individuals corresponding to the var. aborigineus, whose progenitor was treated for outcrossing, and the existence of plants with weedy type characteristics, are indicative of the necessity to preserve this germplasm in order to evaluate its agronomic potential as brief as possible. The population here studied constitutes a wild-weed–crop complex growing in Argentina.     

Hatch size, somatic growth rate and size-dependent survival in the endangered ploughshare tortoise

Successful conservation requires a good understanding of both the direct and indirect causes of any decline in population size. Harvests of wild populations often target the largest, oldest individuals that have the greatest economic value. If these individuals contribute disproportionately more to recruitment than conspecifics, the harvest will cause a greater reduction in population viability than initially anticipated. The ploughshare tortoise, endemic to Madagascar, has been reduced to <600 individuals due to exploitation and habitat loss. A trial release of captive-bred juveniles has been carried out but the relative suitability of individuals for release, in terms of body size, was not considered. Using data from a long-term mark-recapture study of an unharvested wild population of ploughshare tortoises, we found that larger juveniles were significantly more likely to survive their first year of life than smaller tortoises. Juveniles that survived beyond their first year of life generally grew at the same rate, but had a significantly larger hatch size (mean = 41.7 mm), compared with juveniles that did not survive (mean = 39.3 mm). The conservation implications of these results are that release of captive-bred ploughshare tortoises is likely to be more successful if larger individuals are released. Previous studies have reported larger female ploughshare tortoises laying larger eggs which hatch larger juveniles. This study shows individuals with a larger hatch size retain their size advantage over smaller conspecifics and are more likely to survive their first year of life. This suggests a harvest that targets the largest females in the ploughshare tortoise population could be highly detrimental to population viability.     

An analysis of the role of local fishermen in the conservation of the loggerhead turtle (Caretta caretta) in Pontal do Ipiranga, Linhares, ES, Brazil

The role of researchers and fishermen in the clutch management of loggerhead sea turtles was evaluated for 10 nesting seasons at Pontal do Ipiranga TAMAR station, Linhares, ES. The comparison of nests transferred by researchers and locals (carebeiros) showed that clutches transferred by researchers presented higher clutch size. Clutch size between nests transferred by carebeiros and left in situ did not show significant differences. Hatching success was significantly higher for clutches left in situ than for those transferred to other places in the beach or to hatcheries. The clutches transferred exclusively by researchers achieved a higher hatching success than those transferred partially or totally by carebeiros. The relocation time of clutches collected by carebeiros and handed to researchers affected hatching success. It is recommended that clutches be left in situ, provided they have adequate conditions for monitoring, but careful clutch translocation, independent of the interval elapsed after laying, may also constitute a viable technique for the conservation of sea turtles in the region.     

Improving density estimates for elusive carnivores: Accounting for sex-specific detection and movements using spatial capture–recapture models for jaguars in central Brazil

Owing to habitat conversion and conflict with humans, many carnivores are of conservation concern. Because of their elusive nature, camera trapping is a standard tool for studying carnivores. In many vertebrates, sex-specific differences in movements – and therefore detection by cameras – are likely. We used camera trapping data and spatially explicit sex-specific capture–recapture models to estimate jaguar density in Emas National Park in the central Brazilian Cerrado grassland, an ecological hotspot of international importance. Our spatially explicit model considered differences in movements and trap encounter rate between genders and the location of camera traps (on/off road). We compared results with estimates from a sex-specific non-spatial capture–recapture model. The spatial model estimated a density of 0.29 jaguars 100 km⁻² and showed that males moved larger distances and had higher trap encounter rates than females. Encounter rates with off-road traps were one tenth of those for on-road traps. In the non-spatial model, males had a higher capture probability than females; density was estimated at 0.62 individuals 100 km⁻². The non-spatial model likely overestimated density because it did not adequately account for animal movements. The spatial model probably underestimated density because it assumed a uniform distribution of jaguars within and outside the reserve. Overall, the spatial model is preferable because it explicitly considers animal movements and allows incorporating site-specific and individual covariates. With both methods, jaguar density was lower than reported from most other study sites. For rare species such as grassland jaguars, spatially explicit capture–recapture models present an important advance for informed conservation planning.     

Management implications of capybara (Hydrochoerus hydrochaeris) social behavior

Capybaras (Hydrochoerus hydrochaeris) are the world’s largest rodent. Free-living populations are commercially harvested for their meat and leather in Colombia, Venezuela and Argentina; however, there is concern that legal and illegal harvesting is not sustainable. Since capybaras are considered an economic resource, there have been several attempts to explore the effect of different hunting strategies on its population dynamics. Two previous population models have been developed with this goal; however neither included capybara social behavior that may affect population dynamics. We developed an age-structured, density-dependent model of capybara herd dynamics to explore the demographic consequences of different hunting strategies. We then added infanticide and female reproductive suppression to explore the demographic consequences of such behavior. We conducted five different simulations and used ANOVA to estimate the effect of hunting females, hunting males, hunting both males and females, and the independent effects of reproductive suppression and infanticide on population size after 50 years. Our model suggests that suppression has the largest effect on population size, followed by hunting females and males hunting, female hunting, male hunting and infanticide. Thus, to develop more realistic harvesting models, managers should determine the degree of reproductive suppression and the frequency of infanticide by males.     

Polygyny and female breeding failure reduce effective population size in the lekking Gunnison sage-grouse

Populations with small effective sizes are at risk for inbreeding depression and loss of adaptive potential. Variance in reproductive success is one of several factors reducing effective population size (N_e) below the actual population size (N). Here, we investigate the effects of polygynous (skewed) mating and variation in female breeding success on the effective size of a small population of the Gunnison sage-grouse (Centrocercus minimus), a ground nesting bird with a lek mating system. During a two-year field study, we recorded attendance of marked birds at leks, male mating success, the reproductive success of radio-tagged females, and annual survival. We developed simulations to estimate the distribution of male reproductive success. Using these data, we estimated population size () and effective population size N_e for the study population. We also simulated the effects of population size, skewed vs. random mating, and female breeding failure on N_e. In our study population, the standardized variance in seasonal reproductive success was almost as high in females as in males, primarily due to a high rate of nest failure (73%). Estimated N_e (42) was 19% of in our population, below the level at which inbreeding depression is observed in captive breeding studies. A high hatching failure rate (28%) was also consistent with ongoing inbreeding depression. In the simulations, N_e was reduced by skewed male mating success, especially at larger population sizes, and by female breeding failure. Extrapolation of our results suggests that six of the seven extant populations of this species may have effective sizes low enough to induce inbreeding depression and hence that translocations may be needed to supplement genetic diversity.     

Captive-reared burrowing owls show higher site-affinity, survival, and reproductive performance when reintroduced using a soft-release

A remaining challenge for animal reintroductions is how best to aid individuals transitioning from captivity into the wild. We tested two techniques – ‘soft’ vs. ‘hard’ release – in a 7-year study on endangered Western burrowing owls (Athene cunicularia hypugaea) in British Columbia, Canada. Traditionally, captive-reared yearling adults were released into the wild, as pairs, directly from artificial burrows (hard-release). Only 46% of 201 owls released in this way (2001–2004) stayed at release sites. To test for improved success, soft-releases were performed in the same manner as concurrent hard-releases (2005–2007), except above-ground enclosures confined each soft-release pair to the vicinity of their burrow for 2 weeks before complete release. Of 140 soft-released owls, 86% stayed at release sites; whereas, 66% of 100 hard-released owls stayed. Breeding-season survival was 70% for soft-released owls vs. 50% for hard-released owls. On average, soft-released owls produced 50% more fledglings than concurrently hard-released owls. Post-fledging survival was 69% and first-year return rate was 7.0% for fledglings from soft-released parents, compared to 50% and 4.0% for fledglings from hard-release parents. Ultimately, 0.17 offspring were recruited into the local wild breeding population per soft-released pair, compared to 0.05 local recruits per hard-released pair. We recommend that enclosure-based soft-releases be used for reintroduction of burrowing owls in British Columbia and elsewhere in North America. We encourage other tests for improved release success with soft-releases, particularly for species with a high tendency for dispersal or those likely to experience significant predation pressure.     

Assessing translocation outcome: Comparing behavioral and physiological aspects of translocated and resident African elephants (Loxodonta africana)

Evaluating translocation outcomes is important for improving wildlife management and conservation actions. Often, when quick decisions need to be made and long-lived animals with slow reproduction rates are translocated, traditional assessment methods such as long-term survival and reproductive success cannot be used for assessing translocation outcomes. Thus, alternative, seldom used, measures such as comparing the behavior and physiology of translocated animals to those of local residents should be employed to assess the translocated animals’ acclimation to their new home. Here we monitored the survival, physiology, and behavior of translocated African elephants (Loxodonta africana) and compared these measures to the local resident population at the release site. Adult male and female translocated elephants’ death rates were higher than those of the local population. Furthermore, the mortality rate of translocated adult males and calves was greater than expected based on their proportion in the translocated elephant population. No difference was found in stress hormone levels between the two populations, but the body condition of the translocated elephants was significantly poorer than that of the local population throughout the study period. The behavioral time budgets of the translocated elephants converged with those of the local population over time. Finally, translocated elephants utilized habitat that was similar to their source site (hills and permanent rivers) more than did the local population. Based on these findings we recommend careful consideration of timing, release location, and individuals targeted in future elephant translocations. More broadly, we introduce and explore seldom used translocation assessment techniques.     

The use of shell morphometrics for the management of the endangered malagasy radiated tortoise (Geochelone radiata)

The radiated tortoise (Geochelone radiata) is an endangered species endemic to the spiny forest of southern Madagascar. The main threat that this species faces is illegal harvesting for both local consumption and foreign markets. Large numbers of live individuals are often confiscated and their population of origin is in most cases unknown. Previous work has identified three genetically distinct units across the current distribution of the species. The objective of the present study was to develop a method of assigning G. radiata individuals to their original population based on the analysis of morphometric characters. We measured 39 variables on the shell of tortoises sampled at seven locations across the species range in order to assess differentiation. We employed a PCA-based method to analyze morphometric data and found three distinct groups of populations based solely on shape variation. Discriminant functions allowed for the correct classification of 83% of individuals among these groups. The abundance of tortoises, a variable strongly linked to the level of human exploitation at each site, was correlated with the morphometric differentiation of populations. Our results thus suggest that humans may have played a role in the recent evolution of radiated tortoises. The outlined methodology could become a useful tool for the management of radiated tortoises, and could theoretically be applied to any chelonian species.     

Pragmatic management increases a flagship species, the Himalayan brown bears, in Pakistan’s Deosai National Park

Deosai Plateau in Northern Pakistan was designated a national park to protect the largest remnant population of brown bears in Pakistan. The natural resources of this high elevation (3500-5000 m) park make a significant contribution to the livelihood of local and nomad communities. The present legislation excludes people from a park, which increases conflicts between management and local people. However, a pragmatic approach was adopted to involve people in conservation in Deosai. Community participation, achieved by recognizing rights and introducing incentives, reduced resistance against the conservation efforts, reduced grazing pressure in bear habitat and helped reduce poaching. The size of the brown bear population was set as an indicator of park success, and was monitored annually from 1993 through 2006. We observed a 5% annual growth of the brown bear population, suggesting that the conservation program has been successful due to a successful cooperation between an NGO, people, and the park management.The increase of the bear population is significant, because we observed an extremely low reproductive rate, due to late age of first reproduction (8.25 years), a long reproductive interval (5.7 years), and a small litter size (1.33). The reproductive rate of the Deosai population is the lowest yet documented for brown bear populations. Poor habitat quality, low quality food, high seasonality, and extreme weather conditions in the Himalaya probably explain the poor reproductive performance. Considering such low reproduction and known exchange of individuals with neighboring populations, we believe that the observed growth was a sum of reproduction and immigration. Brown bears are declining throughout South Asia and often have low-productive rates. Therefore, conservation efforts for brown bears in this region must target reducing human-caused mortalities, particularly of adult females. Involvement of people can increase efficiency in conservation, in addition to reducing cost and conflicts.