Laboratory methods in equine research encompass a variety of techniques and procedures used to analyze biological samples from horses to study health, disease, genetics, and physiology. These methods include hematological analyses, biochemical assays, molecular biology techniques, and microbiological cultures. Commonly utilized laboratory techniques involve blood tests for complete blood count (CBC) and serum chemistry, polymerase chain reaction (PCR) for genetic and infectious disease studies, and enzyme-linked immunosorbent assays (ELISA) for detecting specific proteins or antibodies. These methods provide valuable data that contribute to understanding equine health and disease mechanisms. This page compiles peer-reviewed research studies and scholarly articles that explore the application, development, and outcomes of laboratory methods in the context of equine research.
Beretta C, Fadini L, Stracciari JM, Montesissa C.Febantel and one of its main metabolites, febantel sulphoxide, are chemically modified to only a slight extent when incubated in vitro with sheep and cattle ruminal fluids; other major metabolites, fenbendazole and oxfendazole, are respectively, oxidized to oxfendazole and reduced to fenbendazole. Febantel is negligibly metabolized by hepatic cytosol fractions but microsome preparations effect more extensive metabolic transformations. Important differences in this respect were found between microsome preparations from rat, horse, pig, cattle, sheep, chicken and trout livers.
Matthews SM, Joysey VC.Fifteen equine leucocyte antigens were defined by absorption and titration analysis of alloantisera obtained by natural sensitisation through pregnancy and by planned experimental immunisation. Definitive sera were tested on the cells of 90 unrelated horses and members of eight equine families. The family data suggested that 13 specificities were coded by a single locus (first locus) and one specificity (Eq 14) was coded by a second linked locus. The remaining specificity (Eq 7) was controlled by a third locus unlinked to the first or second loci. Tests on the cells of unrelated horses showed ...
Varner DD, Ward CR, Storey BT, Kenney RM.Equine spermatozoa were incubated in a chemically defined medium for 8 hours. The medium preserved spermatozoal viability, as assessed by total spermatozoal motility, progressive spermatozoal motility, and spermatozoal exclusion of eosin stain. Effects of time and divalent cation ionophore, A23187, on the occurrence and character of the spermatozoal acrosome reaction were determined. Two light microscopic assays, a triple-stain technique and a chlortetracycline fluorescence assay, were calibrated with transmission electron microscopy for detection of the acrosome reaction. Incubation time and ...
Clarke AF, Madelin T.This paper describes and compares three techniques of categorisation of hay, straw and other feeds and beddings collected from stables. A hand-held sampler was used to categorise samples according to the presence of plant material, fungal spores and dust mites. An Andersen sampler was used to categorise samples according to the thermotolerances of fungi and actinomycetes. An aerodynamic particle sizer was used to categorise samples according to respirable particle release rates. The highest burden of respirable particles was associated with the presence of thermophilic and thermotolerant actin...
Fukushima JG, Cascone O, Santomé JA, Biscoglio de Jimenez Bonino MJ.Reactivity of histidine residues in equine growth hormone to ethoxyformic anhydride was studied. The existence of two kinetically different sets was demonstrated: one of them including only the slow reacting histidine 169 (k = 0.164 min-1) and the other containing fast reacting histidines 19 and 21 (k = 0.892 min-1). A correlation between the decrease in the capacity to compete with 125I-labeled hormone for rat liver binding sites and the degree of ethoxyformylation of the fast group was found. Circular dichroism studies indicated no significant conformational changes in the protein with all t...
McDonald J, Gall R, Wiedenbach P, Bass VD, DeLeon B, Brockus C, Stobert D, Wie S, Prange CA, Yang JM.We investigated the use of particle concentration fluorescence immunoassay (PCFIA) as a technique for drug detection in racing horses. The test was constructed from an antiserum to a carboxyfentanyl-BSA conjugate and carboxyfentanyl linked to b-Phycoerythrin. Using these reagents and a PCFIA apparatus levels of fentanyl as low as 0.1 ng/ml could be detected by the assay. In addition, cross-reactivity studies on this assay showed that the anti-serum cross-reacted well with carfentanil, sufentanil and the methylated analogs of fentanyl. We therefore evaluated the ability of these agents to produ...
Singh AK, Mishra U, Ashraf M, Abdennebi EH, Granley K, Dombrovskis D, Hewetson D, Stowe CM.Chemical ionization- and electron impact ionization-selective ion monitoring provided a simple and sensitive method for measuring detomidine (Domosedan), a potent sedative-analgesic drug for horses and cattle. Chemical ionization was at least 10 times more sensitive than electron impact ionization. By using propranolol as an internal standard, we found that the recovery of detomidine from the extraction procedure used in this study was greater than 75% for plasma, whole blood, or urine samples. Approximately 68% of detomidine was bound to plasma protein and 53% was bound to red blood cells.
Cooper HM, Jemmerson R, Hunt DF, Griffin PR, Yates JR, Shabanowitz J, Zhu NZ, Paterson Y.Comparative binding studies with peptide fragments of the whole antigen, or with evolutionarily related intact proteins with varying degrees of sequence homology, have been used extensively to map antigenic sites on proteins to the resolution of single amino acid residues. These methods are limited, however, since high affinity antibodies will often not react with peptides and evolutionarily related proteins are available for only a few antigens. In this study we use site-directed chemical modification of horse cytochrome c to identify residues involved in the binding sites of four monoclonal ...
Magneson GR, Puvathingal JM, Ray WJ.The enzyme phosphoglucomutase can be used as a metal ion indicator to measure the concentrations of free Mg2+ and free Zn2+ in physiological fluids. In horse plasma, the concentration of free Mg2+ is close to 0.5 mM, whereas that of free Zn2+ is about 2 X 10(-10) M, although numerous physiological roles for Zn2+ have been postulated that would require free Zn2+ concentration orders of magnitude higher than this. A titration of plasma with Zn2+ shows that the fractional increase in free Zn2+ is essentially the same as the fractional increase in total exchangeable Zn2+, and the results are consi...
Gadsby PM, Peterson J, Foote N, Greenwood C, Thomson AJ.Magnetic-circular-dichroism (m.c.d.) spectra over the wavelength range 300-2000 nm at room temperature and at 4.2K of horse heart cytochrome c are reported at a series of pH values between 7.8 and 11.0, encompassing the alkaline transition. The effect of glassing agents on the e.p.r. spectrum at various pH values is also reported. Comparison of these results with spectra obtained for the n-butylamine adduct of soybean leghaemoglobin support the hypothesis that lysine is the sixth ligand in the alkaline form of horse heart cytochrome c. The m.c.d. and e.p.r. spectra of horse heart cytochrome c ...
Fincham DA, Mason DK, Paterson JY, Young JD.1. Thoroughbred horses were divisible into five distinct amino acid transport subgroups on the basis of their erythrocyte permeability to L-alanine, measured uptake rates ranging from 5 to 625 mumol l cells-1 h-1 (0.2 mM-extracellular L-alanine, 37 degrees C). 2. Erythrocytes from animals belonging to the lowest L-alanine permeability subgroup (5-15 mumol l cells-1 h-1) (transport-deficient type) exhibited slow nonsaturable transport of this amino acid. In contrast, cells from horses of the four transport-positive subgroups possessed additional high-affinity (apparent L-alanine Km (Michaelis c...
Clark KE, Squires EL, McKinnon AO, Seidel GE.Equine embryos were recovered nonsurgically 6.5 d after ovulation (Exp. 1) and those greater than 200 microns were stored in one of three media: 1) Ham's F10 + 10% fetal calf serum (FCS) under 5% CO2, 5% O2 and 90% N2 at 24 C (Ham's F10); 2) Minimal Essential Medium with Hank's balanced salts + 10% FCS in air (MEM) at 24 C or 3) MEM at 5 C n = 10/treatment). Embryos less than or equal to 200 micron (n = 10) were bisected microsurgically; one-half of each embryo was stored in Ham's F10 and the other half in either Dulbecco's phosphate-buffered saline + 10% FCS in air at 24 C (DPBS), or MEM in a...
Martens RJ, Martens JG, Renshaw HW, Hietala SK.The opsonic capacity of serum containing R. equi-specific antibody was compared with antibody-deficient sera using luminol-dependent chemilumenscence (LDCL) and bactericidal assays. These assays incorporated peripheral blood polymorphonuclear neutrophilic leukocytes (PMNL) exposed to R. equi opsonized with neonatal equine pre-colostral serum (control) or serum from foals with R. equi infections (principal). All sera were complement inactivated at 56 degrees C for 30 min. Bacteria were obtained from the lung of a foal with R. equi pneumonia. Neutrophils were obtained from one adult horse for LD...
Hayes TE.The examination of cerebrospinal fluid (CSF) is often part of the diagnostic work-up of a patient exhibiting signs of disease involving the central nervous system. Awareness of the capabilities and limitations of these laboratory tests is important in assessing the benefit-to-risk ratio of performing such procedures. Collection of CSF is a relatively simple procedure, and together with a thorough history, physical examination, and other diagnostic tests, may be a valuable aid in arriving at a diagnosis or prognosis.
McIntyre JC, Hundley P, Behnke WD.Fluorescence techniques have been employed to study the interaction of porcine and equine colipase with pure taurodeoxycholate and mixed micelles. Nitrotyrosine-55 of porcine colipase is obtained by modification with tetranitromethane (low excess, in the presence of taurodeoxycholate) of the protein followed by gel filtration and ion-exchange chromatography. Verification of the residue modified was obtained by h.p.l.c. peptide purification and sequence analysis. Reduction and quantitative reaction with dansyl chloride yields a fluorescent derivative that is twice as active in conjunction with ...
Hall J, Zha XH, Yu L, Yu CA, Millett F.The interaction of the Rhodobacter sphaeroides cytochrome bc1 complex with Rb. sphaeroides cytochrome c2 and horse cytochrome c was studied by using specific lysine modification and ionic strength dependence methods. The rate of the reactions with both cytochrome c and cytochrome c2 decreased rapidly with increasing ionic strength above 0.2 M NaCl. The ionic strength dependence suggested that electrostatic interactions were equally important to the reactions of the two cytochromes, even though they have opposite net charges at pH 7.0. In order to define the interaction domain on horse cytochro...
Sletten K, Husebekk A, Husby G.The amino acid sequence of the amyloid fibril protein AA from horse was established from characterization of cyanogen bromide fragments, tryptic peptides, and a peptide derived from a digest with Staphylococcus aureus V8 proteinase. The protein was found to consist of 80 amino acid residues. Sequence homologies with protein AA from other species were very striking, and revealed an insertion of two amino acid residues between positions 72 and 73. In position 44, two amino acid residues were found which provide further evidence for a polymorphism in the amyloid fibril protein AA.
Waelchli RO, Winder NC.Endometrial biopsies were obtained from four mares during consecutive oestrous cycles on the first day of oestrus, on the day when ovulation was detected, and four and eight days after ovulation. Cycle stages were confirmed by means of rectal palpation, ultrasonography and plasma progesterone determination. Immunohistochemical evaluation of the formalin fixed biopsy specimens was performed using a peroxidase anti-peroxidase technique. Immunoglobulin (Ig)A-, IgM-, IgG(Fc)- and IgG(T)-containing cells were detected in all biopsies; with IgA- and IgG(Fc)-containing cells generally predominating. ...
Satterlee JD, Moench S.Proton hyperfine resonance assignments for cytochromes c from several species are currently being successfully pursued by several laboratories. These efforts focus mostly on the ferrous forms. In contrast to that work, we have pursued assignments of the proton hyperfine shifted resonances for horse and tuna ferricytochromes c. Our results indicate that assignments are nearly identical in those two proteins. Using the pre-steady state nuclear Overhauser effect, several additional assignments have been made for the tuna protein, whereas for the horse protein, the following protons have been assi...
Higgins AJ, Lees P, Sharma SC, Taylor JB.An accurate and reliable method for the separation of flunixin from, and measurement in, equine inflammatory exudate and plasma by high performance liquid chromatography has been developed. Flunixin can be detected in concentrations as low as 0.05 micrograms/ml using an ultraviolet spectrophotometric detector at 285 nm. Samples were acidified with 2M hydrochloric acid and extracted with dichloromethane. The extract was evaporated and reconstituted in acetonitrile. Iminodibenzyl was used as internal standard. The mean recovery of flunixin from plasma was 97.6 +/- 3.9 per cent. Particular advant...
Magnuson NS, Perryman LE, Wyatt CR, Mason PH, Talmadge JE.Peripheral blood mononuclear cells from foals with hereditary severe combined immunodeficiency (SCID) have morphologic characteristics of large granular lymphocytes (LGL). Attempts to demonstrate cytotoxic activity were without success unless the LGL were incubated with 100 U of human recombinant interleukin 2 (rIL 2)/ml for 24 hr. With rIL 2 incubation, low effector to target ratios (10:1) consistently yielded high levels of cytotoxic activity (30 to 50%) in a standard 4-hr 51Cr-release assay using YAC-1 lymphoma or K562 erythroleukemia cell lines as targets. Monoclonal antibody EqT12 reacted...
Sedgwick AD, Dawson J, Lees P.Subpopulations of equine leucocytes, polymorphonuclear and mononuclear cells, were separated from whole blood on a discontinuous Percoll gradient and used in studies of chemokinesis and chemotaxis. Polymorphonuclear cells responded to the chemo-attractant properties of zymosan-activated plasma in Boyden chamber and agarose microdroplet assays but they responded only slightly (Boyden chamber) or not at all (agarose microdroplet) to the peptide N-formyl-methionyl-leucyl-phenylalanine (FMLP). Equine mononuclear cell movement was increased by FMLP in both assay systems and these cells also respond...
Hardee GE, Tshabalala MA, Moore JN, Gokhales RD.Separate high performance liquid chromatographic methods were developed for thiabendazole (TBZ) and 5-hydroxy thiabendazole (5-OH-TBZ) determination in horse plasma using 1-methyl-2-phenyl benzimidazole (MPBZ) as an internal standard. In both methods TBZ and 5-OH-TBZ were extracted from plasma using organic solvents, injected on to a C-18 column, and eluents monitored by a fluorescence detector. However, mobile phase composition, extraction solvent as well as detector wavelength differed in the two methods. The linear range for TBZ was 0.02 to 0.77 microgram ml-1 while that for 5-OH-TBZ was 0....
Chirino-Trejo JM, Prescott JF.The antigens extracted from strains belonging to seven capsular serotypes of Rhodococcus equi, as well as from two wild strains isolated from pneumonic foals, were examined. Whole-cell antigens and soluble products present in broth culture supernatants were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, electroblotted onto nitrocellulose, and stained with serum from hyperimmunized rabbits or foals. Foal sera used included sera from pneumonic animals with known titer to equi factors; from animals bled monthly on a farm with enzootic pneumonia, and from animals bled mont...
Sugino H, Bousfield GR, Moore WT, Ward DN.The complete amino acid sequence of the beta-subunit of equine chorionic gonadotropin (eCG beta) has been established by both automated Edman and manual 5-dimethylaminonaphthalene-1-sulfonyl-Edman degradations. Specific fragments were produced by cleavage with Staphylococcus aureus V8 protease, trypsin, or dilute HCl. For the sequence analyses of the heavily glycosylated COOH-terminal portion, a chemical deglycosylation procedure with trifluoromethanesulfonic acid was employed. The peptide chain of eCG beta consists of 149 amino acid residues. Five or more oligosaccharide chains are attached t...
Bousfield GR, Liu WK, Sugino H, Ward DN.The amino acid sequence was determined for equine lutropin beta (eLH beta). Large fragments were derived from reduced, carboxymethylated eLH beta by digestion with Staphylococcus aureus V8 protease, by cyanogen bromide cleavage, and by cleavage of acid-labile Asp-Pro bonds. The fragments were purified by gel filtration and high performance liquid chromatography (HPLC). The fragments were sequenced by automated Edman degradation to establish the primary structure of eLH beta. Some peptides were further digested with chymotrypsin and the resulting peptides purified by HPLC. In addition to sequen...
Gross MK, Toscano DG, Toscano WA.Calmodulin (CaM), the calcium binding protein that modulates the activity of a number of key regulatory enzymes, is present at high levels in sperm. To determine whether CaM regulates adenylate cyclase in mammalian sperm, the actions of EGTA and selected CaM antagonists on a solubilized adenylate cyclase from mature equine sperm were examined. The activity of equine sperm adenylate cyclase was inhibited by EGTA in a concentration-dependent manner with a half-maximal inhibitory concentration (IC50) of 2 mM. Equine sperm adenylate cyclase was also inhibited in a concentration-dependent manner by...
Migliorisi A, Hassel DM, Moore AR, Blair BW, Wilkins PA.To characterize coagulopathies before and after antivenom administration in rattlesnake-envenomated equids. Unassigned: Client-owned envenomated equids treated with antivenom at Colorado State University (2023 to 2024). Rattlesnake bite severity score (RBSS), platelet count, prothrombin time, activated thromboplastin time, D-dimers, antithrombin, and viscoelastic testing variables were assessed before and after antivenom administration. Paired comparisons of hemostatic variables between admission and approximately 24 hours, and correlation analysis with RBSS, were performed. Length of hospital...
Krücken J, Diekmann I, Andreotti S, Bredtmann CM, Mbedi S, Sparmann S, Schmidt JS, de Almeida Borges F, de Freitas MG, Sallé G, Hofer H....Equines are parasitized by complex communities of Strongylidae (Nematoda) comprising multi-species infections. Currently, Cyathostominae are most prevalent, while Strongylus species are only rarely detected. Since eggs and, in most cases, infective larvae cannot be differentiated to species level, except for Strongylus spp., species-specific knowledge of the pathology, epidemiology and ecology of these parasitic nematodes is limited. Reference sequence data for several cyathostomin species are limited or missing. Deep amplicon sequencing of internal transcribed spacer 2 (ITS-2) regions of nema...
Weingarten A, Häusler M, Serangeli J, Verheijen I, Reiter E, Radzevičiūtė R, Stoessel A, Krause J, Spyrou MA, Conard NJ, Nieselt K, Posth C.Deep-time palaeogenomics offers rare insights into macroevolutionary events for both extant and extinct species. Aside from a Middle Pleistocene genome from North American permafrost (780-560 ka) and a number of Late Pleistocene specimens, most ancient horse DNA studies have focused on tracing the origins of domestication and subsequent periods. Here we present mitochondrial genomes from two Equus mosbachensis specimens from Schöningen, Germany, a Middle Pleistocene archaeological site complex with direct and repeated evidence of hominin-horse interactions on the shore of a palaeolake. Usin...
Oztas T, Akar M, Virkanen J, Beier C, Goericke-Pesch S, Peltoniemi O, Kareskoski M, Björkman S.Heavy metal contamination poses significant environmental and health risks to both humans and animals. This study investigates the concentrations of arsenic (As), cadmium (Cd), and lead (Pb) in blood, hair, and semen samples collected from 16 breeding stallions in Western Finland to assess whether concentrations in hair and serum samples represent concentrations in semen. The analysis was conducted using an inductively coupled plasma mass spectrometry (ICP-MS) system. Results showed that hair samples exhibited the highest concentrations of heavy metals, particularly Pb and As, while semen show...
Núñez-Montero K, Leal K, Rojas-Villalta D, Castro M, Larronde C, Wagenknecht L, Contreras MJ.Studies on the bacterial composition of seminal samples have primarily focused on species isolated from semen and their effects on fertility and reproductive health. Culture-independent techniques, such as 16S rRNA gene sequencing and shotgun metagenomics, have revolutionized our ability to identify unculturable bacteria, which comprise >90% of the microbiome. These techniques allow for comprehensive analysis of microbial communities in seminal samples, shedding light on their interactions and roles. In this study, we characterized the taxonomic diversity of seminal microbial communities in he...
Hifumi T, Tanaka T, Suzu I, Sato M, Akioka K, Fujimata C, Shinkai R, Maeda T, Kusakisako K, Ikadai H, Miyoshi N.Alveolar echinococcosis is a zoonotic disease caused by a larval-stage infection. Geographical haplotyping targeting the parasite's mitochondrial cytochrome () gene has been reported for isolates from definitive and intermediate hosts (wild canids and rodents); however, there are limited reports on strain typing for the dead-end host, the horse, which could act as a sentinel for E. Accordingly, we investigated the diversity of in isolates obtained from slaughtered Japanese and Canadian horses originating from the Iburi and Hidaka regions in Hokkaido and from Alberta, respectively, with PCR ...
McCreight KA, Barbosa LN, Odoi A, Reed P, Rajeev S.We estimated the seroprevalence in dogs, cats, and horses from Tennessee, USA, using the microscopic agglutination test (MAT) against 12 serovars. We observed seropositivity in 110 of 374 (29.4%) dogs, 21 of 170 (12.4%) cats, and 42 of 88 (47.7%) horses. The highest seroprevalence was observed for serovars Autumnalis (74.6%) in dogs, and Bratislava in cats (42.9%) and horses (95.2%). We found a significant level of potential cross-reactivity between multiple serovars tested, with highest cross-reactivity to serovar Autumnalis in dogs. seroprevalence was significantly higher in vaccinated ...
Ichikawa Y, Iinuma Y, Okagawa T, Shimbo R, Enkhtuul B, Khurtsbaatar O, Kinoshita Y, Niwa H, Aoshima K, Kobayashi A, Batbaatar V, Ohashi K, Kimura T.Glanders is a World Organization for Animal Health (WOAH)-notifiable equine disease caused by the infection of Burkholderia mallei, and is endemic in Mongolia, South Asia, Africa, and South America. While the complement fixation test (CFT) has been widely used for serodiagnosis of glanders and is considered a standard serological test, it has several limitations. These limitations include poor specificity, labor intensive techniques, variability in antigen and protocol. Consequently, indirect enzyme-linked immunosorbent assays (iELISAs) based on recombinant proteins have been developed as alte...
Touareb D, Latifi S, Saoiabi S, Habraji L, Hammani O, Azzaoui K, Jodeh S, Yaghi S, Sabbahi R, Hammouti B, Saoiabi S.Lead (Pb²⁺) contamination in aquatic environments represents a serious global threat due to its toxicity, persistence, and non-biodegradable nature. In this study, hydroxyapatite (HA) was synthesized from equine scapula bone and investigated as a low-cost, sustainable adsorbent for Pb²⁺ removal from aqueous media. HA was thermally treated at 100 °C, 500 °C, and 900 °C to assess the effect of calcination on physicochemical and adsorption properties. Characterization techniques including XRD, FTIR, SEM/EDX, TGA and XRF confirmed structural and compositional evolution with temperature...
Ostuni A, Frontoso R, Crudele MA, Barca L, Amati M, Boni R, De Vendel J, Raimondi P, Bavoso A.Equine Infectious Anemia Virus (EIAV), a lentivirus marked by considerable genetic variability, poses significant diagnostic challenges. Existing diagnostic tools encompass the Agar Gel Immunodiffusion Assay (AGID), enzyme-linked immunosorbent assay (ELISA), and Western blotting (WB). ELISA and AGID mainly utilize the p26 capsid protein, often sourced from the Wyoming reference strain. To broaden the range of viral proteins and strains employed in these immunoassays, we previously developed a novel p26/double-strain gp45 indirect ELISA. In this study, we evaluated the performance of this ELISA...
Khaing Y, Htun LL, Linn KS, Kyaw WO, Nwae TT, Chel HM, Win SY, Murata S, Nakao R, Nonaka N, Bawm S.The study aimed to determine the prevalence of blood parasites in horses and identify risk factors and molecular detection of piroplasm species (Theileria equi and Babesia caballi) of horses in Myanmar. Blood samples (n = 302) were collected from five regions of Myanmar. Blood smears were screened for presence of piroplasms. Samples positive for piroplasms were subjected to molecular identification using primers specific to the small subunit ribosomal RNA (SSU rRNA) of piroplasms. The overall prevalence of blood parasites (piroplasms, Trypanosoma spp., and microfilaria of Setaria equina) w...
An T, Dugarjaviin M, Han H.The Mongolian horse, one of the oldest and most genetically diverse breeds, exhibits a wide variety of coat colors and patterns, including both wild-type and unique features. A notable characteristic of dun Mongolian horses is the presence of Bider markings-symmetrical, black-mottled patterns observed on the shoulder blades. These markings are also seen in Przewalski's horses. The dun coat color, a common wild-type phenotype in domestic horses, is characterized by pigment dilution with distinct dark areas and is regulated by mutations in the gene. This study aimed to investigate the role of ...
Köhne M, Diel E, Packeiser EM, Böttcher D, Tönissen A, Unruh C, Goericke-Pesch S, Ulrich R, Sieme H.In the past, most research in equine reproduction has been performed in vivo but the use of in vitro and ex vivo models has recently increased. This study aimed to evaluate the functional stability of an ex vivo hemoperfused model for equine uteri with molecular characterization of marker genes and their proteins. In addition, the study validated the respective protein expression and the aptness of the software QuPath for identifying and scoring immunohistochemically stained equine endometrium. After collection, uteri (n = 12) were flushed with preservation solution, transported to the labor...
Palmisano M, Kulp J, Bender S, Stefanovski D, Robinson M, Johnson A.Eight-hydroxy-2'-deoxyguanosine (8-OHdG), a biomarker of oxidative damage evaluated in human neurodegenerative disease, has potential to correlate with postmortem diagnosis of neuroaxonal dystrophy/degenerative myeloencephalopathy (NAD/DM) in horses. Objective: We hypothesized that 8-OHdG will be higher in CSF and serum from NAD/DM horses compared with horses with other neurologic diseases (CVSM, EPM) and a control group of neurologically normal horses. We also hypothesized that 8-OHdG will be higher in CSF compared with serum from NAD/DM horses. Methods: Fifty client-owned horses with postmor...
Hickman E, Carberry V, Carberry C, Cooper B, Mordant AL, Mills A, Sokolsky M, Herring LE, Alexis NE, Rebuli ME, Jaspers I, Sheats K, Rager JE.Growing evidence supports the importance of extracellular vesicle (EV) as mediators of communication in pathological processes, including those underlying respiratory disease. However, establishing methods for isolating and characterizing EVs remains challenging, particularly for respiratory samples. This study set out to address this challenge by comparing different EV isolation methods and evaluating their impacts on EV yield, markers of purity, and proteomic signatures, utilizing equine/horse bronchoalveolar lavage samples. Horses can serve as effective translational animal models for respi...
Nakase M, Thapa J, Batbaatar V, Khurtsbaatar O, Enkhtuul B, Unenbat J, Lkham B, Fujita S, Koshikawa A, Tuanyok A, Saechan V, Higashi H, Hayashida K....Glanders and melioidosis are contagious zoonotic diseases caused by Burkholderia mallei and B. pseudomallei, respectively. Bacterial isolation and polymerase chain reaction (PCR) have been used to detect these bacteria in animals suspected of infection; however, both methods require skilled experimental techniques and expensive equipment. These obstacles make it difficult to diagnose B. mallei and B. pseudomallei infections in areas where reagents and equipment are difficult to procure. To solve this problem, we developed an easy and ready-to-use dried-format diagnostic tool based on loop-medi...
Cutarelli A, De Falco F, Serpe F, Izzo S, Fusco G, Catoi C, Roperto S.BPV1, BPV2, BPV13, and BPV14 are all genotypes of bovine delta papillomaviruses (δPV), of which the first three cause infections in horses and are associated with equine sarcoids. However, BPV14 infection has never been reported in equine species. In this study, we examined 58 fresh and thawed commercial semen samples from healthy stallions. In 34 (58.6%), bovine δPV DNA was detected and quantified using droplet digital polymerase chain reaction (ddPCR). Real time quantitative PCR (qPCR) was able to identify bovine δPV DNA in 5 samples (8.6%). Of the BPV-infected semen samples, 15 were posi...
Walter J, Colleoni S, Lazzari G, Fortes C, Grossmann J, Roschitzki B, Laczko E, Naegeli H, Bleul U, Galli C.Assisted reproductive technologies are an emerging field in equine reproduction, with species-dependent peculiarities, such as the low success rate of conventional IVF. Here, the 'cumulome' was related to the developmental capacity of its corresponding oocyte. Cumulus-oocyte complexes collected from slaughterhouse ovaries were individually matured, fertilized by ICSI, and cultured. After maturation, the cumulus was collected for proteomics analysis using label-free mass spectrometry (MS)-based protein profiling by nano-HPLC MS/MS and metabolomics analysis by UPLC-nanoESI MS. Overall, a total o...
Spicer LJ, Maylem ERS, Schütz LF.Ovarian granulosa cells produce a variety of biologically active compounds in addition to steroid hormones that include numerous families of growth factors, cytokines and adipokines. Many of these function as endocrine, paracrine and autocrine hormones to regulate ovarian activity. The goal of this review is to provide an update on the evidence in domestic animals on how FSH, insulin and IGF1 regulate the function of granulosa cells with a focus on ovarian steroidogenesis and cell proliferation with comparisons across six domestic animals: pigs, cattle, horses, water buffalo, goats and sheep. ...
Ma X, Zhang Y, Na L, Qi T, Ma W, Guo X, Wang XF, Wang X.African horse sickness (AHS) is an acute, fatal, contagious disease of animals of the family Equidae and is caused by infection with the African horse sickness virus (AHSV). Based on the outer capsid protein VP2, AHSV is classified into nine serotypes (AHSV-1 to -9) with little or no serological cross-reactivity between them. In 2020, AHS outbreaks caused by AHSV-1 were reported in Thailand and Malaysia, marking the first occurrences of AHS in Southeast Asia. However, little is known about the antigenic profile of AHSV-1 VP2. In this study, a recombinant VP2 protein was expressed in and used ...
Harkin EE, Browne JA, Murphy BA.Hair follicles provide an easily accessible tissue for interrogating gene expression for multiple purposes in mammals. RNAlater® is a liquid storage solution that stabilises and preserves cellular RNA, eliminating the need to immediately process or freeze tissue specimens. The manufacturer advises storage of samples at 2-8°C overnight before transfer to -20°C. This study aimed to evaluate RNA integrity in hair follicle samples collected from horses, stabilized in RNAlater®, and stored under three short-term storage conditions. Mane hair samples complete with follicles were collected from f...
Szkolnicka K, Mituniewicz-Małek A, Dmytrów I, Bogusławska-Wąs E.During the last years, growing interest in the use of mare's milk in food production is observed. The subject of the study was to evaluate the feasibility of mare's milk for the production of yogurt ice cream and synbiotic ice cream. Four variants of mare's milk ice cream were developed: ice cream with yogurt bacteria without inulin (YO) and with 2% of inulin (YO+I), synbiotic ice cream with 2% inulin and Lacticaseibacillus rhamnosus (LCR+I) and with Lactiplantibacillus plantarum (LP+I). Ice creams were enriched with inulin in order to evaluate its influence on the viability of LAB and on the ...
Sánchez A, Durán G, Segura Á, Herrera M, Vargas M, Villalta M, Arguedas M, Moscoso E, Umaña D, Gómez A, Gutiérrez JM, León G.Snakebite envenomation inflicts a high burden of mortality and morbidity in sub-Saharan Africa. Antivenoms are the mainstay in the therapy of envenomation, and there is an urgent need to develop antivenoms of broad neutralizing efficacy for this region. The venoms used as immunogens to manufacture snake antivenoms are normally selected considering their medical importance and availability. Additionally, their ability to induce antibody responses with high neutralizing capability should be considered, an issue that involves the immunization scheme and the animal species being immunized. Results...
Kovarikova S, Blahova J, Vanova I, Marsalek P.Determination of the urinary protein-to-creatinine ratio (UPC) is an important tool in the quantification of proteinuria in animals. However, the result may be affected by the different methods of determining the urinary protein concentration. The aim of this study was to compare the turbidimetric method using benzethonium chloride and the colorimetric method using pyrogallol red in the measurement of the urinary protein concentration in dogs, cats, guinea pigs and horses. A total of 464, 192, 216 and 119 urine samples from dogs, cats, guinea pigs and horses were examined in the study, respect...
Brandon J, Reider H, Pabilonia KL, Moore AR.The bromocresol green albumin assay (ALB) has been used in birds and reportedly is noncomparable with electrophoretic albumin (ALB) in many species. It is accepted for use in some species and rejected in others. Objective: We aimed to compare the performance of ALB and ALB methods within backyard chickens and compare the performance of ALB in chickens with other veterinary species where the ALB method is accepted and used clinically. Methods: Chicken plasma collected during reference interval development and samples submitted for diagnostic biochemistry profile were evaluated using the ALB and...
Gupta S, Sethi K, Vohra S, Gupta S, Rani R, Kumar S, Kumar R.We report on the in vitro and in vivo success of Imidocarb dipropionate (IDP), an aromatic diamidine compound, as an effective drug against Surra-causing organism, Trypanosoma evansi. IDP is a clinically acclaimed drug for the treatment of babesiosis and anaplasmosis. We have also investigated cytotoxic effect of IDP on equine Peripheral mononuclear cells (PBMCs) and Vero cell line. The data generated indicate that the IC of IDP (2.73 µM) was significantly lower than its cytotoxic concentration (32.66 and 80.70 µM against PBMCs and Vero cell line, respectively). We have recorded signific...
Kholik K, Sukri A, Riwu KHP, Kurniawan SC, Khairullah AR.Bacterial identification can be done using various testing techniques. Molecular techniques are often used to research dangerous diseases, an approach using genetic information on the pathogenic agent. The enterohemorrhagic invasive species 0157:H7 was identified from the feces of working horses on the island of Sumbawa. Another advance in molecular technology is genome amplification with qPCR which is the gold standard for detecting Unassigned: This study aims to detect and identify the invasive species 0157:H7 using the gene encoding with the qPCR method sourced from horse feces. Unassig...
Ren T, Liu M, Zhou L, Zhang L, Qin Y, Ouyang K, Chen Y, Huang W, Wei Z.Getah virus (GETV) is a re-emerging mosquito-borne RNA virus that induces fever, hind limb edema, swollen submandibular lymph nodes, and urticaria in horses. In pigs, the virus often results in stillbirths among pregnant sows, and neurological symptoms leading to death in piglets. Currently, there are no specific treatments or drugs available for GETV infection. The use of reporter viruses to monitor viral replication and spread in real-time within infected cells and animals provides a powerful tool for targeting antiviral drugs throughout the viral life cycle. Their fluorescence-tracked chara...
