An Advax-adjuvanted Inactivated Cell-culture Japanese Encephalitis Vaccine Induces Broadly Neutralising Anti-flavivirus Antibodies and T-cell Immunity and Provides Single Dose Protection

ccJE+Advax is an inactivated cell culture Japanese encephalitis (JE) vaccine formulated with Advax™, a novel polysaccharide adjuvant based on delta inulin. This vaccine has previously shown promise in murine and equine studies and the current study sought to better understand its mechanism of action and assess the feasibility of single dose vaccine protection. Mice immunised with ccJE-Advax had higher serum neutralisation titres than those immunised with ccJE alone or with alum adjuvant. ccJE+Advax induced extraordinarily broad cross-neutralising antibodies against multiple flaviviruses including West Nile virus (WNV), Murray Valley Encephalitis Virus (MVEV), St Louis Encephalitis virus (SLE) and Dengue-1 and -2 viruses. Notably, the DENV-2 cross-neutralising antibodies from ccJE+Advax immunised mice uniquely had no DENV-2 antibody dependent enhancement (ADE) activity, by contrast to high ADE activity seen with DENV-1 cross-reactive antibodies induced by mbJE or ccJE alone or with alum adjuvant. JEV-stimulated splenocytes from ccJE+Advax immunised mice showed increased IL-17 and IFN-γ production, consistent with a mixed Th1 and Th17 response, whereas ccJE-alum was associated with production of mainly Th2 cytokines. There is an ongoing lack of human vaccines against particular flaviviruses, including WNV, SLE and MVEV. Given its ability to provide single-dose JEV protection as well as to induce broadly neutralising antibodies free of ADE activity, ccJE+Advax vaccine could be highly useful in all situations where rapid protection is desirable but ADE needs to be avoided, e.g. during a local outbreak or for use in travellers or the military requiring rapid travel to JEV endemic regions.

Species-Level Gut Microbiota Analysis after Antibiotic-Induced Dysbiosis in Horses

All current studies have used Illumina short-read sequencing to characterize the equine intestinal microbiota. Long-read sequencing can classify bacteria at the species level. The objectives of this study were to characterize the gut microbiota of horses at the species level before and after trimethoprim sulfadiazine (TMS) administration and to compare results with Illumina sequencing. Nine horses received TMS (30 mg/kg) orally for 5 days twice a day to induce dysbiosis. Illumina sequencing of the V4 region or full-length PacBio sequencing of the 16S rRNA gene was performed in fecal samples collected before and after antibiotic administration. The relative abundance and alpha diversity were compared between the two technologies. PacBio failed to classify the equine intestinal microbiota at the species level but confirmed Bacteroidetes as the most abundant bacteria in the feces of the studied horses, followed by Firmicutes and Fibrobacteres. An unknown species of the Bacteroidales order was highly abundant (13%) and deserves further investigation. In conclusion, PacBio was not suitable to classify the equine microbiota species but detected greater richness and less unclassified bacteria. Further efforts in improving current databanks to be used in equine studies are necessary.

The horse as a model for translational orthopedic research: examples of studies on the regeneration of cartilage and bone conducted in Costa Rica

The paper provides results published or to be published of long-term in vivo equine studies to evaluate techniques of possible regenerative matrices of cartilage and bone, by means of cell-free implants or stimulation of the bone marrow. From the fixation techniques analyzed, it can be concluded that the best alternatives are the pressure technique for subchondral defects and a novel hydrogel with self-adhesive capacity for chondral defects. The equine coxal tuberosity was used for the first time as a model for regeneration studies of bone defects, analyzing scaffolds based on tricalcium phosphate, polymers and nanoparticles, by means of 3-D printing. Osteoconductivity, osteoinductivity, and the importance of microporosity were documented.Given that decellularized materials do not always give significant desired results in the regeneration of cartilage, it is important to conduct long-term studies. The technique of nanofracture and a novel self-adhesive hydrogel in the knee of the equine showed promising preliminary results in the regeneration of cartilage.The knee and the coxal tuberosity of the horse represent models of studying cartilage and bone regeneration in a true translational sense as a source of highly valuable information for clinical studies, for both horses and humans.

Functionally Annotating Regulatory Elements in the Equine Genome Using Histone Mark ChIP-Seq

One of the primary aims of the Functional Annotation of ANimal Genomes (FAANG) initiative is to characterize tissue-specific regulation within animal genomes. To this end, we used chromatin immunoprecipitation followed by sequencing (ChIP-Seq) to map four histone modifications (H3K4me1, H3K4me3, H3K27ac, and H3K27me3) in eight prioritized tissues collected as part of the FAANG equine biobank from two thoroughbred mares. Data were generated according to optimized experimental parameters developed during quality control testing. To ensure that we obtained sufficient ChIP and successful peak-calling, data and peak-calls were assessed using six quality metrics, replicate comparisons, and site-specific evaluations. Tissue specificity was explored by identifying binding motifs within unique active regions, and motifs were further characterized by gene ontology (GO) and protein–protein interaction analyses. The histone marks identified in this study represent some of the first resources for tissue-specific regulation within the equine genome. As such, these publicly available annotation data can be used to advance equine studies investigating health, performance, reproduction, and other traits of economic interest in the horse.

Humanizing Horses: Transitions in Perception and Perspective

In Tibetan history and culture, horses were among the most important animals, if not the most important of all. Horses were the mounts that provided transport, particularly for the nobility and kings, allowing them to travel more quickly and comfortably. Horses were also used for hunting, postal services, and to build a cavalry for warfare. In addition, they played a role in various entertainments, including horse racing, games, and parades. The unusually large number of manuscripts on horses attests to the value of horses in the Tibetan imaginaire compared to other animals that lived in the company of the people on the High Plateau, in Tibet itself, and in Tibetan cultural areas. This article begins with an outline of the uses and benefits of horses in Tibetan culture. It touches upon the animal’s role as the mount of Tibetan kings and debates regarding horses’ mental faculties. Then it presents a survey of the content of various manuscripts on equine studies based on sources from three stages: (1) the earliest Tibetan sources from Dunhuang; (2) translations from Indian texts; and (3) extensive compendia that merges all of the knowledge on horses available at the time of their composition. It analyzes the style and content of books that indicate the approach of the authors to the topic of “horse” and points to their view of horses in relation to Tibetan culture and Buddhism. Moreover, the books’ content mirrors the various functions and applications of horses in Tibet and India. It reveals the purpose of these books in general and illustrates the relation between textuality and orality. The study demonstrates the link between hippology and hippiatry, and the development of equine studies in Tibet. It shows the influence of humans on horse medicine and, moreover, contributes to an improved understanding of the development of Tibetan medical sciences in general.

Fetal heart rate and fetal heart rate variability in Lipizzaner broodmares

Monitoring fetal heart rate (FHR) and fetal heart rate variability (FHRV) helps to understand and evaluate normal and pathological conditions in the foal. The aim of this study was to establish normal heart rate reference values for the ongoing equine pregnancy and to perform a heart rate variability (HRV) time-domain analysis in Lipizzaner mares. Seventeen middle- and late-term (days 121–333) pregnant Lipizzaner mares were examined using fetomaternal electrocardiography (ECG). The mean FHR (P = 0.004) and the standard deviation of FHR (P = 0.012) significantly decreased during the pregnancy. FHR ± SD values decreased from 115 ± 35 to 79 ± 9 bpm between months 5 and 11. Our data showed that HRV in the foal decreased as the pregnancy progressed, which is in contrast with the findings of earlier equine studies. The standard deviation of normal-normal intervals (SDNN) was higher (70 ± 25 to 166 ± 108 msec) than described previously. The root mean square of successive differences (RMSSD) decreased from 105 ± 69 to 77 ± 37 msec between the 5th and 11th month of gestation. Using telemetric ECG equipment, we could detect equine fetal heartbeat on day 121 for the first time. In addition, the large differences observed in the HR values of four mare-fetus pairs in four consecutive months support the assumption that there might be ‘high-HR’ and ‘low-HR’ fetuses in horses. It can be concluded that the analysis of FHR and FHRV is a promising tool for the assessment of fetal well-being but the applicability of these parameters in the clinical setting and in studs requires further investigation.

The use of cardiac biomarkers in veterinary medicine: the equine perspective

In human medicine, cardiac biomarkers, such as natriuretic peptides and troponins, are routinely used for the diagnosis, prognosis and monitoring of heart diseases. Similarly, these biomarkers are determined in small animals to differentiate non-cardiac from cardiac diseases. Knowledge about these biomarkers in horses is limited and requires further investigation. The first equine studies about atrial natriuretic peptide (ANP) and N-terminal ANP (NT-proANP) are promising, and show a clear correlation with atrial dimension size. Equine brain natriuretic peptides assays are still unavailable. The troponins, in particular troponin I, have been more extensively studied in horses, and their use for the diagnosis of myocardial damage has been fully demonstrated. They have replaced the less specific lactate dehydrogenase and creatine kinase isoenzymes, which makes the use of the last mentioned no longer legitimate. A final possible equine biomarker is aldosterone. Reference values in horses have been established. However, in only one study, a correlation between aldosterone and cardiac disease has been reported.