Effective Number Of Founder Genomes Research Articles

Evaluation of inbreeding and genetic diversity in Japanese Shorthorn cattle by pedigree analysis.

The Japanese Shorthorn is a Japanese Wagyu breed maintained at a small population size. We assessed the degree of inbreeding and genetic diversity among Japanese Shorthorn cattle using pedigree analysis. We analyzed the pedigree records of registered Japanese Shorthorn born between 1980 and 2018, after evaluating the pedigree completeness. The average of the actual inbreeding coefficients increased at the same rates annually from approximately 1.5% in 1980 to 4.2% in 2018 and was higher than the expected inbreeding coefficients over time. The effective population size based on the individual coancestry rate largely decreased from 127.8 in 1980 to 82.6 in 1999, and then remained almost constant at approximately 90. Three effective numbers of ancestors decreased over time until 1995, then remained almost constant. In particular, the effective number of founder genomes (N ge ) decreased from 43.8 in 1980 to 11.9 in 2018. The index of genetic diversity based on N ge decreased from 0.99 in 1980 to 0.96 in 2018 due to genetic drift in non‐founder generations. Changes in inbreeding and genetic diversity parameters were similar between Japanese Shorthorn and other Japanese Wagyu breeds, but the magnitude of the changes was lower in the Japanese Shorthorn.

Assessment of the genetic diversity of Two Czech autochthonous sheep breeds

Based on pedigree information, the population structure, within-breed genetic variability and causes of genetic variability losses in Wallachian sheep (WS), and Sumava sheep (SS) were studied. The maximum number of generations observed in both WS and SS was 16, and the equivalent number of complete generations 5.66 and 4.35, respectively. The average generation intervals were in WS and SS 4.33 and 4.72 years, respectively. A span of four years, 2015–2018, was used as the reference population (animals from the last generation). The average inbreeding coefficient and average relatedness coefficient were 5 % and 9 % for WS, while both were 3 % for SS. The rate of the inbreeding coefficient was 0.5 % in WS and 1 % in SS. The effective number of founders, effective number of ancestors and effective number of founder genomes were 104, 50 and 22.8 for SS and 39, 21 and 10.28 for WS. The effective population size based on an increase in the inbreeding coefficient and computed based on average coancestry were 50.61 and 64.5 in WS and 99.56 and 166.14 in SS. These results indicate a low value of genetic diversity in Wallachian sheep. The loss of variability was caused by the bottleneck effect and partly also genetic drift and the unequal contribution of the founders.

Genetic diversity evolution in the Mexican Charolais cattle population.

ObjectiveThe aim was to characterize the genetic diversity evolution of the registered Mexican Charolais cattle population by pedigree analysis.MethodsData consisted of 331,390 pedigree records of animals born from 1934 to 2018. Average complete generation equivalent, generation interval, effective population size (Ne), and effective numbers of founders (fe), ancestors (fa), and founder genomes (Ng) were calculated for seven five-year periods. The inbreeding coefficient was calculated per year of birth, from 1984 to 2018, whereas the gene contribution of the most influential ancestors was calculated for the latter period.ResultsAverage complete generation equivalent consistently increased across periods, from 4.76, for the first period (1984 through 1988), to 7.86, for the last period (2014 through 2018). The inbreeding coefficient showed a relative steadiness across the last seventeen years, oscillating from 0.0110 to 0.0145. During the last period, the average generation interval for the father-offspring pathways was nearly 1 yr. longer than that of the mother-offspring pathways. The effective population size increased steadily since 1984 (105.0) and until 2013 (237.1), but showed a minor decline from 2013 to 2018 (233.2). The population displayed an increase in the fa since 1984 and until 2008; however, showed a small decrease during the last decade. The effective number of founder genomes increased from 1984 to 2003, but revealed loss of genetic variability during the last fifteen years (from 136.4 to 127.7). The fa:fe ratio suggests that the genetic diversity loss was partially caused by formation of genetic bottlenecks in the pedigree; in addition, the Ng:fa ratio indicates loss of founder alleles due to genetic drift. The most influential ancestor explained 1.8% of the total genetic variability in the progeny born from 2014 to 2018.ConclusionInbreeding, Ne, fa, and Ng are rather beyond critical levels; therefore, the current genetic status of the population is not at risk.

Assessing genetic diversity of various Canadian sheep breeds through pedigree analyses

The loss of genetic variability in a population will drastically affect the success of a breeding program by reducing selection response and fitness and, consequently, affecting reproduction, resilience, and production efficiency. The objective of this study was to perform an in-depth analysis of the pedigree of the Canadian sheep breeds to assess the levels of inbreeding, effective population size, and other metrics of genetic diversity, which included the five most important sheep breeds in Canada: Dorset, Polypay (PO), Rideau-Arcott, Romanov (RV), and Suffolk, using a large dataset (1 336 926 animals). As measures of genetic diversity, effective population size, inbreeding coefficient, effective number of founders, effective number of founder genomes, effective number of nonfounders, and effective number of ancestors were estimated. The completeness and depth of the Canadian sheep pedigree datasets were reasonably high, with <20% parental information missing. More attention should be given to PO breed, which was found to have the smallest effective population size (55), and RV breed, which had the highest average level of inbreeding (4.8%). Techniques such as optimum contribution selection and minimum coancestry mating could be used to minimize the inbreeding of future generations, while maintaining genetic progress at a desirable level.

Genetic Diversity of Lusitano Horse in Brazil Using Pedigree Information

This study aimed to evaluate population parameters and to describe the genetic diversity of the Lusitano breed in Brazil using pedigree data. Two populations were evaluated: total population (TP) containing 18,922 animals, and reference population (RP) composed of a part of TP containing 8,329 animals, representing the last generation. The generation interval (10.1 ± 5.1 years) was in the range for horse populations. Pedigree completeness in RP shows almost 100% filling in the three most recent generations, indicating improvement in the pedigree data and accuracy of the results, and the inbreeding coefficient (4.46%) and average relatedness (5.97%) for RP, indicating control on the part of breeders. The effective population size was 89 (TP) and 90 (RP). The effective number of founders (fe) were 33 and 29, effective number of ancestors (fa) were 30 and 26, and effective number of founder genomes (fg) were 19 and 15 for TP and RP, respectively, indicating a reduction of genetic variability in the last generations. The total number of ancestors that explains 100% of the genetic diversity in the Lusitano breed in Brazil was 427 (TP) and 341 (RP). The reproductive parameters, probabilities of gene origin showing loss of variability in the last generations, and the genetic contributions of ancestors suggest the need to monitor genetic diversity over time in breeding programs to allow control of the next generations and to increase their variability.

Inbreeding and Mortality Risk in a Captive Population of Mhorr Gazelle (Nanger dama mhorr)

This study aimed to evaluate the genetic variation and examine the association of inbreeding level on mortality risk (at days 7, 30 and 180 – weaning age) of Mhorr gazelle in captivity for the year 1969–2000 as recorded in the studbook record kept by Australasian Regional Association of Zoological Parks and Aquaria (ARAZPA). The effective number of founders, ancestors and founder genomes was found to be 3.42, 3, and 1.44 for the studied reference population. The reference population is composed of animals which are alive, with known parents and known sex. Animals that are less than 10 years old (based on birth dates up to 2008) with no remarks on its death are considered alive. The population may not have experienced a severe bottleneck, as the values on the effective number of founders and ancestors are almost equal. However, the effective number of founder genomes is low, which demonstrates gene loss due to genetic drift. The mean inbreeding coefficients of the individual, sire and dam were found to be 0.2971 ± 0.1043, 0.2300 ± 0.1141 0.2339 ± 0.1070, respectively. The maximum inbreeding level of the population is 0.5247 (52.47%). This means that parent–offspring or full-sib mating must have happened. The increase in inbreeding level of an individual was found to be significantly associated (p<0.10) with an increase in p mortality risk at day 180 or weaning age. Increasing inbreeding level of sires was found to be significantly associated with increasing risk in mortality at day 30, which indicates that inbred parents also can influence the survival of an offspring. Efficient breeding programs are as important for decreasing mortality in captive populations, as the provision of optimum zoo management practices.

Pedigree analysis of a highly fragmented population, the Lidia cattle breed

The aim of the study was to analyze the pedigree information of the Lidia bovine breed based on animals registered in the Herdbook and belonging to those lineages recognized as conforming to the official breed standard. Pedigree records of 272,574 animals belonging to 83 herds classified in 30 lineages were used. The average number of equivalent generations known was 4.5 (varying among lineages from 4 in Braganza to 5.2 in Baltasar Iban). The generation interval (7.5 years) was longer than that estimated in other cattle breeds. The effective size was less than 50 and consequently the estimated increase in inbreeding per generation was greater than 1% in all the lineages analyzed. The increase in inbreeding level expected for the next 50 years varied from 7.4% in Braganza to 31.3% in Diego Garrido. The ratios among the effective number of founders, the effective number of ancestors and the effective number of founder genomes was considered evidence that genetic drift explained most of the loss of genetic variability in the Lidia bovine breed due to the reduced effective population sizes of the lineages, more than bottlenecks did, as they have been less dramatic. The lineage allele loss due to the genetic drift and the effect of inbreeding are the major concerns in managing the genetic diversity of the Lidia bovine breed. The analysis of pedigree information still remains as the main useful resource to establish genetic diversity conservation guidelines in the Lidia bovine breed. Minimizing inbreeding increase within lineages in the sub-divided Lidia breed must be the major concern in managing the genetic diversity of this breed.

Joint genealogical analysis as a tool for diversity evaluation in Pinzgau cattle populations

Abstract. The presented paper gives an overview about genetic diversity in Pinzgau populations in Austria and Slovakia. In total, 12 442 animals were included in the study representing the reference population. All living herdbook cows, dual purpose and beef, from both countries with a maximum foreign gene proportion of 12.5 % were considered. Thus, four subpopulations were analysed. Parameters of diversity based on probability of identity-by-descent and probability of gene origin were computed by the methods of pedigree analysis. The average inbreeding coefficient (five generations taken into account) was 0.0186 (ΔF=0.0041), 0.0242 (ΔF=0.0047), 0.0151 (ΔF=0.0026), and 0.0126 (ΔF=0.0006) in the Austrian dairy (AD), the Austrian beef (AB), the Slovak dairy (SD) and the Slovak beef (SB) subpopulation, respectively. Effective population size ranged from 122.5 (AD) to 809.4 (SB). Effective number of founders was 67.3 and 106.6 in AD and SD and 73 and 212.3 in the AB and SB subpopulations. Effective number of ancestors was 29.59; 34.42; 42.36 and 73.81 in AD, AB, SD and SB, respectively. Effective number of founder genomes was 15.32; 18.15; 23.41 and 41 in AD, AB, SD and SB subpopulations, respectively. Similarities between subpopulations were calculated based on a cosine approach. The highest similarity was found between AD and AB (cosine value 0.9954). SD and AD subpopulations were very similar (cosine value 0.9181).

Pedigree analysis of Iran-Black sheep and inbreeding effects on growth and reproduction traits

Pedigree information and data on growth and reproduction traits in Iran-Black sheep breed, collected during a period of 28 years (1980–2008) in Abbasabad breeding station, north-east of Iran, were used to genealogical characterization of the breed using pedigree analysis and investigation the effect of inbreeding on growth related traits including birth weight (BW) and weaning weight (WW) and reproduction traits including litter size at birth per ewe lambing (LSB), litter size at weaning per ewe lambing (LSW), total litter weight at birth per ewe lambing (TLWB) and total litter weight at weaning per ewe lambing (TLWW). The Iran-Black is a composite breed; the first cross of Chios rams with Iranian Baluchi ewes. Inbreeding depression was investigated for all the studied traits by fitting individual increase in inbreeding coefficient (ΔFi) of the lambs and the ewes as linear covariates under univariate animal models. The average inbreeding coefficient during the study period for all and inbred animals were 8.08% and 9.90%, respectively. The animals born in the last 3 years (2006–2008) were considered as reference population and genealogical analysis was performed. The values of effective population size estimated from the individual increase rate in coancestry and the individual increase in inbreeding were 28 and 27, respectively. Genealogical parameters estimated based on the probabilities of gene origin including the effective numbers of founders, the effective numbers of ancestors, the effective numbers of founder genomes (founder genome equivalents) and the effective numbers of non-founder genomes were estimated as 13, 11, 6 and 12, respectively. Generation interval (in years) and the average equivalent complete generation were 3.39 and 7.10, respectively. Significant individual inbreeding depression was only found for BW (P<0.05) as −7g per 1% ΔFi. Maternal inbreeding depressions were estimated as −17g, −37g, −346g and −62g per 1% ΔFi for BW, TLWB and TLWW (P<0.01) and WW (P<0.05), respectively.

Genetic diversity in Kermani sheep assessed from pedigree analysis

Genetic diversity in Kermani sheep was assessed using the pedigree records by quantifying demographic parameters under pedigree analysis via studbook information collected from 1990 to 2011 in the breeding station of Kermani sheep, Sharebabak, south-east of Iran. Lambs born during 2008–2011 were considered as reference population. The mean generation interval from four pathways was 4.48 years. The generation interval in sire-progeny pathway was longer than dam-progeny. Mean inbreeding and average coancestry for reference population were computed as 0.51% and 1.04%, respectively. Average equivalent complete generation, as a measure of pedigree completeness, was 2.22. Effective population size was estimated to be 100 from the individual rate in coancestry and 120 from the individual increase in inbreeding. Genealogical parameters estimated based on probabilities of gene origin including the effective number of founders, the effective number of ancestors, the effective number of founder genomes (founder genome equivalents) and the effective number of non-founder genomes for reference population were estimated as 149, 72, 48 and 71, respectively. Approximately, 50% of total genetic variation was explained by the 33 most influential ancestors, with a maximum individual contribution of 4.8%. The average relationship coefficient among active rams and the average relationship coefficient between active rams and ewes during the last 4 years (2008–2011) were 0.11 and 0.05, respectively. The results indicated that although some evidences on bottlenecks and genetic drift during recent years were identified in the studied population, a relatively considerable genetic variability exists in this population.

Pedigree analysis in the German Paint Horse: Genetic variability and the influence of pedigree quality

Pedigree information of the German Paint Horse was analysed in order to assess genetic variability within this population. A data set consisting of 14,313 horses was provided by the German Paint Horse Breeding Association. Three reference populations of registered Paint horses born in Germany between the years 2000 and 2009 (1), between 2000 and 2009 based on a manually improved pedigree (2) and between 1990 and 1999 (3) were defined. For all three reference populations the quality of pedigree information, average inbreeding coefficients, effective number of founders, effective number of ancestors, effective number of founder genomes and most important ancestors were analysed. One aim of this study was to compare the results before and after the pedigree was improved. The current Paint Horse population in Germany (reference population 2) has an average number of complete generation-equivalent of 4.77. Average inbreeding coefficients are 0.49% for all and 0.41% for five considered generations. The number of founders for the current population is 4341. The effective number of founders is 963, the effective number of ancestors is 186.1 and the effective number of remaining genomes in the reference population is 118.5. Half of the gene pool of the current German population is explained by 120 animals. These rather high values indicate a good genetic diversity within the breed. However, results have to be interpreted carefully as depth of pedigree and the rather low average generation equivalent of the German Paint Horse have to be taken into account.

Using pedigree information to study genetic diversity and re-evaluating a selection program in an experimental flock of Afshari sheep

Abstract. The purpose of this paper was to evaluate the inbreeding consequences of a short-term selection experiment which was initiated in 1998 in an Afshari sheep flock. Moreover, the conducted selection experiment was re-evaluated through assessing change in ranking of the first 10 influential ancestors when their genetic contributions were replaced with their breeding values. A total of 1714 animals were registered in the herdbook with a founder population comprised of 243 animals. The average coancestry (f) and inbreeding (F) in the reference population were 2.1% and 1.2%, respectively. Estimated value of the effective population size (Ne) was 50. The effective number of founders (fe) was estimated to be 40 and the effective number of ancestors (fa) was 34. Estimates of breeding values revealed that owing to phenotypic selection some ancestors with lower breeding values had greater contribution to the reference genome than those with greater breeding values and, for this reason, the population has been deprived from the maximum genetic improvement that could be achieved if selection was based on breeding values. The effective number of founder genomes (fg) was computed to be 23 and the effective number of non-founder genomes (fne) was 55. The index of genetic diversity decreased by almost 2.2% over the period studied. In general, decrease in genetic variability was low and Ne was not very low for a small-closed population under selection, indicating even in populations which undergo selection, besides achieving genetic gain, the rate of inbreeding is controllable if matings carefully planned.

Analysis of genetic diversity in a close population of Zandi sheep using genealogical information

In a Fisher–Wright idealized population (Fisher 1930; Wright 1931), characterized by Poisson distribution of family size, large number of breeders, random mating, equal sex ratio, absence of selection, mutation, or migration, and discrete generations, the rate of loss of heterozygosity is inversely proportional to population size. In real populations, however, genetic diversity is affected by several ongoing natural processes of mutation, migration, genetic drift and selection. For this reason, Wright (1931) proposed the concept of the effective population size (Ne) for extending population genetics theory developed for ideal populations to real populations. This parameter has been useful in designing strategies for conservation of endangered animal species because of its direct relationship with the level of inbreeding (Caballero and Toro 2000). However, when Ne is estimated from pedigree data, the parameter accuracy may be affected by several factors, as extensively reviewed by Boichard et al. (1997). Consequently, in recent decades, new complementary techniques have been developed for the analysis of genealogies. Lacy (1989) introduced the concept of effective number of founders ( fe) in order to measure the overall founder representation in a managed population accounting for the loss of genetic diversity from unequal founder contributions, and the concept of founder genome equivalents ( fg), also called effective number of founder genomes, to account the effects of bottlenecks, unequal founder contributions, and genetic drift. Boichard et al. (1997) developed the concept of effective number of ancestors ( fa) to supplement the effective number of founders. The numbers obtained from

Pedigree structure and genetic variation in the two endangered Norwegian horse breeds: Døle and Nordland/Lyngen

To examine the genetic variation in two endangered Norwegian horse breeds, the pedigree structures were investigated, and key parameters such as inbreeding coefficients, effective population size, effective number of founders, effective number of ancestors and effective number of founder genomes were calculated. The data consisted of 31,142 individuals of the Døle horse and 1973 individuals of the Nordland/Lyngen horse, for which the complete generation equivalent was 10.5 and 7.2, respectively. In both breeds, the pedigree data were more than 98.5% complete in the fourth generation, allowing the rate of inbreeding and the effective population size to be precisely estimated, actually measuring the fractional loss of heterozygosity, comparable across generations (not so for the other measures). The level of inbreeding was about 12% in both breeds, with a rather wavy pattern during the past 50 years in the Døle. Considering the last generations only, the effective population size was found to be 152 in the Døle and 42 in the Nordland/Lyngen. For both populations selection in the future should be based on optimal contribution.

Change in genetic size of small-closed populations: lessons from a domestic mammal population

The aim of this study was to monitor changes in genetic size of a small-closed population of Iranian Zandi sheep, by using pedigree information from animals born between 1991 and 2005. The genetic size was assessed by using measures based on the probability of identity-by-descend of genes (coancestry, f, and effective population size, N(e) ), as well as measures based on probability of gene origin (effective number of founders, f(e) , effective number of founder genomes, f(g) , and effective number of non-founder genomes, f(ne) ). Average coancestry, or the degree of genetic similarity of individuals, increased from 0.81% to 1.44% during the period 1993 to 2005, at the same time that N(e) decreased from 263 to 93. The observed trend for f(e) was irregular throughout the experiment in a way that f(e) was 68, 87, 77, 92, and 80 in 1993, 1996, 1999, 2002, and 2005, respectively. Simultaneously, f(g) , the most informative effective number, decreased from 61 to 35. The index of genetic diversity (GD) which was obtained from estimates of f(g) , decreased about 2% throughout the period studied. In addition, a noticeable reduction was observed in the estimates of f(ne) from 595 in 1993 to 61 in 2005. The higher than 1 ratio of f(e) to f(g) indicated the presence of bottlenecks and genetic drift in the development of this population of Zandi sheep. From 1993 to 1999, f(ne) was much higher than f(e) , thereby indicating that with respect to loss of genetic diversity, the unequal contribution of founders was more important than the random genetic drift in non-founder generations. Subsequently, random genetic drift in non-founder generations was the major reason for f(e) > f(ne) . The minimization of average coancestry in new reproductive individuals was recommended as a means of preserving the population against a further loss in genetic diversity.

Inbreeding trends and pedigree analysis of Bavarian mountain hounds, Hanoverian hounds and Tyrolean hounds

The objective of this study was to analyse genetic diversity for the three scent-hound breeds Bavarian mountain hound (BMH), Hanoverian hound (HH) and Tyrolean hound (TH) using all available pedigree information from scent-hound kennel clubs for these three breeds throughout Europe. The pedigree data of the BMH and the HH date back to 1912 and 1894, respectively. Pedigree data of the TH were available from the 1960s onwards. The reference populations included all BMH (n = 3231), HH (n = 1371) and TH (n = 1167) dogs registered between 1992 and 2004. Average generation intervals were 5.3 years for the BMH and 5.0 years for the HH and TH. Average inbreeding coefficients for the reference populations were 4.5%, 6.8% and 9.5% for the BMH, HH and TH. The effective numbers of founders, ancestors and founder genomes were lowest for the TH and highest for the BMH. The effective numbers of founder genomes were 10.9, 5.6 and 4.3 for the BMH, HH and TH. Effective population size was largest for the BMH with 72.7 effective breeding animals, followed by the HH with 50.9 and TH with 26.5. The most important ten ancestors had genetic contributions to the reference populations of 54.4%, 65.2% and 77.9% in the BMH, HH and TH. The results of our study indicate the need for careful breed management in these highly specialized hound breeds to maintain genetic diversity. European stud books should be established for these dog breeds in order to avoid inbreeding due to missing pedigree records.

Genetic variability of a Criollo cattle population under conservation

The active germplasm Bank of the Integrated Unit (Facultad de Ciencias Agrarias, Unversidad Nacional de Mar del Plata and the Experimental Station of INTA) at Balcarce, keeps both in vivo (live herd) and in vitro Criollo Cattle (samples of semen and embryos cryogenically), with the aim of preserving genetic variability for potential future use. Parameters based on probable origin of genes, on population size and on the estimation of components of (co)variances for birth and weaning weights were estimated to evaluate the genetic variability of the material contained in the Bank. The information available came from 667 individuals, of which 84 had unknown parents, and 142 half-founders (only one parent known), having a mean number of births per year of 24 calves from four different sires. AI was performed during the mating season almost every year. The evolution of effective number of founders (fe), effective number of ancestors (fa), effective number of founder genomes (Ng) and the ratio fe/fa over different periods from the creation of the Bank, were analysed. Annual increase of inbreeding coefficient (F) was 0·2% and the effective population size (Ne) was estimated as 23, 14 and 25 individuals for the periods defined by years 1978–82, 1983–87 and 1993–97, respectively. The ratio fe/fa was higher in the years where the ratio number of sires/number of dams was lower, indicating that the impact of the effective number of sires is more important than the effective number of females in generating genetic bottlenecks. When the reference population was taken for the period 1989–97, 16·45 out of the 85·63 founders of the population made contributions not explained by other ancestors, but if possible losses of genomes due to genetic drift are considered, the number of genomes still present was reduced to 9·09 indicating a possible narrow genetic bottleneck in the last period. The Ng/fe ratio was 0·11, 0·10 and 0·11 for the reference populations defined by years 1989–92, 1993–97 and 1989–97, respectively. The main objective in germplasm conservation programmes should be the maintenance of genetic variability. Thus, matings in the population under conservation should be designed in order to mate those animals that maximize the effective number of genome founders (Ng) to avoid losses due to genetic drift.

Inbreeding in Danish Dairy Cattle Breeds

The purpose of this study was to monitor current and predict future rates of inbreeding in the Danish dairy breeds. Calves born from 1999 until 2003 and registered as Danish Holstein (1,883,983), Danish Jersey (336,966), or Danish Red (261,047) were reference populations. Average complete generation equivalent was approximately 7. For calves born in 2003, average inbreeding was 3.9, 3.4, and 1.4% for Holstein, Jersey, and Danish Red, respectively. In recent years, effective population sizes were 49, 53, and 47, respectively. Based on coancestry statistics, future effective population sizes will be 43, 42, and 51, respectively. The effective number of founders, effective number of ancestors, and effective number of founder genomes were calculated. These measures of genetic diversity were all low for Holstein and Jersey and somewhat larger for Danish Red. The most important ancestors of Danish Holstein were Elevation (13.8%), Chief (10.9%), and Bell (8.5%). The most important ancestor of Danish Red was Momentum (9.4%), a Red Holstein-Friesian. The most important ancestor for Danish Jersey was FYN Lemvig (12.1%) with a large number of progeny in the reference population. The results of this study indicate the necessity for active management of the rate of inbreeding in the future.