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Translational Animal Science 2024The use of tissue specimens for undergraduate instruction is a very valuable tool. However, fresh tissue specimens are not always available and many common preservation...
The use of tissue specimens for undergraduate instruction is a very valuable tool. However, fresh tissue specimens are not always available and many common preservation techniques can result in discoloration, offensive odors, and/or dangerous chemical residues. The Elnady Technique was developed as a means to produce tissue specimens that "are realistic, durable, have no offensive odor, and are dry, soft and flexible" (Elnady, F.A. 2016 The Elnady Technique: An innovative, new method for tissue preservation. 33:237-242. doi:10.14573/altex.1511091). Briefly for soft tissue, specimens were preserved by fixing in formalin. The tissue specimen was then dehydrated with a series of acetone baths. Once the tissue was fully dehydrated, the specimen was impregnated in glycerin. Excess glycerin was then removed by draining followed by immersion in cornstarch. Cornstarch residue was removed with a soft brush, and the specimen was stored in a plastic bag. Multiple specimens (including the female reproductive tract of the cat, goat, horse, and sow; digestive tract of cat, chicken, and dog; 1-day-old lamb stomach; goat rumen, reticulum, omasum, and abomasum; and sheep heart and kidney) have been preserved and used in various animal science course laboratories (126 laboratory sections and 1,696 students at Berry College). Some of the specimens have been in use for seven years and are still in usable condition. Anonymously surveyed Berry College Animal Science Faculty strongly agreed or agreed that Elnady preserved tissues are a useful teaching aid ( = 5). The Elnady Technique has proven to be a useful means of preserving tissue samples used in undergraduate animal science courses.
PubMed: 38800102
DOI: 10.1093/tas/txae077 -
International Journal of Biological... Mar 2024Chondroitin sulfate (CS) + dermatan sulfate (DS) and hyaluronan (HA) concentrations and the sulfation patterns of CS-DS in the cartilaginous tissues and alimentary...
Quantitative, compositional, and immunohistochemical analyses of chondroitin sulfate, dermatan sulfate, and hyaluronan in internal organs of deer (Cervus nippon centralis and C. n. yesoensis) and cattle (Bos taurus).
Chondroitin sulfate (CS) + dermatan sulfate (DS) and hyaluronan (HA) concentrations and the sulfation patterns of CS-DS in the cartilaginous tissues and alimentary canals of Honshu Sika deer, Hokkaido Sika deer, and cattle were investigated in the present study. CS + DS concentrations were high in cartilaginous tissues, namely, the trachea and scapular cartilage region (5- 12 g*), and low in the alimentary canal (~0.3 g*). HA concentrations were low in cartilaginous tissues and the alimentary canal (~0.2 g*). All tissues mainly contained A-type [HexAGalNAc(4-sulfate)] and C-type [HexAGalNAc(6-sulfate)] CS + DS. The ratios of A-type/C-type CS + DS were 1.2- 3.1 and 0.9- 16.4 in cartilaginous tissues and the alimentary canal, respectively. CS + DS predominantly comprised β-D-GlcA and α-L-IdoA in cartilaginous tissues and the alimentary canal, respectively. The alimentary canal characteristically contained up to 14 % highly sulfated E-type [HexAGalNAc(4,6-disulfate)] and D-type [HexA(2-sulfate)GalNAc(6-sulfate)] CS + DS. The specific distributions of CS and DS were immunohistochemically confirmed using CS + DS-specific antibodies. Although the omasum of cattle is more likely to have higher concentrations of CS + DS and HA, no significant species differences were observed in the concentrations or sulfation patterns of CS + DS among species for Honshu Sika deer, Hokkaido Sika deer, and cattle. (*per 100 g of defatted dry tissue).
Topics: Cattle; Animals; Chondroitin Sulfates; Dermatan Sulfate; Hyaluronic Acid; Deer; Sulfates
PubMed: 38281521
DOI: 10.1016/j.ijbiomac.2024.129680 -
Animal Bioscience Feb 2024Ruminants possess a specialized four-compartment forestomach, consisting of the reticulum, rumen, omasum, and abomasum. The rumen, the primary fermentative chamber,...
Ruminants possess a specialized four-compartment forestomach, consisting of the reticulum, rumen, omasum, and abomasum. The rumen, the primary fermentative chamber, harbours a dynamic ecosystem comprising bacteria, protozoa, fungi, archaea, and bacteriophages. These microorganisms engage in diverse ecological interactions within the rumen microbiome, primarily benefiting the host animal by deriving energy from plant material breakdown. These interactions encompass symbiosis, such as mutualism and commensalism, as well as parasitism, predation, and competition. These ecological interactions are dependent on many factors, including the production of diverse molecules, such as those involved in quorum sensing (QS). QS is a density-dependent signalling mechanism involving the release of autoinducer (AIs) compounds, when cell density increases AIs bind to receptors causing the altered expression of certain genes. These AIs are classified as mainly being N-acyl-homoserine lactones (AHL; commonly used by Gram-negative bacteria) or autoinducer-2 based systems (AI-2; used by Gram-positive and Gram-negative bacteria); although other less common AI systems exist. Most of our understanding of QS at a gene-level comes from pure culture in vitro studies using bacterial pathogens, with much being unknown on a commensal bacterial and ecosystem level, especially in the context of the rumen microbiome. A small number of studies have explored QS in the rumen using 'omic' technologies, revealing a prevalence of AI-2 QS systems among rumen bacteria. Nevertheless, the implications of these signalling systems on gene regulation, rumen ecology, and ruminant characteristics are largely uncharted territory. Metatranscriptome data tracking the colonization of perennial ryegrass by rumen microbes suggest that these chemicals may influence transitions in bacterial diversity during colonization. The likelihood of undiscovered chemicals within the rumen microbial arsenal is high, with the identified chemicals representing only the tip of the iceberg. A comprehensive grasp of rumen microbial chemical signalling is crucial for addressing the challenges of food security and climate targets.
PubMed: 38186253
DOI: 10.5713/ab.23.0374 -
Gene Mar 2024Phosphodiesterase 9A (PDE9A) plays a crucial role in activating the cGMP-dependent signaling pathway and may have important effects on the growth and development of the...
Phosphodiesterase 9A (PDE9A) plays a crucial role in activating the cGMP-dependent signaling pathway and may have important effects on the growth and development of the gastrointestinal tract in Hu sheep. In this study, we analyzed the single nucleotide polymorphisms of PDE9A in 988 Hu sheep and their correlation with growth performance, feed efficiency, and gastrointestinal development. Additionally, we examined the expression level of different PDE9A genotypes in the gastrointestinal tract of Hu sheep by using fluorescence quantitative PCR. The results revealed a moderate level of polymorphism (0.25 < PIC < 0.50) at the g.286248617 T > C mutation site located in the first intron of PDE9A in Hu sheep, with three genotypes: CC, CT, and TT. The weights of the omasum, colon, and cecum were significantly greater in the CC genotype than in the TT genotype (P < 0.05), and the expression level of PDE9A in the tissues of the rumen, ileum, cecum, and colon was notably lower in the CC genotype individuals (P < 0.05). These findings suggest that the polymorphism of PDE9A affects the weight of the stomach, colon, and cecum in Hu sheep through expression regulation. Overall, the results of this study suggest that the g.286248617 T > C mutation site in the first intron of PDE9A can serve as a potential molecular marker for breeding practices related to the gastrointestinal weight of Hu sheep.
Topics: Animals; 3',5'-Cyclic-AMP Phosphodiesterases; Breeding; Gastrointestinal Tract; Genotype; Polymorphism, Single Nucleotide; Sheep
PubMed: 38184018
DOI: 10.1016/j.gene.2024.148137 -
Frontiers in Veterinary Science 2023This study aimed to evaluate the implications of supplementary nutrition during the mid-to-late pregnancy on various parameters in Hanwoo cows and their subsequent...
Effects of mid-to-late prepartum feed supplementation in Hanwoo beef cows on their performance, blood metabolites, and the carcass characteristics and metabolites of their neonatal calves.
INTRODUCTION
This study aimed to evaluate the implications of supplementary nutrition during the mid-to-late pregnancy on various parameters in Hanwoo cows and their subsequent neonatal calves.
MATERIALS AND METHODS
Eight Hanwoo cows in their first parity were divided into two groups. The control group (C, 100%) received 3kg of concentrate and 5kg of rice straw throughout the pregnancy period, while the treatment group (T, 150%) increased their diet during mid-to-late pregnancy. Both performance assessments and blood metabolite analyses were performed for the pregnant cows. Neonatal calves were subjected to morphometric evaluations, blood sampling, and detailed morphometric analyses of carcasses and gastrointestinal components.
RESULTS
Performance indices of the cows showed that both Pregnancy Period (PregP) and Body Condition Score (BCS) were significantly improved with supplemental feeding ( <0.05). Improvements in Body Weight (BW) and Feed Conversion Ratio (FCR) were not statistically significant. Blood metabolite analysis for the cows revealed decreased levels of triglycerides (TGLate), non-esterified fatty acids (NEFALate), and progesterone (P4Late), with a notable increase in glucose (GluLate) levels ( <0.01). In the neonatal calves, anatomical metrics of the gastrointestinal tissues showed increased Omasum Width (OmasWdth) values in the supplemented group ( =0.053). There was significant increase of papillae and villus lengths in the rumen and small intestine ( <0.01 and <0.05, respectively). Morphometric evaluations displayed longer body lengths (BLnth) and larger chest width (ChestWdth) in the treated calves ( <0.05 and <0.01, respectively). Carcass characteristics showed no substantial variations between the groups, while blood analysis in the calves revealed decreased GPT levels in the nutritionally supplemented group (<0.05).
DISCUSSION
The findings indicate that supplementing the diets of Hanwoo cows during mid-to-late pregnancy leads to significant changes in select maternal blood metabolites and influences specific anatomical and morphometric features in neonatal calves, all without significant shifts in carcass attributes.
PubMed: 38076568
DOI: 10.3389/fvets.2023.1287119 -
Animal Bioscience May 2024The present study was executed to explore the molecular mechanism of fibroblast growth factor 10 (FGF10) gene in bovine adipogenesis.
OBJECTIVE
The present study was executed to explore the molecular mechanism of fibroblast growth factor 10 (FGF10) gene in bovine adipogenesis.
METHODS
The bovine FGF10 gene was overexpressed through Ad-FGF10 or inhibited through siFGF10 and their negative control (NC) in bovine adipocytes, and the multiplicity of infection, transfection efficiency, interference efficiency were evaluated through quantitative real-time polymerase chain reaction, western blotting and fluorescence microscopy. The lipid droplets, triglycerides (TG) content and the expression levels of adipogenic marker genes were measured during preadipocytes differentiation. The differentially expressed genes were explored through deep RNA sequencing.
RESULTS
The highest mRNA level was found in omasum, subcutaneous fat, and intramuscular fat. Moreover, the highest mRNA level was found in adipocytes at day 4 of differentiation. The results of red-oil o staining showed that overexpression (Ad-FGF10) of the FGF10 gene significantly (p<0.05) reduced the lipid droplets and TG content, and their downregulation (siFGF10) increased the measurement of lipid droplets and TG in differentiated bovine adipocytes. Furthermore, the overexpression of the FGF10 gene down regulated the mRNA levels of adipogenic marker genes such as CCAAT enhancer binding protein alpha (C/EBPα), fatty acid binding protein (FABP4), peroxisome proliferator-activated receptor-γ (PPARγ), lipoprotein lipase (LPL), and Fas cell surface death receptor (FAS), similarly, down-regulation of the FGF10 gene enriched the mRNA levels of C/EBPα, PPARγ, FABP4, and LPL genes (p<0.01). Additionally, the protein levels of PPARγ and FABP4 were reduced (p<0.05) in adipocytes infected with Ad-FGF10 gene and enriched in adipocytes transfected with siFGF10. Moreover, a total of 1,774 differentially expressed genes (DEGs) including 157 up regulated and 1,617 down regulated genes were explored in adipocytes infected with Ad-FGF10 or Ad-NC through deep RNA-sequencing. The top Kyoto encyclopedia of genes and genomes pathways regulated through DEGs were the PPAR signaling pathway, cell cycle, base excision repair, DNA replication, apoptosis, and regulation of lipolysis in adipocytes.
CONCLUSION
Therefore, we can conclude that the FGF10 gene is a negative regulator of bovine adipogenesis and could be used as a candidate gene in marker-assisted selection.
PubMed: 37946430
DOI: 10.5713/ab.23.0185 -
PloS One 2023Genetic group, age at entry into confinement and at slaughter, are characteristics that have an important influence on lamb performance and carcass. The aim of this...
Genetic group, age at entry into confinement and at slaughter, are characteristics that have an important influence on lamb performance and carcass. The aim of this study was to evaluate the performance, carcass characteristics and non-carcass components from different genetic groups (Santa Inês and ½ Dorper x ½ Santa Inês) sheep, submitted to different feedlot entry and exit strategies. Were used 72 lambs males and castrated; 36 Santa Inês (SI) and 36 crossbred (Dorper x Santa Inês-DSI), with 6 months of average initial age. The groups were established in a completely randomized experimental design, in a 2x3x4 factorial arrangement, from the combination of genetic groups (GG), body weight at the beginning of confinement (WBC) and length of stay in confinement (LSC). The body weight classes at the beginning of confinement were: light (25 kg), intermediate (28 kg) and heavy (31 kg), for Santa Inês and crossbreeds, respectively. Slaughters were carried out every 28 days of confinement, in four LSC: 0, 28, 56 and 84 days. The GG did not influence performance, carcass and non-carcass component traits of lambs (p > 0.05). There was an effect of the WBC on the weights: final (FW), metabolic (MW), body at slaughter (BWS), empty body (EBW), hot carcass (HCY) and cold (CCW), loin, shoulder, leg musculature; loin eye area (LEA) and loin fat (p < 0.05). There was also an effect on LSC, for FW, average daily weight gain (ADG), MW, weight and yield of body components, weight of cuts and tissue ratio components of cuts (p < 0.05). In non-carcass components, effect on full and empty weight of: omasum, rumen-reticulum, small intestine; empty large intestine, liver and kidneys, paws and skin, and perirenal, pelvic and inguinal fat (p < 0.05). Interaction double effect on the tissue muscle/fat:bone ratio (MF:B) and for the full omasal component (p < 0.05). And triple interaction effect for ADG, full omasum and perirenal fat (p < 0.05). Weight at the beginning of confinement and confinement time are the characteristics that most influence performance, quantitative characteristics of carcass and non-carcass components. Regardless of the genetic group and age class, the animals reach the same weight after 84 days of confinement. Thus, the confinement of heavier lambs (31 kg) can be a profitable alternative, as they presented the highest weights for the most commercially valued cuts (shank and loin). The confinement strategy must adapt to market situations.
Topics: Male; Sheep; Animals; Body Composition; Weight Gain; Hybridization, Genetic; Phenotype; Red Meat
PubMed: 37943781
DOI: 10.1371/journal.pone.0293819 -
Journal of Animal Science and... Oct 2023Four-chambered stomach including the forestomachs (rumen, reticulum, and omasum) and abomasum allows ruminants convert plant fiber into high-quality animal products. The...
BACKGROUND
Four-chambered stomach including the forestomachs (rumen, reticulum, and omasum) and abomasum allows ruminants convert plant fiber into high-quality animal products. The early development of this four-chambered stomach is crucial for the health and well-being of young ruminants, especially the immune development. However, the dynamics of immune development are poorly understood.
RESULTS
We investigated the early gene expression patterns across the four-chambered stomach in Hu sheep, at 5, 10, 15, and 25 days of age. We found that forestomachs share similar gene expression patterns, all four stomachs underwent widespread activation of both innate and adaptive immune responses from d 5 to 25, whereas the metabolic function were significantly downregulated with age. We constructed a cell landscape of the four-chambered stomach using single-cell sequencing. Integrating transcriptomic and single-cell transcriptomic analyses revealed that the immune-associated module hub genes were highly expressed in T cells, monocytes and macrophages, as well as the defense-associated module hub genes were highly expressed in endothelial cells in the four-stomach tissues. Moreover, the non-immune cells such as epithelial cells play key roles in immune maturation. Cell communication analysis predicted that in addition to immune cells, non-immune cells recruit immune cells through macrophage migration inhibitory factor signaling in the forestomachs.
CONCLUSIONS
Our results demonstrate that the immune and defense responses of four stomachs are quickly developing with age in lamb's early life. We also identified the gene expression patterns and functional cells associated with immune development. Additionally, we identified some key receptors and signaling involved in immune regulation. These results help to understand the early life immune development at single-cell resolution, which has implications to develop nutritional manipulation and health management strategies based on specific targets including key receptors and signaling pathways.
PubMed: 37821933
DOI: 10.1186/s40104-023-00933-1 -
Frontiers in Veterinary Science 2023The development of the four stomachs of yak is closely related to its health and performance, however the underlying molecular mechanisms are largely unknown. Here, we...
The development of the four stomachs of yak is closely related to its health and performance, however the underlying molecular mechanisms are largely unknown. Here, we systematically analyzed mRNAs of four stomachs in five growth time points [0 day, 20 days, 60 days, 15 months and 3 years (adult)] of yaks. Overall, the expression patterns of DEmRNAs were unique at 0 d, similar at 20 d and 60 d, and similar at 15 m and adult in four stomachs. The expression pattern in abomasum was markedly different from that in rumen, reticulum and omasum. Short Time-series Expression Miner (STEM) analysis demonstrated that multi-model spectra are drastically enriched over time in four stomachs. All the identified mRNAs in rumen, reticulum, omasum and abomasum were classified into 6, 4, 7, and 5 cluster profiles, respectively. Modules 9, 38, and 41 were the most significant three colored modules. By weighted gene co-expression network analysis (WGCNA), a total of 5,486 genes were categorized into 10 modules. , , , and were the hub genes of the turquoise module, and , , , , and were the hub genes of the blue module. Furthermore, functional KEGG enrichment analysis suggested that the turquoise module was involved in gastric acid secretion, sphingolipid metabolism, ether lipid metabolism, etc., and the blue module was enriched in pancreatic secretion, pantothenate and CoA biosynthesis, and starch and sucrose metabolism, etc. Our study aims to lay a molecular basis for the study of the physiological functions of rumen, reticulum, omasum and abomasum in yaks. It can further elucidate the important roles of these mRNAs in regulation of growth, development and metabolism in yaks, and to provide a theoretical basis for age-appropriate weaning and supplementary feeding in yaks.
PubMed: 37808112
DOI: 10.3389/fvets.2023.1204706 -
Analytical Chemistry Aug 2023Gastric cancer is one of the most common malignant digestive cancers, and its diagnostic has still faced challenges based on metabolic analysis due to complex sample...
Gastric cancer is one of the most common malignant digestive cancers, and its diagnostic has still faced challenges based on metabolic analysis due to complex sample pretreatment and low metabolite abundance. In this study, inspired by the structure of bovine omasum, we synthesized a novel interfacial carbon-based nanocomposite of graphene supported nickel nanoparticles-encapsulated in the nitrogen-doped carbon nanotube (Ni/N-CNT/rGO), which was served as a novel matrix with enhanced ionization efficiency for the matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) saliva metabolic analysis of gastric cancer. Benefiting from its high sp graphitic degree, large surface area, strong UV absorption, and rich active sites, Ni/N-CNT/rGO matrix exhibited excellent performances of reproducibility, coverage, salt-tolerance, sensitivity, and adsorption ability in MALDI-TOF MS. The differential scanning calorimetry (DSC) and thermal conversion behaviors explained the highly efficient LDI mechanism. Based on saliva metabolic fingerprints, Ni/N-CNT/rGO assisted LDI MS with cross-validation analysis could successfully distinguish gastric cancer patients from healthy controls through the screening of four potential biomarkers with an accuracy of 92.50%, specificity of 88.03%, and sensitivity of 97.12%. This work provided a fast and sensitive MS sensing platform for the metabolomics characterization of gastric cancer and might have potential value for precision medicine in the future.
PubMed: 37458487
DOI: 10.1021/acs.analchem.3c01358