In vitro research involving horses refers to the study of equine cells, tissues, or biological molecules outside their normal biological context, typically in controlled laboratory environments. This research approach allows scientists to investigate cellular processes, molecular interactions, and the effects of various treatments without the ethical and logistical complexities of in vivo studies. In vitro studies contribute to understanding equine physiology, pathology, and pharmacology by providing insights into cellular responses to pathogens, drugs, and other stimuli. This page compiles peer-reviewed research studies and scholarly articles that explore various in vitro methodologies and their applications in equine science, including cell culture techniques, molecular assays, and drug efficacy testing.
RANDALL CC, ORR MF, SCHELL FG.Intracellular organisms resembling Histoplasma capsulatum have been observed in apparently normal amnio-allantoic membrane and adult horse spleen maintained in tissue culture.
The significance of this finding and comparison with another fungus Cryptococcus jarciminosus is discussed.
BAER H, KABAT EA, KNAUB V.Blood group substances have been isolated from the saliva of human beings of blood group B and from the linings of individual horse stomachs. The properties of the human B substances are similar to those of hog and human blood group substances previously isolated. The horse substances showed lower hexosamine and reducing sugar and higher total and non-hexosamine nitrogen than do the materials from the other species. Materials isolated from individual horse stomachs possess either A or B activity or both. Certain stomachs yielded products of identical analytical composition but with neither blo...
Olitsky PK, Cox HR, Syverton JT.We have studied certain properties, additional to those previously described (3), of the virus of vesicular stomatitis of horses, and of the characteristic biological reactions of the virus of equine encephalomyelitis. It has been found that the virus of stomatitis, ordinarily dermotropic, can acquire neurotropism and the neurotropic encephalomyelitis virus can, in turn, be rendered dermotropic in its action. The neurotropism in both instances is associated with definite, although not pronounced, viscerotropism. Both viruses can bring about a similar infection in the white mouse, rat, guinea p...
Woods AC, Chesney AM.A filterable agent has been obtained from the humors and tissues of the eyes of horses suffering from active periodic ophthalmia. The intra-vitreous injection of this filtrate produced in normal horses the same clinical and pathological picture observed in the natural disease. This filtrate injected into rabbits produced a different clinical picture, but the essential pathological lesions closely resembled those found in horses. After passage of the filterable agent through six generations of rabbits, it again produced the clinical and pathological picture of the natural disease when injected ...
Parker JT, McCoy MV.1. The serum of horses immunized with increasing doses of certain anaerobically produced autolysates of pneumococci contain potent neutralizing antibodies for the pneumotoxin. 2. The method for the in vitro titration of these horse antipneumotoxic serums is given.
Olitsky PK, Long PH.The virus of vesicular stomatitis is not readily killed by formalin. This chemical is one of a group which coagulates the proteins of the medium in which the virus is usually contained. It has already been found(7) that other reagents of the protein-coagulating group are not actively virucidal) and the effect of formalin in this instance is therefore characteristic of the group. The so called formalinized vaccines which give rise to immunity can be shown to have done so because of the presence of living virus. A single injection of such so called "vaccine," or of other material containing livi...
Friedlander M, Sobotka H, Banzhaf EJ.The precipitin indices for a number of monovalent and polyvalent antipneumococcus sera were determined under known conditions, and found to vary as did the number of protective units. The ratio precipitin index/protective units in monovalent sera was found to lie between 2.8 and 4.8 for Type I and to be about ten times greater for Type III. Lower values were found in polyvalent horses and when mixing heterologous monovalent sera with each other. The influence of the duration of treatment upon the quotient was studied. Several refined and concentrated preparations showed a relative increase in ...
Reimann HA.It is conceivable that a change from the virulent, non-phagocytable S form of Pneumococcus to the avirulent phagocytable R form may take place in pneumococcus disease, but the experiments here reported do not settle the question whether or not this is an important factor in determining the outcome in natural infection. It has been shown experimentally that the degradation from the S form to the R form actually does take place in cultures of Pneumococcus growing in agar subcutaneously embedded in guinea pigs, in agar enclosed in vials subcutaneously embedded in rabbits, and spontaneously in the...
The objective of this study was to enhance the in vitro sperm quality and in vivo fertility of frozen-thawed equine semen by the addition of l-carnitine (LC) to post-thawed semen. Different concentrations of LC were added to thawed samples to obtain four treatments control and 0.5, 1 and 2 mM LC. In the in vitro experiments, sperm motility and kinematics, membrane integrity and intracellular calcium ion concentration ([Ca ] ) were investigated, and the antioxidant bioactivity of LC was assessed by measuring hydrogen peroxide and nitrite concentrations (NO ). The fertility rate was assessed v...
In horses, osteoarthritis (OA) mostly affects metacarpophalangeal and metatarsophalangeal (fetlock) joints. The current modalities used for diagnosis of equine limb disorders lack ability to detect early OA. Here, we propose a new alternative approach to assess experimental cartilage damage in fetlock joint using Acoustic Emissions (AE). Objective: To evaluate the potential of AE technique in diagnosing OA and see how AE signals changes with increasing severity of OA. Methods: An in vitro experimental study. Methods: A total of 16 distal limbs (8 forelimbs and 8 hindlimbs) from six Finn horses...
Sheridan L, Hutton P, Noble G, Nobari B.Dysbiosis of the intestinal microbiome is a precursor to hindgut acidosis. As microbial diversity and fermentation are influenced by diet, including a postbiotic could help maintain healthy gastrointestinal microbes and reduce acidosis risk. Objective: It was hypothesised that including a postbiotic in batch culture would reduce the degree of pH decline and lactate accumulation during fermentation. Methods: In vitro batch cultures with faecal inoculant were prepared with either low starch (LS) or high glucose (G) substrate to represent normal and acidic fermentation. Saccharomyces cerevisiae d...
Otero JC, Neild DM, Ferrante AA, Gambarotta MC, Caldevilla ML.Glycerol, a penetrating cryoprotectant, is most employed for deep freezing spermatozoa. However, it can induce toxic, chemical, and osmotic effects, altering the lipid structure of the sperm membrane. Rapid glycerol addition has been demonstrated to have fewer toxic effects than its removal. Objective: To minimize both the toxic and osmotic effects of glycerol on equine sperm through post-thaw dilution using two different extenders. Methods: Samples of equine semen frozen with 5% glycerol from nine stallions were thawed, re-diluted 1:2, and incubated for 30 minutes at 22°C in Tyrodes (Tyr) an...
Graide H, Duysens J, Frank T, Mouithys-Mickalad A, Niesten A, Sandersen C, Ceusters J, Serteyn D.Musculoskeletal pathologies present challenges in athletic horses, often leading to functional impairment. The slow or limited regenerative capacity of bone, joint, and tendon/ligament injuries, coupled with the limitations of conventional treatments, highlights the need for innovative therapies such as ortho-biologics and mesenchymal stem/stroma cells. Traditional 2D cell culture systems with fetal bovine serum (FBS) fail to replicate the complexity of the in vivo environment, whereas 3D cultures more accurately mimic native tissue architecture and cell-cell interactions. This study describes...
Meeremans M, Devriendt B, Bairiot S, Van Poucke M, Peelman L, Demeyere K, Meyer E, Van Vlierberghe S, De Schauwer C.Macrophages play key roles in tissue homeostasis and regeneration-associated inflammation. Unlike humans, a reliable protocol to obtain and polarise equine monocyte-derived macrophages is lacking. In this study the polarisation of equine macrophages, derived from CD172a peripheral blood monocytes is described. After differentiation, IFN-γ/LPS or IL-4 were used to induce pro- and anti-inflammatory phenotypes, respectively. Evaluation criteria included morphology, mRNA (RT-qPCR) and protein expression (flow cytometry, immunofluorescence), nitric oxide and arginase production, cytokine secretion...
Parker M, Arnade H, Parker JL, Gordon J, Peroni JF.Investigation of alternative antibiotic therapies is critical for the future of medicine. Platelet lysate (PL) is a blood product that has shown antibiotic potential, which could be used to augment or replace current antibiotics. Objective: To compare the effects of equine PL versus antibiotics on growth of clinically isolated antibiotic resistant bacteria. We hypothesized that PL would exert an antimicrobial effect on equine bacterial isolates resistant to antibiotics. Methods: In vitro experimental study. Platelets were collected via apheresis from nine donor horses and underwent two freeze-...
Rychtarova J, Fulka H, Loi P, Fulka J.In the mouse, spermatozoa are highly resistant to DNA damage, even when frozen without cryoprotectants, and can produce offspring when subsequently used for ICSI (intracytoplasmic sperm injection). It is not known whether the same applies to other mammals as well. For example, in the horse, even conventional sperm freezing is still very problematic and frequently leads to sperm immobility. It has, however, never been tested whether sperm immobility also mirrors sperm head DNA damage, and if so, to what extent. In our study, we evaluated the damage to DNA in horse frozen and thawed motile and i...
Rodrigues MF, Neves AP, Ferreira R, Pessoa GA, Araujo DN, de Souza GV.Melatonin has demonstrated beneficial effects in regulating inflammatory processes in various tissues, but its role in mare uterine physiology remains unclear. Objective: To investigate the effects of melatonin on the gene expression of cytokines (IL-1β, IL-6, IL-8, IL-10, and TNF-α) in equine uterine explants cultured for 6, 12, or 24 hours. Methods: The study was divided into two experiments. Part 1 aimed to evaluate the cytotoxicity of melatonin at concentrations of 0, 0.5, 1, 2, 4, and 8 mM on endometrial cells. Part 2 aimed to determine the expression of inflammatory cytokines in uterin...
Wickizer LF, Burk SV, Lehman JS.Red maple toxicosis arises when horses consume wilted maple leaves, resulting in methemoglobinemia and hemolytic anemia due to oxidative damage to erythrocytes. Objective: The objective of this study was to determine how maple species (red versus Freeman maple) and horse characteristics, management, and barn site contributed to in vitro methemoglobin production and hemolysis. Methods: Horse information was obtained through an owner questionnaire. Extracts from red and Freeman maple leaves were separately incubated with washed erythrocytes from each horse (n=120). The in vitro percentage methem...
Rudeekiatthamrong A, Nguyen GT, Kamyingkird K. infection (Surra) remains a major constraint to equine health and productivity in Thailand. The only available trypanocidal drug, diminazene aceturate (DA), has limited efficacy, poor blood-brain barrier penetration, and toxicity in horses. This study aimed to investigate the inhibitory effects of commonly used equine antibiotics, gentamicin (GMC), ceftiofur (CTF), and trimethoprim-sulfamethoxazole (TS), against (Thai strain isolated from dairy cattle number 953; TEDC 953) to identify potential therapeutic alternatives or adjuncts for equine trypanosomosis. Unassigned: An growth inhibition...
Loncar KD, Ortis HA, Foss RR.Production of embryos via intracytoplasmic sperm injection (ICSI) is a common clinical procedure in equine practice. A recent study indicates that treatment of mares with phenylbutazone paste significantly lowers blastocyst production after ICSI of recovered oocytes. The current study was conducted to determine the effect of different methods of phenylbutazone administration on in vitro embryo production of recovered oocytes in mares. Unassigned: In study 1 (July 29 through September 18, 2024), 14 mares received either oral liquid phenylbutazone once daily for 10 days or no treatment. On the l...
Luis-Calero M, Marinaro F, Muñoz-García CC, Fernández-Hernández P, Ortiz-Rodríguez JM, Fernández-González R, Pericuesta E, Gutiérrez-Adán A....In vitro embryo production is widely used in equine clinical reproduction; however, its efficiency remains suboptimal due to the undermined developmental competence of in vitro-matured oocytes. In this work, we evaluated the effect of mare preovulatory follicular fluid (PFF) secretome supplementation during in vitro maturation (IVM) of equine oocytes recovered post-mortem. Secretome was isolated from PFF and was added to the IVM medium at concentrations of 20 µg/ml (S20) or 40 µg/ml (S40). The percentage of mature oocytes, as well as embryo cleavage, blastocyst development, and gene expr...
Cardillo NN, Villarino NF, Kappmeyer LS, Chung CJ, Suarez CE, Bastos RG.Equine piroplasmosis (EP) is a tick-borne disease of equids caused by the intraerythrocytic apicomplexan parasites Theileria equi, Babesia caballi and the recently identified Theileria haneyi. Acute cases can be severe, with anemia, jaundice, abortion or sudden death. Survivors remain lifelong carriers, serving as reservoirs for tick-borne and iatrogenic transmission. No vaccines are currently available, and control strategies rely heavily on accurate diagnostics and chemotherapeutic intervention. Imidocarb dipropionate (ID) is the current standard of care for both acute treatment and radical ...
Stolle LM, Oltmanns H, Meißner J, Heun F, Schieder AK, Wolff HT, Ohnesorge B, Busse C.To determine the in vitro antimicrobial activity of specific antiseptics against common equine ocular surface pathogens. Methods: Staphylococcus aureus (S. aureus) (n = 12), Streptococcus equi subspecies zooepidemicus (S. zooepidemicus) (n = 9), Enterobacter hormaechei (E. hormaechei) (n = 6), and Bacillus cereus (B. cereus) (n = 5) were collected from corneal samples of horses with ulcerative keratitis. Reference strains were included. Minimum bactericidal concentrations (MBC) of polyhexanide, povidone-iodine, and hypochlorous acid were tested using the microdilution method. ...
Pinto-Bravo P, Rebordão MR, Amaral AS, Szóstek-Mioduchowska A, Fernandes C, Galvão AM, Silva E, Alpoim-Moreira J, da Costa RPR, Skarzynski DJ....Oviduct glycoprotein 1 (OVGP1) is a key protein involved in oviductal functions. β-estradiol (E) and progesterone (P), oxytocin (OXT) and tumor necrosis factor-α (TNFα) modulate the equine oviduct function, through prostaglandin regulation. The objective was to evaluate OVGP1 expression within each equine oviduct segment (infundibulum, ampulla isthmus), throughout the estrous cycle. The in vitro effect of (i) E, P, OXT, TNFα; and (ii) spermatozoa, on oviduct OVGP1 transcription and secretion was studied. Gene transcription was assessed by real-time PCR; protein expression by western blot; ...
Sadowska A, Molcan T, Słyszewska M, Skarzynski DJ, Ferreira-Dias G, Szóstek-Mioduchowska A.Equine endometrosis is a chronic degenerative condition of the endometrium. A hallmark feature of endometrosis is endometrial fibrosis accompanied by degenerative changes in the adjacent tissue structure. Tumor necrosis factor α (TNFα), a pleiotropic cytokine produced by various immune cells, plays a dual role in coordinating immune responses and regulating cell/tissue homeostasis, making it an important regulator of fibrotic-related disorders. However, the exact role of TNFα in the development of equine endometrial fibrosis remains to be discovered and explained. Therefore, the main aim of...
Gugole PM, Zannoni A, Iacono E, Merlo B.Cryopreservation of equine mature oocytes remains a major challenge in assisted reproduction, mainly due to the limited availability of material and logistical constraints requiring a holding phase. To address these issues, this study evaluated whether naloxone (NX), an opioid receptor antagonist with reported antioxidant properties, could improve vitrification outcomes and whether bovine oocytes could serve as a suitable preliminary and supportive model. Two experiments were conducted. In experiment 1, immature (bGV) and mature (bMII) bovine oocytes were vitrified without (VIT) or with NX (VI...
Barvelink M, Brok B, Djajadiningrat-Laanen SC, Vernooij JCM, Broens EM, Slenter IJM.To assess the antibacterial activity of manuka honey against bacterial isolates commonly associated with infected corneal ulcerations in horses, and to investigate possible combined effects of manuka honey and commonly prescribed topical antibiotics. Methods: Four Staphylococcus aureus, including three methicillin-resistant (MRSA), two methicillin-resistant coagulase-negative staphylococci (S. sciuri and S. haemolyticus), and two Streptococcus equi subspecies zooepidemicus isolates from horses with stromal ulcerative keratitis were selected. Minimum inhibitory concentrations (MICs) and minimu...
Beaumont RE, Flood C, Guest DJ.Interleukin 1 beta (IL-1β) is upregulated following a tendon injury and in vitro studies have shown that it leads to numerous negative effects on tendon cell function and gene expression. IL-1β activates nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and we hypothesised that inhibiting NF-κB activation would mediate the negative effects of IL-1β on equine tendon cells in 3-dimensional (3D) cultures. Results: Here, we tested three inhibitors of NF-κB signalling (Bortezomib, BAY11-7082 and Wedelolactone) along withTJ-M2010-5, an inhibitor of MyD88, which is a critic...
Wójtowicz A, Sadowska A, Molcan T, Słyszewska M, Drzewiecka EM, Tobolski D, Ferreira-Dias G, Szóstek-Mioduchowska A.Mare endometrosis is a chronic degenerative condition of the endometrium, primarily characterized by fibrosis, involving interactions among fibroblasts, immune cells, and epithelial cells regulated by cytokines and growth factors. T helper (Th)1 and Th2 cells seem to play a pivotal role in fibrosis. However, their roles in equine endometrial fibrosis remain unknown. This study explores Th1 and Th2 cell distribution across different stages of endometrium histopathological Kenney and Doig categories; and evaluated their secretome effects on non-fibrotic endometrium derived fibroblast functional ...
Beshir A, Kemal J, Abraha B, Tola EH.Wounds on the skin are a common health issue affecting working equines. This study aimed to evaluate the in-vitro antibacterial properties of crude methanolic extracts from selected medicinal plants against pathogens isolated from equine skin wounds in Merti district. Agar well and disc diffusion tests were used to determine the mean zone of inhibition, while broth dilution methods were used to determine the minimum inhibitory concentrations and minimum bactericidal concentrations (MBCs), further confirming the potent antibacterial effects of the selected medicinal plant extracts. One way anal...
Muñoz-García CC, Soriano-Campos MP, Luis-Calero M, Gallardo-Soler A, González-Fernández L, Macías-García B.The use of mesenchymal stromal cells (MSCs) in equine reproduction is increasing its interest in the treatment of specific pathologies. MSCs have been isolated from follicular aspirates obtained during transvaginal oocyte aspiration in women, offering a novel source for autologous therapies in reproductive treatments. However, this approach has not been tested in mares despite the common use of transvaginal oocyte aspiration for oocyte collection to produce equine embryos in vitro. Our study aimed to investigate the feasibility of isolating MSCs from equine ovarian follicular aspirates obtaine...
Wottlin LR, Anderson RC, Bhatta R, Leatherwood J, Olson EG, Callaway TR, Ricke SC.Environmental impacts of animal agriculture are under intense scrutiny, thus strategies to decrease greenhouse gas emissions and volatile nitrogen waste are valuable. As dietary inclusion of tannins has been shown to decrease methanogenesis and improve nitrogen retention in ruminants, this study was designed to determine the effects of graded levels of hydrolysable tannin (chestnut) and condensed tannin (mimosa) on equine in vitro cecal fermentation. Methods: Two in vitro experiments were conducted, each with three replications per treatment. Treatments were 0.2 g alfalfa without or with 5, 1...
Del Prete C, Gaspari G, Kosior MA, Merlo B, Iacono E, Longobardi C, Martino NA, Dell'Aquila ME, Damiano S, Cocchia N, Gasparrini B, Lange-Consiglio A.Over the years, regenerative therapies have emerged as promising alternatives for persistent breeding-induced endometritis. In vitro studies testing the effects of these therapies on equine endometrial cells are still scarce. This study aimed to evaluate in vitro the effect of Wharton's jelly (WJ) mesenchymal stromal/stem cell (MSCs)-derived conditioned medium (WJ-CM) and platelet-rich plasma (PRP) on equine endometrial cells, with or without lipopolysaccharide (LPS)-induced inflammation. The WJ-CM was obtained after 24 h of starvation in Ringer's lactate of WJ-MSCs and PRP was prepared using...
de la Fuente A, Scoggin C, Bradecamp E, Martin-Pelaez S, van Heule M, Troedsson M, Daels P, Meyers S, Dini P.Maturation is a critical step in the development of an oocyte, and it is during this time that the oocyte advances to metaphase II (MII) of the meiotic cycle and acquires developmental competence to be fertilized and become an embryo. However, in vitro maturation (IVM) remains one of the limiting steps in the in vitro production of embryos (IVP), with a variable percentage of oocytes reaching the MII stage and unpredictable levels of developmental competence. Understanding the dynamics of oocyte maturation is essential for the optimization of IVM culture conditions and subsequent IVP outcomes....
Pereira Lopes A, Emanuel da Silva R, Sousa Santos L, de Morais Nobre ML, de Araujo Sousa PS, Almeida Rocha J, Veras Quelemes P, de Araujo-Nobre AR....The aim of this study was to investigate the antibacterial, antibiofilm and modulating activities of trans-trans-farnesol (tt-farnesol) on two strains of Streptococcus equi subsp. equi and two strains of S. equi subsp. zooepidemicus, pathogenic or commensal bacteria of horses. Tt-farnesol showed bactericidal activity against all tested strains, with a minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) between 8 and 4 μg/mL. The sesquiterpene caused significant damage to the bacterial membrane, and its antibacterial activity did not appear to depend on oxidat...
DI Stefano DA, Coccoluto L, Panina-Bordignon P, Brambilla E, Ruffini F, Murtaj V, Orlando F, Colombo M, Frigerio C, DI Bona A, Recupero D, Morroni M....Bone grafting is a surgical procedure that involves the use of bone tissue or bone substitutes to repair damaged bone. In dentistry and maxillofacial surgery, bone graft substitutes from various sources are commonly used. Given their critical role in clinical outcomes, it is essential to thoroughly investigate the biological and mechanical properties of these materials. Methods: In this in vitro study, we evaluated the biological properties of two equine-derived bone graft substitutes in comparison to β-tricalcium phosphate. The materials included one equine-derived graft containing hydrolyze...
Koshy SA, Ajeebsanu MM, Karakka Kal AK, Subhahar MB, Karatt TK, Philip M.Effective pain management remains a persistent challenge, with opioids limited by tolerance, dependence, respiratory depression, and misuse. This has created demand for safer, nonaddictive alternatives. Suzetrigine (VX-548, Journavx) is a novel analgesic with promising efficacy, but its potential misuse in competitive racing requires investigation. Limited information is available on its metabolism and detection, emphasizing the need for metabolic characterization to aid antidoping strategies. Methods: In vitro metabolism of suzetrigine was evaluated using equine liver microsomes and homogeniz...
