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Topic:Equine Health

Equine health encompasses the study and management of diseases, disorders, and overall well-being of horses. It involves understanding various physiological systems, preventive care, and treatment strategies to maintain optimal health in equine populations. Common areas of focus include nutrition, infectious diseases, orthopedic conditions, and reproductive health. Research in equine health aims to advance knowledge on diagnostic methods, therapeutic interventions, and management practices that improve horse welfare and performance. This page collects peer-reviewed research studies and scholarly articles that explore the diverse aspects of equine health, offering insights into current findings and advancements in the field.
Optimisation of a serum albumin removal protocol for use in a proteomic study to identify the protein biomarkers for silent gastric ulceration in horses.
Journal of equine science    September 19, 2018   Volume 29, Issue 3 53-60 doi: 10.1294/jes.29.53
Poltep K, Tesena P, Yingchutrakul Y, Taylor J, Wongtawan T.Silent gastric ulceration occurs without evidence of clinical signs and is common in horses. There is currently no a simple and effective method to diagnose this disease. Proteomics can be used to identify serum biomarkers, but the most abundant serum protein, albumin, could conceal candidate biomarkers. Therefore, it is recommended to remove albumin before a proteomic study; however, there is no specific albumin depletion kit or standard protocol available for horse samples. The objectives of this study were to optimise a protocol to remove equine serum albumin and to use albumin-depleted ser...
Evaluating continuous blood coagulopathy in assessing the severity of acute colitis in Thoroughbred racehorses.
Journal of equine science    September 19, 2018   Volume 29, Issue 3 79-85 doi: 10.1294/jes.29.79
Nomura M, Mizobe F, Kato T, Kuroda T, Urayama S, Muranaka M.Although severe blood coagulopathy in horses with acute colitis causes multiple organ failure, which may be fatal, few studies have focused on the correlation between the fluctuations of coagulation parameters and severity of colitis. In this study, we evaluated the fluctuations of coagulation parameters in 14 Thoroughbred racehorses with acute colitis for 5 days from the day of hospitalization and compared them between 5 survivors and 9 non-survivors. Noteworthy features in the non-survivors were that antithrombin activity and fibrin degradation products continuously decreased and increased, ...
Anatomy of the vestibulum esophagi and surgical implications during prosthetic laryngoplasty in horses.
Veterinary surgery : VS    September 19, 2018   Volume 47, Issue 7 942-950 doi: 10.1111/vsu.12944
Brandenberger O, Martens A, Robert C, Wiemer P, Pamela H, Vlaminck L, Barankova K, Haspeslagh M, Perkins JD, Ducharme N, Rossignol F.To describe the anatomy of the entry to the equine esophagus (vestibulum esophagi) and to assess the risk of penetrating its adventitia and/or lumen during laryngoplasty. Methods: Ex vivo cadaveric study. Methods: Five isolated equine larynges and 39 equine head and neck specimens. Methods: The anatomy of the vestibulum esophagi was studied by dissection of 5 cadaver specimens. Then, a bilateral laryngoplasty was performed, including 5 suture placements through the muscular processes, caudal, rostral, and sagittal, with straight and curved needles. Two of the 3 surgeons performing the implanta...
Changes of lying behavior in Thoroughbred foals influenced by age, pasturing time, and weather conditions.
Journal of equine science    September 19, 2018   Volume 29, Issue 3 61-66 doi: 10.1294/jes.29.61
Murase H, Matsui A, Endo Y, Sato F, Hada T.The lying behavior of Thoroughbred foals on breeding farms was continuously measured using triaxial accelerometers. Accelerometers were attached on the side of the hind limb cannon and under the halter of six foals to record triaxial angle data every 10 sec for a period of 24 hr. Lying behavior was divided into sternal lying and lateral lying based on head angle. Sampling was performed for two consecutive days each week until weaning. Sampling periods were divided into two periods on the basis of pasturing time: daytime pasturing (period A: 7-hr pasturing period, 2-60 days of age) and overnigh...
Equine suspensory ligament and tendon explants cultured with platelet-rich gel supernatants release different anti-inflammatory and anabolic mediators.
Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie    September 19, 2018   Volume 108 476-485 doi: 10.1016/j.biopha.2018.09.065
Bonilla-Gutiérrez AF, Castillo-Franz C, López C, Álvarez ME, Giraldo CE, Carmona JU.The aim of this study was to evaluate the release of pro- and anti-inflammatory as well as anabolic mediators stimulated by a leukocyte-reduced platelet-rich gel supernatant (Lr-PRGS) and a leukocyte-reduced plasma supernatant (Lr-PL) at two concentrations (25 and 50%) on normal equine suspensory ligament explants (SLEs) and tendon explants (TEs). SLEs and TEs from six horses were independently incubated for 48 h with Lr-PRGS and Lr-PL at concentrations of 25 and 50%, respectively. Samples were collected from the incubated tissues at 1 h and 48 h, which were employed for ELISA determinat...
How to score sedation and adjust the administration rate of sedatives in horses: a literature review and introduction of the Ghent Sedation Algorithm.
Veterinary anaesthesia and analgesia    September 19, 2018   Volume 46, Issue 1 4-13 doi: 10.1016/j.vaa.2018.08.005
Schauvliege S, Cuypers C, Michielsen A, Gasthuys F, Gozalo-Marcilla M.To summarize the different methods used to assess sedation and/or adjust the dose or administration rate of alpha-2 agonists in horses and to propose an algorithm to adjust the administration rate of a constant rate infusion of an alpha-2 agonist in horses. Methods: PubMed and Web of Science; search terms: horse, sedation and score. Conclusions: Most authors distinguish between sedation depth, sedation quality and degree of ataxia. These three features are evaluated using scoring systems similar to those classically used to assess pain, i.e. simple descriptive scales, numerical rating scales (...
Ultrasound tissue characterisation of the superficial digital flexor tendons in juvenile Thoroughbred racehorses during early race training.
Equine veterinary journal    September 18, 2018   Volume 51, Issue 3 349-355 doi: 10.1111/evj.13006
Plevin S, McLellan J, van Schie H, Parkin T.Injuries to the superficial digital flexor tendon (SDFT) are one of the leading causes of Thoroughbred (TB) wastage. Increasingly, the aim is to prevent injury rather than treat it. Conventional ultrasonography is not sufficiently sensitive to accurately monitor tendon and predict injury. Ultrasound tissue characterisation (UTC) is a relatively new technique, which improves tendon characterisation by providing a 3-dimensional (3D) SDFT reconstruction and objective calculation of fibre alignment by classifying fibres into one of 4 echo-types. Objective: To report a reference range of echo-types...
Does BMP2 play a role in the pathogenesis of equine degenerative suspensory ligament desmitis?
BMC research notes    September 18, 2018   Volume 11, Issue 1 672 doi: 10.1186/s13104-018-3776-9
Young M, Moshood O, Zhang J, Sarbacher CA, Mueller POE, Halper J.Horses afflicted with degenerative suspensory ligament desmitis (DSLD) suffer from progressive leg pain and lameness without history of trauma. DSLD is a systemic disorder caused by abnormal accumulation of proteoglycans in many connective tissues. One proteoglycan found in higher quantities in DSLD is decorin. The accumulated decorin has an abnormally glycosylated glycosaminoglycan chain in DSLD. In addition to acellular accumulations of proteoglycans foci of active fibroblasts/tenoblasts were observed in some tendons and suspensory ligaments (SLs) from DSLD cases We have hypothesized that th...
The magnitude of respiratory sinus arrhythmia of a large mammal (the horse) is like that of humans.
Respiratory physiology & neurobiology    September 18, 2018   Volume 259 170-172 doi: 10.1016/j.resp.2018.09.006
Piccione G, Giudice E, Giannetto C, Mortola JP.Heart rate (FH) accelerates in inspiration and decelerates in expiration, a phenomenon known as Respiratory Sinus Arrhythmia (RSA). Although the presence of RSA has been documented in many species, how its magnitude compares among species is unknown. We asked whether the magnitude of RSA in a large mammal, the horse, differed from that of previously measured humans. From electrocardiogram and pneumography, the peaks and troughs of FH were identified breath-by-breath in four horses (Italian Saddlebred geldings) during resting wakefulness. RSA was computed as the peak-trough FH difference, in pe...
Ultrastructural and histological characteristics of the endometrium during early embryo development in mares.
Theriogenology    September 18, 2018   Volume 123 1-10 doi: 10.1016/j.theriogenology.2018.09.018
Camozzato GC, Martinez MN, Bastos HBA, Fiala-Rechsteiner S, Meikle A, Jobim MIM, Gregory RM, Mattos RC.The aim of this study was to evaluate ultrastructural and histological changes in the endometrium on days 7, 10 and 13 post-ovulation in pregnant and cyclic mares. Mares were routinely examined by transrectal palpation and ultrasonographic examination of the reproductive tract until estrus was detected. In the first cycle, endometrial biopsies from 30 cyclic mares (Cyclic group) were collected on days 7, 10 and 13 post-ovulation. In the second cycle, the same mares were bred by a fertile stallion. At days 7, 10 and 13 post-ovulation intrauterine biopsies were collected. Immediately after sampl...
Effect of valacyclovir on EHV-5 viral kinetics in horses with equine multinodular pulmonary fibrosis.
Journal of veterinary internal medicine    September 17, 2018   Volume 32, Issue 5 1763-1767 doi: 10.1111/jvim.15230
Easton-Jones CA, Madigan JE, Barnum S, Maxwell LK, Taylor SD, Arnesen T, Pusterla N.Equine herpesvirus-5 is commonly isolated from the lungs of horses with EMPF, suggesting an etiological link. Valacyclovir is used empirically to treat EMPF; however, no data is available concerning its impact on EHV-5 viral kinetics. Objective: To determine the effect of oral administration of valacyclovir on EHV-5 viral load measured by qPCR in blood, nasal secretions (NS) and BALF in horses with EMPF. Methods: Six horses diagnosed with EMPF. Methods: A prospective clinical trial was performed. Horses received 10 days of PO administered valacyclovir (loading dose 30 mg/kg, maintenance dose 2...
An observational study of environmental exposures, airway cytology, and performance in racing thoroughbreds.
Journal of veterinary internal medicine    September 17, 2018   Volume 32, Issue 5 1754-1762 doi: 10.1111/jvim.15226
Ivester KM, Couëtil LL, Moore GE.Mild equine asthma is presumed to arise in response to environmental exposures but the relative impact of differing inflammatory phenotypes upon performance are largely unexplored. Objective: Airway inflammation negatively affects performance and cytological phenotype varies with environmental exposure. Methods: Thoroughbred racehorses in active training and racing. Methods: Thoroughbreds were recruited 24-48 hours before racing. Each horse was eligible for re-enrollment with each race entry. Within one hour of race completion, physical examination, respiratory endoscopy, and BAL were performe...
Non-surgical embryo transfer technique and recipient mare pregnancy rate.
The Veterinary record    September 16, 2018   Volume 183, Issue 10 320-322 doi: 10.1136/vr.k3700
Card C.No abstract available
Utility of C-reactive protein and serum amyloid A in the diagnosis of equine protozoal myeloencephalitis.
Journal of veterinary internal medicine    September 14, 2018   Volume 32, Issue 5 1726-1730 doi: 10.1111/jvim.15254
Mittelman NS, Stefanovski D, Johnson AL.Accurate antemortem EPM diagnosis requires evidence of intrathecal antibody production. Some advocate the use of acute phase proteins in addition to serology, which alone results in substantial false positives. Objective: The purpose of this study was to determine if serum C-reactive protein (CRP) or serum amyloid A (SAA) concentrations were elevated in cases of equine protozoal myeloencephalitis (EPM) compared to other neurological diseases. Methods: 25 clinical cases of equine neurological disease: EPM (10), cervical vertebral stenotic myelopathy (CVSM) (10), neuroborreliosis (2), equine mot...
Newborn foal with atypical myopathy.
Journal of veterinary internal medicine    September 14, 2018   Volume 32, Issue 5 1768-1772 doi: 10.1111/jvim.15236
Karlíková R, Široká J, Mech M, Friedecký D, Janečková H, Mádrová L, Hrdinová F, Drábková Z, Dobešová O, Adam T, Jahn P.The case of atypical myopathy (AM) in newborn Haflinger foal with clinical signs of depression and weakness appearing 6 hours after birth resulting in recumbency 12 hours after birth is described. The foal's dam was diagnosed with AM in the 6th month of gestation based on clinical signs of a myopathy, elevated serum activity of creatine kinase, metabolomic analysis and the presence of methylenecyclopropyl acetyl carnitine (MCPA-carnitine) in the blood. At the time of delivery, the mare was grazing on a pasture near sycamore trees but was free of clinical signs of AM. Metabolomic analysis of th...
Origin and spread of Thoroughbred racehorses inferred from complete mitochondrial genome sequences: Phylogenomic and Bayesian coalescent perspectives.
PloS one    September 14, 2018   Volume 13, Issue 9 e0203917 doi: 10.1371/journal.pone.0203917
Yoon SH, Lee W, Ahn H, Caetano-Anolles K, Park KD, Kim H.The Thoroughbred horse breed was developed primarily for racing, and has a significant contribution to the qualitative improvement of many other horse breeds. Despite the importance of Thoroughbred racehorses in historical, cultural, and economical viewpoints, there was no temporal and spatial dynamics of them using the mitogenome sequences. To explore this topic, the complete mitochondrial genome sequences of 14 Thoroughbreds and two Przewalski's horses were determined. These sequences were analyzed together along with 151 previously published horse mitochondrial genomes from a range of breed...
Survey for Equine Herpesviruses in Polar Bears (Ursus maritimus) and Exotic Equids Housed in US AZA Institutions.
Journal of zoo and wildlife medicine : official publication of the American Association of Zoo Veterinarians    September 14, 2018   Volume 49, Issue 3 599-608 doi: 10.1638/2016-0189.1
Flanders JA, Wack RF, Pusterla N, Mapes SM, Collins D, Gamble KC.Infection by equine herpesvirus (EHV) strains (EHV-1, EHV-9) in ursid species, including polar bears ( Ursus maritimus), has been associated with neurological disease and death. A serosurvey of captive exotic equid and polar bear populations in US Association of Zoos and Aquaria institutions was performed to determine the prevalence of EHV strains using quantitative polymerase chain reaction (qPCR) and enzyme-linked immunosorbent assay (ELISA) tests. Equid species surveyed included zebra ( Equus spp.), Przewalski's wild horse ( Equus ferus przewalskii), Persian onager ( Equus hemionus), and So...
Differences in the intrinsic chondrogenic potential of equine umbilical cord matrix and cord blood mesenchymal stromal/stem cells for cartilage regeneration.
Scientific reports    September 14, 2018   Volume 8, Issue 1 13799 doi: 10.1038/s41598-018-28164-9
Rakic R, Bourdon B, Demoor M, Maddens S, Saulnier N, Galéra P.Umbilical cord blood mesenchymal stromal/stem cells (UCB-MSCs) and umbilical cord matrix MSCs (UCM-MSCs) have chondrogenic potential and are alternative sources to standard surgically derived bone marrow or adipose tissue collection for cartilage engineering. However, the majority of comparative studies explore neonatal MSCs potential only on ISCT benchmark assays accounting for some bias in the reproducibility between in vitro and in clinical studies. Therefore, we characterized equine UCB-MSCs and UCM-MSCs and investigated with particular attention their chondrogenesis potential in 3D cultu...
Conservative versus surgical treatment of 21 sports horses with osseous trauma in the proximal phalangeal sagittal groove diagnosed by low-field MRI.
Veterinary surgery : VS    September 14, 2018   Volume 47, Issue 7 908-915 doi: 10.1111/vsu.12936
Lipreri G, Bladon BM, Giorio ME, Singer ER.To compare the outcome following conservative vs surgical management of sports horses with a diagnosis of subchondral bone trauma of the proximal aspect of the proximal phalanx (PP) by low-field MRI. Methods: Retrospective case series. Methods: Twenty-one mature sports horses with evidence of high water signal within the proximal sagittal groove of the PP according to low-field MRI of the fetlock, with no definitive radiographic evidence of fracture. Methods: Medical records (2010-2017) of horses admitted to 2 referral hospitals were reviewed. Historical, clinical, radiographic, and MRI findin...
Minimally invasive, compartmentalized, modified open castration technique with primary closure in equids.
Journal of the American Veterinary Medical Association    September 14, 2018   Volume 253, Issue 7 897-906 doi: 10.2460/javma.253.7.897
Crosa AT, Desjardins MR.OBJECTIVE To describe use of a modified open castration technique with a scrotal approach and primary closure in equids. DESIGN Prospective case series. ANIMALS 38 client-owned, sexually intact male equids. PROCEDURES With owner consent, elective castration was performed with a modified open technique on patients (1 pony, 1 miniature horse, and 36 horses of other breeds) under general anesthesia. The procedure included minimal dissection into the scrotal region for removal of testes, with cremaster muscles left intact and the parietal vaginal tunic closed in place. Primary closure of surgical ...
Actinomyces denticolens as a causative agent of actinomycosis in animals.
The Journal of veterinary medical science    September 14, 2018   Volume 80, Issue 11 1650-1656 doi: 10.1292/jvms.18-0207
Murakami S, Kobayashi T, Sekigawa Y, Torii Y, Kanesaki Y, Ishige T, Yokoyama E, Ishiwata H, Hamada M, Tamura T.The name "Actinomyces suis" was applied to each actinomycete isolate from swine actinomycosis by Grässer in 1962 and Franke in 1973. Nevertheless, this specific species was not included in the "Approved List of Bacterial Name" due to absence of the type cultures. Therefore, "Actinomyces suis" based on the description of Franke 1973 has been considered as "species incertae sedis". We isolated a number of Actinomyces strains from swine. The representative strains of them was designated as Chiba 101 that was closely similar to the description in "Actinomyces suis" reported by Franke in 1973. Int...
Prevalence and risk factors for IgG antibodies to Neospora spp. in three types of equids from Southern Punjab, Pakistan.
Acta tropica    September 13, 2018   Volume 188 240-243 doi: 10.1016/j.actatropica.2018.09.007
Nazir MM, Ayaz MM, Ahmed AN, Rasheed I, Faraz A, Akram Q, Akhtar S, Maqbool A, Tabassum S, Zheng Y, Lindsay DS.Equine hosts suffer from neurological disease, congenital infection, and reproductive problems associated with Neospora spp. infection. We conducted a cross sectional study using sera from 631 equids (324 horses, 218 donkeys and 89 mules) from the southern region of Punjab province, Pakistan to determine the prevalence of antibodies against Neospora spp. in this diverse group of equines. Fisk factors associated with seropositivity were evaluated statistically based on equine type, breed, age, husbandry, breeding methods, and reproductive history. Prevalence of antibodies to Neospora spp. was d...
Effects of exercise on myokine gene expression in horse skeletal muscles.
Asian-Australasian journal of animal sciences    September 13, 2018   Volume 32, Issue 3 350-356 doi: 10.5713/ajas.18.0375
Lee HG, Choi JY, Park JW, Park TS, Song KD, Shin D, Cho BW.To examine the regulatory effects of exercise on myokine expression in horse skeletal muscle cells, we compared the expression of several myokine genes (interleukin 6 [IL-6], IL-8, chemokine [C-X-C motif] ligand 2 [CXCL2], and chemokine [C-C motif] ligand 4 [CCL4]) after a single bout of exercise in horses. Furthermore, to establish in vitro systems for the validation of exercise effects, we cultured horse skeletal muscle cells and confirmed the expression of these genes after treatment with hydrogen peroxide. Methods: The mRNA expression of IL-6, IL-8, CXCL2, and CCL4 after exercise in skelet...
Age-associated changes in the equine flash visual evoked potential.
Veterinary ophthalmology    September 13, 2018   Volume 22, Issue 4 388-397 doi: 10.1111/vop.12601
Ström L, Michanek M, Ekesten B.To investigate age-associated changes of flash visual evoked potentials (FVEPs) in sedated horses. Methods: Twenty-eight clinically healthy Standardbred Warmblooded trotters, aged 36 hours to 28 years. Methods: Light-adapted FVEPs and FERGs were recorded (An-vision RETIport, Roland-consult, Germany) in response to flash stimuli. Sedation was obtained using alpha-2-agonists intravenously. Akinesia of the eyelids was induced and pupils were dilated. Results: Reproducible FVEPs and FERGs were readily recorded from all foals and horses. The FVEP waveform included up to four positive components (...
The sodium-glucose co-transporter 2 inhibitor velagliflozin reduces hyperinsulinemia and prevents laminitis in insulin-dysregulated ponies.
PloS one    September 13, 2018   Volume 13, Issue 9 e0203655 doi: 10.1371/journal.pone.0203655
Meier A, Reiche D, de Laat M, Pollitt C, Walsh D, Sillence M.There are no registered veterinary drugs for treating insulin dysregulation and preventing insulin-associated laminitis in horses. Velagliflozin is a sodium-glucose co-transport 2 inhibitor that reduces renal glucose reabsorption, promotes glucosuria, and consequently, decreases blood glucose and insulin concentrations. This study aimed to determine if velagliflozin reduced hyperinsulinemia and prevented laminitis in insulin-dysregulated ponies fed a challenge diet high in non-structural carbohydrates (NSC). An oral glucose test (1 g dextrose/kg BW) was used to screen 75 ponies for insulin dys...
Novel mcr-5.3 variant in a CTX-M-8-producing Escherichia coli ST711 isolated from an infected horse.
The Journal of antimicrobial chemotherapy    September 12, 2018   Volume 73, Issue 12 3520-3522 doi: 10.1093/jac/dky341
Fernandes MR, Cerdeira L, Silva MM, Sellera FP, Muñoz M, Junior FG, Azevedo SS, Power P, Gutkind G, Lincopan N.No abstract available
The first report of polymorphisms and genetic characteristics of the prion protein gene (PRNP) in horses.
Prion    September 12, 2018   Volume 12, Issue 3-4 245-252 doi: 10.1080/19336896.2018.1513316
Kim YC, Jeong BH.Prion diseases have a wide host range, but prion-infected cases have never been reported in horses. Genetic polymorphisms that can directly impact the structural stability of horse prion protein have not been investigated thus far. In addition, we noticed that previous studies focusing on horse-specific amino acids and secondary structure predictions of prion protein were performed for limited parts of the protein. In this study, we found genetic polymorphisms in the horse prion protein gene (PRNP) in 201 Thoroughbred horses. The identified polymorphism was assessed to determine whether this p...
Genetic diversity and prevalence of piroplasm species in equids from Turkey.
Comparative immunology, microbiology and infectious diseases    September 12, 2018   Volume 59 47-51 doi: 10.1016/j.cimid.2018.08.005
Ozubek S, Aktas M.Equine piroplasmosis (EP) is a protozoon disease caused by Babesia caballi and Theileria equi transmitted by ticks from the Ixodidae family. This study investigated the genetic heterogeneity and diversity of piroplasm genotypes using the Reverse Line Blotting (RLB) technique for piroplasm species in equids in Turkey. A total of 233 blood samples from 142 horses and 91 donkeys were collected in Şanlıurfa, Tunceli, and Iğdır. The RLB assay was performed for simultaneous detection of piroplasm species. The prevalence of piroplasm infection was 33.5% (95% CI; 27.4-39.9). T. equi was the most c...
Generation of Functional Myocytes from Equine Induced Pluripotent Stem Cells.
Cellular reprogramming    September 12, 2018   Volume 20, Issue 5 275-281 doi: 10.1089/cell.2018.0023
Amilon KR, Cortes-Araya Y, Moore B, Lee S, Lillico S, Breton A, Esteves CL, Donadeu FX.Induced pluripotent stem cells (iPSCs) have revolutionized human biomedicine through their use in disease modeling and therapy. In comparison, little progress has been made toward the application of iPSCs in veterinary species. In that regard, skeletal myocytes from iPSCs would have great potential for understanding muscle function and disease in the equine athlete. In this study, we generated skeletal myotubes by transducing equine iPSC-derived mesenchymal derivatives with an inducible lentiviral vector coding for the human sequence of the myogenic factor, MyoD. Myosin heavy chain-positive my...
Purification of equine IgG3 by lectin affinity and an interaction analysis via microscale thermophoresis.
Analytical biochemistry    September 12, 2018   Volume 561-562 27-31 doi: 10.1016/j.ab.2018.09.010
De-Simone SG, Nascimento HJ, Prado IC, Aguiar AS, Melgarejo AR, Pina JLS, Ferreira PF, Provance DW.The availability of purified antibodies is a prerequisite for many applications and the appropriate choice(s) for antibody-purification is crucial. Numerous methods have been developed for the purification of antibodies from different sources with affinity chromatography-based methods being the most extensively utilized. These methods are based on high specificity, easy reversibility and biological interactions between two molecules (e.g., between receptor and ligand or antibody and antigen). However, no simple techniques have yet been described to characterize and purify subclasses of immunog...