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Topic:Veterinary Research

Veterinary research in horses encompasses the study of diseases, health management, and medical treatments specific to equine species. This field investigates various aspects of horse health, including infectious diseases, metabolic disorders, and musculoskeletal conditions. Researchers focus on understanding the pathophysiology of equine ailments, developing diagnostic tools, and evaluating therapeutic interventions. The study of horse health also involves examining preventive measures such as vaccination protocols and nutritional management to promote overall well-being. This page collects peer-reviewed research studies and scholarly articles that explore the diverse areas of veterinary science related to horses, providing insights into disease mechanisms, treatment strategies, and advancements in equine healthcare.
Stereotypic horses (Equus caballus) are not cognitively impaired.
Animal cognition    October 17, 2018   Volume 22, Issue 1 17-33 doi: 10.1007/s10071-018-1217-8
Briefer Freymond S, Ruet A, Grivaz M, Fuentes C, Zuberbühler K, Bachmann I, Briefer EF.Stereotypies in animals are thought to arise from an interaction between genetic predisposition and sub-optimal housing conditions. In domestic horses, a well-studied stereotypy is crib-biting, an abnormal behaviour that appears to help individuals to cope with stressful situations. One prominent hypothesis states that animals affected by stereotypies are cognitively less flexible compared to healthy controls, due to sensitization of a specific brain area, the basal ganglia. The aim of this study was to test this hypothesis in crib-biting and healthy controls, using a cognitive task, reversa...
Biological variation of routine haematology and biochemistry measurands in the horse.
Equine veterinary journal    October 17, 2018   Volume 51, Issue 3 384-390 doi: 10.1111/evj.13017
Wright ME, Croser EL, Raidal S, Baral RM, Robinson W, Lievaart J, Freeman KP.Clinical pathology results are typically interpreted by referring to population-based reference intervals. The use of individualised (subject-based) reference intervals is more appropriate for measurands with a high degree of variation between individuals. Objective: To determine the biological variation of routinely analysed equine haematology and biochemistry measurands and calculate indices of individuality and reference change values which enable production of individualised reference intervals, in a group of healthy, privately owned horses. Methods: In a prospective cohort study, thirty-n...
Species distribution and seasonal dynamics of equine tick infestation in two Mediterranean climate niches in Israel.
Parasites & vectors    October 16, 2018   Volume 11, Issue 1 546 doi: 10.1186/s13071-018-3093-0
Tirosh-Levy S, Gottlieb Y, Apanaskevich DA, Mumcuoglu KY, Steinman A.Ticks are important ectoparasites of horses that can affect animal welfare and vector several infectious, including zoonotic, diseases. In order to investigate the species distribution, epidemiology and seasonal dynamics of ticks infesting horses in Israel, 3267 ticks were collected from 396 horses in 24 farms across the country from July 2014 to June 2015. Results: Ticks were found on 50% of the farms and on 25% of the horses, with Hyalomma being the most prevalent genus (70% of ticks). Pasture was the most prominent risk factor for tick infestation (99% of ticks, P < 0.001), and is repres...
5′-flanking variants of equine casein genes (CSN1S1, CSN1S2, CSN2, CSN3) and their relationship with gene expression and milk composition.
Journal of applied genetics    October 16, 2018   Volume 60, Issue 1 71-78 doi: 10.1007/s13353-018-0473-2
Cieslak J, Wodas L, Borowska A, Pawlak P, Czyzak-Runowska G, Wojtowski J, Puppel K, Kuczynska B, Mackowski M.Genes encoding casein proteins are important candidates for milk composition traits in mammals. In the case of the domestic horse, our knowledge of casein genes is limited mainly to coding sequence variants. This study involved screening for polymorphism in 5'-flanking regions of four genes encoding equine caseins (CSN1S1, CSN1S2, CSN2, and CSN3) and making a preliminary assessment of their effect on the gene expression (on the mRNA and protein levels) and milk composition traits in selected horse breeds. Altogether, 23 polymorphisms (21 described previously SNPs and two novel InDels) were fou...
A survey of Dictyocaulus arnfieldi (Nematoda) infections in equids in Urmia region, Iran.
Annals of parasitology    October 15, 2018   Volume 64, Issue 3 235-240 doi: 10.17420/ap6403.158
Saadi, A, Tavassoli, M, Dalir-Naghadeh, B, Samiei A.Dictyocaulus arnfieldi is a lungworm commonly found in equids; however, relatively little is known about it. The aim of the present study was to establish the prevalence of equine lungworms in naturally-infected working and sporting horses, donkeys and mules in rural areas of Urmia, northwest Iran. The fecal samples were collected from 299 working horses, 57 sporting horses, 66 donkeys and 37 mules during the period March 2014 to June 2016. The collected fecal samples were processed within 48 hours following rectal sampling. The larval count/g (LPG) of feces was determined using the Baermann t...
Mammals repel mosquitoes with their tails.
The Journal of experimental biology    October 15, 2018   Volume 221, Issue Pt 20 jeb178905 doi: 10.1242/jeb.178905
Matherne ME, Cockerill K, Zhou Y, Bellamkonda M, Hu DL.The swinging of a mammal's tail has long been thought to deter biting insects, which, in cows, can drain up to 0.3 liters of blood per day. How effective is a mammal's tail at repelling insects? In this combined experimental and theoretical study, we filmed horses, zebras, elephants, giraffes and dogs swinging their tails. The tail swings at triple the frequency of a gravity-driven pendulum, and requires 27 times more power input. Tails can also be used like a whip to directly strike at insects. This whip-like effect requires substantial torques from the base of the tail on the order of 10-1...
Diffusion of charged and uncharged contrast agents in equine mandibular condylar cartilage is not affected by an increased level of sugar-induced collagen crosslinking.
Journal of the mechanical behavior of biomedical materials    October 14, 2018   Volume 90 133-139 doi: 10.1016/j.jmbbm.2018.10.022
Mirahmadi F, Koolstra JH, Fazaeli S, Lobbezoo F, van Lenthe GH, Snabel J, Stoop R, Everts V.Nutrition of articular cartilage relies mainly on diffusion and convection of solutes through the interstitial fluid due to the lack of blood vessels. The diffusion is controlled by two factors: steric hindrance and electrostatic interactions between the solutes and the matrix components. Aging comes with changes in the cartilage structure and composition, which can influence the diffusion. In this study, we treated fibrocartilage of mandibular condyle with ribose to induce an aging-like effect by accumulating collagen crosslinks. The effect of steric hindrance or electrostatic forces on the d...
κ Light-chain monoclonal gammopathy and cast nephropathy in a horse with multiple myeloma.
Journal of the American Veterinary Medical Association    October 13, 2018   Volume 253, Issue 9 1177-1183 doi: 10.2460/javma.253.9.1177
Eberhardt C, Malbon A, Riond B, Schoster A.CASE DESCRIPTION A 27-year-old Dutch Warmblood mare was evaluated because of a history of lethargy, reluctance to move, weight loss, persistent hyperproteinemia, and recurrent episodes of mild lameness. CLINICAL FINDINGS Hematologic evaluation revealed anemia (RBC concentration, 3.84 × 106 cells/μL), thrombocytopenia (47 × 103 thrombocytes/μL), and hyperproteinemia (total protein concentration, 11.2 g/dL) with hyperglobulinemia and hypoalbuminemia. Results of protein electrophoresis of serum and urine samples indicated a monoclonal gammopathy; the paraprotein was identified as a κ light c...
Improvement of recombinant-truncated Burkholderia motility protein A (BimA)-based indirect ELISA for equine glanders.
Journal of immunoassay & immunochemistry    October 12, 2018   Volume 39, Issue 5 565-575 doi: 10.1080/15321819.2018.1523800
Singh S, Dohre SK, Kamthan A, Pal V, Karothia BS, Singha HS, Kumar S.Glanders is a contagious and highly fatal disease of equines with zoonotic potential. It is caused by a Gram-negative, nonmotile bacterium Burkholderia mallei. Complement fixation test (CFT) is one of the most commonly used tests for diagnosis of glanders; however, it has some limitations. A recombinant-truncated Burkholderia intracellular motility A (BimA) protein-based indirect enzyme-linked immunosorbent assay (iELISA) was previously reported by us for glanders diagnosis, which has been re-optimized in this study using a panel of glanders positive (n = 75) and glanders negative (n = 227...
Lentiviral-based reporter constructs for profiling chondrogenic activity in primary equine cell populations.
European cells & materials    October 12, 2018   Volume 36 156-170 doi: 10.22203/eCM.v036a12
Martin-Pena A, Porter RM, Plumton G, McCarrel TM, Morton AJ, Guijarro MV, Ghivizzani SC, Sharma B, Palmer GD.Successful clinical translation of mesenchymal stem cell (MSC)-based therapies for cartilage repair will likely require the implementation of standardised protocols and broadly applicable tools to facilitate the comparisons among cell types and chondroinduction methods. The present study investigated the utility of recombinant lentiviral reporter vectors as reliable tools for comparing chondrogenic potential among primary cell populations and distinguishing cellular-level variations of chondrogenic activity in widely used three-dimensional (3D) culture systems. Primary equine MSCs and chondroc...
Generation of an equine biobank to be used for Functional Annotation of Animal Genomes project.
Animal genetics    October 11, 2018   Volume 49, Issue 6 564-570 doi: 10.1111/age.12717
Burns EN, Bordbari MH, Mienaltowski MJ, Affolter VK, Barro MV, Gianino F, Gianino G, Giulotto E, Kalbfleisch TS, Katzman SA, Lassaline M, Leeb T....The Functional Annotation of Animal Genomes (FAANG) project aims to identify genomic regulatory elements in both sexes across multiple stages of development in domesticated animals. This study represents the first stage of the FAANG project for the horse, Equus caballus. A biobank of 80 tissue samples, two cell lines and six body fluids was created from two adult Thoroughbred mares. Ante-mortem assessments included full physical examinations, lameness, ophthalmologic and neurologic evaluations. Complete blood counts and serum biochemistries were also performed. At necropsy, in addition to tiss...
Sequence variant analysis of RNA sequences in severe equine asthma.
PeerJ    October 11, 2018   Volume 6 e5759 doi: 10.7717/peerj.5759
Tessier L, Côté O, Bienzle D.Severe equine asthma is a chronic inflammatory disease of the lung in horses similar to low-Th2 late-onset asthma in humans. This study aimed to determine the utility of RNA-Seq to call gene sequence variants, and to identify sequence variants of potential relevance to the pathogenesis of asthma. Methods: RNA-Seq data were generated from endobronchial biopsies collected from six asthmatic and seven non-asthmatic horses before and after challenge (26 samples total). Sequences were aligned to the equine genome with Spliced Transcripts Alignment to Reference software. Read preparation for sequenc...
Freezing, Vitrification, and Freeze-Drying of Equine Spermatozoa: Impact on Mitochondrial Membrane Potential, Lipid Peroxidation, and DNA Integrity.
Journal of equine veterinary science    October 11, 2018   Volume 72 8-15 doi: 10.1016/j.jevs.2018.10.006
Restrepo G, Varela E, Duque JE, Gómez JE, Rojas M.Maintaining the integrity of equine sperm subjected to preservation protocols is essential for the successful development of assisted reproduction procedures. The aim of this study was to assess the mitochondrial membrane potential, lipid peroxidation, and DNA integrity of equine sperm subjected to freezing, vitrification, and freeze-drying. Eight ejaculates obtained from four Colombian Creole horses were subjected to programmable freezing, vitrification, and freeze-drying. After thawing or rehydration, sperm motility and kinetics were assessed through a CASA system. The mitochondrial membrane...
Digital PCR detection of plasmid DNA administered to the skeletal muscle of a microminipig: a model case study for gene doping detection.
BMC research notes    October 10, 2018   Volume 11, Issue 1 708 doi: 10.1186/s13104-018-3815-6
Tozaki T, Gamo S, Takasu M, Kikuchi M, Kakoi H, Hirota KI, Kusano K, Nagata SI.Doping control is an important and indispensable aspect of fair horse racing; genetic doping has been recently included to this. In this study, we aimed to develop a detection method of gene doping. A plasmid cloned with human erythropoietin gene (p.hEPO, 250 μg/head) was intramuscularly injected into a microminipig. Subsequently, p.hEPO was extracted from 1 mL of plasma and detected by droplet digital polymerase chain reaction. Results: The results confirmed that the maximum amount of plasmid was detected at 15 min after administration and the majority of the plasmid was degraded in the b...
Validation of a new experimental model for assessing drug efficacy against infection with Trypanosoma equiperdum in horses.
Veterinary parasitology    October 9, 2018   Volume 263 27-33 doi: 10.1016/j.vetpar.2018.10.005
Hébert L, Guitton E, Madeline A, Géraud T, Carnicer D, Lakhdar L, Pitel PH, Coste M, Laloy E, Giraudet A, Zientara S, Büscher P, Laugier C, Hans A....Trypanosoma equiperdum, the causative agent of dourine, may affect the central nervous system, leading to neurological signs in infected horses. This location protects the parasite from most (if not all) existing chemotherapies. In this context, the OIE terrestrial code considers dourine as a non-treatable disease and imposes a stamping-out policy for affected animals before a country may achieve its dourine-free status. The use of practices as drastic as euthanasia remains controversial, but the lack of a suitable tool for studying a treatment's efficacy against dourine hampers the developmen...
Genotype imputation accuracy in multiple equine breeds from medium- to high-density genotypes.
Journal of animal breeding and genetics = Zeitschrift fur Tierzuchtung und Zuchtungsbiologie    October 9, 2018   Volume 135, Issue 6 420-431 doi: 10.1111/jbg.12358
Chassier M, Barrey E, Robert C, Duluard A, Danvy S, Ricard A.Genotype imputation is now a key component of genomic analyses as it increases the density of available genotypes within a population. However, many factors can influence imputation accuracy. The aim of this study was to assess and compare the accuracy of imputation of high-density genotypes (Affymetrix Axiom Equine genotyping array, 670,806 SNPs) from two moderate-density genotypes (Illumina Equine SNP50 BeadChip, 54,602 SNPs and Illumina Equine SNP70 BeadChip, 65,157 SNPs), using single-breed or multiple-breed reference sets. Genotypes were available from five groups of horse breeds: Arab (A...
Objective pain assessment in horses (2014-2018).
Veterinary journal (London, England : 1997)    October 9, 2018   Volume 242 1-7 doi: 10.1016/j.tvjl.2018.10.001
van Loon JPAM, Van Dierendonck MC.In recent decades, much effort has been invested in scientific studies of objective and reliable assessment of pain in horses. Various types of pain assessment tools have been described and (partly) validated for different types of pain in horses. Currently, composite pain scales and facial expression-based pain scales seem to be the most promising tools for pain assessment in horses and numerous studies have recently been published on the use of these pain scales in horses. Therefore, this narrative review mainly focusses on these two types of pain scales and on the studies that have appeared...
Phylogenetic analysis of the cytochrome P450 (CYP450) nucleotide sequences of the horse and predicted CYP450s of the white rhinoceros (Ceratotherium simum) and other mammalian species.
PeerJ    October 9, 2018   Volume 6 e5718 doi: 10.7717/peerj.5718
Leiberich M, Marais HJ, Naidoo V.The plight of the white rhinoceros () and the increasing need of treatment options for injured poaching victims led to the necessity to expand the knowledge on applicable drugs in this endangered species. With very little information available on drug pharmacokinetics in rhino, veterinarians have to rely on information generated from other species. The horse being a closely related species, has served as the model for dose extrapolations. However, from recent research on enrofloxacin and carprofen, the white rhino showed considerable differences in the pharmacokinetic properties of these drugs...
Equine bone marrow-derived mesenchymal stem cells: optimization of cell density in primary culture.
Stem cell investigation    October 9, 2018   Volume 5 31 doi: 10.21037/sci.2018.09.01
Zahedi M, Parham A, Dehghani H, Kazemi Mehrjerdi H.The primary cell seeding density of bone marrow-derived mononuclear cells (BM-MNCs) affects several cellular behaviors, including attachment to the culture dish, proliferation, and differentiation. Methods: The aim of this study was to determine the best density of equine BM-MNCs in primary culture (P0) for obtaining the maximum bone marrow-derived mesenchymal stem cell (BM-MSC) yields at the end of P0. Bone marrow samples of two healthy mares were aspirated. The MNCs were isolated and cultured at different densities (1×10, 2×10, 4×10, 8×10, and 1×10 cells/cm). Within the 7 and 14 days af...
Characterization of Apoptosis, Autophagy and Oxidative Stress in Pancreatic Islets Cells and Intestinal Epithelial Cells Isolated from Equine Metabolic Syndrome (EMS) Horses.
International journal of molecular sciences    October 8, 2018   Volume 19, Issue 10 3068 doi: 10.3390/ijms19103068
Kornicka K, Śmieszek A, Szłapka-Kosarzewska J, Irwin Houston JM, Roecken M, Marycz K.Endocrine disorders are becoming an increasing problem in both human and veterinary medicine. In recent years, more and more horses worldwide have been suffering from equine metabolic syndrome (EMS). This metabolic disorder is characterized by pathological obesity, hyperinsulinaemia, hyperglycaemia and insulin resistance. Although metabolic disorders, including diabetes, have been extensively studied, there are still no data on the molecular effects of EMS in horses. Thus, the aim of this study was to evaluate apoptosis, oxidative stress, autophagy and microRNA (miR) expression in multipotent ...
A single dose of African horse sickness virus (AHSV) VP2 based vaccines provides complete clinical protection in a mouse model.
Vaccine    October 8, 2018   Volume 36, Issue 46 7003-7010 doi: 10.1016/j.vaccine.2018.09.065
Aksular M, Calvo-Pinilla E, Marín-López A, Ortego J, Chambers AC, King LA, Castillo-Olivares J.African horse sickness is a severe, often fatal, arboviral disease of equids. The control of African horse sickness virus (AHSV) in endemic countries is based currently on the use of live attenuated vaccines despite some biosafety concerns derived from its biological properties. Thus, experimental vaccination platforms have been developed over the years in order to avoid the biosafety concerns associated with the use of attenuated vaccines. Various studies showed that baculovirus-expressed AHSV-VP2 or modified Vaccinia Ankara virus expressing AHSV-VP2 (MVA-VP2) induced virus neutralising antib...
Correction to: Culture of mesenchymal stem cells derived from equine synovial membrane in alginate hydrogel microcapsules.
Stem cell research & therapy    October 7, 2018   Volume 9, Issue 1 259 doi: 10.1186/s13287-018-0999-6
Santos VH, Pfeifer JPH, de Souza JB, Milani BHG, de Oliveira RA, Assis MG, Deffune E, Moroz A, Alves ALG.The original article [1] contained a minor error regarding the mean diameter of the alginate microcapsules described in relation to Fig. 4 in the Results section. The microcapsules had an actual mean diameter of 3000 μm instead of 1000 μm as mistakenly mentioned in the original article.
Equine asthma: Integrative biologic relevance of a recently proposed nomenclature.
Journal of veterinary internal medicine    October 7, 2018   Volume 32, Issue 6 2088-2098 doi: 10.1111/jvim.15302
Bond S, Léguillette R, Richard EA, Couetil L, Lavoie JP, Martin JG, Pirie RS.The term "equine asthma" has been proposed as a unifying descriptor of inflammatory airway disease (IAD), recurrent airway obstruction (RAO), and summer pasture-associated obstructive airway disease. Whilst the term will increase comprehensibility for both the lay and scientific communities, its biologic relevance must be compared and contrasted to asthma in human medicine, recognizing the limited availability of peer-reviewed equine-derived data, which are largely restricted to clinical signs, measures of airway obstruction and inflammation and response to therapy. Such limitations constrain ...
Celebrating excellence in equine medicine.
The Veterinary record    October 6, 2018   Volume 183, Issue 13 400 doi: 10.1136/vr.k4178
No abstract available
Dysregulation of Cortisol Metabolism in Equine Pituitary Pars Intermedia Dysfunction.
Endocrinology    October 6, 2018   Volume 159, Issue 11 3791-3800 doi: 10.1210/en.2018-00726
Morgan RA, Keen JA, Homer N, Nixon M, McKinnon-Garvin AM, Moses-Williams JA, Davis SR, Hadoke PWF, Walker BR.Equine Cushing disease [pituitary pars intermedia dysfunction (PPID)] is a common condition of older horses, but its pathophysiology is complex and poorly understood. In contrast to pituitary-dependent hyperadrenocorticism in other species, PPID is characterized by elevated plasma ACTH but not elevated plasma cortisol. In this study, we address this paradox and the hypothesis that PPID is a syndrome of ACTH excess in which there is dysregulation of peripheral glucocorticoid metabolism and binding. In 14 horses with PPID compared with 15 healthy controls, we show that in plasma, cortisol levels...
Proteome and transcriptome profiling of equine myofibrillar myopathy identifies diminished peroxiredoxin 6 and altered cysteine metabolic pathways.
Physiological genomics    October 5, 2018   Volume 50, Issue 12 1036-1050 doi: 10.1152/physiolgenomics.00044.2018
Valberg SJ, Perumbakkam S, McKenzie EC, Finno CJ.Equine myofibrillar myopathy (MFM) causes exertional muscle pain and is characterized by myofibrillar disarray and ectopic desmin aggregates of unknown origin. To investigate the pathophysiology of MFM, we compared resting and 3 h postexercise transcriptomes of gluteal muscle and the resting skeletal muscle proteome of MFM and control Arabian horses with RNA sequencing and isobaric tags for relative and absolute quantitation analyses. Three hours after exercise, 191 genes were identified as differentially expressed (DE) in MFM vs. control muscle with >1 log fold change (FC) in genes involve...
Ganglion Cytology: A Novel Rapid Method for the Diagnosis of Equine Dysautonomia.
Veterinary pathology    October 5, 2018   Volume 56, Issue 2 244-247 doi: 10.1177/0300985818806051
Piccinelli C, Jago R, Milne E.Equine dysautonomia (grass sickness) is characterized by autonomic neuronal degeneration and is often fatal. As outbreaks occur, rapid diagnosis is essential but confirmation currently requires histological examination. This study evaluated diagnostic accuracy of cytological examination of cranial cervical ganglion (CCG) scrapings for dysautonomia diagnosis. CCG smears from 20 controls and 16 dysautonomia cases were stained with May-Grünwald Giemsa (MGG), hematoxylin and eosin (HE), and cresyl fast violet (CFV), with HE-stained histological sections of CCG as gold standard for diagnosis. Exam...
Identification of a VapA virulence factor functional homolog in Rhodococcus equi isolates housing the pVAPB plasmid.
PloS one    October 4, 2018   Volume 13, Issue 10 e0204475 doi: 10.1371/journal.pone.0204475
Willingham-Lane JM, Coulson GB, Hondalus MK.Rhodococcus equi is a facultative intracellular bacterium of macrophages and is an important pathogen of animals and immunocompromised people wherein disease results in abcessation of the lungs and other sites. Prior work has shown that the presence of the major virulence determinant, VapA, encoded on the pVAPA-type plasmid, disrupts normal phagosome development and is essential for bacterial replication within macrophages. pVAPA- type plasmids are typical of R. equi strains derived from foals while strains from pigs carry plasmids of the pVAPB-type, lacking vapA, and those from humans harbor ...
Assessment of a Rabies Virus Rapid Diagnostic Test for the Detection of Australian Bat Lyssavirus.
Tropical medicine and infectious disease    October 4, 2018   Volume 3, Issue 4 109 doi: 10.3390/tropicalmed3040109
Certoma A, Lunt RA, Vosloo W, Smith I, Colling A, Williams DT, Tran T, Blacksell SD.Australian bat lyssavirus (ABLV) is closely related to the classical rabies virus and has been associated with three human fatalities and two equine fatalities in Australia. ABLV infection in humans causes encephalomyelitis, resulting in fatal disease, but has no effective therapy. The virus is maintained in enzootic circulation within fruit bats ( spp.) and at least one insectivorous bat variety ( ). Most frequently, laboratory testing is conducted on pteropodid bat brains, either following a potential human exposure through bites, scratches and other direct contacts with bats, or as opportun...
Impact of Mixed Equine Influenza Vaccination on Correlate of Protection in Horses.
Vaccines    October 4, 2018   Volume 6, Issue 4 71 doi: 10.3390/vaccines6040071
Dilai M, Piro M, El Harrak M, Fougerolle S, Dehhaoui M, Dikrallah A, Legrand L, Paillot R, Fassi Fihri O.To evaluate the humoral immune response to mixed Equine Influenza vaccination, a common practice in the field, an experimental study was carried out on 42 unvaccinated thoroughbred weanling foals divided into six groups of seven. Three groups were vaccinated using a non-mixed protocol (Equilis Prequenza-Te, Proteqflu-Te or Calvenza-03) and three other groups were vaccinated using a mix of the three vaccines mentioned previously. Each weanling underwent a primary EI vaccination schedule composed of two primary immunisations (V1 and V2) four weeks apart followed by a third boost immunisation (V3...