Topic:Storage

Encapsulation of copper phenanthroline within horse spleen apoferritin: characterisation, cytotoxic activity and ability to retain temozolomide.
RSC advances    April 29, 2024   Volume 14, Issue 20 14008-14016 doi: 10.1039/d3ra07430g
Cassioli ML, Fay M, Turyanska L, Bradshaw TD, Thomas NR, Pordea A.Protein capsules are promising drug delivery vehicles for cancer research therapies. Apoferritin (AFt) is a self-assembling 12 nm diameter hollow nanocage with many desirable features for drug delivery, however, control of drug retention inside the protein cage remains challenging. Here we report the encapsulation of copper(ii)-1,10-phenanthroline (Cu(phen)) within the horse spleen AFt (HSAFt) nanocage, by diffusion of the metal through the pores between the protein subunits. Transmission electron microscopy revealed the formation of organised copper adducts inside HSAFt, without affecting pro...
Stability and Detection Limit of Avian Influenza, Newcastle Disease Virus, and African Horse Sickness Virus on Flinders Technology Associates Card by Conventional Polymerase Chain Reaction.
Animals : an open access journal from MDPI    April 21, 2024   Volume 14, Issue 8 1242 doi: 10.3390/ani14081242
Taesuji M, Rattanamas K, Yim PB, Ruenphet S.The Flinders Technology Associates (FTA) card, a cotton-based cellulose membrane impregnated with a chaotropic agent, effectively inactivates infectious microorganisms, lyses cellular material, and fixes nucleic acid. The aim of this study is to assess the stability and detection limit of various RNA viruses, especially the avian influenza virus (AIV), Newcastle disease virus (NDV), and African horse sickness virus (AHSV), on the FTA card, which could significantly impact virus storage and transport practices. To achieve this, each virus dilution was inoculated onto an FTA card and stored at r...
A simplified fixed-time insemination protocol using frozen-thawed stallion spermatozoa stored at 17°C for up to 24 h before insemination.
Equine veterinary journal    April 14, 2024   doi: 10.1111/evj.14096
Morris L, Harteveld R, Gibb Z.Insemination of mares with frozen-thawed spermatozoa requires intensive management and results in 40%-60% per cycle pregnancy rates. Objective: To determine if satisfactory fertility is possible for frozen-thawed semen after processing it through a microfluidic device, followed by storage at 17°C for up to 24 h before fixed-time insemination. Methods: Uncontrolled field trials. Methods: A pilot study evaluated the motility of frozen-thawed spermatozoa after centrifugation and storage (17°C) in two different media for up to 48 h. Subsequently, the motility of frozen-thawed semen processed...
Allele frequency of muscular genetic disorders in Bull-Catching (vaquejada) Quarter Horses.
Journal of equine veterinary science    March 24, 2024   105052 doi: 10.1016/j.jevs.2024.105052
Quarter horses (QH), a prominent athletic breed in Brazil, are affected by muscular genetic disorders such as myosin-heavy chain myopathy (MYHM), polysaccharide storage myopathy (PSSM1), hyperkalemic periodic paralysis (HyPP), and malignant hyperthermia (MH). Bull-catching (vaquejada), primarily involving QH, is a significant equestrian sport in Brazil. Since the allele frequencies (AF) of MYHM, PSSM1, HyPP, and MH in vaquejada QH remain unknown, this study evaluated the AF in 129 QH vaquejada athletes, specifically from the Brazilian Northeast. These variants were exclusively observed in hete...
Effect of leukoreduction on the metabolism of equine packed red blood cells during refrigerated storage.
Journal of veterinary internal medicine    February 26, 2024   doi: 10.1111/jvim.17015
Miglio A, Rocconi F, Cremonini V, D'Alessandro A, Reisz JA, Maslanka M, Lacroix IS, Tiscar G, Di Tommaso M, Antognoni MT.Understanding of the biochemical and morphological lesions associated with storage of equine blood is limited. Objective: To demonstrate the temporal sequences of lipid and metabolic profiles of equine fresh and stored (up to 42 days) and leukoreduced packed red blood cells (LR-pRBC) and non-leukoreduced packed RBC (nLR-pRBC). Methods: Packed RBC units were obtained from 6 healthy blood donor horses enrolled in 2 blood banks. Methods: Observational study. Whole blood was collected from each donor using transfusion bags with a LR filter. Leukoreduction pRBC and nLR-pRBC units were obtained an...
Evaluation of blastocyst re-expansion, quality in relation to storage temperature, and sexing using blastocoel fluid after manual perforation with a hand-held needle involving in vivo produced equine embryos.
Theriogenology    February 16, 2024   Volume 219 39-48 doi: 10.1016/j.theriogenology.2024.02.012
Camargo GS, de Barros LD, Oliveira-Filho JP, Bromberger CR, Dias-Melicio LA, Alves Dos Santos L, Bergfelt DR, Ferraz de Andrade ER, Canesin HS....The present study was designed to evaluate equine blastocyst re-expansion rate, quality, and sex following perforation of the blastocoel, collection of blastocoel fluid (BF), and PCR amplification of free DNA. Experiment 1 tested the feasibility of the BF sample collection with a hand-held, small-gauged needle (26g) and subsequent PCR amplification of the TSP-Y gene for males and AMEL-Y gene for males and AMEL-X gene for females. Experiment 2 tested the application of the technique. Equine embryos were collected via uterine flushes 8d after ovulation. Thereafter, embryos (n = 19) were initia...
Pre-analytical stability of selected biochemical analytes in serum of horses and oxen stored at -20°C.
Veterinary medicine and science    February 15, 2024   Volume 10, Issue 2 e1368 doi: 10.1002/vms3.1368
Megerssa YC, Gari FR.Delays between blood collection and analysis are inevitable, and samples are always stored in the refrigerator. The current study aimed to evaluate the stability of serum total cholesterol (TC), triglycerides (TG), total protein (TP), albumin and urea (URA) in horses and oxen after storage at -20°C. Methods: Sera from apparently healthy 20 male horses and 20 oxen were obtained and aliquots of serum were divided into 3 portions. The first tube was used for baseline (T0) measurement of analyte values, whereas the other two tubes, T1 and T2, were stored at -20°C for 1 and 2 months, respectively...
Effect of a 14-Day Period of Heat Acclimation on Horses Using Heated Indoor Arenas in Preparation for Tokyo Olympic Games.
Animals : an open access journal from MDPI    February 6, 2024   Volume 14, Issue 4 546 doi: 10.3390/ani14040546
Munsters C, Siegers E, Sloet van Oldruitenborgh-Oosterbaan M.To optimise the performance and welfare of horses during equestrian competitions in hot climates, it is advised to acclimate them to the heat. The effects of training in a heated indoor arena were studied. Four Olympic horses (13.3 ± 2.2 years; three eventers, one para-dressage horse) were trained for 14 consecutive days in a heated indoor arena (32 ± 1 °C; 50-60% humidity) following their normal training schedule in preparation for the Tokyo Olympic games. Standardised exercise tests (SETs) were performed on Day 1 and Day 14, measuring heart rate (HR; bpm), plasma lactate concentration (LA...
Environmental Management of Equine Asthma.
Animals : an open access journal from MDPI    January 30, 2024   Volume 14, Issue 3 446 doi: 10.3390/ani14030446
Diez de Castro E, Fernandez-Molina JM.Environmental practices related to the inhalation of airborne dust have been identified as the main cause of equine asthma (EA) and reasonably, they are truly relevant in its treatment and control, especially for horses with its severe form. Vast research regarding environmental recommendations has been conducted in recent years. However, no recent exhaustive reviews exist that gather all this new evidence. The aim of this review is to report and compare the most pertinent information concerning the environmental management of EA. The main findings highlight the importance of the type of forag...
Preserving frozen stallion sperm on dry ice using polymers that modulate ice crystalization kinetics.
Cryobiology    January 29, 2024   104852 doi: 10.1016/j.cryobiol.2024.104852
Uhlmannsiek L, Shen H, Eylers H, Martinsson G, Sieme H, Wolkers WF, Oldenhof H.Cryopreserved semen is routinely shipped in liquid nitrogen. Dry ice could serve as an alternative coolant, however, frozen storage above liquid nitrogen temperatures (LN2, -196 °C) may negatively affect shelf-life and cryosurvival. In this study, we determined critical temperatures for storage of cryopreserved stallion sperm. We evaluated: (i) effects of cooling samples to different subzero temperatures (-10 °C to -80 °C) prior to storing in LN2, (ii) stability at different storage temperatures (i.e., in LN2, dry ice, -80 °C and -20 °C freezers, 4 °C refrigerator), and (iii)...
Exhaled breath condensate analysis in horses: A scoping review.
Research in veterinary science    January 22, 2024   Volume 168 105160 doi: 10.1016/j.rvsc.2024.105160
de Freitas Santi T, Barbosa B, Weber SH, Michelotto PV.Exhaled breath condensate (EBC) collection is a non-invasive sampling method that provides valuable information regarding the health status of the respiratory system by measuring inflammatory mediators, such as pH, hydrogen peroxide, and leukotriene B4. This scoping review aimed to provide an update on the collection and analysis of EBC in horses. A systematic search of three electronic databases, PubMed, Google Scholar, Science Direct, identified 40,978 articles, of which 1590 duplicates were excluded. Moreover, 39,388 articles were excluded because of irrelevance to this review, such as stud...
Analysis of the Influence of Lactiplantibacillus plantarum and Lacticaseibacillus rhamnosus Strains on Changes in the Hexachlorobenzene Content in Fermented Mare Milk during Refrigerated Storage.
Molecules (Basel, Switzerland)    January 21, 2024   Volume 29, Issue 2 528 doi: 10.3390/molecules29020528
Witczak A, Mituniewicz-Mau0142ek A, Dmytru00f3w I.(1) Background: Hexachlorobenzene (HCB) is a persistent organic pollutant that is possibly carcinogenic to humans. It is still found in the environment, humans and animals, and in foods, including milk and dairy products; (2) Methods: The influence of the probiotic cultures and subsp. on the possibility of effecting the biodegradation of HCB in dairy products fermented from mare milk was investigated, taking into account the product storage time (maximum 21 days). HCB content was determined using the GC/MS method; (3) Results: A strong negative Pearson correlation ( < 0.05) was found bet...
Acidification is required for calcium and magnesium concentration measurements in equine urine.
BMC veterinary research    January 10, 2024   Volume 20, Issue 1 21 doi: 10.1186/s12917-023-03848-1
Lapsina S, Stirn M, Hofmann-Lehmann R, Schoster A, Riond B.Acidification of equine urine to promote dissociation of ion complexes is a common practice for urine ion concentration measurements. The objective of this study was to evaluate the effect of acidification and storage after acidification on calcium (Ca), magnesium (Mg) and phosphate (P) concentrations and on fractional excretion (FE) of these electrolytes. Thirty-two fresh equine urine samples were analysed between December 2016 and July 2020. Complete urinalysis (stick and sediment) was performed on all samples. Ca, Mg, P and creatinine concentrations were measured in supernatant of centrifug...
Reduced bacterial load in stallion semen by modified single layer centrifugation or sperm washing.
Theriogenology    December 29, 2023   Volume 216 111-117 doi: 10.1016/j.theriogenology.2023.12.034
Malaluang P, Wagner LH, Cojkic A, Spergser J, Aurich C, Morrell JM.The presence of bacteria poses a significant challenge to the quality of stallion semen used in artificial insemination. The bacterial content of insemination doses arises from various sources, such as the healthy stallion, environment, and collection equipment, and is implicated in fertility problems as well as reduced sperm quality during storage. The conventional approach of adding antibiotics to semen extenders raises concerns about antimicrobial resistance and potential negative effects on sperm characteristics, and may not be effective in inhibiting all bacteria. The objective of this st...
Characterization of lyophilized equine colostrum.
Journal of equine veterinary science    November 29, 2023   104975 doi: 10.1016/j.jevs.2023.104975
de-Lima TC, de-Sobral GG, de-Franu00e7a Queiroz AES, Chinelate GCB, Porto TS, Oliveira JTC, Carneiro GF.Foals require maternal colostrum in the first hours of life to prevent failure of transfer of passive immunity (FTIP). Innovative storage methods such as lyophilization may enable conservation of colostrum immunoglobulins by a differentiated process of dehydration. The current study aimed to compare the quality of equine colostrum after freezing and after the lyophilization process. Thirty-one pregnant Quarter Horse mares were used. The IgG concentration of frozen and lyophilized colostrum was determined by simple radial immunodiffusion (SRID) and Brix refractometry. The physical-chemical comp...
Studying longitudinal neutralising antibody levels against Equid herpesvirus 1 in experimentally infected horses using a novel pseudotype based assay.
Virus research    November 17, 2023   Volume 339 199262 doi: 10.1016/j.virusres.2023.199262
Di Genova C, Sutton G, Paillot R, Temperton N, Pronost S, Scott SD.Infection with equid herpesvirus 1 (EHV-1), a DNA virus of the Herpesviridae family represents a significant welfare issue in horses and a great impact on the equine industry. During EHV-1 infection, entry of the virus into different cell types is complex due to the presence of twelve glycoproteins (GPs) on the viral envelope. To investigate virus entry mechanisms, specific combinations of GPs were pseudotyped onto lentiviral vectors. Pseudotyped virus (PV) particles bearing gB, gD, gH and gL were able to transduce several target cell lines (HEK293T/17, RK13, CHO-K1, FHK-Tcl3, MDCK I & II)...
Homemade Nucleic Acid Preservation Buffer Proves Effective in Preserving the Equine Faecal Microbiota over Time at Ambient Temperatures.
Animals : an open access journal from MDPI    October 5, 2023   Volume 13, Issue 19 3107 doi: 10.3390/ani13193107
Ward AB, Harris PA, Argo CM, Watson C, Neacsu M, Russell WR, Ribeiro A, Collie-Duguid E, Heidari Z, Morrison PK.The equine faecal microbiota is often assessed as a proxy of the microbial community in the distal colon, where the microbiome has been linked to states of health and disease in the horse. However, the microbial community structure may change over time if samples are not adequately preserved. This study stored equine faecal samples from = 10 horses in four preservation treatments at room temperature for up to 150 h and assessed the resulting impact on microbial diversity and the differential abundance of taxa. Treatments included "COLD" (samples packaged with a cool pack), "CLX" (2% chlorhexi...
Digitalisation in pre-purchase examination and prospects for an equine health database in Germany-Results of an online survey among equine veterinarians.
Equine veterinary journal    September 12, 2023   doi: 10.1111/evj.14001
Folgmann MS, Kleinsorgen C, Stock KF, Meister D, Hellige M, Feige K, Delling U.Digital recording and storage of health data are becoming increasingly important in the fields of both human and veterinary medicine. There is currently a lack of information on the level of digitalisation among equine veterinarians in Germany and their attitudes towards advanced digitalisation. Objective: To collect opinions of equine veterinarians about (1) digital data collection in pre-purchase examinations (PPEs), and (2) the establishment of an equine health database in Germany. Methods: Cross-sectional survey. Methods: An online survey was developed and distributed via email exclusively...
Effect of pro-inflammatory cytokine priming and storage temperature of the mesenchymal stromal cell (MSC) secretome on equine articular chondrocytes.
Frontiers in bioengineering and biotechnology    August 31, 2023   Volume 11 1204737 doi: 10.3389/fbioe.2023.1204737
Jammes M, Contentin R, Audigiu00e9 F, Cassu00e9 F, Galu00e9ra P. Osteoarthritis (OA) is an invalidating articular disease characterized by cartilage degradation and inflammatory events. In horses, OA is associated with up to 60% of lameness and leads to reduced animal welfare along with extensive economic losses; currently, there are no curative therapies to treat OA. The mesenchymal stromal cell (MSC) secretome exhibits anti-inflammatory properties, making it an attractive candidate for improving the management of OA. In this study, we determined the best storage conditions for conditioned media (CMs) and tested whether priming MSCs with cytokines can enh...
Determination of optimal storage time and temperature for the detection of red blood cell and platelet surface-associated immunoglobulin by flow cytometry in healthy horses.
Veterinary immunology and immunopathology    August 14, 2023   Volume 263 110643 doi: 10.1016/j.vetimm.2023.110643
Frohlich M, Knights K, Springer NL.Differentiating immune-mediated causes from other causes of anemia and thrombocytopenia can be challenging. Flow cytometry can detect surface-associated immunoglobulin (sIg) on red blood cells (RBC) and platelets (PLT) in dogs and horses. Sample storage parameters for ideal assay performance has not been evaluated in horses. The study objective is to identify optimal storage time and temperature of equine whole blood for the detection of RBC-sIg and PLT-sIg via flow cytometry. Both assays were performed on samples at time 0, 4, 24, 48, and 72 h post collection. RBC-sIg samples were stored at ...
Investigation of optimal procedures for storage and use of plasma samples suitable for gene doping tests.
Journal of equine science    June 12, 2023   Volume 34, Issue 2 21-27 doi: 10.1294/jes.34.21
Tozaki T, Ohnuma A, Kikuchi M, Ishige T, Kakoi H, Hirota KI, Takahashi Y, Nagata SI.Gene doping, which is prohibited in horseracing and equestrian sports, can be performed by introducing exogenous genes, known as transgenes, into the bodies of postnatal animals. To detect exogenous genes, a method utilizing quantitative polymerase chain reaction (qPCR) with a hydrolysis probe was developed to test whole blood and plasma samples, thereby protecting the fairness of competition and the rights of stakeholders in horseracing and equestrian sports. Therefore, we aimed to develop sample storage methods suitable for A and B samples in gene doping tests using blood. For sample A, suff...
Sample stability and heparin interference in ionized calcium and ionized magnesium measurements in horses using the Stat Profile Prime Plus co-oximetry electrolyte analyzer.
Veterinary clinical pathology    February 6, 2023   Volume 52, Issue 2 252-260 doi: 10.1111/vcp.13200
Sanmartu00ed J, Robles-Guirado JA, Jose-Cunilleras E, Bassols A.The determination of iCa and iMg is important in veterinary medicine, but their immediate determination in whole blood is not always possible. Their stability in other sample types and the existence of interferences must be evaluated before its use. Objective: We aimed to analyze the effects of storage time on the stability of iCa, iMg, and other analytes in whole blood, plasma, and serum samples in horses and assess the interference of heparin in these measurements. Methods: Whole blood, heparin-plasma, and serum samples from 10 horses were stored at 4°C and analyzed 1, 2, 3, 4, 5, 6, 7, 8, ...
Drying and temperature induced conformational changes of nucleic acids and stallion sperm chromatin in trehalose preservation formulations.
Scientific reports    July 7, 2021   Volume 11, Issue 1 14076 doi: 10.1038/s41598-021-93569-y
Brogna R, Fan J, Sieme H, Wolkers WF, Oldenhof H.Even though dried sperm is not viable, it can be used for fertilization as long as its chromatin remains intact. In this study, we investigated drying- and temperature-induced conformational changes of nucleic acids and stallion sperm chromatin. Sperm was diluted in preservation formulations with and without sugar/albumin and subjected to convective drying at elevated temperatures on glass substrates. Accumulation of reactive oxygen species was studied during storage at different temperatures, and the sperm chromatin structure assay was used to assess DNA damage. Fourier transform infrared spe...
Pre-analytical stability of sorbitol dehydrogenase in equine heparinized plasma.
Veterinary journal (London, England : 1997)    June 9, 2021   Volume 274 105706 doi: 10.1016/j.tvjl.2021.105706
Fouchu00e9 N, Oesch S, Gerber V, Richter H, Howard J, Peters LM.Sorbitol dehydrogenase (SDH) activity is one of the most sensitive and specific markers for hepatocellular injury in horses, but its reported lability makes it impractical for use in many clinical settings. To date, stability of SDH in equine samples has only been evaluated in a limited number of studies in serum samples of horses with activities within reference intervals. The objective of the study was to determine pre-analytical stability of equine SDH activity in heparinized plasma stored at different temperatures for up to 72 h. Twenty client-owned horses admitted to a veterinary teaching...
Impact of Ambient Temperature Sample Storage on the Equine Fecal Microbiota.
Animals : an open access journal from MDPI    March 15, 2021   Volume 11, Issue 3 819 doi: 10.3390/ani11030819
Martin de Bustamante M, Plummer C, MacNicol J, Gomez D.Sample storage conditions are an important factor in fecal microbiota analyses in general. The objective of this study was to investigate the effect of sample storage at room temperature on the equine fecal microbiota composition. Fecal samples were collected from 11 healthy horses. Each sample was divided into 7 sealed aliquots. One aliquot was immediately frozen at -80 °C; the remaining aliquots were stored at room temperature (21 to 22 °C) with one transferred to the freezer at each of the following time points: 6, 12, 24, 48, 72 and 96 h. The Illumina MiSeq sequencer was used for high-th...
Long-term storage of feces at -80 °C versus -20 °C is negligible for 16S rRNA amplicon profiling of the equine bacterial microbiome.
PeerJ    March 9, 2021   Volume 9 e10837 doi: 10.7717/peerj.10837
Gavriliuc S, Stothart MR, Henry A, Poissant J.The development of next-generation sequencing technologies has spurred a surge of research on bacterial microbiome diversity and function. But despite the rapid growth of the field, many uncertainties remain regarding the impact of differing methodologies on downstream results. Sample storage temperature is conventionally thought to be among the most important factors for ensuring reproducibility across marker gene studies, but to date much of the research on this topic has focused on short-term storage in the context of clinical applications. Consequently, it has remained unclear if storage a...
Effects of storage time and temperature on thromboelastographic analysis in dogs and horses.
Veterinary clinical pathology    February 23, 2021   Volume 50, Issue 1 9-19 doi: 10.1111/vcp.12980
Lemon AV, Goddard A, Hooijberg EH.The accessibility of thromboelastography (TEG) to general practitioners is limited by short sample storage times (30 minutes) and storage temperatures (20-23°C). Objective: We aimed to evaluate the stability of canine and equine citrated blood samples when stored for extended periods of time, both at room temperature (RT) (20-23°C) and refrigerator temperature (FT) (2-7.5°C). Methods: Citrated whole blood samples from healthy dogs and horses (n = 10 for each) were stored for 30 minutes (baseline) at RT before TEG analysis. Baseline values for TEG variables R, K, α, MA, LY30, and LY60 w...
In Vitro Evaluation of the Effect of Storage at -20°C and Proximal Gastrointestinal Conditions on Viability of Equine Fecal Microbiota Transplant.
Journal of equine veterinary science    December 18, 2020   Volume 98 103360 doi: 10.1016/j.jevs.2020.103360
Kopper JJ, Alexander TL, Kogan CJ, Berreta AR, Burbick CR.Fecal microbiota transplant (FMT), a technique used to restore normal intestinal microbial communities, has been successful in treating humans with Clostridioides difficile colitis. Subsequently, FMT is being used in veterinary patients with suspected intestinal dysbiosis. Unfortunately, little data are available regarding best practices for FMT in horses. The objective of this study was to evaluate the effects of storing manure prepared for equine FMT (MP-FMT) at -20°C for up to 4 weeks and passage through a simulated proximal gastrointestinal (GI) tract on the viability of MP-FMT. The resu...
Preanalytical variables affecting the measurement of serum paraoxonase-1 activity in horses. Rossi G, Richardson A, Jamaludin H, Secombe C.Paraoxonase-1 (PON-1) activity is a new inflammatory and oxidative marker. Technical effects and biological factors could affect the accuracy of PON-1 activity measurement. We investigated the effects of storage at different temperatures, repeated freeze-thaw cycles, interferences from hemolytic, lipemic, and icteric samples, and seasonal effects on PON-1 activity in horses. We evaluated 2 substrates with an automated spectrophotometer. Ten equine serum samples were stored under different conditions. Although storage at room (21°C) or refrigeration (4°C) temperature induced a statistically s...
The effect of four different freezing conditions and time in frozen storage on the concentration of commonly measured growth factors and enzymes in equine platelet-rich plasma over six months.
BMC veterinary research    August 14, 2019   Volume 15, Issue 1 292 doi: 10.1186/s12917-019-2040-4
McClain AK, McCarrel TM.Platelet-rich plasma (PRP) is a therapeutic biologic that is used for treatment of musculoskeletal pathologies in equine athletes. Due to the expense of PRP kits, and the volumes obtained, freezing aliquots for future dosing is common. Aliquots of PRP are also commonly frozen for later analysis of growth factor concentrations in in vitro research. A variety of freezing methods are used and storage duration until analysis is often not reported. The optimal frozen storage conditions and duration to maintain concentrations of commonly measured growth factors and enzymes in PRP are unknown. Our ob...
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