Effects of Bovine Maternal Nutrient Restriction on Offspring MicroRNA and mRNA Expression and Muscle Fiber Type

For producers in more temperate areas, such as the Intermountain West, poor nutrition during the second trimester of gestation is common due to seasonal changes in forage and nutrient availability. The majority of muscle fibers are formed and adipogenesis is initiated in the second trimester, making it a critical time for skeletal muscle and adipose development in beef cattle. However, the extent to which these changes persist in the offspring postnatally is unknown. In this study, maternal nutrition was restricted during the second trimester in order to analyze the effects of maternal nutrient restriction on offspring skeletal muscle growth. Offspring were monitored throughout production postnatally and skeletal muscle samples were taken at weaning, the beginning of the feedlot phase,and at harvest.We investigated whether calves from cows restricted in the second trimester had a different expression of microRNA (miRNA) or messengerRNA (mRNA) known to be downstream targets of those miRNA. We also analyzed mRNA expression of myosin heavy chain (MHC) isoforms to determine whether maternal nutrition in the second trimester impacts muscle fiber type. There were no changes observed in mRNA or MHC expression between the two different treatments at either time point. Differences in expression of several miRNAs important in development of adipose and skeletal muscle were observed between the treatment groups. The findings of this research indicate that maternal nutrition during the second trimester of gestation alters miRNA expression in the skeletal muscle. However, more research is needed to determine exactly how these miRNA impact growth of skeletal muscle postnatally

Understanding the Influence of Trenbolone Acetate and Polyamines on Proliferation of Bovine Satellite Cells

Approximately 90% of beef cattle on feed in the United States receive at least one anabolic implant, which results in increased growth, efficiency, and economic return to producers. However, the complete molecular mechanism through which anabolic implants function to improve skeletal muscle growth remains unknown. This study had 2 objectives: (1) determine the effect of polyamines and their precursors on proliferation rate in bovine satellite cells (BSC); and (2) understand whether trenbolone acetate (TBA), a testosterone analog, has an impact on the polyamine biosynthetic pathway. To address these, BSC were isolated from 3 finished steers and cultured. Once cultures reached 75% confluency, they were treated in 1% fetal bovine serum (FBS) and/or 10 nM TBA, 10 mM methionine (Met), 8 mM ornithine (Orn), 2 mM putrescine (Put), 1.5 mM spermidine (Spd), or 0.5 mM spermine (Spe). Initially, a range of physiologically relevant concentrations of Met, Orn, Put, Spd, and Spe were tested to determine experimental doses to implement the aforementioned experiments. One, 12, or 24 h after treatment, mRNA was isolated from cultures and abundance of paired box transcription factor 7 (Pax7), Sprouty 1 (Spry), mitogen-activated protein kinase-1 (Mapk), ornithine decarboxylase (Odc), and S adenosylmethionine (Amd1) were determined, and normalized to 18S. No treatment × time interactions were observed (P ≥ 0.05). Treatment with TBA, Met, Orn, Put, Spd, or Spe increased (P ≤ 0.05) BSC proliferation when compared with control cultures. Treatment of cultures with Orn or Met increased (P ≤ 0.01) expression of Odc 1 h after treatment when compared with control cultures. Abundance of Amd1 was increased (P \u3c 0.01) 1 h after treatment in cultures treated with Spd or Spe when compared with 1% FBS controls. Cultures treated with TBA had increased (P \u3c 0.01) abundance of Spry mRNA 12 h after treatment, as well as increased mRNA abundance of Mapk (P \u3c 0.01) 12 h and 24 h after treatment when compared with 1% FBS control cultures. Treatment with Met increased (P \u3c 0.01) mRNA abundance of Pax7 1 h after treatment as compared with 1% FBS controls. These results indicate that treatments of BSC cultures with polyamines and their precursors increase BSC proliferation rate, as well as abundance of mRNA involved in cell proliferation. In addition, treatment of BSC cultures with TBA, polyamines, or polyamine precursors impacts expression of genes related to the polyamine biosynthetic pathway and proliferation

The Influence of Maternal Dietary Intake During Mid-Gestation on Growth, Feedlot Performance, miRNA and mRNA Expression, and Carcass and Meat Quality of Resultant Offspring

This research analyzed how maternal plane of nutrition during mid-gestation impacts growth, blood metabolites, expression of microRNA and messenger RNA in skeletal muscle, feedlot performance, and carcass characteristics of progeny. Thirty-two cows were bred to the same Angus sire and fed to either maintain a body condition score (BCS) of 5.0 to 5.5 (maintenance [MAIN]; n = 15) or to lose 1 BCS (restriction [REST]; n = 17) over an 84-d period of mid-gestation. Following the second trimester, all cows were co-mingled and fed at maintenance for the remainder of gestation. Following the 84-d treatment period, REST cows had a lower (P < 0.01) BCS than MAIN cows. At the end of the third trimester, there was no difference (P = 0.78) in BCS between the treatment groups. There was no difference (P > 0.10) between offspring in birthweight, weaning weight, average daily gain, feed efficiency, dry matter intake, carcass yield, steak quality, or in circulating levels of glucose, cortisol, insulin, or insulin-like growth factor-1. REST offspring expressed more (P < 0.05) miR-133a, miR-133b, miR-181d, miR-214, miR-424 and miR-486 at weaning than MAIN offspring. At harvest, REST offspring expressed more (P < 0.05) miR-133a and less (P < 0.01) miR-486 than MAIN offspring. REST steaks were perceived as more tender (P = 0.05) by a trained sensory panel. These results indicate that maternal nutrient restriction during mid-gestation resulting in a loss of 1 BCS has an effect on microRNA expression in the skeletal muscle but does not alter postnatal growth potential, carcass quality, or end product quality of the offspring. This suggests that moderate restriction in maternal nutrition during the second trimester, which results in a drop in BCS that can be recovered during the third trimester, should not cause alarm for producers when considering future offspring performance

Understanding the Effects of Trenbolone Acetate, Polyamine Precursors, and Polyamines on Proliferation, Protein Synthesis Rates, and the Abundance of Genes Involved in Myoblast Growth, Polyamine Biosynthesis, and Protein Synthesis in Murine Myoblasts

Research suggests that androgens increase skeletal muscle growth by modulating polyamine biosynthesis. As such, the objective of this study was to investigate effects of anabolic hormones, polyamine precursors, and polyamines relative to proliferation, protein synthesis, and the abundance of mRNA involved in polyamine biosynthesis, proliferation, and protein synthesis in C2C12 and Sol8 cells. Cultures were treated with anabolic hormones (trenbolone acetate and/or estradiol), polyamine precursors (methionine or ornithine), or polyamines (putrescine, spermidine, or spermine). Messenger RNA was isolated 0.5 or 1, 12, or 24 h post-treatment. The cell type had no effect (p \u3e 0.10) on proliferation, protein synthesis, or mRNA abundance at any time point. Each treatment increased (p \u3c 0.01) proliferation, and anabolic hormones increased (p = 0.04) protein synthesis. Polyamines increased (p \u3c 0.05) the abundance of mRNA involved in polyamine biosynthesis, proliferation, and protein synthesis. Treatment with polyamine precursors decreased (p \u3c 0.05) the abundance of mRNA involved in proliferation and protein synthesis. Overall, C2C12 and Sol8 myoblasts do not differ (p \u3e 0.10) in proliferation, protein synthesis, or mRNA abundance at the time points assessed. Furthermore, anabolic hormones, polyamines, and polyamine precursors increase proliferation and protein synthesis, and polyamines and their precursors alter the abundance of mRNA involved in growth

Full Viral Genome Sequencing and Phylogenomic Analysis of Feline Herpesvirus Type 1 (FHV-1) in Cheetahs (Acinonyx jubatus).

Feline herpesvirus type 1 (FHV-1) is endemic in captive cheetahs and sporadically causes devastating disease. Modified live vaccines (MLV), intended for use in domestic cats, are used in some captive cheetah populations and have been anecdotally linked to disease in certain subpopulations. Ten FHV-1 isolates from ten captive cheetahs and one isolate from an MLV used to inoculate four of the host animals were analyzed. Viral DNA was extracted for full-genome sequencing by Illumina MiSeq with viral genomes then used for phylogenomic and recombinational analyses. The FHV-1 shed by vaccinated cheetahs were almost identical to the MLV, with few variants among viral genomes. Eight cheetah FHV-1 isolates and the MLV were grouped in a clade along with FHV-1 isolates from domestic cats in the USA. The remaining two cheetah FHV-1 isolates (unknown host vaccine status) were not associated with a clade. The likely ancestral origin of these two isolates involves recombination events between Australian domestic cat and cheetah FHV-1 isolates. Collectively, these data suggest that the MLV is capable of causing clinical disease and viral shedding in some cheetahs and represents evidence of interspecies transmission of virus between domestic and wild cats

Full Viral Genome Sequencing and Phylogenomic Analysis of Feline Herpesvirus Type 1 (FHV-1) in Cheetahs (\u3ci\u3eAcinonyx jubatus\u3c/i\u3e)

Feline herpesvirus type 1 (FHV-1) is endemic in captive cheetahs and sporadically causes devastating disease. Modified live vaccines (MLV), intended for use in domestic cats, are used in some captive cheetah populations and have been anecdotally linked to disease in certain subpopulations. Ten FHV-1 isolates from ten captive cheetahs and one isolate from an MLV used to inoculate four of the host animals were analyzed. Viral DNA was extracted for full-genome sequencing by Illumina MiSeq with viral genomes then used for phylogenomic and recombinational analyses. The FHV-1 shed by vaccinated cheetahs were almost identical to the MLV, with few variants among viral genomes. Eight cheetah FHV-1 isolates and the MLV were grouped in a clade along with FHV-1 isolates from domestic cats in the USA. The remaining two cheetah FHV-1 isolates (unknown host vaccine status) were not associated with a clade. The likely ancestral origin of these two isolates involves recombination events between Australian domestic cat and cheetah FHV-1 isolates. Collectively, these data suggest that the MLV is capable of causing clinical disease and viral shedding in some cheetahs and represents evidence of interspecies transmission of virus between domestic and wild cats

Relative and Quantitative Characterization of the Bovine Bacterial Ocular Surface Microbiome in the Context of Suspected Ocular Squamous Cell Carcinoma

The ocular surface microbiome is altered in certain disease states. The aim of this study was to characterize the bovine bacterial ocular surface microbiome (BBOSM) in the context of ocular squamous cell carcinoma (OSCC). The conjunctiva of normal (n = 28) and OSCC (n = 10) eyes of cows aged 2 to 13 years from two farms in Louisiana and Wyoming were sampled using individual sterile swabs. DNA extraction followed by 16S ribosomal ribonucleic acid (rRNA) gene sequencing and real-time polymerase chain reaction (RT-PCR) were performed to, respectively, assess the relative and absolute BBOSM. Discriminant analysis (DA) was performed using RT-PCR data, and relative abundance analysis was performed using 16S rRNA gene sequencing data. The 11 most abundant phyla in both normal and OSCC-affected cows were identified using 16S rRNA gene sequencing analysis. The relative abundance of Euryarchaeota was found to be significantly lower (p = 0.0372) in OSCC eyes compared to normal eyes. Relative abundance differences within and between geographic locations were also identified. Quadratic DA categorized samples as OSCC or normal with 100% sensitivity and 83.3–100% specificity. Relative abundance analysis identified relative BBOSM phylum alterations in OSCC. Quadratic DA can be used to accurately categorize BBOSM from normal and OSCC ocular surface samples

Alterations to the bovine bacterial ocular surface microbiome in the context of infectious bovine keratoconjunctivitis

Abstract Background Infectious bovine keratoconjunctivitis (IBK) is a common cause of morbidity in cattle, resulting in significant economic losses. This study aimed to characterize the bovine bacterial ocular surface microbiome (OSM) through conjunctival swab samples from Normal eyes and eyes with naturally acquired, active IBK across populations of cattle using a three-part approach, including bacterial culture, relative abundance (RA, 16 S rRNA gene sequencing), and semi-quantitative random forest modeling (real-time polymerase chain reaction (RT-PCR)). Results Conjunctival swab samples were obtained from eyes individually classified as Normal (n = 376) or IBK (n = 228) based on clinical signs. Cattle unaffected by IBK and the unaffected eye in cattle with contralateral IBK were used to obtain Normal eye samples. Moraxella bovis was cultured from similar proportions of IBK (7/228, 3.07%) and Normal eyes (1/159, 0.63%) (p = 0.1481). Moraxella bovoculi was cultured more frequently (p  0.05). Alpha-diversity indices for geographic location (p < 0.001), age (p < 0.0001), sex (p < 0.05) and breed (p < 0.01) and beta-diversity indices for geographic location (p < 0.001), disease status (p < 0.01), age (p < 0.001), sex (p < 0.001) and breed (p < 0.001) were significantly different between groups. Modeling of RT-PCR values reliably categorized the microbiome of IBK and Normal eyes; primers for Moraxella bovoculi, Moraxella bovis, and Staphylococcus spp. were consistently the most significant canonical variables in these models. Conclusions The results provide further evidence that multiple elements of the bovine bacterial OSM are altered in the context of IBK, indicating the involvement of a variety of bacteria in addition to Moraxella bovis, including Moraxella bovoculi and R. nasimurium, among others. Actinobacteriota RA is altered in IBK, providing possible opportunities for novel therapeutic interventions. While RT-PCR modeling provided limited further support for the involvement of Moraxella bovis in IBK, this was not overtly reflected in culture or RA results. Results also highlight the influence of geographic location and breed type (dairy or beef) on the bovine bacterial OSM. RT-PCR modeling reliably categorized samples as IBK or Normal