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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.
Purification of F(ab’)2 anti-snake venom by caprylic acid: a fast method for obtaining IgG fragments with high neutralization activity, purity and yield. Pooled horse plasma containing antibodies against Crotalus durissus terrificus whole venom were digested with pepsin at an enzyme-substrate ratio of 8:1, pH 3.1, for 40 min and the F(ab')2M fragments purified by adding 8.7% caprylic acid (pH 5.0). For comparison, F(ab')2B purified by precipitation with ammonium sulphate and uncleaved IgG purified with caprylic acid were also prepared. Fab' fragments were obtained by reduction and alkylation of F(ab')2B. The anti-whole C.d. terrificus venom titers, determined by Dot-Blot were 12,800 (IgG), 6400 [F(ab')2B], 4800 [F(ab')2M] and 3200 (Fab'B). Immu...
Membrane viscosity of lymphocytes and influence of phytohemagglutinin. The membrane viscosity of peripheral blood lymphocytes (PBLs) of equine, bovine and canine was measured by the use of time-resolved fluorescence depolarization technique with 1, 6-diphenyl-1,3,5-hexatriene (DPH). The viscosity values were 0.55, 0.59 and 0.50 poise for equine, bovine and canine PBLs, respectively. These values were compared with steady-state anisotropies and order parameters measured from electron spin resonance (ESR) of 5-doxyl stearic acid. Both values were increased with increase of viscosity. The fluid property of the membranes stimulated with phytohemagglutinin-P (PHA) was...
Isolation of granulocytes and mononuclear cells from the blood of dogs, cats, horses and cattle. A simple discontinuous Percoll density-gradient technique was adapted for isolation of granulocytes and mononuclear cells from cats, dogs, horses and cattle. Separation was accomplished at low speeds using a standard tabletop centrifuge. Cell purity was 100% for both granulocytes and mononuclear cells and cell viability exceeded 95%. Percent recovery of leukocytes ranged from 69 to 83%.
Biotransformation of 1-dehydrotestosterone in the equine male castrate: identification of the neutral unconjugated and glucuronic acid conjugated metabolites in horse urine. The in vivo biotransformation of (1,2(n)-3H)1-dehydrotestosterone was studied in three equine male castrates and a number of neutral metabolites were identified in the urinary unconjugated and glucuronic acid conjugate fractions by gas chromatography/mass spectrometry. The metabolites were extracted from aliquots of the 0-24 h urine samples by Amberlite XAD-2 and separated into combined unconjugated plus glucuronic acid conjugated and sulphoconjugated fractions by Sephadex LH-20 column chromatography. After enzymatic hydrolysis of the glucuronides, the crude neutral unconjugated steroids plus ...
Purification of specific heterologous F(ab)2 fragments with DEAE-Zeta-Prep cartridges for ion-exchange chromatography. A simple two-step procedure for purifying F(ab)2 fragments of horse immunoglobulins is described. In the first step, the horse plasma is diluted, made up to 12% (w/v) with ammonium sulphate and digested with pepsin. In the second step, the previously dialyzed solution is chromatographed. Instead of a normal ion-exchange resin, a DEAE-cellulose, covalently linked to a synthetic vinyl polymer, was used (DEAE-Zeta-Prep). With this assembly it is possible to perform chromatography at a high flow-rate without the problems related to the use of large columns. The yield and purity of the final produc...
The primary structure of donkey (Equus asinus) lysozyme contains the Ca(II) binding site of alpha-lactalbumin. The complete primary structure of donkey lysozyme has been established by pulsed liquid-phase sequencing of tryptic and chymotryptic peptides isolated by RP-HPLC. The positions of the Cys residues were identified by labeling the Cys residues with DABIA-reagent. Donkey lysozyme is a c-type lysozyme which is 129 amino acids long. It exhibits 50% homology to the human protein. We observe the full Ca(II) binding site suggested for the homologous alpha-lactalbumines. Although horse lysozyme has been reported to contain asparagine in position 61, which was in conflict with the three-dimensional stru...
Ultrasonic transmission velocity and single photon absorptiometric measurement of metacarpal bone strength: an in vitro study in the horse. Ten pairs of third metacarpal bones from Thoroughbred horses aged two to 12 years were used to estimate bone strength. Measurements of transverse cortical ultrasound velocity, cortical cross sectional area and bone mineral content were made using ultrasonic transmission velocity and single photon absorptiometry. These data were used to determine bone mineral density, compact bone density and modulus of elasticity. The results were compared with those measured by direct means or chemical analysis and satisfactory correlations were obtained between estimated and measured values. Single photon ab...
Capillary permeability to endogenous macromolecules in the equine digit. Microvascular permeability characteristics were evaluated in digits of 7 adult horses. After capillaries were isolated and an extracorporeal perfusion circuit for the digit was established, a lymphatic vessel draining the distal portion of the phalangeal region was cannulated at the level of the coronary band. Venous pressure was increased in a stepwise manner, and lymph flow, lymph protein concentration (Cl), and plasma protein concentration (Cp) were determined after measured variables were allowed to reach steady state. Lymph-to-plasma protein concentration ratios (Cl/Cp) and lymph and plas...
Application of cloned fragments of equine herpesvirus type-1 DNA for detection of virus-specific DNA in equine tissues. Tissue specimens obtained from equine herpesvirus-1 (EHV-1), subtype 1-infected aborted foetuses were analysed for the presence of virus DNA by means of Southern blot and dot blot hybridisations. The specificity of the methods was confirmed although the sensitivity was inferior to classical techniques such as virus isolation. However, the possibility of detecting the state of the virus DNA and the ability to distinguish between subtypes were important features, and the dot blot method was shown to have potential for a rapid diagnostic test. This report demonstrates some potential practical app...
Cytochrome c: ion binding and redox properties. Studies on ferri and ferro forms of horse, bovine, and tuna cytochrome c. The ion binding properties of horse, bovine, and tuna cytochrome c (both oxidized and reduced) have been measured using a combination of ultrafiltration, neutron activation, and ion chromatography. The ions investigated were chloride, phosphate, and Tris-cacodylate. Ion chromatography and neutron activation analysis techniques were employed to determine the concentration of free anions. Binding constants are obtained from modified Scatchard plots (in the range of 10-2000 M-1). The redox potentials for cytochrome c at different ionic strengths, pH 7.0, have been determined. In this paper we rep...
Molecular mechanics calculation of geometries of NAD+ derivatives, modified in the nicotinamide group, in a ternary complex with horse liver alcohol dehydrogenase. The geometry of seven NAD+ analogues bound to horse liver alcohol dehydrogenase (LADH) modified only in their nicotinamide group, have been studied using AMBER molecular mechanics energy-minimization procedures. Starting geometries were taken from X-ray crystallographic data for NAD+/Me2SO/LADH reported by Eklund and co-workers. In this study the NAD+ analogues were encaged by the constituent amino acids of the enzyme within a range of 0.6 nm from the initial NAD+/Me2SO/Zn2+ complex. The calculational method used is able to rationalize individual substituent effects and to evaluate the essenti...
Isolation and quantitation of cadmium-, zinc- and copper-metallothioneins by high-performance liquid chromatography-atomic absorption spectrometry. Metallothioneins (MTs) were separated and quantitated by reversed-phase high-performance liquid chromatography (RP-HPLC), in direct combination with atomic absorption spectrometry (AAS) for quantitation of the metal contents in MTs. MTs were eluted from an RP-8 column with a gradient of Tris buffer pH 7.0 and methanol, and were detected by UV absorbance (220 nm). Commercially available purified MTs from horse kidney and rabbit liver were analyzed for purity and metal composition. One lot of horse kidney yielded only 50% of the estimated value. In some cases, the certified metal content differe...
Horse urinary kallikrein, II. Effect of subsite interactions on its catalytic activity. The effect of secondary-subsite interactions on the catalytic efficiency of horse urinary kallikrein was studied using as substrates oligopeptides and peptidyl-4-nitroanilides with L-Arg at P1. The known secondary specificity of tissue kallikreins for hydrophobic residues at P2 was also demonstrated for horse urinary kallikrein and a higher preference of this enzyme for L-Phe over L-Leu at P2 was evident. Interaction of subsites S3 with D-Pro and D-Phe enhanced the catalytic efficiency but tripeptidyl-4-nitroanilides with acetyl-D-Pro, L-Pro and acetyl-L-Pro at P3 were no better substrates tha...
Establishment of equine T-lymphocyte cultures dependent on recombinant human interleukin-2. Long-term equine lymphocyte cultures were initiated and maintained in continuous culture with medium containing recombinant human interleukin-2. Cultures were successfully maintained with lectin activation signals and recombinant human interleukin-2 or with recombinant human interleukin-2 alone. All cell cultures that were characterized had a T-lymphocyte phenotype and had lectin-dependent or -independent cytotoxicity directed to various cell types. These findings demonstrate that long-term equine T-lymphocytes cultures can be initiated and maintained easily.
[Sublimation-dried mare’s milk and the possibility of its use in creating infant and dietary food products]. The main biological parameters were studied in sublimated mare's milk: protein fraction composition, amino-acid spectrum, fatty-acid composition of lipids and the content of vitamins and mineral substances. A high biological value of the sublimated mare's milk has been proved, as well as significant proximity of its composition to human milk. Preliminary data have evidenced expediency of mare's milk use for the development of adapted milk mixtures for baby foods. However, the fatty component of the mixtures needs certain correction by addition of vegetable oil. The sublimated mare's milk can b...
Conserved repetitive DNA sequences (Bkm) in normal equine males and sex-reversed females detected by in situ hybridization. In situ hybridization with a cloned banded krait sex-specific repetitive DNA probe (Bkm) indicates a high concentration of Bkm sequences on the horse Y chromosome in both normal XY males and XY sex-reversed females. Lesser, but still significant, concentrations of Bkm sequences were mapped to horse chromosomes 3, 4, and 30.
Variability of fiber size, capillary density and capillary length related to horse muscle fixation procedures. Muscle samples were taken from the superficial part of the semitendinosus muscle of four standardbred horses to compare the effects of three different fixation procedures on fiber size and capillarity. Infiltration fixation in an extended position of the muscle was compared to immersion fixation and infiltration fixation in a flexed position of the limb. The capillary length density was used to estimate muscle capillarity. Infiltration fixation was found to be a viable alternative to perfusion fixation of muscles for morphometry of fiber size and capillary length density allowing muscle tissue...
Chromosomal localization of the major histocompatibility complex of the horse (ELA) by in situ hybridization. The first gene assignment to a horse chromosome is reported for equine leucocyte antigen (ELA), the major histocompatibility complex of the horse. A cloned DNA sequence derived from a class I gene of the porcine major histocompatibility complex was used as a probe for an in situ hybridization experiment. We present the regional localization of ELA, using this sequence, to equine chromosome 20q14-q22.
Purification of brush border membrane vesicles from horse kidney cortex using Percoll. A rapid method for preparation of brush border membrane vesicles from a large amount of horse kidney cortex is described. Self-orienting Percoll-gradient centrifugation minimized contamination by microsomal membranes. The characteristics of this preparation were checked by electron microscopy and measurement of L-alanine uptake.
Restriction fragment length polymorphisms of horse class II MHC genes observed using various human alpha- and beta-chain cDNA probes. Genomic DNA isolated from 20 horses was digested with up to six restriction endonucleases and subjected to southern blot hybridization analysis using various human class II alpha- and beta-chain cDNA probes. A high degree of restriction fragment length polymorphism (RFLP) was found for the DQ alpha, DP beta, DQ beta and DR beta probes, about 20 polymorphic bands being detected for each. DR alpha showed 2-4 polymorphic bands, whereas no evidence for DP alpha-like genes was found. A number of correlations of RFLPs with individual alloantisera were apparent.
Calibration of the mercury-in-silastic strain gauge in tendon load experiments. A calibration method is presented by which the signals of mercury-in-silastic strain gauges (MISS), implanted in the tendons of in vitro loaded equine hindlegs, were converted to tendon loads. The relationships between MISS-signals and tendon loads were obtained from tensile-force tests applied to the tendons. Special attention was paid to the correction of the MISS-signals for amplitude-shifts resulting from internal repositioning of the MISS after tendon isolation and temperature differences. Shift corrections equivalent to tendon strains up to 2.8% were necessary in the in vitro experiment....
Methods for collecting follicular oocytes from mares. A series of experiments was conducted to develop a procedure for consistent, repeatable collection of oocytes from the preovulatory follicle of the mare. In one experiment, in situ follicular aspiration with a needle and syringe was performed on 19 mares. From 37 aspirations, four oocytes were recovered (10% recovery rate). In a second experiment, ovaries were visualized via standing flank laparotomy during which two different aspiration techniques were used. Use of a needle and syringe as in the first experiment resulted in successful oocyte recovery in one of seven (14%) attempts. Aspiration...
Difference in growth behavior of human, swine, equine, and avian influenza viruses at a high temperature. Growth characteristics of a wide range of influenza A viruses from different mammals and bird species were examined in an established line of canine kidney (MDCK) cells at an ordinary (37 degrees C) and a high temperature (42 degrees C). Although all viruses employed in the present study possessed a capability of replicating at 37 degrees C, virus growth at 42 degrees C showed considerable variation and reflected differences in the natural hosts of the isolates. All reference strains and isolates from bird species grew well in the MDCK cells maintained at 42 degrees C, but human viruses did no...
Antigenic variation of equine infectious anemia virus as detected by virus neutralization. Brief report. The antigenic structure of 16 viruses isolated from four horses which were inoculated with a clone of equine infectious anemia (EIA) virus was compared by the neutralization test. The antigenic structure of viruses isolated after development of neutralizing antibody differed from virus to virus. Back mutation of the antigenic structure was also demonstrated by serial passage of the virus in horses. These results suggest that EIA virus is subject to multidirectional antigenic variation. The possibility that the variants originated in the heterologous virus population in the inoculum seems to be...
A sensitive microtitre plate enzyme immunoassay of oestradiol-17 beta in the cow and mare. Microtitre plates were coated with antiserum against oestradiol-17 beta-6-(O-carboxymethyl)-oxime bovine serum albumin raised in sheep. The plasma samples (0.2-1.0 ml) were extracted with peroxide-free diethyl ether prepared daily by treatment with Al2O3. The enzyme conjugate was prepared by coupling oestradiol-17 beta-6-(O-carboxymethyl)-oxime to horse-radish peroxidase. The conjugate was chromatographed on a Sephadex G-25 column. The standard curve ranged from 0.37 to 18.40 fmol/well of oestradiol-17 beta. The amount of oestradiol-17 beta causing a 50% reduction of maximum binding was 4.4 fm...
[Isolation and purification of proteolytic enzymes on organo-silica sorbents with immobilized gramicidin S]. Biospecific sorbents for affinity chromatography of proteolytic enzymes have been synthesized by attaching cyclopeptide antibiotic gramicidin S to organo-silica supports. It is shown possible to attach gramicidin S to the organo-silica supports using glutaric aldehyde, p-benzoquinone, soluble and insoluble carbodiimides. The sorbents prepared by these methods were successfully applied for the purification of the crude pepsin from horse gastric juice and proteolytic complex produced by Acremonium chrysogenum.
