Putative quantitative trait loci associated with the probability of contracting infectious bovine keratoconjunctivitis.

Infectious bovine keratoconjunctivitis, also known as pinkeye, is an economically important disease in cattle. The objective of this study was to detect QTL associated with infectious bovine keratoconjunctivitis in offspring from a Brahman x Hereford sire. The sire was mated to Hereford, Angus, and F1 cows to produce 288 offspring in 1994 and mated to MARC III ((1/4) Hereford, (1/4) Angus, (1/4) Red Poll, and (1/4) Pinzgauer) cows in 1996 to produce 259 offspring (547 animals total). Infectious bovine keratoconjunctivitis was diagnosed by physical examination in 36 animals of the family. Records included unilateral and bilateral frequency, but not severity. Records were binary: 0 for unaffected and 1 for affected cattle. A putative QTL for infectious bovine keratoconjunctivitis was identified on chromosome 1, with a maximum F-statistic (F = 10.15; P = 0.0015) at centimorgan 79 of the linkage group. The support interval spanned centimorgans 66 to 110. There was also evidence suggesting the presence of a QTL for infectious bovine keratoconjunctivitis on chromosome 20, with a maximum F-statistic (F = 10.35; P = 0.0014) at centimorgan 16 of the linkage group. The support interval ranged from centimorgan 2 to 35. This report provides the initial evidence of QTL for infectious bovine keratoconjunctivitis. Although a candidate gene was identified for one of the regions of interest, further studies are needed to identify the genetic basis of resistance to the disease.

Identification of genetic markers for fat deposition and meat tenderness on bovine chromosome 5: development of a low-density single nucleotide polymorphism map.

As genetic markers, SNP are well suited for the development of genetic tests for production traits in livestock. They are stable through many generations and can provide direct assessment of individual animal's genetic merit if they are in linkage disequilibrium and phase with functional genetic variation. Bovine chromosome 5 has been shown to harbor genetic variation affecting production traits in multiple cattle populations; thus, this chromosome was targeted for SNP-based marker development and subsequent association analysis with carcass and growth phenotypes. Discovery of SNP was performed in a panel of 16 sires representing two sires from each of seven beef breeds and two Holstein sires by PCR amplification and sequencing using primers designed from genomic sequence obtained by low-coverage sequencing of bacterial artificial chromosome (BAC) clones. From 550 SNP, 296 (54%) were tentatively identified as having a minor allele frequency >10%. Forty-five SNP derived from 15 BAC were chosen based on minor allele frequency and were genotyped in 564 steers and their sires. Production and carcass data were collected on the steers as a part of the Germplasm Evaluation (GPE), Cycle VII Project at the U.S. Meat Animal Research Center (Clay Center, NE), which involves of the evaluation of sires from seven of the most popular U.S. breeds. Haplotypes based on seven SNP derived from a BAC containing the bovine genes HEM1 and PDE1B were associated with traits related to carcass fat. Steers homozygous for the major haplotype had 0.15 +/- 0.04 cm less subcutaneous fat, 0.57 +/- 0.18 kg less rib fat, 0.18 +/- 0.07 lower yield grade, 1.11 +/- 0.35% less predicted fat yield, and 0.79 +/- 0.3% greater predicted retail product yield than heterozygotes. The frequency of the major haplotype was 0.70 in the steers, and it ranged from 0.44 (Limousin) to 0.98 (Simmental and Gelbvieh) in a panel consisting of an average of 20 purebred sires from each of the seven breeds. A second set of haplotypes based on four SNP derived from a BAC containing the genes NOL1 and CHD4 was associated with Warner-Bratzler shear force. Steers homozygous for the major haplotype had 0.27 +/- 0.11 kg greater shear force than those heterozygous for the major haplotype and one of two minor haplotypes. The frequency of the major haplotype was 0.59 in the steers and ranged from 0.27 (Hereford) to approximately 0.95 (Angus and Red Angus) in the panel of purebred sires. These results demonstrate the feasibility of targeting QTL regions for SNP-based marker development and that a low level of coverage can identify markers associated with phenotypic traits.

Quantitative trait loci for male reproductive traits in beef cattle.

The objective of the present study was to detect quantitative trait loci (QTL) for male reproductive traits in a half-sib family from a Bos indicus (Brahman) x Bos taurus (Hereford) sire. The sire was mated with MARC III (1/4 Hereford, 1/4 Angus, 1/4 Red Poll and 1/4 Pinzgauer) cows. Testicular traits were measured from 126 male offspring born in 1996 and castrated at 8.5 months. Traits analysed were concentration of follicle stimulating hormone in peripheral blood at castration (FSH), paired testicular weight (PTW) and paired testicular volume (PTV) adjusted for age of dam, calculated age at puberty (AGE), and body weight at castration (BYW). A putative QTL was observed for FSH on chromosome 5. The maximum F-statistic was detected at 70 cM from the beginning of the linkage group. Animals inheriting the Hereford allele had a 2.47-ng/ml higher concentration of FSH than those inheriting the Brahman allele. Evidence also suggests the existence of a putative QTL on chromosome 29 for PTW, PTV, AGE and BYW. The maximum F-statistic was detected at cM 44 from the beginning of the linkage group for PTW, PTV and AGE, and at cM 52 for BYW. Animals that inherited the Brahman allele at this chromosomal region had a 45-g heavier PTW, a 42-cm(3) greater PTV, a 39-day younger AGE and a 22.8-kg heavier BYW, compared with those inheriting the Hereford allele. This is the first report of QTL for male reproductive traits in cattle.

Identification of quantitative trait loci for growth and carcass composition in cattle.

A genomic screening to detect quantitative trait loci (QTL) affecting growth, carcass composition and meat quality traits was pursued. Two hundred nineteen microsatellite markers were genotyped on 176 of 620 (28%) progeny from a Brahman x Angus sire mated to mostly MARC III dams. Selective genotyping, based on retail product yield (%) and fat yield (%), was used to select individuals to be genotyped. Traits included in the study were birth weight (kg), hot carcass weight (kg), retail product yield, fat yield, marbling score (400 = slight00 and 500 = small00), USDA yield grade, and estimated kidney, heart and pelvic fat (%). The QTL were classified as significant when the expected number of false positives (ENFP) was less than 0.05 (F-statistic greater than 17.3), and suggestive when the ENFP was <1 (F-statistic between 10.2 and 17.3). A significant QTL (F = 19; ENFP = 0.02) was detected for marbling score at centimorgan (cM) 54 on chromosome 2. Suggestive QTL were detected for fat yield at 50 cM, for retail product yield at 53 cM, and for USDA yield grade at 63 cM on chromosome 1, for marbling score at 56 cM, for retail product yield at 70 cM, and for estimated kidney, heart and pelvic fat at 79 cM on chromosome 3, for marbling score at 44 cM, for hot carcass weight at 49 cM, and for estimated kidney, heart and pelvic fat at 62 cM on chromosome 16, and for fat yield at 35 cM on chromosome 17. Two suggestive QTL for birth weight were identified, one at 12 cM on chromosome 20 and the other at 56 cM on chromosome 21. An additional suggestive QTL was detected for retail product yield, for fat yield, and for USDA yield grade at 26 cM on chromosome 26. Results presented here represent the initial search for quantitative trait loci in this family. Validation of detected QTL in other populations will be necessary.

Detection of quantitative trait loci for growth and carcass composition in cattle.

The objective of the present study was to detect quantitative trait loci for economically important traits in a family from a Bos indicus x Bos taurus sire. A Brahman x Hereford sire was used to develop a half-sib family (n = 547). The sire was mated to Bos taurus cows. Traits analyzed were birth (kg) and weaning weights (kg); hot carcass weight (kg); marbling score; longissimus area (cm2); USDA yield grade; estimated kidney, pelvic, and heart fat (%); fat thickness (cm); fat yield (%); and retail product yield (%). Meat tenderness was measured as Warner-Bratzler shear force (kg) at 3 and 14 d postmortem. Two hundred and thirty-eight markers were genotyped in 185 offspring. One hundred and thirty markers were used to genotype the remaining 362 offspring. A total of 312 markers were used in the final analysis. Seventy-four markers were common to both groups. Significant QTL (expected number of false-positives < 0.05) were observed for birth weight and longissimus area on chromosome 5, for longissimus area on chromosome 6, for retail product yield on chromosome 9, for birth weight on chromosome 21, and for marbling score on chromosome 23. Evidence suggesting (expected number of false-positives < 1) the presence of QTL was detected for several traits. Putative QTL for birth weight were detected on chromosomes 1, 2, and 3, and for weaning weight on chromosome 29. For hot carcass weight, QTL were detected on chromosomes 10, 18, and 29. Four QTL for yield grade were identified on chromosomes 2, 11, 14, and 19. Three QTL for fat thickness were detected on chromosomes 2, 3, 7, and 14. For marbling score, QTL were identified on chromosomes 3, 10, 14, and 27. Four QTL were identified for retail product yield on chromosomes 12, 18, 19, and 29. A QTL for estimated kidney, pelvic, and heart fat was detected on chromosome 15, and a QTL for meat tenderness measured as Warner-Bratzler shear force at 3 d postmortem was identified on chromosome 20. Two QTL were detected for meat tenderness measured as Warner-Bratzler shear force at 14 d postmortem on chromosomes 20 and 29. These results present a complete scan in all available progeny in this family. Regions underlying QTL need to be assessed in other populations.

An enhanced linkage map of the sheep genome comprising more than 1000 loci.

A medium-density linkage map of the ovine genome has been developed. Marker data for 550 new loci were generated and merged with the previous sheep linkage map. The new map comprises 1093 markers representing 1062 unique loci (941 anonymous loci, 121 genes) and spans 3500 cM (sex-averaged) for the autosomes and 132 cM (female) on the X chromosome. There is an average spacing of 3.4 cM between autosomal loci and 8.3 cM between highly polymorphic [polymorphic information content (PIC) > or = 0.7] autosomal loci. The largest gap between markers is 32.5 cM, and the number of gaps of > 20 cM between loci, or regions where loci are missing from chromosome ends, has been reduced from 40 in the previous map to 6. Five hundred and seventy-three of the loci can be ordered on a framework map with odds of > 1000 : 1. The sheep linkage map contains strong links to both the cattle and goat maps. Five hundred and seventy-two of the loci positioned on the sheep linkage map have also been mapped by linkage analysis in cattle, and 209 of the loci mapped on the sheep linkage map have also been placed on the goat linkage map. Inspection of ruminant linkage maps indicates that the genomic coverage by the current sheep linkage map is comparable to that of the available cattle maps. The sheep map provides a valuable resource to the international sheep, cattle, and goat gene mapping community.

Comparative mapping of BTA15 and HSA11 including a region containing a QTL for meat tenderness.

The starting point of the present study was the reported identification of a chromosomal region on bovine Chromosome (Chr) 15 (BTA15) carrying loci affecting meat tenderness. A comparative linkage map of BTA15 and human Chr 11 (HSA11) was constructed to identify potential positional candidate genes and to provide a resource of genetic markers to support marker-assisted selection (MAS). Relative rearrangements between the bovine and human genomes for these chromosomes are the most complex observed in comparative mapping between the two species, with nine alternating blocks of conserved synteny between HSA11 and bovine Chrs 15 and 29. The results of this study were the addition of nine genes to the HSA11/BTA15 comparative linkage map, and development of five microsatellite markers within the quantitative trait locus (QTL) interval. One gene with known effects on muscle development (MYOD1) was mapped to the interval. A second gene (CALCA) involved in regulation of calcium levels, a key factor in postmortem tenderization, also mapped within the interval. Refinement of the comparative map and QTL position will reduce the interval on the human transcription map to be scanned in search of candidates, reducing the effort and resources required to identify the allelic variation responsible for the genetic effect.

Sequence evaluation of four pooled-tissue normalized bovine cDNA libraries and construction of a gene index for cattle.

An essential component of functional genomics studies is the sequence of DNA expressed in tissues of interest. To provide a resource of bovine-specific expressed sequence data and facilitate this powerful approach in cattle research, four normalized cDNA libraries were produced and arrayed for high-throughput sequencing. The libraries were made with RNA pooled from multiple tissues to increase efficiency of normalization and maximize the number of independent genes for which sequence data were obtained. Target tissues included those with highest likelihood to have impact on production parameters of animal health, growth, reproductive efficiency, and carcass merit. Success of normalization and inter- and intralibrary redundancy were assessed by collecting 6000-23,000 sequences from each of the libraries (68,520 total sequences deposited in GenBank). Sequence comparison and assembly of these sequences was performed in combination with 56,500 other bovine EST sequences present in the GenBank dbEST database to construct a cattle Gene Index (available from The Institute for Genomic Research at http://www.tigr.org/tdb/tgi.shtml). The 124,381 bovine ESTs present in GenBank at the time of the analysis form 16,740 assemblies that are listed and annotated on the Web site. Analysis of individual library sequence data indicates that the pooled-tissue approach was highly effective in preparing libraries for efficient deep sequencing.

A comprehensive search for quantitative trait loci affecting growth and carcass composition of cattle segregating alternative forms of the myostatin gene.

The objective of this study was to identify quantitative trait loci for economically important traits in two families segregating an inactive copy of the myostatin gene. Two half-sib families were developed from a Belgian Blue x MARC III (n = 246) and a Piedmontese x Angus (n = 209) sire. Traits analyzed were birth, weaning, and yearling weight (kg); preweaning average daily gain (kg/d); postweaning average daily gain (kg/d); hot carcass weight (kg); fat depth (cm); marbling score; longissimus muscle area (cm2); estimated kidney, pelvic, and heart fat (%); USDA yield grade; retail product yield (%); fat yield (%); and wholesale rib-fat yield (%). Meat tenderness was measured as Warner-Bratzler shear force at 3 and 14 d postmortem. The effect of the myostatin gene was removed using phase information from six microsatellite markers flanking the locus. Interactions of the myostatin gene with other loci throughout the genome were also evaluated: The objective was to use markers in each family, scanning the genome approximately every 25 to 30 centimorgans (cM) on 18 autosomal chromosomes, excluding 11 autosomal chromosomes previously analyzed. A total of 89 markers, informative in both families, were used to identify genomic regions potentially associated with each trait. In the family of Belgian Blue inheritance, a significant QTL (expected number of false-positives = 0.025) was identified for marbling score on chromosome 3. Suggestive QTL for the same family (expected number of false-positives = 0.5) were identified for retail product yield on chromosome 3, for hot carcass weight and postweaning average daily gain on chromosome 4, for fat depth and marbling score on chromosome 8, for 14-d Warner-Bratzler shear force on chromosome 9, and for marbling score on chromosome 10. Evidence suggesting the presence of an interaction for 3-d Warner-Bratzler shear force between the myostatin gene and a QTL on chromosome 4 was detected. In the family of Piedmontese and Angus inheritance, evidence indicates the presence of an interaction for fat depth between the myostatin gene and chromosome 8, in a similar position where the evidence suggests the presence of a QTL for fat depth in the family with Belgian Blue inheritance. Regions identified underlying QTL need to be assessed in other populations. Although the myostatin gene has a considerable effect, other loci with more subtle effects are involved in the expression of the phenotype.

Consensus and comprehensive linkage maps of bovine chromosome 7.

The objective of this project was to integrate the currently available linkage maps for bovine chromosome 7 (BTA7) by combining data sets from eight research groups. A total of 54 unique markers were typed in eight pedigrees. Multilocus linkage analysis with CRI-MAP produced a bovine chromosome 7 consensus framework map of 27 loci ordered with odds greater than 1000:1. Furthermore, we present a bovine chromosome 7 comprehensive map integrating 54 loci. The locus order is in general agreement with the recently published linkage maps except for one discrepancy. The order of loci BM9289, BMS713, and ILSTS001 was reversed in the consensus framework map relative to the published USDA-MARC bovine chromosome 7 linkage map.

Quantitative trait loci affecting growth and carcass composition of cattle segregating alternate forms of myostatin.

The effects of the bovine myostatin gene on chromosome 2 on birth and carcass traits have been previously assessed. The objective of this study was to identify additional quantitative trait loci (QTL) for economically important traits in two families segregating an inactive copy of myostatin. Two half-sib families were developed from Belgian Blue x MARC III (n = 246) and Piedmontese x Angus (n = 209) sires. Traits analyzed were birth (kg) and yearling weight (kg); hot carcass weight (kg); fat depth (cm); marbling score; longissimus muscle area (cm2); estimated kidney, pelvic, and heart fat (%); USDA yield grade; retail product yield (%); fat yield (%); and wholesale rib-fat yield (%). Meat tenderness was measured as Warner-Bratzler shear force at 3 and 14 d postmortem. The effect of myostatin on these traits was removed by using phase information obtained from the previous study with six microsatellite markers flanking the locus. Selective genotyping was done on 92 animals from both families to identify genomic regions potentially associated with retail product yield and fat depth, using a total of 150 informative markers in each family. Regions in which selective genotyping indicated the presence of QTL were evaluated further by genotyping the entire population and additional markers. For the family with Belgian Blue inheritance (n = 246), a significant QTL for birth and yearling weight was identified on chromosome 6. Suggestive QTL were identified for longissimus muscle area and hot carcass weight on chromosome 6 and for marbling on chromosomes 17 and 27. For the family with Piedmontese inheritance (n = 209), suggestive QTL on chromosome 5 were identified for fat depth, retail product yield, and USDA yield grade and on chromosome 29 for Warner-Bratzler shear force at 3 and 14 d postmortem. Interactions suggesting the presence of QTL were observed between myostatin and chromosome 5 for Warner-Bratzler shear force at 14 d postmortem and between myostatin and chromosome 14 for fat depth. Thus, in families segregating an inactive copy of myostatin in cattle, other loci influencing quantitative traits can be detected. These results are the initial effort to identify and characterize QTL affecting carcass and growth traits in families segregating myostatin.

Bovine UCP2 and UCP3 map to BTA15.

Two recently described uncoupling proteins, UCP2 and UCP3, may have important roles in determining feed conversion and/or maintenance energy requirements in livestock. Sequence was determined for the entire coding region and four of the six introns for bovine UCP3. A partial cDNA sequence was obtained for bovine UCP2. Radiation hybrid mapping was used to place UCP3 on BTA15, a chromosome previously shown to harbor a locus influencing meat tenderness. In order to place UCP3 in the existing linkage map, five single nucleotide polymorphisms (SNPs) were developed. Linkage analysis using one of five SNPs was used to place UCP3 between 53.1 and 53.5 CM. No recombination was detected between UCP3 and IDVGA10, IDVGA32, and INRA145. The sequence of PCR products from a single BAC amplified with primers specific for either UCP2 or UCP3 indicate that UCP2 and UCP3 are separated by < 200 kb.

Quantitative analysis of birth, weaning, and yearling weights and calving difficulty in Piedmontese crossbreds segregating an inactive myostatin allele.

The Piedmontese breed has a high frequency of double-muscling. Animals tested in this breed are homozygous for a guanine to adenine transition in exon 3 (C313Y) of the myostatin (MSTN) gene. This transition seems to be responsible for the double-muscling phenotype. The objective of this study was to compare effects of alternative MSTN genotypes on proportion of assisted calving and weights at birth, weaning, and 1 yr of age. Reciprocal backcross and F2 calves out of Piedmontese-Angus (PA) and Piedmontese-Hereford (PH) dams born in 1995 (n = 82), 1996 (n = 75), and 1997 (n = 144) were evaluated for birth (BWT, kg), adjusted weaning (W200, kg), and yearling (W365, kg) weights and calving difficulty expressed as a proportion of assisted calving (CD). The number of copies of C313Y was assessed in each calf. Data were analyzed with a model that included effects of year, sex, subclasses of proportion Piedmontese (.25, .5, .75) by number of C313Y copies (0 = +/+, 1 = mh/+, 2 = mh/mh), and age of dam as covariate. For BWT, heterozygous mh/+ animals were 3.2 +/- .8 kg heavier than +/+ animals. Homozygous mh/mh animals increased .19 +/- .06 in proportion of CD compared with mh/+ animals. Differences between homozygous animals (mh/mh - +/+) were 5.2 +/- 1 kg for BWT and .21 +/- .06 for CD. Heterozygous mh/+ animals were 9.1 +/- 4 kg heavier at W200 than homozygous +/+ animals. Homozygous +/+ and heterozygous animals were 20 +/- 8 and 24.5 +/- 8 kg, respectively, heavier at W365 than mh/mh animals. Differences between mh/+ and the mean of mh/mh and +/+ genotypes for W200 and W365 were 8.8 +/- 3 and 18 +/- 5 kg, respectively, suggesting dominance effects on postnatal growth. Production of heterozygous animals, to take advantage of the positive impact of one copy of C313Y on carcass traits, may be a viable option when the value of increased retail product yield is greater than the increased cost associated with calving difficulty.

A region on bovine chromosome 15 influences beef longissimus tenderness in steers.

A genome scan was conducted using 196 microsatellite DNA markers spanning 29 autosomal bovine chromosomes and Warner-Bratzler shear force collected at d 2 and 14 postmortem on steaks from the longissimus muscle of 294 progeny from one Brahman x Hereford bull mated to Bos taurus cows to identify QTL for beef tenderness. One QTL was identified and located 28 cM (95% confidence interval is 17 to 40 cM) from the most centromeric marker on BTA15. The QTL interacted significantly with slaughter group. The difference in shear force of steaks aged 14 d postmortem between progeny with the Brahman paternally inherited allele vs those with Hereford was 1.19 phenotypic standard deviations (explained 26% of phenotypic variance) for one slaughter group and was not significant for three other slaughter groups. Apparently, unknown environmental factors present for three of the four slaughter groups were capable of masking the effect of this QTL. The sensitivity of the QTL effect to environmental factors may complicate utilization of markers for genetic improvement. Future research to elucidate the cause of the QTL x slaughter group interaction may lead to improved strategies for controlling variation in meat tenderness via marker-assisted selection, postmortem processing, or live animal management.

A primary screen of the bovine genome for quantitative trait loci affecting carcass and growth traits.

A primary genomic screen for quantitative trait loci (QTL) affecting carcass and growth traits was performed by genotyping 238 microsatellite markers on 185 out of 300 total progeny from a Bos indicus x Bos taurus sire mated to Bos taurus cows. The following traits were analyzed for QTL effects: birth weight (BWT), weaning weight (WW), yearling weight (YW), hot carcass weight (HCW), dressing percentage (DP), fat thickness (FT), marbling score (MAR), longissimus muscle area (LMA), rib bone (RibB), rib fat (RibF), and rib muscle (RibM), and the predicted whole carcass traits, retail product yield (RPYD), fat trim yield (FATYD), bone yield (BOYD), retail product weight (RPWT), fat weight (FATWT), and bone weight (BOWT). Data were analyzed by generating an F-statistic profile computed at 1-cM intervals for each chromosome by the regression of phenotype on the conditional probability of receiving the Brahman allele from the sire. There was compelling evidence for a QTL allele of Brahman origin affecting an increase in RibB and a decrease in DP on chromosome 5 (BTA5). Putative QTL at or just below the threshold for genome-wide significance were as follows: an increase in RPYD and component traits on BTA2 and BTA13, an increase in LMA on BTA14, and an increase in BWT on BTA1. Results provided represent a portion of our efforts to identify and characterize QTL affecting carcass and growth traits.