Analysis of properdin (BF) genotypes associated with litter size in a commercial pig cross population

Properdin (BF) was investigated as a candidate gene influencing litter size in a commercial pig cross population. The BF gene was chosen because of its integral role in influencing uterine epithelium growth and because several quantitative trait loci (QTL) with impact on reproductive traits have been detected near the centromere of porcine chromosome 7. A total of 123 F₂ (Large White × Landrace) × Leicoma sows were genotyped using polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method. The sows were divided into two extreme performance groups, one with a high litter size (n = 61, ≥14.3 piglets per litter) and the other with a low litter size (n = 62, ≤11.3 piglets per litter). Although genotype and allele frequencies were uneven with 2.4% (AA), 16.3% (AB), 81.3% (BB) and 0.11 (A): 0.89 (B), the allele A was the unfavourable one, leading to less offspring. With regard to the level of significance at p < 0.05, the total number of born (TNB) and number of born alive (NBA) piglets were associated with BF genotypes. The genotype AA led to 10.55 TNB and 10.00 NBA, whereas the genotype BB led to 13.19 TNB and 12.11 NBA. The genotype AB was intermediate. In future, a systematic mating test is necessary in order to obtain more balanced genotype frequencies. Furthermore, it should be taken into consideration that the investigated polymorphism is located in an intronic region and the causative mutation is not clear yet.     

Genome-wide association study of swine farrowing traits. Part I: Genetic and genomic parameter estimates

The primary objective of this study was to determine genetic and genomic parameters among swine (Sus scrofa) farrowing traits. Genetic parameters were obtained using MTDFREML. Genomic parameters were obtained using GENSEL. Genetic and residual variances obtained from MTDFREML were used as priors for the Bayes C analysis of GENSEL. Farrowing traits included total number born (TNB), number born alive (NBA), number born dead (NBD), number stillborn (NSB), number of mummies (MUM), litter birth weight (LBW), and average piglet birth weight (ABW). Statistically significant heritabilities included TNB (0.09, P = 0.048), NBA (0.09, P = 0.041), LBW (0.20, P = 0.002), and ABW (0.26, P < 0.0001). Statistically significant genetic correlations included TNB-NBA (0.97, P < 0.0001), TNB-LBW (0.74, P < 0.0001), NBA-LBW (0.56, P < 0.0017), NSB-LBW (0.87, P < 0.0395), and LBW-ABW (0.63, P < 0.0002). Genetic parameters are similar to others found in the literature. The proportion of phenotypic variance explained by genomic markers (GP) generated by GENSEL was TNB (0.04), NBA (0.06), NBD (0.00), NSB (0.01), MUM (0.00), LBW (0.11), and ABW (0.31). Limited information is available in the literature about genomic parameters. Only the GP estimate for NSB is significantly lower than what has been published. The GP estimate for ABW is greater than the estimate for heritability found in this study. Other traits with significant heritability had GP estimates half the value of heritability. This research indicates that significant genetic markers will be found for TNB, NBA, LBW, and ABW that will have either immediate use in industry or provide a roadmap to further research with fine mapping or sequencing of areas of significance. Furthermore, these results indicate that genomic selection implemented at an early age would have similar annual progress as traditional selection, and could be incorporated along with traditional selection procedures to improve genetic progress of litter traits.     

Effect of candidate gene polymorphisms on reproductive traits in a Large White pig population

The objective of this study was to test for association of candidate single nucleotide polymorphisms (SNPs) with sow prolificacy reproductive traits, such as litter size, ovulation rate and lifetime performance, in gilts of a Large White pig population. Preliminary research on 25 animals selected from the high‐ and low‐performance groups of 347 animals with case‐control studies indicated that seven genes were associated with total number of piglets born (TNB). Six of the seven genes were associated with reproductive traits, including TNB, number of piglets born alive (NBA) and average weight of piglet weaning (AWW). A MBL2 SNP was significantly associated with TNB and NBA in first parity. A CFB SNP was associated with TNB in first parity. An ACE SNP was associated with TNB in first and second parities. An EGF polymorphism was associated with TNB, NBA and AWW in second parity. A KCNC2 polymorphism was significantly associated with TNB and NBA in second parity. A SLC22A5 SNP was associated with TNB and NBA in second parity. Six candidate SNPs were associated with TNB; the only exception was a PRKAG3 polymorphism. A candidate gene approach enables some of these polymorphisms to be used in genetic improvement programs based on marker‐assisted selection.     

陆川猪和大白猪LEPR基因对产仔数的效应分析

为了比较LEPR基因对猪产仔数效应大小,采用PCR-SSCP技术在陆川猪和大白猪群中进行了LEPR基因的基因型频率检测及不同基因型的总产仔数和产活仔数效应分析。结果表明,LEPR基因对陆川猪头胎和头4胎平均总产仔数（TNB）及产活仔数（NBA）影响差异显著(P<0.05),对大白猪头胎的总产仔数及产活仔数影响差异显著(P<0.05),而对大白猪头4胎平均总产仔数及产活仔数影响差异不显著(P>0.05)。     

Oestrogen receptor genotypes and litter size in Hungarian Large White pigs

A total of 869 litter records of 226 Hungarian Large White sows have been analysed to investigate the possible use of the oestrogen receptor gene (ESR) as marker to improve litter size. First, second and later parities have been evaluated separately. Frequencies of A = 0.55 and B =0.45 have been calculated for the two ESR alleles and the observed/expected number of the three genotypes were as follows: AA: 71/69.1, AB: 108/111.8 and BB: 47/45.1. BB type first and later parity sows were superior to AB and AA sows for number born alive (NBA), total number of born (TNB) and the corrected number of weaned piglets (CNW), respectively.     

Selection potential of different prolificacy traits in the finnish landrace and large white populations

Heritabilities and genetic correlations for different prolificacy traits were estimated to assess possibilities of selection for high number of piglets weaned. Three litter-size traits: total number of piglets born (TNB), number of piglets born alive (NBA), number of piglets weaned (NW); four piglet survival traits: number of stillborn piglets (NSB), percent of stillborn piglets (NSB%), piglet mortality between birth and weaning (PM), percent of dead piglets during suckling (PM%); and three traits measuring time intervals: age at first farrowing (AFF), first farrowing interval (FFI), and gestation length (GL) were analysed. The Finnish national litter recording scheme provided data on the first parity litters of 11 329 Landrace and 8 362 Large White pigs born between 1986 and 2000. The heritabilitiy estimates were moderate for AFF and GL (0.24–0.37), and low for all the other traits (0.03–0.11). The genetic correlations between TNB and PM (0.68 in Landrace and 0.43 in Large White) and between NBA and PM (0.64 in Landrace and 0.31 in Large White) suggest that selection only for high TNB or NBA will lead to increased PM. The results showed further that GL will increase indirectly if the selection pressure is for low PM (r _g =?0.050 in Landrace and ?0.43 in Large White.     

大白猪哺乳期背膘损失对繁殖性能的影响

为了研究大白母猪哺乳期背膘损失对其繁殖性能的影响,试验选取大白母猪1 178头,统计6个胎次的总产仔数、产活仔数、健仔数、断奶窝重、断奶发情间隔、哺乳期背膘损失等性状,根据哺乳期的背膘损失情况将母猪分为6组：<0、0~1、1~2、3~4、5~6、>6 mm,以断奶窝重为协变量,利用最小二乘检验开展组间总产仔数、产活仔数、健仔数、断奶发情间隔等性状的差异显著性分析。结果显示,大白母猪6个胎次平均总产仔数、产活仔数、健仔数、断奶窝重、断奶发情间隔和哺乳期背膘损失分别为13.67头、11.51头、10.32头、65.90 kg、4.83 d和2.89 mm,大白母猪1~6胎产仔数在各组间存在显著差异（P<0.05）。综合全部6个胎次的结果可知,第4、5、6组间差异不显著（P>0.05）,但均显著高于第1、2、3组（P<0.05）;第4组总产仔数最高,达13.54头,比第1、2、3组分别高出1.90、2.29和1.63头（P<0.05）。虽然各胎次中各组间产活仔数和健仔数性状有时也出现显著差异,但综合分析6个胎次组间并未出现显著差异（P>0.05）。从断奶发情间隔性状来看,各组间均未出现显著差异（P>0.05）。结果表明,在大白猪生产中,将哺乳期母猪的背膘损失控制在3~4 mm可以获得更高的总产仔数。     

Comparison of genetic improvement for litter size at birth by direct and indirect selection in swine herd

Responses to selection for number of piglets born alive (NBA) by the total number of piglets born (TNB), the NBA, and the NBA plus number of piglets born dead (NBD) were compared using the accuracy of selection and expected genetic gain calculated from the selection index with family information and the real response to selection, using data generated by Monte Carlo computer simulation. The accuracy of selection for NBA selected by TNB was higher than that by NBA only if the genetic correlation between TNB and NBA was close to 1.0, or the value of heritability for the TNB was much larger than that for the NBA. The accuracy of selection for the NBA selected by the combination of the TNB and the NBA was generally highest in the three selection methods in each family structure. Selection by the TNB resulted in the greatest expected genetic gain for the TNB among the selection methods. In the best linear unbiased prediction (BLUP) selection, the genetic gain for the NBA accumulated by the NBA tended to be similar to that accumulated by the combination of the NBA and the NBD, and both genetic gains at generation 10 were significantly larger than that by the TNB (P < 0.001). The accumulated responses selected by the two‐trait animal model BLUP estimated from genetic parameters with errors were similar to those estimated from the true parameters, and there was no significant difference between them. These results indicate that selection by the NBA or by the NBA and the NBD gives more genetic improvement in the NBA than that by the TNB.     

大白猪、长白猪和杜洛克猪RBP4基因多态性及其与繁殖性能的相关分析

本试验通过PCR-RFLP技术对大白猪、长白猪和杜洛克猪的视黄醇结合蛋白4(retinol binding protein 4,RBP4)基因第4外显子Msp Ⅰ酶切位点进行多态性分析,并对各基因型母猪的第2、3胎次产仔数、产活仔数、断奶仔猪数、初生窝重、21日龄的窝重及断奶成活率进行测定,进而分析RBP4基因多态性与繁殖性能的关系。结果表明,RBP4基因在3个品种中均存在多态位点;χ²适合性检验表明大白猪在该位点偏离Hardy-Weinberg平衡状态(P<0.05),而长白猪和杜洛克猪在该位点处于Hardy-Weinberg平衡状态(P>0.05)。其中,大白猪中优势基因型为BB基因型,而长白猪和杜洛克猪中优势基因型为AA基因型。在繁殖性能上,长白猪BB基因型群体第2胎总产仔数、产活仔数、断奶仔猪数、初生窝重、21日龄断奶窝重分别高于AA基因型0.97头、1.28头、0.92头、1.74 kg和4.42 kg,高于AB基因型0.92头、1.02头、0.98头、1.43 kg和4.98 kg;第3胎次中总体上BB基因型也为有利基因型。     

The contribution of dominance and inbreeding depression in estimating variance components for litter size in Pannon White rabbits

In a synthetic closed population of Pannon White rabbits, additive (V_A), dominance (V_D) and permanent environmental (V_Pe) variance components as well as doe (b_Fd) and litter (b_Fl) inbreeding depression were estimated for the number of kits born alive (NBA), number of kits born dead (NBD) and total number of kits born (TNB). The data set consisted of 18,398 kindling records of 3883 does collected from 1992 to 2009. Six models were used to estimate dominance and inbreeding effects. The most complete model estimated V_A and V_D to contribute 5.5 ± 1.1% and 4.8 ± 2.4%, respectively, to total phenotypic variance (V_P) for NBA; the corresponding values for NBD were 1.9 ± 0.6% and 5.3 ± 2.4%, for TNB, 6.2 ± 1.0% and 8.1 ± 3.2% respectively. These results indicate the presence of considerable V_D. Including dominance in the model generally reduced V_A and V_Pe estimates, and had only a very small effect on inbreeding depression estimates. Including inbreeding covariates did not affect estimates of any variance component. A 10% increase in doe inbreeding significantly increased NBD (b_Fd = 0.18 ± 0.07), while a 10% increase in litter inbreeding significantly reduced NBA (b_Fl = ?0.41 ± 0.11) and TNB (b_Fl = ?0.34 ± 0.10). These findings argue for including dominance effects in models of litter size traits in populations that exhibit significant dominance relationships.     

Effect of the polymorphism of prolactin receptor (PRLR) and leptin (LEP) genes on litter size in Polish pigs

The aim of the experiment was to use the DNA mutations in the PRLR and LEP genes to determine associations between the genotype and litter size in Polish Large White × Landrace sows. Reproductive traits investigated were: total number of piglets born (TNB), number of piglets born alive (NBA) and number of piglets weaned. The polymorphism in PRLR and LEP genes was detected using the polymerase chain reaction–restriction fragment‐length polymorphism (PCR‐RFLP) method, with specific primers and the restriction enzymes AluI and HinfI respectively. Two different alleles of PRLR and LEP gene were identified: alleles A (0.62) and B (0.38) of the PRLR gene and alleles C (0.10) and T (0.90) of the LEP gene. The relationships between the PRLR and LEP genotypes and TBN, NBA and NW were analysed. The analysis showed, in first parity sows, statistically significant (p ≤ 0.01) differences between sows carrying different PRLR genotypes. In later parities, sows with the AA genotype still had the largest litter size compared with AB and BB sows, but the difference was statistically not significant. Analysis of the interaction PARITY × PRLR showed small and statistically not significant differences. The analysis of relationship between different LEP genotypes and TNB, NBA, NW showed small and statistically non‐significant differences.     

应用动物模型REML法估计金华猪繁殖性状遗传参数

试验资料取自金华猪场 1 979～1 999年金华猪繁殖性能的生产记录。采用动物模型约束性最大似然法 (REML)对该猪群的遗传参数进行估计 ,测定 1 1 4 4头纯繁金华母猪的产仔哺育记录。结果表明 ,产仔数 (TNB)、产活仔数 (NBA)、初生重 (PBW)、初生窝重 (LBW)、 2 0日龄头数(NFC)、 2 0日龄窝重 (LWFC)、 2 0日龄重 (PWFC)、 60日龄头数 (NW)、 60日龄窝重 (LWW)、 60日龄重 (PWW)、乳头数 (TN)的遗传力分别为0 1 2± 0 0 9、 0 1 8± 0 1 1、 0 1 5± 0 0 9、 0 0 2± 0 0 9、 0 1 0± 0 1 1、 0 0 8± 0 1 3、 0 1 8± 0 1 0、 0 2 3± 0 1 1、 0 31± 0 1 3、 0 35± 0 1 1和 0 1 9± 0 1 0。TNB与NBA、LBW、NW、LWW的遗传相关范围为 0 66～ 0 88,TNB与PBW的遗传相关为 - 0 43。NBA与LBW、NW、LWW的遗传相关范围为 0 60～ 0 64,NBA与PBW遗传相关为 - 0 63。PBW与PWW的遗传相关为 0 51。     

Divergent selection for uterine capacity in rabbits. II. Correlated response in litter size and its components estimated with a cryopreserved control population

Our objective was to evaluate the correlated responses to selection for litter size and its components after 10 generations of divergent selection for uterine capacity (UC). A total of 294 intact females from the 11th and 12th generations of divergent selection for high and low UC and from a cryopreserved control population was used (139, 112, and 43 females, respectively). Uterine capacity was assessed as litter size in unilaterally ovariectomized females. Traits recorded on females for up to five parities were litter size (LS) and number born alive (NBA). Laparoscopy was performed in all females at d 12 of their second parity, and the ovulation rate (OR) and number of implanted embryos (IE) were recorded in these females. Embryo survival (ES = IE/OR), fetal survival (FS = LS/IE), and prenatal survival (PS = LS/OR) were computed. Correlated responses in LS and in its components were inferred using Bayesian methods. Correlated responses in LS were asymmetric. The divergence between high and low lines was 2.35 kits, mainly because of a higher correlated response in the low line (1.88 kits). The lower LS in the low line was associated with a lower PS (control - low = 0.14), because of decreases in ES and FS.     

撒坝猪专门化母系繁殖性状的相关与通径分析

利用撒坝猪专门化母系 7个世代共 576头母猪的繁殖性能数据 ,对总产仔数 (x1 )、活产仔数 (x2 )、初生窝重 (x3)、 2 0日龄窝重 (x4 )、断奶窝重 (x5)、育成率 (x6 )和断奶仔数 (y) 7个繁殖性状进行了相关和通径分析 ,并在此基础上建立了估计断奶仔数的最优回归方程。结果表明 ,撒坝猪专门化母系的繁殖性能在经过 7个世代的选育后 ,获得了明显提高。各繁殖性状间在表型值上都存在不同程度的相关 ,其中以总产仔数与活产仔数的相关程度最高 (r=0 92 0 1 ) ,其次为 2 0日龄窝重与断奶仔数 (r=0 82 88) ,而初生窝重与育成率间的相关系数最低 (r=0 0 381 )。在所分析的 7个性状中 ,除了总产仔数和活产仔数与育成率间为负相关外 ,其余性状间均为正相关。所建立的估计断奶仔数的多元回归方程为 :y=- 6 1 1 90 +0 6986x2 +0 0 33 7x4 +0 0 0 4 8x5+0 0 71 8x6 。     

不同配种模式对二元母猪繁殖性能的影响

为探讨不同配种方式对二元母猪产仔数的影响,以杜洛克猪、长白猪、大白猪、皮特兰猪四个品种公猪精液采用人工授精的方法对11654头长大二元母猪配种,对所产11654窝总产仔数、窝产活仔数、窝损失等进行统计和分析,分析不同配种模式对二元母猪繁殖性能的影响。结果表明,配种次数、不同品种公猪组合配种对二元母猪总产仔数、产活仔数以及窝损失均无显著影响（P〉0.05）,但杜洛克公猪所配二元母猪的总产仔数和窝产仔数均高于大白和长白猪（P〈0.05）;采用长×杜×大组合配种的二元母猪总产仔数和窝产活仔数均高于其它三种组合。试验结果表明,在生产中,可在长白、大白、杜洛克猪中任选两个品种,通过二次配种的方式提高母猪的产仔数。     

Estimation of genetic parameters for farrowing traits in purebred Landrace and Large White pigs

Genetic parameters were estimated for six reproductive traits related to farrowing events in Landrace and Large White pigs; total number born (TNB), number born alive (NBA), number stillborn (NSB), total litter weight at birth (LWB), mean litter weight at birth (MWB), and gestation length (GL). We analyzed 62,534 farrowing records for 10,637 Landrace dams and 49,817 farrowing records for 8,649 Large White dams. Estimated heritabilities of TNB, NBA, NSB, LWB, MWB, and GL by single‐trait repeatability model analyses were 0.12, 0.12, 0.08, 0.18, 0.19, and 0.29, respectively, in Landrace, and 0.12, 0.10, 0.08, 0.18, 0.16, and 0.34, respectively, in Large White. Genetic correlation between NBA and NSB was unfavorable: 0.20 in Landrace and 0.33 in Large White. Genetic correlations of GL with the other five traits were weak: from ?0.18 with NSB to ?0.03 with NBA in Landrace, and from ?0.22 with NSB to ?0.07 with NBA in Large White. LWB had a highly favorable genetic correlation with NBA (0.74 in both breeds), indicating the possibility of using LWB for the genetic improvement of NBA.     

Genetic parameters for litter traits at different parities in purebred Landrace and Yorkshire pigs

Comparison of the multi‐trait animal model and the traditional repeatability model was carried out using data obtained from 6,424 Landrace and 20,835 Yorkshire sows farrowed from January 2000 to April 2018 in order to estimate genetic parameters for litter traits at different parities. Specifically, records of the total number born (TNB), number born alive (NBA), total number of mortality (MORT), number of stillborn (NSB) and number of mummified pigs (MUM) were used. Although results showed the heterogeneity of heritability for litter traits at different parities, the mean heritability estimates from the multi‐trait model were found to be higher than those of the repeatability model for all traits in both pig breeds. In terms of genetic correlation between parities, a slight difference in genetic control in the first parity was noted for TNB and NBA in Landrace and Yorkshire pigs. The correlation between the first parity and later parities ranged from 0.48 to 0.74 for TNB and NBA in both breeds. Moreover, genetic correlation between parities for MORT and NSB was observed to be high for parities higher than 2 in Yorkshire pigs. For MUM, genetic correlation between the first and other parities was generally low in both breeds, indicating that culling pigs on the basis of MUM at the first parity could probably be unreasonable. Overall, the results of this study suggest that the multi‐trait approach for litter size traits is useful for the accurate estimation of genetic parameters.     

Selection for litter size at day five to improve litter size at weaning and piglet survival rate

Selection for total number of piglets born (TNB) since 1992 has led to a significant increase in this trait in Danish Landrace and Danish Yorkshire but has also been accompanied by an increase in piglet mortality. The objective of this study was to estimate the genetic and phenotypic parameters for litter size and survival to find alternative selection criteria to improve litter size at weaning. Data from Landrace (9,300 litters) and Yorkshire (6,861 litters) were analyzed using REML based on a linear model including genetic effects of sow and service-sire. The estimates of heritability (based on the sow component) for TNB, number born alive (NBA), and number alive at d 5 after birth (N5D) and at weaning (about 3 wk, N3W) ranged from 0.066 to 0.090 in Landrace and 0.050 to 0.070 in Yorkshire. Genetic correlations between TNB and N3W were 0.289 in Landrace and 0.561 in Yorkshire, but between N5D and N3W the estimated genetic correlation was 0.995 in both populations. The approximate estimates of heritability for survival rate per litter at birth (SVB = NBA/TNB), from birth to d 5 (SV5 = N5D/NBA), and from d 5 to weaning (SVW = N3W/N5D) were 0.130, 0.131, and 0.023, respectively, in Landrace, and 0.095, 0.043, and 0.009, respectively, in Yorkshire. Genetic correlations between TNB and survival rates at different stages were negative. On the other hand, genetic correlations between N5D and survival rates and between N3W and survival rates were strongly or moderately positive, except for the correlations with SVW in Yorkshire. The results suggest that selection for N5D could be an interesting alternative to improve litter size at weaning and piglet survival for Danish Landrace and Danish Yorkshire.     

荣昌猪初产繁殖性状的全基因组关联研究

旨在鉴定荣昌猪初产繁殖性状的重要变异位点和基因,为荣昌猪繁殖性状的遗传改良提供重要的分子标记和基因资源。本研究选取429头荣昌母猪进行猪50K芯片基因分型,经过质量控制和基因型填充后,保留35 046个SNPs用于分析。采用主成分分析法研究群体结构,利用混合线性模型(mixed-linear model, MLM)将出生年、出生月作为固定效应,将主成分值作为协变量对总产仔数、活产仔数、死胎数和初生窝重性状进行全基因组关联分析(GWAS)。结果显示,在全基因组显著水平上鉴定出2个影响荣昌猪初生窝重的SNPs和1个影响荣昌猪死胎数的SNP;在潜在显著水平上鉴定到5个影响荣昌猪总产仔数的SNPs, 3个影响荣昌猪活产仔数的SNPs和10个影响荣昌猪死胎数的SNPs。通过全基因组关联分析筛选到1个显著的SNP(SSC17:57 315 180 bp)同时影响荣昌猪总产仔数、活产仔数和初生窝重,1个显著的SNP(SSC1:279 214 647 bp)同时影响荣昌猪活产仔数和总产仔数,暗示基因在不同性状间具有一因多效性。本研究根据候选基因的相关分子生物学功能,确定BMP7基因为影响荣昌猪总产仔数...     

猪催乳素受体基因第8内含子多态性及其对产仔性能的影响

采用PCR-SSCP技术检测了大白猪、长白猪、杜洛克猪、马身猪、山西黑猪和山西白猪等6个品种364头个体催乳素受体基因(prolactin,PRLR)第8内含子的多态性,并分析了多态位点对猪头胎产仔性能的影响。结果表明,猪PRLR基因第8内含子存在多态性,共检测到A、B、C 3个等位基因和AA、AB、AC、BB、BC和CC 6种基因型。在杜洛克猪、长白猪、山西黑猪和山西白猪群体中,A等位基因的频率最高,分别为0.8276、0.5439、0.5345和0.7669;而在大白猪和马身猪群体中,B等位基因的频率最高,分别为0.6894和0.7000;仅在大白猪、山西黑猪和山西白猪中检测到C等位基因,且频率较低,在0.0379~0.1385之间。PRLR基因型对母猪头胎产仔数有显著影响(P<0.05),对产活仔数的影响未达显著水平。AC基因型母猪的头胎产仔数和产活仔数显著高于BC基因型个体,而与AA、AB和BB基因型个体差异不显著。含有A等位基因的母猪的产仔性能优于不带有A等位基因的个体,表明A等位基因是有利于母猪产仔性能提高的等位基因,可应用于标记辅助选择来提高母猪的繁殖性能。