Semi-automated technique for bovine skeletal muscle fiber cross-sectional area and myosin heavy chain determination.

Myosin heavy chain (MyHC) type and muscle fiber size are informative but time-consuming variables of interest for livestock growth, muscle biology, and meat science. The objective of this study was to validate a semi-automated protocol for determining MyHC type and size of muscle fibers. Muscle fibers obtained from the longissimus and semitendinosus of fed beef carcasses were embedded and frozen within 45 min of harvest. Immunohistochemistry was used to distinguish MyHC type I, IIA, and IIX proteins, dystrophin, and nuclei in transverse sections of frozen muscle samples. Stained muscle cross sections were imaged and analyzed using two workflows: 1) Nikon workflow which used Nikon Eclipse inverted microscope and NIS Elements software and 2) Cytation5 workflow consisting of Agilent BioTek Cytation5 imaging reader and Gen5 software. With the Cytation5 workflow, approximately six times more muscle fibers were evaluated compared to the Nikon workflow within both the longissimus (P < 0.01; 768 vs. 129 fibers evaluated) and semitendinosus (P < 0.01; 593 vs. 96 fibers evaluated). Combined imaging and analysis took approximately 1 h per sample with the Nikon workflow and 10 min with the Cytation5 workflow. When muscle fibers were evaluated by the objective thresholds of the Cytation5 workflow, a greater proportion of fibers were classified as glycolytic MyHC types, regardless of muscle (P < 0.01). Overall mean myofiber cross-sectional area was 14% smaller (P < 0.01; 3,248 vs. 3,780) when determined by Cytation5 workflow than when determined by Nikon workflow. Regardless, Pearson correlation of mean muscle fiber cross-sectional areas determined by Nikon and Cytation5 workflows was 0.73 (P < 0.01). In both workflows cross-sectional area of MyHC type I fibers was the smallest and area of MyHC type IIX fibers was the largest. These results validated the Cytation5 workflow as an efficient and biologically relevant tool to expedite data capture of muscle fiber characteristics while using objective thresholds for muscle fiber classification.

Properties of muscle tissue are affected by cellular-level changes in the isoform of myosin, a protein involved in muscle contraction. The heavy chain subunit of myosin (MyHC) is affected by breed type, changes as animals mature, and interacts with muscle fiber size when growth-promoting technologies are used in meat animals. While MyHC type and muscle fiber size are important for growth potential and meat quality of livestock, measurement of these variables is time consuming. The objective of this study was to validate a semi-automated workflow for identification of MyHC type and measurement of muscle fibers compared to a previously published manual technique. The semi-automated workflow evaluated approximately six times more myofibers in one-sixth of the time compared to the manual workflow. While the semi-automated technique identified the muscle profile with greater relative abundance of glycolytic muscle fibers and 14% smaller fibers, results from both techniques were strongly correlated and found similar biological results. An additional benefit of the semi-automated workflow was the use of objective thresholds to classify MyHC types as opposed to subjective human judgement of the manual workflow. This study demonstrated that the semi-automated workflow efficiently and objectively imaged, classified, and measured muscle fibers.

Effects of the F94L myostatin gene mutation in beef × dairy crossed cattle on muscle fiber type, live performance, carcass characteristics, and boxed beef and retail cut yields.

Producer live performance data and carcasses from steers (n = 116) resulting from the mating of four Limousin/Angus sires heterozygous for the F94L myostatin mutation to Jersey/Holstein dams were utilized to evaluate the effects of one copy of the F94L allele on live performance, carcass traits and USDA grades, and boxed beef and retail yields. Slaughter data were collected at time of harvest and carcass data were collected 48 hours postmortem. One side from each of the 58 carcasses was fabricated into boxed beef and retail cuts by experienced lab personnel 5-8 d postmortem. One copy of the F94L allele did not affect gestation length, birth weight, percentage of unassisted births, feedlot average daily gain, live weight at harvest, hot carcass weight, or dressing percentage (P > 0.05). Muscle fiber analysis indicated that the increase in muscularity by the F94L allele in the semitendinosus and longissimus was likely due to hyperplasia as there was a 19% increase in the quantity of myosin heavy chain type IIA and IIX fibers in the semitendinosus (P < 0.05) with no effect on muscle fiber size (P > 0.05). Carcasses from steers with one F94L allele had larger ribeye areas (99.2 vs. 92.3 sq.cm.), greater ribeye width:length ratios (0.498 vs. 0.479), lower USDA yield grades (2.21 vs. 2.66), and lower marbling scores (438 vs. 480) (P < 0.05). Additionally, for boxed beef yields, one F94L allele, vs. zero F94L alleles, increased (P < 0.05) 85/15 trimmings (+0.59%), top round (+0.28%), strip loin (+0.12%), eye round (+0.11%), tenderloin (+0.07%), boneless foreshank (+0.07%), cap/wedge (+0.06%), and tri-tip (+0.04%). Overall, carcasses from steers with one F94L allele had a greater boxed beef yield (+1.06%), boxed beef plus 85/15 trimmings yield (+1.65%), and total retail cuts plus ground beef 85/15 yield (+1.78%) than carcasses from steers with zero F94L alleles (P < 0.05). One copy of the F94L allele utilized in beef-on-dairy breeding system had no significant impact on live performance traits but resulted in lower marbling scores and increased muscularity as evidenced through larger, more beef-shaped ribeyes, lower USDA yield grades, and greater carcass cutout yields (both boxed beef and retail yields).

In a beef-on-dairy system, one copy of the F94L myostatin allele caused increased muscling, resulting in larger, more beef-shaped ribeyes, more desirable yield grades, and greater boxed beef and retail yields, all of which address inherent deficiencies in dairy and dairy-cross carcasses. These improvements were realized with no negative effects on calving ease or live performance. The F94L did cause a significant and meaningful reduction in marbling score; therefore, marbling ability should be paramount in sire selection if F94L sires are utilized. Using a beef sire homozygous for F94L myostatin in a beef-on-dairy system would ensure that all resulting progenies have exactly one copy of the F94L allele, meaning that this genetic tool could be rapidly implemented in the beef-on-dairy industry segment. When selecting sires for beef-on-dairy programs, accurate EPDs should remain the primary evaluation tool as the F94L effects are reflected in accurate EPDs; however, using a sire homozygous for F94L (2 or 0 copies) should result in more consistent progeny.

Beef embryos in dairy cows: calfhood growth of Angus-sired calves from Holstein, Jersey, and crossbred beef dams.

Improved reproductive management has allowed dairy cow pregnancies to be optimized for beef production. The objective of this sire-controlled study was to characterize the effects of beef or dairy maternal genetics and the dairy management system on calf growth. Pregnancies were created with a 2 × 2 factorial arrangement of dam breed (Holstein or Jersey) and mating type (artificial insemination or implantation of an in vitro produced embryo from a commercial beef cow oocyte). Resulting calves were reared in a calf ranch. Additionally, commercial beef cows were inseminated and reared resulting calves on range. Therefore, the five treatments were Angus × Holstein (A × H; n = 19), Angus × Jersey (A × J; n = 22), Angus × beef gestated by Holstein (H ET; n = 18), Angus × beef gestated by Jersey (J ET; n = 8), and Angus × beef raised by beef (A × B; n = 20). Beginning at birth, calf body weight, cannon circumference, forearm circumference, top width, hip width, and hip height were measured approximately every 28 d until ~196 d of age. At birth, A × J calves weighed the least (P < 0.01). At 150 d of age, body weight was greatest (P < 0.05) among A × B calves, intermediate among H ET and A × H calves, and least among J ET and A × J calves (P < 0.05). Morphometric differences were detected between treatments (multivariate analysis of variance, P < 0.01). Primary discriminant function scores identified A × B calves having lesser values than A × J or A × H calves (analysis of variance [ANOVA], P < 0.01); A × B calves had greater cannon circumference, greater top width, and less hip height (standardized loadings of -0.47, -0.48, and 0.63, respectively). Secondary discriminant function scores identified J ET and H ET to have greater forearm circumference-a key indicator of muscling-than A × J or A × H (ANOVA, P < 0.01; standardized loading of 0.99). The dairy management system limited growth rate of beef genetics compared to the beef management system. In addition, Holstein dams transmitted greater growth potential than Jersey dams. Replacing maternal dairy genetics with beef genetics moderated frame size and created a more muscular phenotype.

Beef embryos in dairy cows: feedlot performance, mechanistic responses, and carcass characteristics of straightbred Holstein calves and Angus-sired calves from Holstein, Jersey, or crossbred beef dams.

Improved reproductive management has allowed dairy cow pregnancies to be optimized for beef production. The objective of this sire-controlled study was to test the feedlot performance of straightbred beef calves raised on a calf ranch and to compare finishing growth performance, carcass characteristics, and mechanistic responses relative to beef × dairy crossbreds and straightbred beef cattle raised in a traditional beef cow/calf system. Tested treatment groups included straightbred beef steers and heifers reared on range (A × B; n = 14), straightbred beef steers and heifers born following embryo transfer to Holstein dams (H ET; n = 15) and Jersey dams (J ET; n = 16) The finishing trial began when cattle weighed 301 ±â€…32.0 kg and concluded after 195 ±â€…1.4 d. Individual intake was recorded from day 28 until shipment for slaughter. All cattle were weighed every 28 d; serum was collected from a subset of steers every 56 d. Cattle of straightbred beef genetics (A × B, H ET, and J ET) and A × H were similar in final shrunk body weight, dry matter intake, and carcass weight (P > 0.05 for each variable). Compared with A × J cattle, J ET was 42 d younger at slaughter with 42 kg more carcass weight (P < 0.05 for both variables). No difference was observed in longissimus muscle area between all treatments (P = 0.40). Fat thickness was greatest for straightbred beef cattle, least for A × J cattle, and intermediate for A × H cattle (P < 0.05). When adjusted for percentage of adjusted final body weight, feed efficiency was greater for straightbred beef cattle compared with beef × dairy crossbred cattle (P = 0.04). A treatment × day interaction was observed for circulating insulin-like growth factor I (IGF-I; P < 0.01); 112 d after being implanted, beef × dairy crossbred cattle had greater circulating IGF-I concentration than cattle of straightbred beef genetics (P < 0.05). Straightbred beef calves born to Jersey cows had more efficient feedlot and carcass performance than A × J crossbreds. Calves of straightbred beef genetics raised traditionally or in a calf ranch performed similarly in the feedlot.

Improved reproductive management has allowed dairy cow pregnancies to be optimized for beef production. The objectives of this study were to use an embryo transfer model 1) to investigate the effect of the dairy management system on beef genetics and 2) to directly compare the merit of Holstein and Jersey genetics for feedlot and carcass performance with modern beef genetics. Feedlot and carcass performance of straightbred beef cattle were similar regardless if the calf was raised in the traditional beef cow/calf system or if the calf was raised at a calf ranch. Based on greater daily live gain and carcass weight, Holstein maternal genetics had greater terminal merit than Jersey maternal genetics. Regardless of dam breed, dairy genetics increased carcass leanness. Minimal differences were detected between adjusted feed efficiency of beef and beef × dairy cattle, but underestimation of mature size of beef × dairy could have overestimated efficiency. Genetic differences were more impactful than differences between the conventional beef and dairy calfhood management systems on feedlot and carcass performance.

Body, carcass, and steak dimensions of straightbred Holstein calves and Angus-sired calves from Holstein, Jersey, and crossbred beef dams.

Beef genetics are used with increasing frequency on commercial dairies. Although use of beef genetics improves calf value, variability has been reported in beef × dairy calf phenotype for traits related to muscularity and carcass composition. The objective of this study was to characterize morphometric and compositional differences between beef, beef × dairy, and dairy-fed cattle. Tested treatment groups included Angus-sired straightbred beef steers and heifers (A × B; n = 45), Angus × Holstein crossbreds (A × H; n = 15), Angus × Jersey crossbreds (A × J; n = 16), and straightbred Holsteins (H, n = 16). Cattle were started on trial at mean BW of 302 ±â€…29.9 kg and then fed at 196 ±â€…3.4 d. Morphometric measures were recorded every 28 d during the finishing period, ultrasound measures were recorded every 56 d, and morphometric carcass measures were recorded upon slaughter. Muscle biopsies were collected from the longissimus thoracis of a subset of steers (n = 43) every 56 d. Strip loins were collected from carcasses (n = 78) for further evaluation. Frame size measured as hip height, hip width, and body length was greatest for H cattle (P < 0.05), and A × H cattle had greater hip height than A × J cattle (P < 0.05). Relative to BW as a percentage of mature size, ribeye area of all cattle increased at a decreasing rate (negative quadratic term: P < 0.01), and all ultrasound measures of fat depots increased at an increasing rate (positive quadratic term: P < 0.01). Although no difference was observed in muscle fiber area across the finishing period from the longissimus thoracis (P = 0.80), H cattle had a more oxidative muscle phenotype than A × B cattle (P < 0.05). Additionally, H cattle had the smallest area of longissimus lumborum in the posterior strip loin, greatest length-to-width ratio of longissimus lumborum in the posterior strip loin, and least round circumference relative to round length (P < 0.05). Beef genetics improved muscularity in portions of the carcass distal to the longissimus thoracis.

Divergent selection of beef and dairy breeds has caused differences in skeletal size and muscularity. When calves from dairy systems enter the beef supply chain, variability in mature size and carcass composition are introduced. The objective of this study was to characterize morphometric differences in cattle populations with different proportions of beef and dairy genetics. Body measurements confirmed differences in mature size of beef-type cattle, dairy-type cattle, and beef × dairy cattle; Holstein influence was associated with greater skeletal growth. With advancing maturity, the rate of muscle accretion decreased quadratically while the rate of fat accretion increased quadratically. Although muscularity across all cattle types was similar in the longissimus near the last rib, differences were observed in the posterior end of the strip loin, the forearm, and the round. Differences in mature size, muscularity, and steak dimensions were observed between beef-type cattle, dairy-type cattle, and beef × dairy cattle.

Cecal microbiota of feedlot cattle fed a four-species Bacillus supplement.

As commercial fed cattle consume large amounts of concentrate feedstuffs, hindgut health can be challenged. The objective of this study was to evaluate the effects of a commercially available Bacillus feed additive on cattle health outcomes and cecal microbiota of fed cattle at the time of harvest. Commercial cattle from a single feedlot were identified for characterization of cecal microbial communities using 16S ribosomal ribonucleic acid gene sequencing. All cattle were fed a common corn-based finishing diet. Control cattle (CON) were administered no treatment while treated cattle (TRT) were supplemented daily with 0.050 g of MicroSaf 4C 40 (2 billion colony forming units of Bacillus spp.; Phileo by Lesaffre, Milwaukee, WI). Immediately after harvest and evisceration, the cecal contents of cattle were sampled. After DNA extraction, amplification, and sequencing, reads from CON samples (N = 12) and TRT samples (N = 12) were assigned taxonomy using the SILVA 138 database. Total morbidity, first treatment of atypical interstitial pneumonia, and early shipments for harvest were decreased among TRT cattle compared to CON cattle (P ≤ 0.021). On average, cecal microbiota from TRT cattle had greater alpha diversity than microbiota from CON cattle as measured by Shannon diversity, Pielou's evenness, and feature richness (P < 0.010). Additionally, TRT microbial communities were different (P = 0.001) and less variable (P < 0.001) than CON microbial communities when evaluated by unweighted UniFrac distances. By relative abundance across all samples, the most prevalent phyla were Firmicutes (55.40%, SD = 15.97) and Bacteroidetes (28.17%, SD = 17.74) followed by Proteobacteria (6.75%, SD = 10.98), Spirochaetes (4.54%, SD = 4.85), and Euryarchaeota (1.77%, SD = 3.00). Spirochaetes relative abundance in TRT communities was greater than that in CON communities and was differentially abundant between treatments by ANCOM testing (W = 11); Monoglobaceae was the only family-level taxon identified as differentially abundant (W = 59; greater mean relative abundance in TRT group by 2.12 percentage points). Half (N = 6) of the CON samples clustered away from all other samples based on principal coordinates and represented cecal dysbiosis among CON cattle. The results of this study indicated that administering a four-species blend of Bacillus positively supported the cecal microbial communities of finishing cattle. Further research is needed to explore potential mechanisms of action of Bacillus DFM products in feedlot cattle.

Microbes in the rumen break down fiber and complex nutrients into energy that cattle can absorb. Rumen microbes are becoming well studied, but the microbes of the hindgut­specifically of the cecum and large intestine­are less well-studied. As feedlot cattle eat large amounts of grain, maintaining health and balance of microbes in the hindgut is important. Overconsumption of a meal causes a greater proportion of digestion to occur in the hindgut, causing greater acid production that damages the gastrointestinal lining. If dietary microbial supplements support a more diverse microbial population, the challenges caused by greater hindgut digestion could be mitigated. To test this, cecal microbes were characterized after feedlot cattle were fed a conventional diet, with or without a supplement of Bacillus bacteria. Cecal samples from cattle that were fed Bacillus had greater microbial diversity. Approximately half of the cecal samples from cattle that were not fed Bacillus had disrupted microbial balance. Based on taxonomic assignment, bacteria observed in these disrupted samples indicated greater energy density of digesta and increased methane production. Supplementing feedlot cattle with Bacillus could improve hindgut microbial diversity.

A Pilot Study: the Development of a Facility-Associated Microbiome and Its Association with the Presence of Listeria Spp. in One Small Meat Processing Facility.

Microbial communities which persist in food processing facilities may have a detrimental impact on food safety and spoilage. In meat processing, Listeria monocytogenes is an organism of concern due to its ability to cause significant human illnesses and persist in refrigerated environments. The microbial ecology of Listeria spp. in small meat processing facilities has not been well characterized. Therefore, we collected samples from a newly constructed meat processing facility as an opportunity to investigate several research objectives: (i) to determine whether a stable, consistent microbiome develops in a small meat processing facility during the first 18 months of operation, (ii) to evaluate the environmental factors that drive microbial community formation, and (iii) to elucidate the relationship between microbial communities and the presence of Listeria species. We evaluated microbiomes using 16S rRNA gene sequencing and Listeria presence using quantitative PCR. We demonstrated that microbial communities differentiate by the functional room type, which is representative of several environmental differences such as temperature, sources of microbes, and activity. Temperature was an especially important factor; in rooms with low temperatures, communities were dominated by psychotrophs, especially Pseudomonas, while warmer rooms supported greater diversity. A stable core community formed in facility drains, indicating that mechanisms which cause persistence are present in the communities. The overall presence of Listeria in the facility was low but could be tied to specific organisms within a room, and the species of Listeria could be stratified by room function. IMPORTANCE This study provides critical knowledge to improve meat safety and quality from small meat processing facilities. Principally, it demonstrates the importance of facility design and room condition to the development of important microbial communities; temperature, sanitation regimen, and physical barriers all influence the ability of microorganisms to join the stable core community. It also demonstrates a relationship between the microbial community and Listeria presence in the facility, showing the importance of managing facility sanitation plans for not only pathogens, but also the general facility microbiome.

Liver abscess microbiota of feedlot steers finished in natural and traditional management programs.

Liver abscess etiology in feedlot steers involves the escape of bacteria from the digestive tract to form a polymicrobial abscess within or on the external surface of the liver. However, little is known about the effects of feedlot finishing systems on the microbial composition of the liver abscess purulent material. Liver abscesses were collected at the time of harvest from steers originating from a single feedlot managed in either a traditional program (which included tylosin phosphate supplementation) or a natural program (without tylosin phosphate supplementation). The purulent material of liver abscesses from traditionally managed steers (N = 53 abscesses) and that of naturally managed steers (N = 62 abscesses) was characterized using the V4 region of the 16S rRNA gene. Two phyla and three genera were found in greater than 1% relative abundance across all abscesses. The genus Fusobacterium was identified in all liver abscess samples and accounted for 64% of sequencing reads. Bacteroides and Porphyromonas genera accounted for 33% and 1% of reads, respectively. Trueperella was more likely to be found in the liver abscesses of naturally managed steers than traditionally managed steers (P = 0.022). Over 99% of the genus-level bacterial sequences observed across all liver abscesses belonged to Gram-negative genera. Bacteria known to colonize both the rumen and hindgut were identified within liver abscesses. No differences in alpha diversity or beta diversity were detected between liver abscess communities (between the two management programs or individual pens) when tested as richness, Shannon Diversity Index, or weighted UniFrac distances (P > 0.05). These results were consistent with previous identification of Fusobacterium necrophorum as the primary bacteriologic agent within liver abscesses and emphasized the relationship between the gastrointestinal microbiota and liver abscess formation. Though the microbiota of the liver abscess purulent material was similar between steers fed an antibiotic-free diet and those fed an antibiotic-containing diet from the same feedlot, divergence was detected in liver abscess communities with some being dominated by Fusobacterium and others being dominated by Bacteroides.

As feedlot cattle consume grain, the rumen becomes more acidic. If the lining of the digestive tract is damaged, bacteria that normally remain in the digestive tract can enter the body. Certain bacteria like Fusobacterium necrophorum are involved in the formation of liver abscesses. Feedlot cattle are commonly fed an antibiotic (tylosin phosphate) to reduce the occurrence of liver abscesses, but increasing scrutiny is placed on the antibiotic use. However, the effect of eliminating the antibiotic used to prevent liver abscesses on the bacterial communities involved in liver abscess formation is unknown. This study compared the bacteria found within liver abscesses of cattle fed tylosin phosphate with that of cattle not fed tylosin phosphate. All liver abscesses contained F. necrophorum, and Bacteroides was the second most commonly identified bacterium. Trace amounts of bacteria known to colonize the mouth and digestive tract were observed. Trueperella, a bacteria targeted by tylosin phosphate, was found more frequently in liver abscesses from cattle that received no antibiotic. While the core bacterial composition of the liver abscess was unaffected by antibiotic supplementation to feedlot steers, reduced Trueperella in liver abscesses from cattle-fed tylosin phosphate could be related to a reduction in liver abscess prevalence.