Quantitative trait loci for milk production and functional traits in two Danish Cattle breeds

Quantitative trait loci (QTL) in Danish Jersey and Danish Red cattle were independently mapped by least squares regression analysis. For Jersey breed, five grandsire families were genotyped for 186 markers on 16 chromosomes (BTAs). Eight traits analysed were milk yield (MY), fat percentage (FP), protein percentage (PP), clinical mastitis (CM), somatic cell score (SCS), maternal stillbirth, maternal calf size (MCS) and maternal calving difficulty. For Red breed, nine grandsire families were genotyped for 166 markers on 18 BTAs. Six traits analysed were MY, FP, PP, CM, SCS and female fertility. Nine and five QTL were detected in Jersey and Red breed, respectively, in across family tests. In Jersey breed, the results indicate QTL for CM and MCS on BTA 3. Additionally, there is an indication of QTL for MCS and FP on BTA 1 and a tentative evidence for a QTL for MY on BTA 26. There is a high risk of detected QTL being false positives. The detected QTL in Jersey breed indicate interesting results from a breeding perspective, but a practical application should await genome-wide association studies.     

Quantitative trait loci for leg weakness traits in a Landrace purebred population

Leg weakness in pigs is a serious problem in the pig industry. We performed a whole genome quantitative trait locus (QTL) analysis to find QTLs affecting leg weakness traits in the Landrace population. Half-sib progeny ( n  = 522) with five sires were measured for leg weakness traits. Whole genome QTL mapping was performed using a half-sib regression-based method using 190 microsatellite markers. No experiment-wide significant QTLs affecting leg weakness traits were detected. However, at the 5% chromosome-wide level, QTLs affecting leg weakness traits were detected on chromosomes 1, 3, 10 and 11 with QTL effects ranging from 0.07 to 0.11 of the phenotypic variance. At the 1% chromosome-wide level, QTLs affecting rear feet score and total leg score were detected on chromosomes 2 and 3 with QTL effects of 0.11 and 0.13 of the phenotypic variance, respectively. On chromosome 3 and 10, some QTLs found in this study were located at nearby positions. The present study is one of the first reports of QTLs affecting fitness related traits such as leg weakness traits, that segregate within the Landrace population. The study also provides useful information for studying QTLs in purebred populations.     

Detection and fine mapping of quantitative trait loci for bone traits on chicken chromosome one

In broiler chickens, bone problems are an important welfare issue that has been linked to genetic selection for rapid growth. The objectives of this study were to identify and fine map quantitative trait loci (QTL) associated with bone traits. The Northeast Agricultural University resource population (NEAURP) being an F(2) population was used in this study, and a total of 17 bone traits were measured. In primary genome scan, the linkage map was constructed with 23 microsatellite markers across the entire chicken chromosome 1. Seventeen QTLs for bone traits were identified and 12 of these were found between LEI0079 and ROS0025 (50.8 cM apart). To fine map the QTLs located between LEI0079 and ROS0025, more markers and more individuals were used and a new partial linkage map was constructed. The confidence intervals for QTLs were sharply narrowed down from 24.5～52.6 to 2.7～17.0 Mb. This study identified chromosome regions harbouring significant QTLs affecting bone traits and showed that the use of more markers and individuals could decrease the confidence interval of QTL effectively. The results provide a useful reference for further candidate gene research and MAS for bone traits.     

Molecular genetics of behaviour: research strategies and perspectives for animal production

Genetic factors are undoubtedly involved in inter-individual variability of the behaviours that may be important for livestock production, as shown by pedigree studies, comparison of genetic stocks raised in the same environment, and selection experiments. The knowledge of gene polymorphisms responsible for genetic variability would increase the efficiency of selection, as shown for instance by the identification of the ryanodine receptor gene that harbours the mutations responsible for the porcine stress syndrome, that allows the eradication of the susceptibility allele. One strategy is to screen systematically the genes that are known to be involved in regulation of behaviour (functional candidate genes). This strategy is however very difficult for most behavioural traits, since behaviour is an emerging function from the whole brain/body and the molecular pathways involved in genetic variability are very poorly understood. Another strategy is to investigate linkage between trait variation and genetic markers in a segregating population (usually an intercross or backcross between two strains or breeds contrasting for the trait under study). It allows the detection of genomic regions influencing that trait (quantitative trait loci or QTL), and further investigation aims at the identification of the gene(s) located in each of these regions and the molecular polymorphisms involved in phenotypic variation. Although many QTL have been published for behavioural traits in experimental animals, very few examples are available where strong candidate genes have been identified. Further progress will be very much dependent upon the careful definition of behavioural traits to be studied (including their importance for animal production), on the reliability of their measurement in a large number of animals and on the efficient mastering of environmental factors of variability. The fast increase in the knowledge of genome sequence in several species will undoubtedly facilitate the application to farm animal species of the knowledge obtained in model organisms, as well as the use of model organisms to explore candidate genes detected by QTL studies in farm animals.     

猪数量性状基因及其标记研究进展

对猪中有关生长、胴体、肉质和繁殖性状的数量性状位点及其标记的研究现状与前景作了综述。     

Fine mapping quantitative trait loci under selective phenotyping strategies based on linkage and linkage disequilibrium criteria

In fine mapping of a large‐scale experimental population where collection of phenotypes are very expensive, difficult to record or time‐demanding, selective phenotyping could be used to phenotype the most informative individuals. Linkage analyses based sampling criteria (LAC) and linkage disequilibrium‐based sampling criteria (LDC) for selecting individuals to phenotype are compared to random phenotyping in a quantitative trait loci (QTL) verification experiment using stochastic simulation. Several strategies based on LAC and LDC for selecting the most informative 30%, 40% or 50% of individuals for phenotyping to extract maximum power and precision in a QTL fine mapping experiment were developed and assessed. Linkage analyses for the mapping was performed for individuals sampled on LAC within families and combined linkage disequilibrium and linkage analyses was performed for individuals sampled across the whole population based on LDC. The results showed that selecting individuals with similar haplotypes to the paternal haplotypes (minimum recombination criterion) using LAC compared to random phenotyping gave at least the same power to detect a QTL but decreased the accuracy of the QTL position. However, in order to estimate unbiased QTL parameters based on LAC in a large half‐sib family, prior information on QTL position was required. The LDC improved the accuracy to estimate the QTL position but not significantly compared to random phenotyping with the same sample size. When applying LDC (all phenotyping levels), the estimated QTL effect were closer to the true value in comparison to LAC. The results showed that the LDC were better than the LAC to select individuals for phenotyping and contributed to detection of the QTL.     

Detection of quantitative trait loci affecting non-return rate in French dairy cattle

The purpose of this study was to map quantitative trait loci (QTL) influencing female fertility estimated by non-return rate (NRR) in the French dairy cattle breeds Prim'Holstein, Normande and Montbeliarde. The first step was a QTL detection study on NRR at 281 days after artificial insemination on 78 half-sib families including 4993 progeny tested bulls. In Prim'Holstein, three QTL were identified on Bos taurus chromosomes BTA01, BTA02 and BTA03 (p < 0.01), whereas one QTL was identified in Normande on BTA01 (p < 0.05). The second step aimed at confirming these three QTL and refining their location by selecting and genotyping additional microsatellite markers on a sub-sample of 41 families from the three breeds using NRR within 56, 90 and 281 days after AI. Only the three QTL initially detected in Prim'Holstein were confirmed. Moreover, the analysis of NRR within 56, 90 and 281 days after AI allowed us to distinguish two FF QTL on BTA02 in Prim'Holstein, one for NRR56 and one for NRR90. Estimated QTL variance was 18%, 14%, 11.5% and 14% of the total genetic variance, respectively, for QTL mapping to BTA01, BTA02 (NRR90 and NRR56) and BTA03.     

Identification of candidate markers on bovine chromosome 14 (BTA14) under milk production trait quantitative trait loci in Holstein

The objective of this study was to identify single-nucleotide polymorphisms using a bovine chromosome 14 high-density SNP panel after accounting for the effect of DGAT1. Linkage disequilibrium information and sire heterozygosity were used to select markers for linkage analysis on bovine chromosome 14 for milk production traits in 321 Holstein animals. Results show putative milk peaks at 42 and 61 cM, both at p<0.10, a fat yield peak at 42 and 63 cM, both at p<0.05; a protein yield peak at 42 (p<0.01) and 84 cM (p<0.05); fat per cent peaks at 3 (p<0.01) and 29 cM (p<0.05), and a protein per cent peak at 4 cM (p<0.05). Once quantitative trait loci positions were established, allele substitution effects for all markers were evaluated using the same statistical model. Overlaying information between quantitative trait loci (QTL) and allele effect analysis enabled the identification (p<0.01) of 20 SNPs under the milk yield QTL, 2 under both of the fat yield peaks, 8 and 9 under the protein yield peaks, 2 and 6 for the fat per cent peaks and 5 for the protein per cent peak. One SNP in particular, ss61514555:A>C, showed association with 3 of the 5 traits: milk (p=1.59E-04), fat (p=6.88E-05) and protein yields (p=5.76E-05). Overall, combining information from linkage disequilibrium, sire heterozygosity and genetic knowledge of traits enabled the characterization of additional markers with significant associations with milk production traits.     

多年生黑麦草叶片长度数量性状位点(QTLs)研究

选择固定有益等位基因,研究多年生黑麦草叶片长度数量性状位点,结果表明:与叶长最为相关的3个标记所表达的变异占表型总变异的50.9%(r2);在选择中如能将这3个标记最优化地组合在一起,其综合共同作用有望使叶片长度延长和缩短26 mm;鉴于供试材料在最初3个世代的群体选择中所估测到叶片长度的遗传力为0.32,这样在今后的选择工作中,即使只使用现有的两个标记,其效果亦优于传统的表型选择法.     

Power of a genome scan to detect and locate quantitative trait loci in cattle using dense single nucleotide polymorphisms

There is increasing use of dense single nucleotide polymorphisms (SNPs) for whole‐genome association studies (WGAS) in livestock to map and identify quantitative trait loci (QTL). These studies rely on linkage disequilibrium (LD) to detect an association between SNP genotypes and phenotypes. The power and precision of these WGAS are unknown, and will depend on the extent of LD in the experimental population. One complication for WGAS in livestock populations is that they typically consist of many paternal half‐sib families, and in some cases full‐sib families; unless this subtle population stratification is accounted for, many spurious associations may be reported. Our aim was to investigate the power, precision and false discovery rates of WGAS for QTL discovery, with a commercial SNP array, given existing patterns of LD in cattle. We also tested the efficiency of selective genotyping animals. A total of 365 cattle were genotyped for 9232 SNPs. We simulated a QTL effect as well as polygenic and environmental effects for all animals. One QTL was simulated on a randomly chosen SNP and accounted for 5%, 10% or 18% of the total variance. The power to detect a moderate‐sized additive QTL (5% of the phenotypic variance) with 365 animals genotyped was 37% (p < 0.001). Most importantly, if pedigree structure was not accounted for, the number of false positives significantly increased above those expected by chance alone. Selective genotyping also resulted in a significant increase in false positives, even when pedigree structure was accounted for.     

A genome scan for quantitative trait loci affecting the length of small intestine in a White Duroc × Chinese Erhualian intercross resource population

The small intestine is a vital organ in animal gastrointestinal system, in which a large variety of nutrients are absorbed. To identify quantitative trait loci (QTL) for the length of porcine small intestine, phenotypic values were measured in 1034 individuals at 240 d from a White Duroc × Chinese Erhualian intercross F₂ population. The length of small intestine showed strong correlation with growth traits and carcass length in the F2 population. A whole‐genome scan was performed based on 183 microsatellites covering the pig genome in the F₂ population. A total of 10 QTL for this trait were identified on 8 pig chromosomes (SSC), including four 1% genome‐wide significant QTL on SSC2, 4, 7 and 8, one 5% genome‐wide significant QTL on SSC12, and five 5% chromosome‐wide significant QTL on SSC5, 7, 13 and 14. The Erhualian alleles were generally associated with shorter length of the small intestine except the alleles on SSC7 and 13. The QTL on SSC4 overlapped with the previously reported QTL for the length of small intestine. Several significant QTL on SSC2, 8, and 12 were consistent with previous reports. The significant QTL detected on SSC7 was reported for the first time. All QTL identified in this study corresponded to the known region significantly associated with growth traits, supporting the important role of the length of small intestine in pig growth.     

QTL detection for milk production traits in goats using a longitudinal model.

Eight paternal half-sib families were used to identify chromosomal regions associated with variation in the lactation curves of dairy goats. DNA samples from 162 animals were amplified by PCR for 37 microsatellite markers, from Capra hircus autosomes CHI3, CHI6, CHI14 and CHI20. Milk samples were collected during 6 years, and there were 897 records for milk yield (MY) and 814 for fat (FP) and protein percentage (PP). The analysis was conducted in two stages. First, a random regression model with several fixed effects was fitted to describe the lactation function, using a scale (alpha) plus four shape parameters: beta and gamma, both associated with a decrease in the slope of the curve, and delta and phi that are related to the increase in slope. Predictions of alpha, beta, gamma, delta and phi were regressed using an interval mapping model, and F-tests were used to test for quantitative trait loci (QTL) effects. Significant (p < 0.05) QTLs were found for: (i) MY: CHI6 at 70-80 cM for all parameters; CHI14 at 14 cM for delta and phi; (ii) FP: CHI14, at 63 cM was associated with beta; CHI20, at 72 cM, showed association with alpha; (iii) PP: chromosomal regions associated with beta were found at 59 cM in CHI3 and at 55 cM in CHI20 with alpha and gamma. Analyses using more families and more animals will be useful to confirm or to reject these findings.     

Interval mapping of growth quantitative trait loci in Japanese Black beef cattle using microsatellite DNA markers and half-sib regression analysis

A confirmatory scan for the regions of bovine chromosome 1 segregating the quantitative trait loci (QTL) influencing birthweight, weaning weight, yearling weight, and preweaning and postweaning average daily gains was performed by genotyping half‐sib progeny of four Japanese Black sires using microsatellite DNA markers. Data were analyzed by generating an F‐statistic every 1 cM on a linkage map by the regression of phenotype on the probabilities of inheriting an allele from the sire after adjusting for the fixed effects of sire, sex, parity and season of birth as well as age as a covariate. Permutation tests at chromosome‐wide significance thresholds were carried out over 10 000 iterations. A significant QTL for birthweight at 114 cM was detected in the sire 2 family. This identification of a birthweight QTL in Japanese Black cattle may be useful for the implementation of marker‐assisted selection.     

Mapping the quantitative trait loci (QTL) for body shape and conformation measurements on BTA1 in Japanese Black cattle

The detection and mapping of segregating quantitative trait loci (QTL) that influence withers height, hip height, hip width, body length, chest width, chest depth, shoulder width, lumbar width, thurl width, pin bone width, rump length, cannon circumference, chest girth, abdominal width and abdominal girth at weaning was conducted on chromosomal regions of bovine chromosome one. The QTL analysis was performed by genotyping half‐sib progeny of five Japanese Black sires using microsatellite DNA markers. Probability coefficients of inheriting allele 1 or 2 from the sire at specific chromosomal locations were computed. The phenotypic data of progeny were regressed on these probability coefficients in a within‐common‐parent regression analysis using a linear model that included fixed effects of sex, parity and season of birth, as well as age as a covariate. F‐statistics were calculated every 1 cM on a linkage map. Permutation tests of 10 000 iterations were conducted to obtain chromosome‐wide significance thresholds. A significant QTL for chest width was detected at 91 cM in family 3. The detection of this QTL boosts the prospects of implementing marker‐assisted selection for body conformation traits in Japanese Black beef cattle.     

利用24个微卫星进行猪数量性状座位定位及其遗传效应分析

以 3头英系大白公猪与 7头梅山母猪杂交产生的三代资源家系用来检测猪重要经济性状的数量性状座位(QTL) ,2 0 0 0年下半年随机选留 140头F2代个体 ,进行屠宰测定 ,记录了包括生长、胴体组成等 43个性状 ;从已定位于家猪 3、4和 7号染色体上的遗传标记中选用 2 4个微卫星标记对所有个体进行基因型检测。采用最小二乘回归区间定位法进行QTL检测 ,通过置换实验来确定显著性阈值。在所研究的 32个生长和胴体性状中 ,3条染色体总共 16个QTL达到染色体显著水平 (P <0 0 5 ) ,其中 4个达到染色体极显著性水平 (P <0 0 1) ;同时在 4号和 7号染色体上还检测到了影响器官重性状的 3个QTL ,达到了染色体显著水平 (P <0 0 5 )。在某些QTL座位 ,其有利等位基因来源于具有较低性状平均值的品种。 2QTL模型分析下 ,在 4号染色体上检测到影响板油重的 2个QTL ,并且它们的效应方向相反。     

Quantitative trait loci analysis for growth and carcass traits in a Meishan x Duroc F2 resource population

We constructed a pig F2 resource population by crossing a Meishan sow and a Duroc boar to locate economically important trait loci. The F2 generation was composed of 865 animals (450 males and 415 females) from four F1 males and 24 F1 females and was genotyped for 180 informative microsatellite markers spanning 2,263.6 cM of the whole pig genome. Results of the genome scan showed evidence for significant quantitative trait loci (<1% genomewise error rate) affecting weight at 30 d and average daily gain on Sus scrofa chromosome (SSC) 6, carcass yield on SSC 7, backfat thickness on SSC 7 and SSC X, vertebra number on SSC 1 and SSC 7, loin muscle area on SSC 1 and SSC 7, moisture on SSC 13, intramuscular fat content on SSC 7, and testicular weight on SSC 3 and SSC X. Moreover, 5% genomewise significant QTL were found for birth weight on SSC 7, average daily gain on SSC 4, carcass length on SSC 6, SSC 7, and SSC X and lightness (L value) on SSC 3. We identified 38 QTL for 28 traits at the 5% genomewise level. Of the 38 QTL, 24 QTL for 17 traits were significant at the 1% genomewise level. Analysis of marker genotypes supported the breed of origin results and provided further evidence that a suggestive QTL for circumference of cannon bone also was segregating within the Meishan parent. We identified genomic regions related with growth and meat quality traits. Fine mapping will be required for their application in introgression programs and gene cloning.     

Crossbreeding parameters of general immune response traits in White Leghorn chickens

Crossbreeding parameters of immune response traits were estimated from a set of well characterized crossbred populations derived from three chicken lines selected over 12 generations for three different general immune response traits and their F1, F2 and backcrosses. The three traits investigated were the selection criteria from each of the lines, i.e. antibody response to the Newcastle disease virus vaccine 3 weeks after vaccination (ND3), cell-mediated immune response (response to phytohemagglutinin, PHA) and phagocytic activity measured as carbon clearance (CC). Crossbreeding parameters included direct and maternal additive line effects, direct and maternal heterosis as well as direct epistatic recombination loss. They were estimated as linear combinations of genetic group effects estimated using animal model methodology. Significant line differences were obtained for ND3 and, to a lesser extent, CC. They were mainly due to direct effects, maternal effects being significant for none of the 3 traits. Significantly negative direct heterosis effects were also observed for ND3 and CC, but not for PHA. Maternal heterosis effects were not estimated for CC. They were non significant for PHA, and negative and significant (− 0.78 ± 0.24) for ND3. The significant favourable recombination gain estimated for ND3 (3.21 ± 0.88) indicates that epistatic interactions could be important for this trait.The present work shows that it was worthwhile to complete second generation crosses to be able to assess to what extent immunity gained by selection is maintained in advanced crossbred generations, and to compare the transmission of immune traits implicated in different aspects of immunity.     

Quantitative trait loci mapping for meat quality and muscle fiber traits in a Japanese wild boar x Large White intercross

Three generations of a swine family produced by crossing a Japanese wild boar and three Large White female pigs were used to map QTL for various production traits. Here we report the results of QTL analyses for skeletal muscle fiber composition and meat quality traits based on phenotypic data of 353 F(2) animals and genotypic data of 225 markers covering almost the entire pig genome for all of the F(2) animals as well as their F(1) parents and F(0) grandparents. The results of a genome scan using least squares regression interval mapping provided evidence that QTL (<1% genome-wise error rate) affected the proportion of the number of type IIA muscle fibers on SSC2, the number of type IIB on SSC14, the relative area (RA) of type I on SSCX, the RA of type IIA on SSC6, the RA of type IIB on SSC6 and SSC14, the Minolta a* values of loin on SSC4 and SSC6, the Minolta b* value of loin on SSC15, and the hematin content of the LM on SSC6. Quantitative trait loci (<5% genome-wise error rate) were found for the number of type I on SSC1, SSC14, and SSCX, for the number of type IIA on SSC14, for the number of type IIB on SSC2, for the RA of type IIA on SSC2, for the Minolta b* value of loin on SSC3, for the pH of loin on SSC15, and for the i.m. fat content on SSC15. Twenty-four QTL were detected for 11 traits at the 5% genome-wise level. Some traits were associated with each other, so the 24 QTL were located on 11 genomic regions. In five QTL located on SSC2, SSC6, and SSC14, each wild boar allele had the effect of increasing types I and IIA muscle fibers and decreasing type IIB muscle fibers. These effects are expected to improve meat quality.     

Quantitative trait loci mapping in an F2 Duroc x Pietrain resource population: I. Growth traits

Pigs from the F(2) generation of a Duroc x Pietrain resource population were evaluated to discover QTL affecting growth and composition traits. Body weight and ultrasound estimates of 10th-rib backfat, last-rib backfat, and LM area were serially measured throughout development. Estimates of fat-free total lean, total body fat, empty body protein, empty body lipid, and ADG from 10 to 22 wk of age were calculated, and random regression analyses were performed to estimate individual animal phenotypes representing intercept and linear rates of increase in these serial traits. A total of 510 F(2) animals were genotyped for 124 micro-satellite markers evenly spaced across the genome. Data were analyzed with line cross, least squares regression, interval mapping methods using sex and litter as fixed effects. Significance thresholds of the F-statistic for single QTL with additive, dominance, or imprinted effects were determined at the chromosome- and genome-wise levels by permutation tests. A total of 43 QTL for 22 of the 29 measured traits were found to be significant at the 5% chromosome-wise level. Of these 43 QTL, 20 were significant at the 1% chromosome-wise significance threshold, 14 of these 20 were also significant at the 5% genome-wise significance threshold, and 10 of these 14 were also significant at the 1% genome-wise significance threshold. A total of 22 QTL for the animal random regression terms were found to be significant at the 5% chromosome-wise level. Of these 22 QTL, 6 were significant at the 1% chromosome-wise significance threshold, 4 of these 6 were also significant at the 5% genome-wise significance threshold, and 3 of these 4 were also significant at the 1% genome-wise significance threshold. Putative QTL were discovered for 10th-rib and last-rib backfat on SSC 6, body composition traits on SSC 9, backfat and lipid composition traits on SSC 11, 10th-rib backfat and total body fat tissue on SSC 12, and linear regression of last-rib backfat and total body fat tissue on SSC 8. These results will facilitate fine-mapping efforts to identify genes controlling growth and body composition of pigs that can be incorporated into marker-assisted selection programs to accelerate genetic improvement in pig populations.     

Identification of recombination hotspots and quantitative trait loci for recombination rate in layer chickens

Background: The frequency of recombination events varies across the genome and between individuals, which may be related to some genomic features. The objective of this study was to assess the frequency of recombination events and to identify QTL(quantitative trait loci) for recombination rate in two purebred layer chicken lines.Methods: A total of 1200 white-egg layers(WL) were genotyped with 580 K SNPs and 5108 brown-egg layers(BL)were genotyped with 42 K SNPs(single nucleotide polymorphisms). Recombination events were identified within half-sib families and both the number of recombination events and the recombination rate was calculated within each0.5 Mb window of the genome. The 10% of windows with the highest recombination rate on each chromosome were considered to be recombination hotspots. A BayesB model was used separately for each line to identify genomic regions associated with the genome-wide number of recombination event per meiosis. Regions that explained more than 0.8% of genetic variance of recombination rate were considered to harbor QTL.Results: Heritability of recombination rate was estimated at 0.17 in WL and 0.16 in BL. On average, 11.3 and 23.2 recombination events were detected per individual across the genome in 1301 and 9292 meioses in the WL and BL,respectively. The estimated recombination rates differed significantly between the lines, which could be due to differences in inbreeding levels, and haplotype structures. Dams had about 5% to 20% higher recombination rates per meiosis than sires in both lines. Recombination rate per 0.5 Mb window had a strong negative correlation with chromosome size and a strong positive correlation with GC content and with CpG island density across the genome in both lines. Different QTL for recombination rate were identified in the two lines. There were 190 and 199 non-overlapping recombination hotspots detected in WL and BL respectively, 28 of which were common to both lines.Conclusions: Differences in the recombination rates, hotspot locations, and QTL regions associated with genomewide recombination were observed between lines, indicating the breed-specific feature of detected recombination events and the control of recombination events is a complex polygenic trait.