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Topic:Biotechnology
Biotechnology in horses encompasses the application of biological techniques and tools to enhance equine health, performance, and reproduction. This field includes genetic engineering, cloning, and the development of vaccines and therapeutics tailored to equine physiology. Techniques such as gene editing and stem cell therapy are explored for their potential to address genetic disorders, improve tissue regeneration, and enhance disease resistance in horses. Additionally, advancements in reproductive biotechnology, such as artificial insemination and embryo transfer, contribute to genetic diversity and breeding efficiency. This page compiles peer-reviewed research studies and scholarly articles that investigate the applications, methodologies, and implications of biotechnology in equine science.
Allometric relationships of cell numbers and size in the mammalian lung. Allometric studies have shown that lung volume, alveolar surface area, and diffusing capacity increase proportionally with body weight across a broad range of mammalian species. Changes in the number of cells and in average cell size and surface areas with increasing body weight have not been defined. We speculated that cell size is determined more by cell function than by species and body weight. To test this hypothesis, nine species ranging in size from shrew (2 to 3 g) to horse (510 kg) were studied. Random sites from the distal alveolar region of each species were analyzed using morphometr...
Cloning of highly polymorphic microsatellites in the horse. We have isolated equine microsatellites by screening a genomic library with (TG)n and (TC)n probes. TG microsatellites were found to be more abundant than TC repeats, with an estimated frequency of one per 100,000bp. Sequence analysis of eight TG-positive clones revealed varying structures of the repeat regions; perfect stretches of TG repeats, imperfect stretches of TG repeats and compound regions of TG and TC repeats. Five loci were analysed by PCR and showed extensive polymorphism; three to seven alleles and heterozygosities of 0.40-0.76 were observed when screening 20-30 unrelated individu...
Identification of equine chromosomes in horse x mouse somatic cell hybrids. Giemsa-11 (G-11) staining and in situ hybridization were used to identify the equine chromosome complement of horse x mouse somatic cell hybrids. The presence of horse chromosomes in somatic cell hybrids was determined by differential G-11 staining. The slides were then destained and hybridized with biotinylated total horse (Equus caballus) genomic DNA without suppression. Fluorescence detection permitted rapid confirmation of horse chromosomal DNA in the hybrid cells.
Stabilization of the structure of horse plasma vitamin D binding protein by disulfide bonds. Vitamin D binding protein (DBP) was isolated from horse plasma in a four-step procedure that involved Affi-Gel Blue affinity chromatography, gel filtration, hydroxylapatite chromatography, and anion exchange high-pressure liquid chromatography. The yield of DBP from 80 mL of plasma was 6-7 mg. Horse plasma DBP closely resembles other plasma DBPs, being a tryptophan-free protein of Mr 53,000. It is able to bind to and block the polymerization of monomeric actin. The secondary structure of DBP was calculated from circular dichroism measurements to be 39% alpha-helix, 42% beta-sheet, and 19% rand...
DNA fingerprinting in horses using a simple (TG)n probe and its application to population comparisons. A synthetic polynucleotide (TG)n was hybridized to equine DNA digested with HinfI and hypervariable hybridization patterns were obtained. Mendelian inheritance of these DNA fingerprinting patterns was confirmed by pedigree analysis. Estimates of the probabilities of identical band patterns in unrelated individuals of different breeds (Swedish Trotters, North Swedish Trotters, Thoroughbreds and Arabians) were in the range 1 x 10(-4) - 7 x 10(-6). The variability derived with the (TG)n probe in horses was higher than what we obtained with several other commonly used probes for DNA fingerprinting...
In vivo measurement of bone quality in the horse: estimates of precision for ultrasound velocity measurement and single photon absorptiometry. The in vivo precision of ultrasound velocity measurement and single photon absorptiometry for the assessment of equine bone quality is discussed. In vivo precisions for ultrasound velocity measurements were less than 0.5 per cent, whereas cortical cross-sectional area, compact bone density and modulus of elasticity were around 1 per cent, and bone mineral content and density were just over 2 per cent. Except for ultrasound velocity, substantial improvements could be achieved by taking the mean of five readings for each measurement. The long-term precision of the techniques was also high, with ...
Artificial respiration in the anesthetized horse using bilateral, percutaneous, cervical phrenic-nerve stimulation with needle electrodes: a preliminary report. In this preliminary study, artificial respiration was produced in four anesthetized horses using trains of stimuli applied to long needle electrodes inserted bilaterally at the base of the neck. The needles were insulated to within 1 cm of the tips. The frequency of the stimuli (0.1 msec) was 35/sec and the train duration (duration of inspiration) was 1 sec. Inspired volume increased with increasing stimulus intensity. In two animals, inspired volumes of 6 liters were achieved. In another animal 4.5 liters was achieved and in another, 2.5 liters. This lower value probably represented less-than...
Capillary supply and fibre area in locomotor muscles of horse and steer–a comparison between histochemistry and electron microscopy. In order to investigate possible differences in variables defining capillary supply of skeletal muscle derived from two methods, the electron-microscopical and the amylase-PAS histochemical methods were applied in a study of horse and steer muscles. Samples from several locomoter muscles were taken at slaughter from one horse and one steer, divided into two and prepared separately for the two techniques. It was found that there was no difference between the two methods in the values for the capillary-to-fibre ratio. Values for mean fibre area, on the other hand, were significantly higher in th...
Linkage of hyperkalaemic periodic paralysis in quarter horses to the horse adult skeletal muscle sodium channel gene. A genetic disease observed in certain Quarter horses is hyperkalaemic periodic paralysis (HYPP). This disease causes attacks of paralysis which can be induced by ingestion of potassium. Recent studies have shown that HYPP in humans is due to single base changes within the adult skeletal muscle sodium channel gene. A large Quarter horse pedigree segregating dominant HYPP was studied to determine if mutations of the sodium channel gene are similarly responsible for HYPP in horses. We used cross-species, PCR-mediated, cDNA cloning and sequencing of the horse adult skeletal muscle sodium channel a...
An ultrasonographic off-set system for examination of equine tendons and ligaments. In a dorsal plane, an improved ultrasonographic off-set system was used to obtain serial ultrasonographic images with enhanced anatomic and pathologic detail of the tendons, ligaments, and associated structures of the limbs of 100 horses. The off-set provided good acoustic coupling between a linear array ultrasonographic transducer and the horse's skin. A water-soluble gel contained within the off-set had acoustic properties similar to those of mammalian soft tissues.
An estimate of melanosome concentration in pigment tissues. Concentration of melanosomes in various tissues has been unknown because of the impracticability of their direct quantification. Using an indirect approach comprising the estimation of melanin both in freeze-dried tissue samples and in isolated melanosomes, we obtained data on the amount of melanosomes in various pigment tissues. The concentrations of melanosomes found in the tissues were relatively high, not only reflecting the dark color of pigment tissues but also explaining their capacity to perform various functions ascribed to the presence of melanin.
Cloning and characterization of gene TNF alpha encoding equine tumor necrosis factor alpha. We report the molecular cloning and nucleotide sequence of the equine gene encoding tumor necrosis factor alpha. The 2610-bp genomic sequence was derived from three overlapping polymerase chain reaction products.
Stability of equine lysozyme. I. Thermal unfolding behaviour. The thermal denaturation of Ca(2+)- and apo-forms of equine lysozyme was followed by using far and near UV circular dichroism and intrinsic fluorescence methods. The difference found between the temperature dependence of the ellipticity at 222 nm and 287 nm, which show two stages in the thermal transition, and those at 228 nm and 294 nm, which indicate only one stage over a wide range of temperatures reflects that different subdivisions of the protein molecule are characterized by a different stability, cooperativity and pathway of denaturation. The first transition, reflected in the increase ...
Co-culture of day-5 to day-7 equine embryos in medium with oviductal tissue. Oviductal and uterine embryos were collected from mares at 5 to 7 days following ovulation 1) to evaluate the effects of oviductal tissue explants on in vitro growth and development of equine embryos and 2) to study the morphologic development of equine embryos in culture. Embryos were incubated for 5 days in a medium (control group) or in medium supplemented with oviductal tissue explants (co-culture group). Embryos were evaluated and the media changed daily. Following 5 days in culture, 10 10 (100%) control embryos and 27 29 (93%) co-cultured embryos had doubled in diameter. All embryos that...
Influence of polysulfated glycosaminoglycan on equine articular cartilage in explant culture. Articular cartilage explants from 3 horses were maintained in tissue culture to test the effects of a polysulfated glycosaminoglycan on proteoglycan biosynthesis. Cultures were exposed to concentrations of 0, 50, or 200 micrograms of the drug/ml for either 2 days or 6 days, and labeled with 35S, before measuring the content of sulfated proteoglycan in the culture media and in extracts of cartilage. In a second experiment, the explants were incubated with the isotope and subsequently exposed to the same concentrations of the polysulfated glycosaminoglycan for 4 days. Subsequently, the amount of...
Three monoclonal antibodies identifying antigens on all equine T lymphocytes, and two mutually exclusive T-lymphocyte subsets. The aim of this study was to produce monoclonal antibodies (mAb) recognizing equine lymphocyte surface antigens. Fusions were conducted using BALB/c mice hyperimmunized with equine thymocytes. Hybridoma supernatants were screened by flow cytometry and positive hybridomas were cloned twice by limiting dilution. These mAb were then characterized for tissue distribution by immunohistology and flow cytometry, and by precipitation and analysis of the lymphocyte antigens which they recognized. Three mAb (CVS5, CVS4 and CVS8) are described which recognize only T lymphocytes in peripheral blood. Two-c...
Characterization of variable regions in the envelope and S3 open reading frame of equine infectious anemia virus. The polymerase chain reaction was used to amplify and clone parts of the envelope gene and overlapping S3 open reading frame, thought to encode rev, of the virulent in vivo-derived Th-1 isolate of equine infectious anemia virus (EIAV). The results indicated that EIAV consists of a heterogeneous mixture of genotypes present at the first febrile cycle after initial infection. We showed that the Th-1 isolate apparently contains nondefective genotypes as well as types which have transmembrane protein truncations or are rev deficient. Furthermore, we could confirm the presence of a hypervariable re...
DNA probes for the detection of Babesia caballi. A genomic library of Babesia caballi DNA was constructed in the plasmid vector pUC13. The specificity of the clones for B. caballi was established by the lack of hybridization to Babesia equi, Babesia bovis, Babesia bigemina and equine DNA. Two probes, pBC11 and pBC191, were isolated that could detect 0.25 ng and 0.125 ng of B. caballi DNA, corresponding to a parasitaemia of 0.12% and 0.06% respectively. pBC191 could detect B. caballi parasites in the blood of an experimentally infected horse as well as in naturally infected horses.
Components of electrogenic transport in unstimulated equine tracheal epithelium. Basic components of unstimulated electrolyte transport across equine tracheal mucosa were characterized. After the tissue was mounted in Ussing chambers, both current and tissue resistance gradually increased for approximately 60 min before reaching stable values. Thereafter, under open-circuit conditions, the tissue had a resistance of 250 +/- 14 omega.cm2, generated a transepithelial potential difference of -34 +/- 1.7 (SE) mV (referenced to the serosal side) and an equivalent short-circuit current (Ieqsc) of -149 +/- 10.2 microA/cm2. Even though 10(-5) M amiloride reduced the current by app...
Expression in Escherichia coli of a synthetic gene coding for horse heart myoglobin. A gene for expression of horse heart myoglobin in Escherichia coli has been constructed in one step from long synthetic oligonucleotides. The synthetic gene contains an efficient translation initiation signal and used codons that are commonly found in E. coli. Unique restriction sites are placed throughout the gene. It has been inserted in a phagemid vector and is expressed from the lac promoter in E. coli at high efficiency, the soluble heme protein representing approximately 10% of soluble protein. Two versions of horse heart myoglobin were produced with aspartic acid or asparagine at residu...
Cloning the cDNA for horse growth hormone and expression in Escherichia coli. A 514 bp cDNA transcript coding for 78% of horse (Equus caballus.) GH has been cloned and sequenced. The deduced amino acid sequence corresponded precisely to that previously obtained by protein sequencing and, in addition, provided new sequence information for the signal peptide. The missing 3' fragment of the cDNA was reconstructed using synthetic oligonucleotides and site-specific directed mutagenesis. The complete cDNA sequence was then inserted into an expression vector (PIN-III-lppp-5) which utilizes a bacterial signal peptide to secrete the expressed product into the periplasmic space o...
1H NMR resonance assignments in a paramagnetic heme protein by two-dimensional spectroscopy: heme resonances in equine met-azido myoglobin. Specific heme protons for the majority of resonances in the downfield resolved region of equine met-azido myoglobin have been assigned using solely the two-dimensional 1H NMR experiments NOESY and COSY. Metazido myoglobin provides a useful test case for the applicability of these techniques to paramagnetic proteins for the following reasons. First met-azido myoglobin is a mixed spin-state protein, with significantly shorter relaxation times and broadened lines relative to pure low-spin systems (eg., met-cyano myoglobin). Second, met-azido hemoglobin and met-azido myoglobin are important as mod...
