Inclusion of Beef Heart in Ground Beef Patties Alters Quality Characteristics and Consumer Acceptability as Assessed by the Application of Electronic Nose and Tongue Technology.

Consumer purchasing of beef is often driven by the trinity of flavor, palatability, and convenience. Currently, beef patties in the United States are manufactured with fat and lean trimmings derived from skeletal muscles. A reduction in total beef supply may require the use of animal by-product utilization such as variety meats to achieve patty formulations. The current study aimed to assess textural, color, and flavor characteristics in addition to volatile compounds through electronic technology, e-nose and e-tongue, of ground beef patties formulated with beef heart. Ground beef patties were manufactured with 0%, 6%, 12%, or 18% beef heart, with the remainder of the meat block being shoulder clod-derived ground beef. Patties (n = 65/batch/treatment) within each batch (n = 3) with each treatment were randomly allocated to cooked color (n = 17/batch/treatment), Allo-Kramer shear force (AKSF; n = 17/batch/treatment), texture profile analysis (TPA; n = 6/batch/treatment), cooking loss (n = 17/batch/treatment), consumer panel (n = 3/batch/treatment), e-nose (n = 1/batch/treatment), and e-tongue (n = 1/batch/treatment) analysis groups. Patties containing beef heart did not require additional cooking time (p = 0.1325) nor exhibit greater cooking loss (p = 0.0803). Additionally, inclusion rates of beef heart increased hardness (p = 0.0030) and chewiness values (p = 0.0316) in TPA, were internally redder (p = 0.0001), and reduced overall liking by consumer panelists (p = 0.0367). Lastly, patties containing beef heart exhibited greater red-to-brown (p = 0.0003) and hue angle (p = 0.0001) values than control patties. The results suggest that beef heart inclusion does alter ground beef quality characteristics and consumer acceptability.

A conserved interdomain microbial network underpins cadaver decomposition despite environmental variables.

Microbial breakdown of organic matter is one of the most important processes on Earth, yet the controls of decomposition are poorly understood. Here we track 36 terrestrial human cadavers in three locations and show that a phylogenetically distinct, interdomain microbial network assembles during decomposition despite selection effects of location, climate and season. We generated a metagenome-assembled genome library from cadaver-associated soils and integrated it with metabolomics data to identify links between taxonomy and function. This universal network of microbial decomposers is characterized by cross-feeding to metabolize labile decomposition products. The key bacterial and fungal decomposers are rare across non-decomposition environments and appear unique to the breakdown of terrestrial decaying flesh, including humans, swine, mice and cattle, with insects as likely important vectors for dispersal. The observed lockstep of microbial interactions further underlies a robust microbial forensic tool with the potential to aid predictions of the time since death.

Influence of Sous Vide Cooking on Ground Beef Patties.

With rising consumer demand for fast-food options, quick-service restaurants are constantly developing new menu items to attract consumers. Sous vide cookery has become popular for the in-home and fine dining consumer but has not been considered the first cooking option for quick service applications. Therefore, ground beef patties were manufactured to measure the influence of sous vide cooking time on the patty characteristics of moisture, color, and objective tenderness. Patties were randomly assigned a sous vide cooking time of 30, 60, or 90 min and then grilled to an internal temperature of 71.1 °C. Patties sous vide cooked for 30 min exhibited the greatest (p < 0.05) cook loss, Allo-Kramer Shear Force (AKSF) and were darker (L*) than patties sous vide cooked for 60 or 90 min. Additionally, neither internal redness, calculated spectral values of chroma, hue angle, or red-to-brown differed (p > 0.05) regardless of sous vide cooking time. Sous vide cooking duration prior to grilling the ground beef patties altered the moisture, color, and objective texture characteristics of ground beef patties.

Influence of Hempseed Meal on Fresh Goat Meat Characteristics Stored in Vacuum Packaging.

The objective of this study was to determine the influence of hempseed meal (HSM) on goat meat characteristics. Goats (N = 10/treatment) were allocated to a diet concentration (0, 10, 20, or 30%) of HSM, fed for 60 days, and harvested. Carcass measurements were collected after chilling, and subsequently fabricated into wholesale subprimals. From the subprimals of the shoulder and leg, steaks were cut 2.54 cm thick, vacuum packaged, and assigned to laboratory methods: cook yield, instrumental color, lipid oxidation, microbial spoilage, and instrumental tenderness. HSM did not alter (p > 0.05) carcass characteristics, microbial spoilage, cook loss, or the thiobarbituric acid reactive substance (TBARS). However, a decrease in objective tenderness measurements (p < 0.05) was observed with greater concentrations of HSM supplementation in the diet. Instrumental surface color values for lightness (L*) indicated that steaks became lighter and less red (a*) as storage time increased (p < 0.05). Results suggest that HSM and storage time do not alter some goat meat traits, but HSM or storage time separately may influence goat meat quality. HSM may be an effective feed ingredient that does not alter carcass quality or meat yield.

Extended Storage of Beef Steaks Using Thermoforming Vacuum Packaging.

Extended storage duration often results in negative quality attributes of fresh or frozen beef steaks. This study focused on evaluating the fresh and cooked meat quality of beef steaks stored using vacuum packaging for 63 days. Steaks 2.54 cm thick were packaged into one of three thermoforming films VPA (250 µ nylon/EVOH/enhanced polyethylene coextrusion), VPB (250 µ nylon/EVOH/enhanced polyethylene coextrusion), or VPC (125 µ nylon/EVOH/enhanced/polyethylene coextrusion). Steaks placed in VPA were lighter (L*) and redder (a*) in surface color (p < 0.05) as the display period increased, whereas steaks packaged in VPB and VPC became darker. Yellowness, hue angle (Hue°), and chroma (C*) values were greater (p < 0.05) in steaks using VPC film as the storage period increased. Calculated spectral values of red to brown were greater (p < 0.05) for steaks in VPA and VPB than in VPC. However, steaks placed in VPC films contained greater (p < 0.05) forms of metmyoglobin and oxymyoglobin and lower calculated relative values of deoxymyoglobin. In addition, packaging treatment altered (p > 0.05) lipid oxidation, but storage time had a greater (p < 0.05) influence on purge loss, cook loss, and Warner-Bratzler shear force (WBSF). Current results suggest that the use of vacuum packaging for extended storage of beef steaks (>60) days is plausible.

Vacuum Packaging Maintains Fresh Characteristics of Previously Frozen Beef Steaks during Simulated Retail Display.

The impact of frozen storage on beef steaks prior to the retail setting may result in changes to the quality and safety of the packaged meat. Therefore, the objective of the current study was to evaluate fresh characteristics on previously frozen steaks during a simulated retail display. Steaks were allocated to one of three packaging treatments (MB, MDF, MFS) and stored frozen (−13 °C) for 25 days in the absence of light. After thawing, steaks were stored in a lighted retail case at 3 °C and evaluated for instrumental surface color, pH, purge loss, lipid oxidation, and microbial spoilage organisms throughout the 25-day fresh display period. There was an increase (p < 0.05) for aerobic plate counts and lipid oxidation from day 20 through 25 on steaks packaged in MFS and MDF, respectively. Steaks packaged in MB were redder (p < 0.05) and more vivid (C*) as storage time increased. Whereas lipid oxidation was greater (p < 0.05) throughout the entire display for steaks packaged in MFS and MDF. It is evident that barrier properties of MB limiting oxygen exposure of the steak preserved fresh meat characteristics after frozen storage. Results from the current study suggest that vacuum packaging films can aid in retarding detrimental effects caused by frozen storage after placing the steaks in fresh retail conditions.

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.

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.

Considerations and best practices in animal science 16S ribosomal RNA gene sequencing microbiome studies.

Microbiome studies in animal science using 16S rRNA gene sequencing have become increasingly common in recent years as sequencing costs continue to fall and bioinformatic tools become more powerful and user-friendly. The combination of molecular biology, microbiology, microbial ecology, computer science, and bioinformatics-in addition to the traditional considerations when conducting an animal science study-makes microbiome studies sometimes intimidating due to the intersection of different fields. The objective of this review is to serve as a jumping-off point for those animal scientists less familiar with 16S rRNA gene sequencing and analyses and to bring up common issues and concerns that arise when planning an animal microbiome study from design through analysis. This review includes an overview of 16S rRNA gene sequencing, its advantages, and its limitations; experimental design considerations such as study design, sample size, sample pooling, and sample locations; wet lab considerations such as field handing, microbial cell lysis, low biomass samples, library preparation, and sequencing controls; and computational considerations such as identification of contamination, accounting for uneven sequencing depth, constructing diversity metrics, assigning taxonomy, differential abundance testing, and, finally, data availability. In addition to general considerations, we highlight some special considerations by species and sample type.

Experiences and lessons learned from two virtual, hands-on microbiome bioinformatics workshops.

In October of 2020, in response to the Coronavirus Disease 2019 (COVID-19) pandemic, our team hosted our first fully online workshop teaching the QIIME 2 microbiome bioinformatics platform. We had 75 enrolled participants who joined from at least 25 different countries on 6 continents, and we had 22 instructors on 4 continents. In the 5-day workshop, participants worked hands-on with a cloud-based shared compute cluster that we deployed for this course. The event was well received, and participants provided feedback and suggestions in a postworkshop questionnaire. In January of 2021, we followed this workshop with a second fully online workshop, incorporating lessons from the first. Here, we present details on the technology and protocols that we used to run these workshops, focusing on the first workshop and then introducing changes made for the second workshop. We discuss what worked well, what didn't work well, and what we plan to do differently in future workshops.

Air versus Water Chilling of Chicken: a Pilot Study of Quality, Shelf-Life, Microbial Ecology, and Economics.

The United States' large-scale poultry meat industry is energy and water intensive, and opportunities may exist to improve sustainability during the broiler chilling process. By USDA regulation, after harvest the internal temperature of the chicken must be reduced to 40°F or less within 16 h to inhibit bacterial growth that would otherwise compromise the safety of the product. This step is accomplished most commonly by water immersion chilling in the United States, while air chilling methods dominate other global markets. A comprehensive understanding of the differences between these chilling methods is lacking. Therefore, we assessed the meat quality, shelf-life, microbial ecology, and techno-economic impacts of chilling methods on chicken broilers in a university meat laboratory setting. We discovered that air chilling methods resulted in superior chicken odor and shelf-life, especially prior to 14 days of dark storage. Moreover, we demonstrated that air chilling resulted in a more diverse microbiome that we hypothesize may delay the dominance of the spoilage organism Pseudomonas Finally, a techno-economic analysis highlighted potential economic advantages to air chilling compared to water chilling in facility locations where water costs are a more significant factor than energy costs.IMPORTANCE As the poultry industry works to become more sustainable and to reduce the volume of food waste, it is critical to consider points in the processing system that can be altered to make the process more efficient. In this study, we demonstrate that the method used during chilling (air versus water chilling) influences the final product microbial community, quality, and physiochemistry. Notably, the use of air chilling appears to delay the bloom of Pseudomonas spp. that are the primary spoilers in packaged meat products. By using air chilling to reduce carcass temperatures instead of water chilling, producers may extend the time until spoilage of the products and, depending on the cost of water in the area, may have economic and sustainability advantages. As a next step, a similar experiment should be done in an industrial setting to confirm these results generated in a small-scale university lab facility.

Microbiome Data Accurately Predicts the Postmortem Interval Using Random Forest Regression Models.

Death investigations often include an effort to establish the postmortem interval (PMI) in cases in which the time of death is uncertain. The postmortem interval can lead to the identification of the deceased and the validation of witness statements and suspect alibis. Recent research has demonstrated that microbes provide an accurate clock that starts at death and relies on ecological change in the microbial communities that normally inhabit a body and its surrounding environment. Here, we explore how to build the most robust Random Forest regression models for prediction of PMI by testing models built on different sample types (gravesoil, skin of the torso, skin of the head), gene markers (16S ribosomal RNA (rRNA), 18S rRNA, internal transcribed spacer regions (ITS)), and taxonomic levels (sequence variants, species, genus, etc.). We also tested whether particular suites of indicator microbes were informative across different datasets. Generally, results indicate that the most accurate models for predicting PMI were built using gravesoil and skin data using the 16S rRNA genetic marker at the taxonomic level of phyla. Additionally, several phyla consistently contributed highly to model accuracy and may be candidate indicators of PMI.

Circulating miRNAs as Potential Alternative Cell Signaling Associated with Maternal Recognition of Pregnancy in the Mare.

During early pregnancy, the conceptus and mare communicate to establish pregnancy. Cell-secreted vesicles (e.g., exosomes) have been reported in serum. Exosomes contain bioactive materials, such as miRNA, that can mediate cell responses. We hypothesized that a) exosomes are present in mare circulation and quantity varies with pregnancy status, b) exosomes contain miRNAs unique to pregnancy status, and c) miRNAs target pathways in endometrium based upon pregnancy status of the mare. First, serum samples were obtained from mares in a crossover design, with each mare providing samples from a pregnant and nonmated control cycle (n = 3/sample day) on Days 12, 14, 16, and 18 postovulation. Flow cytometry revealed the presence of serum microvesicles in mares in two different-sized populations (greater than or less than 100 nm), validated by transmission electron microscopy. Second, serum was collected on Days 9, 11, and 13 (n = 4/day), and endometrial biopsies were collected on Days 11 and 13 (n = 3/day) from pregnant and nonmated mares. Total RNA from serum exosomes was evaluated with quantitative RT-PCR using equine-specific miRNA sequences. A total of 12 miRNAs were found in different quantities on the specified days. Pathway analysis suggested that miRNAs targeted focal adhesion molecules (FAMs). Transcripts corresponding to FAMs were evaluated in endometrial biopsies. Protein levels and localization for PAK6 and RAF1 were further evaluated. Our data suggest that serum exosomes contain miRNA that differ based upon pregnancy status, and may affect mRNA expression related to focal adhesion pathway in the endometrium, with a potential role in maternal recognition of pregnancy.