Bovine DNA Research Articles

New copper(II) thiohydantoin complexes: Synthesis, characterization, and assessment of their interaction with bovine serum albumin and DNA

New copper(II) complexes of 2-alkylthio-5-arylmethylene-4H-imidazolin-4-ones: (5Z)-2-(methylsulfanyl)-3-(prop-2-en-1-yl)-5-(pyridin-2-ylmethylidene)-3,5-dihydro-4H-imidazol-4-one) (1a), (5Z,5′Z)-2,2′-(ethan-1,2-diyldisulfanyldiyl)bis(5-(2-pyridilmethylen)-3-allyl-3,5-dihydo-4Н-imidazole-4-one) (2a) and (5Z,5′Z)-3,3′-hexan-1,6-diylbis[5-(2-pyridilmethylen)-2-methylthiotetrahydro-4Н-imidazole-4-one)] (3a) were synthesized as possible anticancer drugs. Their structures were characterized by 1H NMR spectroscopy, elemental analysis, and X-ray crystallography. The composition of the complexes were found for 1a (Cu:L=1:1), 2a (Cu:L=2:1), and 3a (Cu:L=2:1). The chelation constants were found by competitive complexation with ethylenediamine tetraacetate: 1a (6.7±0.6)×1015M−1, 2a=(4.9±0.4)×1019M−2, and 3a (5.7±0.5)×1019M−2. Supramolecular binding with calf thymus DNA by competitive ethidium bromide quenching was made for complex 2a as the most promising anticancer model, the Stern–Volmer constants were found to be KSV=(8.0±0.4)×106M−1, Kq=(6.5±0.4)×105M−1. The binding of the complex 2a to BSA was made by the Scatchard method, the value of the constant is Kb=(1.9±0.2)×106M−1.

Detection of ovine milk adulteration using taqman real-time pcr assay

Food safety, quality and composition have become the subjects of increasing public concern. To prevent fraud and enhance quality assurance, credible analysis of dairy products is crucial. Bovine milk is more widely available and cheaper than milk of sheep and goat. Bovine milk is also processed in large quantities to produce a range of dairy produce. DNA-based methods have proven to be more reliable, because of the stability of DNA under the conditions of high temperature, high pressure, and chemical treatment used during the processing of some food products. The commercial InnuDETECT cheese assay based on the principle TaqMan real-time PCR systems have been tested for the identification and quantification of bovine DNA in ovine milk samples. DNA was extracted using the InnuPREP DNA Mini Kit and quantified by the QuantiFluor dsDNA system. The assay showed good linearity, with correlation coefficient of R2 = 0.983 and efficiency of 86%. The internal control amplified fragment from different mammalian species (cow, sheep and goat), with similar CT values. Detection of bovine DNA in milk mixtures was achieved even in samples containing 0.5% of bovine milk. The InnuDETECT cheese assay has been successfully used to measure bovine DNA in ovine milk, and will prove useful for bovine species identification and quantitative authentication of animal-derived products.

Simultaneous detection of bovine and porcine DNA in pharmaceutical gelatin capsules by duplex PCR assay for Halal authentication

BackgroundIn the pharmaceutical industry, hard- and soft-shelled capsules are typically made from gelatin, commonly derived from bovine and porcine sources. To ensure that pharmaceutical products comply with halal regulations in Muslim countries (no porcine products allowed), development of a valid, reliable, quick, and most importantly, cost-effective tests are of utmost importance.MethodsWe developed a species-specific duplex polymerase chain reaction (PCR) assay targeting 149 bp porcine and 271 bp bovine mitochondrial DNA (mtDNA) to simultaneously detect both porcine and bovine DNA (in one reaction at the same time) in gelatin. Some additional simplex PCR tests (targeting 126 bp bovine and 212 bp porcine mtDNA) and real-time PCR using a commercially available kit (for identification of porcine DNA) were used to verify the selectivity and sensitivity of our duplex PCR. After optimization of DNA extraction and PCR methods, hard/soft pharmaceutical gelatin capsules (containing drug) were tested for the presence of porcine and/or bovine DNA.ResultsDuplex PCR detected the presence of as little as 0.1% porcine DNA, which was more accurate than the commercially available kit. Of all gelatin capsules tested (n = 24), 50% contained porcine DNA (pure porcine gelatin alone or in combination with bovine gelatin).ConclusionsDuplex PCR presents an easy-to-follow, quick, low-cost and reliable method to simultaneously detect porcine and bovine DNAs (>100 bp) in minute amounts in highly processed gelatin-containing pharmaceutical products (with a 0.1% sensitivity for porcine DNA) which may be used for halal authentication.Graphical abstractSimultaneous detection of porcine and bovine DNA in gelatin capsules by duplex PCR

Molecular characterization and biological activity of bovine interferon-omega3

Bovine interferon-omega3 (BoIFN-ω3) gene was amplified from bovine liver genomic DNA, which encodes a 195-amino acid protein containing a 23-amino acid signal peptide. Analysis of the molecular characteristics revealed that BoIFN-ω3 evolving from IFN-ω, contained four cysteine residues and five alpha helices, showing that BoIFN-ω3 presented the typical molecular characteristics of type I interferon. BoIFN-ω3 exhibited antiviral and antiproliferative activities, which exerted a protective effect against VSV in several mammalian cell lines, as well as against BEV, IBRV, and BVDV in MDBK cell. Moreover, BoIFN-ω3 was shown to be highly sensitive to trypsin, but remaining stable despite changes in pH and temperature. Additionally, BoIFN-ω3 induced the transcription of Mx1, ISG15, and ISG56 genes, as well as the expression of Mx1 protein in a time-dependent manner. These findings will be useful to further study BoIFN-ω in host's defence against infectious diseases, particularly viral infections. Furthermore, results will facilitate further research on the bovine interferon family.

Nanofiltration applications of tough MWNT buckypaper membranes containing biopolymers

The ability of biopolymers (bovine serum albumin, lysozyme, chitosan, gellan gum and DNA) to facilitate formation of aqueous dispersions of MWNTs was investigated using a combination of absorption spectrophotometry and optical microscopy. Subsequently, self-supporting carbon nanotube membranes, known as buckypapers (BPs), were prepared by vacuum filtration of the dispersions. Microanalytical data obtained from the BPs confirmed the retention of biopolymers within their structures. Tensile test measurements performed on the BPs showed that incorporation of the biopolymers resulted in significant improvements in mechanical properties, compared to analogous BPs containing MWNTs and the low molecular mass dispersant Triton X-100. For example, MWNT/CHT BPs (CHT=chitosan) exhibited values for tensile strength, ductility, Young's modulus and toughness of 28±2MPa, 5.3±2.7%, 0.9±0.3GPa and 1.7±0.3J g−1, respectively. Each of these values are significantly greater than those obtained for MWNT/Trix BPs, prepared using a low molecular weight dispersant (6±3MPa, 1.3±0.2%, 0.6±0.3GPa and 0.10±0.06J g−1, respectively). This significant improvement in mechanical properties is attributed to the ability of the long biopolymer molecules to act as flexible bridges between the short CNTs. All BPs possessed hydrophilic surfaces, with contact angles ranging from 29±2° to 57±5°. Nitrogen gas porosimetry showed that the BPs have highly porous internal structures, while scanning electron microscopy (SEM) showed their surface morphologies have numerous pore openings. The permeability of the BPs towards water, inorganic salts, and dissolved trace organic contaminants (TrOCs), such as pharmaceuticals, personal care products, and pesticides, was investigated through filtration experiments. Of the twelve TrOCs investigated in this study, nine were rejected by more than 95% by BPs composed of MWNTs and chitosan. The latter BPs also demonstrated good rejection of both NaCl (30–55%) and MgSO4 (40–70%).

PCR-based Detection of Bovine Papillomavirus DNA from the Cutaneous Papillomas and Surrounding Environments in the Korean Native Cattle, Hanwoo

Two 1-year old calves of Korean Native cattle (Hanwoo) presented cutaneous papillomas on the face and neck. Type 2 bovine papillomavirus (BPV-2) was identified in the cutaneous papillomas based on BPV-specific PCR and subsequent DNA sequencing analysis results. Using DNA samples extracted from two affected calves and unaffected animals reared in the same stable, BPV-2 was not only detected in the cutaneous papillomas of affected animals based on BPV-specific PCR analysis, but also detected in normal skins, hairs, and their environments based on nested PCR analysis. BPV-2 was also detected in DNA samples isolated from animals and environments of that distinct stable with affected calves. However, no BPV-2 was detected in the drinking water of both stables (infected and unaffected). These findings concluded that BPV-2 was transmitted by direct or indirect contact, not by drinking water. This is the first report to show molecular evidence of BPV-2 infection. Rapid and precise molecular identification can be used to screen BPV-2 in cattle farms to understand the biological roles of BPV in animal diseases.

Molecular cloning, polymorphism, and functional activity of the bovine and water buffalo Mx2 gene promoter region.

BackgroundBovine Mx2 gene sequences were already reported, but further information about the gene properties is not yet available. The objective of the current study was to elucidate the structural properties of the bovine Mx2 gene mainly the promoter region and its possible functional role. If available, such information would help in assessing the functional properties of the gene, which was reported to confer antiviral action against recombinant VSV.ResultsExaminations on the bovine genomic BAC clone—confirmed to contain the Mx2 gene—revealed 883-bp sequences. A computer scan unequivocally identified a 788-bp promoter region containing a typical TATA box, three ISREs and other promoter-specific motifs. Comparative analysis of nine bovine genomic DNA samples showed 19 nucleotide substitutions suggesting the existence of five different genotypes in the promoter region. The water buffalo Mx2 promoter region was determined by using primers based on the bovine Mx2 promoter region disclosing 893-bp, with 56 substitutions, two insertions, 9 and 1 nt at two different sites. A functional analysis of the putative ISRE indicated that ISRE played a synergetic role in the activation of bovine Mx2 gene transcription.ConclusionBovine and water buffalo Mx2 promoter region was identified disclosing, the conserved ISRE, located in the proximal end of the promoter region like other members of the antiviral family, suggesting functional activity under interferon stimulation.

Unmapped reads from cattle RNAseq data: A source for missing and misassembled sequences in the reference assemblies and for detection of pathogens in the host

Usually, reads from transcriptome sequencing data unmapped to the target species' reference genome are disregarded. A recent RNAseq project on the new fatal disease Bovine Neonatal Pancytopenia had indicated an unexplained immune response signature to a double-stranded RNA virus. To unravel its background, contigs were de novo assembled from unmapped RNAseq reads and aligned against the bovine genome assemblies and multispecies NCBI databases. Lack of genuine virus sequence contigs rejected the hypothesis of a live virus being causal for the unexplained immune response. Alignment data also demonstrated incomplete bovine reference genome assemblies. In addition, we found that several parasite and virus genome reference assemblies in NCBI were contaminated with bovine DNA and confirmed recombination of bovine DNA into BVD virus strains. Exploring unmapped reads can extract useful biological information regarding the presence of microorganisms and can highlight issues with reference genome assemblies of host and pathogen species.

Whole genome sequencing of 51 breast cancers reveals that tumors are devoid of bovine leukemia virus DNA.

Controversy exists regarding the association of bovine leukemia virus (BLV) and breast cancer. PCR-based experimental evidence indicates that BLV DNA is present in breast tissue and that as many as 37% of cancer cases may be attributable to viral exposure. Since this association might have major consequences for human health, we evaluated 51 whole genomes of breast cancer samples for the presence of BLV DNA. Among 32 billion sequencing reads retrieved from the NCBI database of genotype and phenotype, none mapped on different strains of the BLV genome. Controls for sequence divergence and proviral loads further validated the approach. This unbiased analysis thus excludes a clonal insertion of BLV in breast tumor cells and strongly argues against an association between BLV and breast cancer.

Reduced representation bisulphite sequencing of ten bovine somatic tissues reveals DNA methylation patterns and their impacts on gene expression.

BackgroundAs a major epigenetic component, DNA methylation plays important functions in individual development and various diseases. DNA methylation has been well studied in human and model organisms, but only limited data exist in economically important animals like cattle.ResultsUsing reduced representation bisulphite sequencing (RRBS), we obtained single-base-resolution maps of bovine DNA methylation from ten somatic tissues. In total, we evaluated 1,868,049 cytosines in CG-enriched regions. While we found slightly low methylation levels (29.87 to 38.06 %) in cattle, the methylation contexts (CGs and non-CGs) of cattle showed similar methylation patterns to other species. Non-CG methylation was detected but methylation levels in somatic tissues were significantly lower than in pluripotent cells. To study the potential function of the methylation, we detected 10,794 differentially methylated cytosines (DMCs) and 836 differentially methylated CG islands (DMIs). Further analyses in the same tissues revealed many DMCs (including non-CGs) and DMIs, which were highly correlated with the expression of genes involved in tissue development.ConclusionsIn summary, our study provides a baseline dataset and essential information for DNA methylation profiles of cattle.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-016-3116-1) contains supplementary material, which is available to authorized users.

Bovine Papillomavirus DNA and S100 Profiles in Sarcoids and Other Cutaneous Spindle Cell Tumors in Horses.

Histopathologic differentiation between deep dermal or subcuticular equine sarcoids (ie, nodular sarcoids) and other spindle cell tumors in the dermis and subcutis such as peripheral nerve sheath tumors (PNSTs) can be challenging based on morphologic criteria alone. It has been proposed that polymerase chain reaction (PCR) for bovine papillomavirus (BPV) DNA and S100 immunohistochemistry be used as diagnostic tests to separate equine sarcoids from PNSTs. We reviewed 197 skin-associated spindle cell tumors (ie, soft tissue sarcomas), including PNSTs and sarcoids, received at the University of Florida between 1995 and 2013 and performed BPV PCR and S100 immunohistochemistry on archived paraffin-embedded tissues. We found that BPV DNA was demonstrable in 70% of the sarcoids, 59% of the PNSTs, 37% of the fibrosarcomas, and 22% of other tumors (myxosarcomas, fibromas, and other sarcomas) diagnosed on histomorphologic characteristics. Positive S100 staining was only seen in 12 tumors in the study (5 fibrosarcomas, 3 sarcoids, 2 PNSTs, and 2 other sarcomas). The results demonstrate that BPV is associated with many skin-associated spindle cell soft tissue tumors in horses in addition to sarcoids. S100 was rarely expressed in equine soft tissue sarcomas in the skin but was expressed in many tumor types, including PNSTs and sarcoids. Because 41% of the PNSTs classified by histomorphology in this study were BPV negative and 94% were S100 negative, it is reasonable to classify these as soft tissue sarcomas with nerve sheath tumor histomorphology rather than as either PNSTs or sarcoids.

Presence and Degree of Morphological Alterations in Equine Hoof Canker Tissues in Relation to the Presence of Bovine Papillomaviral DNA

Presence and Degree of Morphological Alterations in Equine Hoof Canker Tissues in Relation to the Presence of Bovine Papillomaviral DNA

Biomimetic nucleus pulposus scaffold created from bovine caudal intervertebral disc tissue utilizing an optimal decellularization procedure.

Intervertebral disc (IVD) degeneration (IDD) and herniation (IDH) can result in low back pain and impart significant socioeconomic burden. These pathologies involve detrimental alteration to the nucleus pulposus (NP) either via biochemical degradation or extrusion from the IVD, respectively. Thus, engineering living NP tissue utilizing biomaterial scaffolds that recapitulate native NP microarchitecture, biochemistry, mechanical properties, and which support cell viability represents an approach to aiding patients with IDD and IDH. To date, an ideal biomaterial to support NP regeneration has yet to be developed; however, one promising approach to generating biomimetic materials is to employ the decellularization (decell) of xenogeneic NP tissue to remove host DNA while maintaining critical native extracellular matrix (ECM) components. Herein, 13 different procedures were evaluated in an attempt to decell bovine caudal IVD NP tissue. An optimal method was identified which was confirmed to effectively remove bovine DNA, while maintaining physiologically relevant amounts of glycosaminoglycan (GAG) and type II collagen. Unconfined static and dynamic compressive mechanical properties of scaffolds approached values reported for human NP and viability of human amniotic stem cells (hAMSCs) was maintained on noncrosslinked and EDC/NHS treated scaffolds for up to 14 days in culture. Taken together, NP tissue obtained from bovine caudal IVDs can be successfully decelled in order to generate a biomimetic scaffold for NP tissue regeneration. © 2016 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 3093-3106, 2016.

A Multiscale TiO2 Nanorod Array for Ultrasensitive Capture of Circulating Tumor Cells.

In this work, a uniform multiscale TiO2 nanorod array is fabricated to provide a "multi-scale interacting platform" for cell capture, which exhibits excellent capture specificity and sensitivity of the target cells after modification with bovine serum albumin (BSA) and DNA aptamer. After studying the capture performance of the BSA-aptamer TiO2 nanorod substrates and other nanostructured substrates, we can conclude that the multisacle TiO2 nanorod substrates could indeed effectively enhance the capture yields of target cancer cells. The capture yield of artificial blood samples on the BSA-aptamer TiO2 nanorod substrates is up to 85%-95%, revealing the potential application of the TiO2 nanorods on efficient and sensitive capture of rare circulating tumor cells.

Specific PCR assays to determine bovine, porcine, fish and plant origin of gelatin capsules of dietary supplements

Gelatin, a purified protein derived mostly from pig skin and bovine tissue, is used widely in both food and pharmaceutical industries. Here, to determine the species of origin of capsule gelatin, we developed a sensitive and reliable test using the polymerase chain reaction (PCR) method, which included 1) species-specific or universal primer sets, designed to detect short 16S ribosomal RNA (rRNA) gene sequences from cow, pig, and fish (tilapia) as well as genes encoding the large subunit of plant ribulose-1,5-bisphosphate carboxylase oxygenase and 2) species-specific PCR coupled with whole-genome amplification. This method was used to verify manufacturing label claims of 28 gelatin capsule samples sold as dietary supplements. The results from 27 samples were consistent with gelatin-related information on the manufacturer label, while one sample that mentioned tilapia gelatin was found to contain only bovine DNA. This rapid method can therefore be used to verify the authenticity of gelatin capsules.

Interaction of biosynthesized gold nanoparticles with BSA and CTDNA: A multi-spectroscopic approach

The interaction between biosynthesized gold nanoparticles (GNP) with bovine serum albumin (BSA) and calf thymus DNA (CTDNA) was investigated from a multi-spectroscopic approach. The apparent binding constant (K) were 2.69×104L/mol and 15.84×104L/mol at 293K, respectively for BSA and CTDNA, and the number of binding sites were ∼1. According to the Van’t Hoff equation, the thermodynamic parameters were calculated (ΔH=−119.496kJ/mol, ΔS=−320.92J/mol/K for BSA and ΔH=−225.89kJ/mol; ΔS=673J/mol/K for CTDNA) and the results indicated hydrogen bonds and van der Waals forces are the main stabilizing force in both of the BSA-GNP and CTDNA-GNP complex. The average binding distance (r=4.45nm) and the critical energy transfer distance (R0=2.94nm) between GNP and BSA were also evaluated according to Förster’s non-radiative energy transfer theory. What is more, UV–Visible, and circular dichroism spectra showed that the addition of GNP changed the secondary structure of BSA and led to a decrease in α-helix. Circular dichroism spectra also indicated conformational changes of CTDNA in the presence of the GNP. Furthermore, the GNP induces small changes in DNA viscosity and melting temperature which is indicative of groove binding mode of GNP with CTDNA.

Mycoplasma bovis MBOV_RS02825 Encodes a Secretory Nuclease Associated with Cytotoxicity.

This study aimed to determine the activity of one Mycoplasma bovis nuclease encoded by MBOV_RS02825 and its association with cytotoxicity. The bioinformatics analysis predicted that it encodes a Ca2+-dependent nuclease based on existence of enzymatic sites in a TNASE_3 domain derived from a Staphylococcus aureus thermonuclease (SNc). We cloned and purified the recombinant MbovNase (rMbovNase), and demonstrated its nuclease activity by digesting bovine macrophage linear DNA and RNA, and closed circular plasmid DNA in the presence of 10 mM Ca2+ at 22–65 °C. In addition, this MbovNase was localized in membrane and rMbovNase able to degrade DNA matrix of neutrophil extracellular traps (NETs). When incubated with macrophages, rMbovNase bound to and invaded the cells localizing to both the cytoplasm and nuclei. These cells experienced apoptosis and the viability was significantly reduced. The apoptosis was confirmed by activated expression of phosphorylated NF-κB p65 and Bax, and inhibition of Iκβα and Bcl-2. In contrast, rMbovNaseΔ181–342 without TNASE_3 domain exhibited deficiency in all the biological functions. Furthermore, rMbovNase was also demonstrated to be secreted. In conclusion, it is a first report that MbovNase is an active nuclease, both secretory and membrane protein with ability to degrade NETs and induce apoptosis.

Simultaneous identification and verification of gelatin type in capsule shells by electrophoresis and polymerase chain reaction

Pharmaceutical dosage forms containing gelatin are recently of high attention in regard to its porcine content. In Muslim countries, halal products assurance is strictly regulated, not only for food but also for pharmaceutical products. This study aimed to develop gelatin identification and verification method in capsule shells by performing simple simultaneous protein- and nucleic acid based-identification methods. Identification and verification performing polyacrylamide gel electrophoresis (PAGE)—coupled with principle component analysis, and polymerase-chain-reaction (PCR)-restriction fragment length polymorphism (RFLP) were carried out. Gelatin was obtained by extracting the capsules using cold acetone; the precipitate was used directly for PAGE, and was used as well for DNA extraction. The results showed that the porcine gelatin reference showed 12 major bands of peptide profile, with predominant bands at ±200 and ±100–135 kDa on 8 % Tris-glycine gel, while bovine gelatin only showed four major bands. Further, PCR-RFLP result showed two specific bands of the porcine gelatin DNA at 228 and 131 bp, whereas the bovine gelatin DNA bands occurred at 316 and 44 bp. The results demonstrated that the capsule shell samples used in this study contained the following: sample 1 was verified contain porcine gelatin, whereas the three other samples identified contain bovine gelatin. Although the simultaneous approaches seemed need to be improved, it is crucial to maintain their low cost and simple protocol.

Spectroscopic, electrochemical and molecular docking studies of dothiepin and doxepin with bovine serum albumin and DNA base.

The interaction of dothiepin (DOT) and doxepin (DOX) with bovine serum albumin (BSA) and a DNA base (adenine) was studied using UV-visible, fluorescence, attenuated total reflection-infra-red (ATR-IR), cyclic voltammetry and molecular docking methods. Strong fluorescence quenching was observed upon interaction of DOT and DOX with BSA/adenine and the mechanism suggested static quenching. Hydrophobic and hydrogen bonding interactions were the predominant intermolecular forces needed to stabilize the copolymer. Upon addition of the drugs: (i) the tautomeric equilibrium structure of the adenine was changed; and (ii) the oxidation and the reduction peaks of the adenine/BSA interaction shifted towards high and low potentials, respectively. In ATR-IR, the band shift of amides I and II indicated a change in secondary structure of BSA upon binding to DOT and DOX drugs. The reduction in voltammetric current in the presence of BSA/adenine was attributed to slow diffusion of BSA/adenine binding with DOX/DOT. The docking method indicated that the drug moiety interacted with the BSA molecule. Copyright © 2016 John Wiley & Sons, Ltd.

Coagulansin-A has beneficial effects on the development of bovine embryos invitro via HSP70 induction.

Coagulansin-A (withanolide) is the steroidal lactone obtained from Withania coagulans which belong to Solanaceae family. The present study investigated the effects of coagulansin-A on bovine oocyte maturation and embryo development invitro. All these oocytes were aspirated from the ovaries obtained from Korean Hanwoo cows at a local abattoir. To determine whether coagulansin-A has beneficial effects on bovine oocyte maturation invitro, 355 oocytes per group (control and treated) in seven replicates were subjected with different concentrations (1, 2.5, 5, 7.5 and 10μM) of coagulansin-A. The coagulansin-A was added in the invitro maturation (IVM) media followed by invitro fertilization (IVF) and then invitro culture (IVC). Only treatment with 5μM coagulansin-A remarkably (P<0.05) improved embryos development (Day 8 blastocyst) having 27.30 and 40.01% for control and coagulansin-A treated groups respectively. Treatment with 5μM coagulansin-A significantly induced activation of heat shock protein 70 (HSP70) (P<0.05). Immunofluorescence analysis revealed that 5μM coagulansin-A treatment also significantly inhibited oxidative stress and inflammation during bovine embryo development invitro by decreasing 8-oxoguanosine (8-OxoG) (P<0.05) and nuclear factor-κB (NF-κB) (P<0.05). The expressions of HSP70 and NF-κB were also conformed through real-time PCR (RT-PCR). Additionally, the terminal deoxynucleotidyl transferase dUTP nick-end labelling (TUNEL) assay confirmed that coagulansin-A treatment significantly improved the embryo quality and reduced bovine embryo DNA damage (P<0.05). The present study provides new information regarding the mechanisms by which coagulansin-A promotes bovine embryo development invitro.