The behavior and welfare of neglected species: some examples from fish and mammals.

Animal welfare is the state of an animal's body and mind and the level to which its requirements are satisfied. Animal welfare is affected by human decisions and actions. Numerous decisions concerning animals are driven by human desires to enhance their own lives, and some of these decisions may be influenced by self-interest or a strong emphasis on economic factors. How to assess the welfare state of animals is a central issue in animal welfare science. Two critical questions can be used to address animal welfare: first, is the animal healthy, and second, does the animal have what it needs? Both of these questions can potentially be answered using the study of animal behavior. The development of behavioral methodologies is crucial for evaluating welfare in contexts where concern for animal welfare is often highest, such as on intensive modern farms and sites where working animals are used. Herein, we discuss animal welfare by focusing on some of its major concepts and explanations. Later, to illustrate key aspects of animal welfare, we chose to examine the information that is available for some 'neglected' livestock species, which are commercially important on a global basis and found in large numbers: buffaloes (Bubalus bubalis), camels (Camelus dromedarius), donkeys (Equus asinus), mules (Equus asinus × Equus caballus), and lumpfish (Cyclopterus lumpus). We chose these species because there are major ongoing concerns about their welfare, and more research is required to help solve the various problems. Overall, there are strong imbalances in terms of the species that are usually studied in terms of animal welfare research, and we call for greater attention to those that have traditionally been neglected.

Severe orbital hematoma with third eyelid swelling and prolapse following a blind maxillary nerve block in a horse.

A 20-year-old quarter horse gelding was presented for routine dental examination. Periodontal disease and luxation of tooth 108 was diagnosed and oral extraction was planned. After an unsuccessful blind maxillary nerve block, it was elected to perform the procedure under total intravenous anesthesia. Following recovery, a focal superficial corneal ulcer, severe retrobulbar swelling, mild exophthalmos, and marked swelling and prolapse of the third eyelid (nictitating membrane) were observed. Clinical signs persisted beyond 48 h despite the use of systemic anti-inflammatories and topical ocular anti-inflammatories and antibiotics. A temporary tarsorrhaphy was subsequently done at 48 h and the horse was discharged after 5 d of hospitalization and regression of clinical signs. Although it is very useful for easing dental extractions, the blind maxillary nerve block is associated with potential complications due to inadvertent vascular puncture. This case report describes a rare complication of prolapse of the third eyelid in a horse after a maxillary nerve block and successful treatment with a temporary tarsorrhaphy. Key clinical message: This case report explains how nictitating membrane swelling and prolapse can occur following a blind maxillary nerve block in the horse and describes treatment with a temporary tarsorrhaphy.

Hématome orbitaire sévère avec gonflement de la troisième paupière et prolapsus à la suite d'un bloc du nerf maxillaire à l'aveugle chez un cheval. Un hongre quarter horse de 20 ans a été présenté pour un examen dentaire de routine. Une maladie parodontale et une luxation de la dent 108 ont été diagnostiquées et une extraction orale a été planifiée. Après l'échec d'un bloc du nerf maxillaire à l'aveugle, il a été décidé d'effectuer la procédure sous anesthésie intraveineuse. Après la guérison, un ulcère cornéen superficiel focal, un gonflement rétrobulbaire sévère, une légère exophtalmie ainsi qu'un gonflement et un prolapsus marqués de la troisième paupière (membrane nictitante) ont été observés. Les signes cliniques ont persisté au-delà de 48 heures malgré l'utilisation d'anti-inflammatoires systémiques et d'anti-inflammatoires oculaires topiques et d'antibiotiques. Une tarsorraphie temporaire a ensuite été réalisée à 48 heures et le cheval est sorti après 5 jours d'hospitalisation et de régression des signes cliniques. Bien qu'il soit très utile pour faciliter les extractions dentaires, le bloc du nerf maxillaire à l'aveugle est associé à des complications potentielles dues à une ponction vasculaire involontaire. Ce rapport de cas décrit une complication rare de prolapsus de la troisième paupière chez un cheval après un bloc nerveux maxillaire et un traitement réussi par tarsorraphie temporaire.Message clinique clé:Ce rapport de cas explique comment un gonflement et un prolapsus de la membrane nictitante peuvent survenir à la suite d'un bloc du nerf maxillaire à l'aveugle chez le cheval et décrit le traitement par tarsorraphie temporaire.(Traduit par Dr Serge Messier).

Spring and fall blood concentrations of adrenocorticotropic hormone, insulin, and thyroxine in healthy horses in Saskatchewan.

Objective: To characterize concentrations of adrenocorticotropic hormone (ACTH), insulin, and thyroxine (T4) at 2 times of the year in healthy horses in Saskatchewan. Animals and procedure: A prospective, observational study was carried out in 52 healthy, client-owned horses. Inclusion criteria were no recent travel outside of Saskatchewan, normal physical examination findings, and no history or evidence of ongoing illness. Blood concentrations of ACTH, insulin and T4 were determined by chemiluminescence. Samples were collected in spring and fall and compared using the paired Student's t-test or Wilcoxon signed-rank test. Generalized estimating equations were used to assess the associations between ACTH, T4, and insulin concentrations and gender, age, season, body condition score, glucose concentration, and breed. Results: There were increases in both ACTH and insulin concentrations in the fall compared to spring (P < 0.001 and P = 0.001, respectively). Other than season, insulin concentration was associated with breed, whereas ACTH concentration was associated with age. Finally, T4 concentration was associated with breed and glucose concentration, but not with season. Conclusion and clinical relevance: These results highlighted the differences between spring and fall concentrations of both ACTH and insulin in healthy horses residing in the Canadian prairie provinces, which are known for extreme differences in summer and winter temperatures as well as day length. Geographically adjusted reference values are necessary to account for these variations, to improve diagnostic accuracy. This is the first published Canadian study evaluating these factors and their associations with hormone concentrations in clinically healthy animals.

Concentrations sanguines printanières et automnales d'hormone adrénocorticotrope, d'insuline et de thyroxine chez des chevaux en bonne santé en Saskatchewan. Objectif: Caractériser les concentrations d'hormone adrénocorticotrope (ACTH), d'insuline et de thyroxine (T4) à 2 moments de l'année chez des chevaux en bonne santé en Saskatchewan. Animaux et procédure: Une étude observationnelle prospective a été réalisée auprès de 52 chevaux en bonne santé appartenant à des clients. Les critères d'inclusion étaient l'absence de voyage récent à l'extérieur de la Saskatchewan, les résultats normaux de l'examen physique et l'absence d'antécédents ou de signes de maladie persistante. Les concentrations sanguines d'ACTH, d'insuline et de T4 ont été déterminées par chimiluminescence. Des échantillons ont été collectés au printemps et à l'automne et comparés à l'aide du test de Student apparié ou du test du rang de signe de Wilcoxon. Des équations d'estimation généralisées ont été utilisées pour évaluer les associations entre les concentrations d'ACTH, de T4 et d'insuline et le sexe, l'âge, la saison, l'état corporel, la concentration de glucose et la race. Résultats: Il y avait une augmentation des concentrations d'ACTH et d'insuline à l'automne par rapport au printemps (P < 0,001 et P = 0,001, respectivement). Hormis la saison, la concentration d'insuline était associée à la race, alors que la concentration d'ACTH était associée à l'âge. Enfin, la concentration de T4 était associée à la race et à la concentration en glucose, mais pas à la saison. Conclusion et pertinence clinique: Ces résultats ont mis en évidence les différences entre les concentrations printanières et automnales d'ACTH et d'insuline chez les chevaux en bonne santé résidant dans les provinces des Prairies canadiennes, reconnues pour leurs différences extrêmes de températures estivales et hivernales ainsi que de durée du jour. Des valeurs de référence géographiquement ajustées sont nécessaires pour tenir compte de ces variations et améliorer la précision du diagnostic. Il s'agit de la première étude canadienne publiée évaluant ces facteurs et leurs associations avec les concentrations d'hormones chez des animaux cliniquement sains.(Traduit par Dr Serge Messier).

Anaesthesia-related morbidity associated with recumbent, low-field magnetic resonance imaging of horses.

CASE HISTORY: Medical records from 2009 to 2021 from a private equine referral hospital in Rochester, NH, USA were analysed for cases that underwent general anaesthesia for low-field MRI of the distal limb. These were used to determine peri-anaesthetic morbidity and mortality. CLINICAL FINDINGS AND OUTCOME: Two hundred and forty-three anaesthetic episodes were recorded in horses undergoing low-field MRI. The peri-anaesthetic complication rate prior to discharge was 6.2% (15/243). No patients experienced a fatal complication. Ninety two of the 243 patients had multiple sites imaged, 90/243 received pre-anaesthetic dantrolene, 134/243 received intra-anaesthetic dobutamine, and 15/243 were positioned in dorsal recumbency. Complications included: abdominal discomfort ("colic"; 9/243), myopathy (4/243), hyphaema (1/243) and carpal fracture (1/243). At the time of discharge, 14/15 complications had resolved. Of 135 horses for which data were available 55 became hypotensive during the procedure (lowest mean arterial pressure < 65 mmHg). Median body weight was 553 (min 363, max 771) kg. Horses were anaesthetised for a median of 150 (min 45, max 210) minutes. There was no evidence of an association between higher body weight (p = 0.051) or longer duration of anaesthesia (p = 0.421) and development of an anaesthetic complication. For categorical variables (dantrolene administration pre-anaesthesia, dobutamine administration during anaesthesia, hypotension (mean < 65 mmHg) during anaesthesia, dorsal vs. lateral recumbency, and imaging of single vs. multiple sites), the 95% CI for the OR included 1, indicating a lack of effect of the variable on the odds of complication. CLINICAL RELEVANCE: The cases included in this series suggest that low-field MRI under general anaesthesia is a viable option for diagnostic imaging in otherwise healthy horses. Complications occur, but most resolve before discharge.

Equine-assisted services for people living with dementia: a systematic review.

BACKGROUND: Dementia has a significant impact on the social, physical, and psychological wellbeing of people living with dementia, their families and society. Animal-assisted interventions can have positive effects on the health and wellbeing of people living with dementia. Equine-assisted services are animal-assisted non-pharmacological interventions which have improved the health and wellbeing of diverse populations. The impact of participating in equine-assisted services on the health and wellbeing of people with dementia is unclear. A systematic review was conducted to synthesise evidence investigating the effects of participating in equine-assisted services on the health and wellbeing of people living with dementia. DESIGN: Systematic review following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. METHODS: The databases CINAHL, EMBASE, MEDLINE, and Web of Science were searched for any research published prior to 14 June 2023. Peer-reviewed publications in the English language utilizing methods deriving quantitative and/or qualitative data were eligible. Methodological quality of included studies was assessed using the Mixed Methods Appraisal Tool. Findings from studies were synthesised using a deductive approach. RESULTS: Of the 223 articles screened, six met the inclusion criteria: four quantitative and two qualitative studies. The six studies represent four separate equine interventions. Studies were of moderate to strong quality. Participants were people living with dementia (n = 44, mean age range 70-83 years), dementia care partners (n = 5, mean age 58), and equine-assisted services providers (n = 5). Interventions varied in duration, activities conducted, outcomes measured, and measurement tools used. Studies found a favourable impact of participating in equine-assisted services on the neuropsychiatric symptoms and quality of life of people living with dementia. Participating in equine-assisted services improved well-being, functional abilities, social participation, and communication, while also having a positive effect on social, emotional, and behavioural outcomes, and physical health. CONCLUSIONS: The limited but high-quality literature investigating the impact of equine-assisted services among people living with dementia suggests that equine-assisted services can have a positive impact on the health and wellbeing of people living with dementia. Additional robust studies contributing to the evidence base are warranted; such studies can support the development of programs and further elucidate the impact of participation.

Spatial clusters of human and livestock anthrax define high-risk areas requiring intervention in Lao Cai Province, Vietnam 1991-2022.

Anthrax, a widespread zoonosis in low and middle-income countries with low disease awareness and insufficient livestock vaccination coverage, has been known in Lao Cai Province in northern Vietnam for years before its apparent absence in 2009, which requires investigation as this infection is frequently reported from neighbouring provinces and countries. We aimed to describe the seasonal patterns of anthrax (1991-2008), compare livestock anthrax vaccine coverage to disease occurrence (1991- 2022), and delineate the high-risk areas to inform local disease surveillance in the province. We illustrated the seasonal pattern of anthrax and provided a comparison between livestock vaccine coverage and disease occurrence by purely spatial SaTScan (Poisson model, 25% population at risk) to detect spatial clusters of human and livestock anthrax using population derived from zonal statistics routines. The number of cases, crude cumulative incidence, and spatial clusters of human and livestock anthrax were mapped in QGIS. Results indicate peak anthrax incidence from May to October. Buffalo, domestic cattle, and horses accounted for 75% of total animal cases. Horse anthrax was more common in Lao Cai than in its neighbours and often occurred in years with human mortality. Vaccination covered less than 30% of the livestock population. We found an apparent pattern where anthrax was controlled from 1998-2003 with higher vaccine coverage (>20%) and identified spatial clusters of human and livestock anthrax in Muong Khuong, Bao Thang, and Bac Ha districts of Lao Cai. The local public health and veterinary agencies are recommended to revisit the high-risk areas and communicate with neighbouring provinces for a regional approach to anthrax surveillance and control.

Equine skeletal scintigraphy: Comparing lesion detection ability of methylene diphosphonate and hydroxymethylene diphosphonate in the caudal cervical and proximal metacarpal/metatarsal regions.

BACKGROUND: Data regarding the lesion detection ability of different radiotracers are lacking in equine bone scintigraphy. METHODS: In this prospective study, hydroxymethylene diphosphonate (HMDP) and methylene diphosphonate (MDP) were compared in horses with increased radiopharmaceutical uptake either in the caudal cervical region (CS group) or in the proximal metacarpal/metatarsal region (PMR group). Region of interest analysis was used to determine normal bone-to-soft tissue ratios, lesion-to-normal bone ratios and lesion-to-soft tissue ratios. Qualitative scoring and total count rates were recorded for each image. RESULTS: A total of 213 scintigrams were included. Within the PMR group, there were significantly higher lesion-to-normal bone ratios for MDP compared with HMDP (p = 0.02). In the CS group, normal bone-to-soft tissue ratios were significantly higher for HMDP (p = 0.01). The interobserver agreement with regard to the qualitative assessment of the scintigrams was poor. LIMITATION: Paired studies, comparing the different radiotracers in the same patient, were not feasible. CONCLUSION: This study revealed minor differences between the two radiotracers, although these have no practical implications. Both radiopharmaceuticals are well suited for detecting lesions at the investigated sites using equine bone scintigraphy.

Novel 6-alkyl-bridged 4-arylalkylpiperazin-1-yl derivatives of azepino[4,3-b]indol-1(2H)-one as potent BChE-selective inhibitors showing protective effects against neurodegenerative insults.

Due to the putative role of butyrylcholinesterase (BChE) in regulation of acetylcholine levels and functions in the late stages of the Alzheimer's disease (AD), the potential of selective inhibitors (BChEIs) has been envisaged as an alternative to administration of acetylcholinesterase inhibitors (AChEIs). Starting from our recent findings, herein the synthesis and in vitro evaluation of cholinesterase (ChE) inhibition of a novel series of some twenty 3,4,5,6-tetrahydroazepino[4,3-b]indol-1(2H)-one derivatives, bearing at the indole nitrogen diverse alkyl-bridged 4-arylalkylpiperazin-1-yl chains, are reported. The length of the spacers, as well as the type of arylalkyl group affected the enzyme inhibition potency and BChE/AChE selectivity. Two compounds, namely 14c (IC50 = 163 nM) and 14d (IC50 = 65 nM), bearing at the nitrogen atom in position 6 a n-pentyl- or n-heptyl-bridged 4-phenethylpiperazin-1-yl chains, respectively, proved to be highly potent mixed-type inhibitors of both equine and human BChE isoforms, showing more than two order magnitude of selectivity over AChE. The study of binding kinetics through surface plasmon resonance (SPR) highlighted differences in their BChE residence times (8 and 47 s for 14c and 14d, respectively). Moreover, 14c and 14d proved to hit other mechanisms known to trigger neurodegeneration underlying AD and other CNS disorders. Unlike 14c, compound 14d proved also capable of inhibiting by more than 60% the in vitro self-induced aggregation of neurotoxic amyloid-ß (Aß) peptide at 100 µM concentration. On the other hand, 14c was slightly better than 14d in counteracting, at 1 and 10 µM concentration, glutamate excitotoxicity, due to over-excitation of NMDA receptors, and hydrogen peroxide-induced oxidative stress assessed in neuroblastoma cell line SH-SY5Y. This paper is dedicated to Prof. Marcello Ferappi, former dean of the Faculty of Pharmacy of the University of Bari, in the occasion of his 90th birthday.

Expression and functional characterization of chimeric recombinant bovine follicle-stimulating hormone produced in Leishmania tarentolae.

Assisted reproductive techniques are routinely used in livestock species to increase and enhance productivity. Ovarian hyperstimulation is a process that currently relies on administering pituitary-derived follicle-stimulating hormone (FSH) or equine chorionic gonadotropin in combination with other hormones to promote the maturation of multiple follicles and thereby achieve superovulation. The use of partially purified preparations of FSH extracted from natural sources is associated with suboptimal and variable results. Recombinant FSH (rFSH) has been produced in a variety of heterologous organisms. However, attaining a bioactive rFSH of high quality and at low cost for use in livestock remains challenging. Here we report the production and characterization of a single chain bovine rFSH consisting of the ß- and α-subunit fused by a polypeptide linker (scbFSH) using Leishmania tarentolae as heterologous expression system. This unicellular eukaryote is non-pathogenic to mammals, can be grown in bioreactors using simple and inexpensive semisynthetic media at 26°C and does not require CO2 or bovine serum supplementation. Stable cell lines expressing scbFSH in an inducible fashion were generated and characterized for their productivity. Different culture conditions and purification procedures were evaluated, and the recombinant product was biochemically and biologically characterized, including bioassays in an animal model. The results demonstrate that L. tarentolae is a suitable host for producing a homogeneous, glycosylated and biologically active form of scbFSH with a reasonable yield.

Use of thermography in the long-term evaluation of scrotal surface temperature and its impact on seminal quality in stallions.

Scrotal surface thermography is a non-invasive method for assessing testicular thermoregulation in stallions; however, few studies have explored the application of this technique concerning the thermal physiology of equine reproductive systems. This study aimed to evaluate the consistency of testicular thermoregulation in stallions over a year using thermography to measure the scrotal surface temperature (SST). Moreover, we assessed the best region for measuring the surface body temperature compared with the SST. Ten light-breed stallions were used in the experiment. Thermographic images of the scrotal and body surfaces (neck and abdomen) were captured. Fresh, cooled and frozen-thawed semen samples were evaluated to verify the impact of thermoregulation on semen quality. Testicular thermoregulation was maintained throughout the year in stallions amidst changes in the external temperature, as evidenced by the weak correlation between the SST and ambient temperature. A lower correlation was observed between the environmental temperature and body surface temperature (BTS) obtained from the abdomen (BTS-A; R = .4772; p < .0001) than with that obtained from the neck (BTS-N; R = .7259; p < .0001). Moreover, both BTS-A and SST were simultaneously captured in a single image. The consistent quality of the fresh, cooled and frozen semen suggests efficient thermoregulation in stallions throughout the year.

Gross, organoleptic and histologic assessment of cadaveric equine heads preserved using chemical methods for veterinary surgical teaching.

BACKGROUND: Preservation of biological tissues has been used since ancient times. Regardless of the method employed, tissue preservation is thought to be a vital step in veterinary surgery teaching and learning. OBJECTIVES: This study was designed to determine the usability of chemically preserved cadaveric equine heads for surgical teaching in veterinary medicine. METHODS: Six cadaveric equine heads were collected immediately after death or euthanasia and frozen until fixation. Fixation was achieved by using a hypertonic solution consisting of sodium chloride, sodium nitrite and sodium nitrate, and an alcoholic solution containing ethanol and glycerin. Chemically preserved specimens were stored at low temperatures (2°C to 6°C) in a conventional refrigerator. The specimens were submitted to gross and organoleptic assessment right after fixative solution injection (D0) and within 10, 20, and 30 days of fixation (D10, D20, and D30, respectively). Samples of tissue from skin, tongue, oral vestibule, and masseter muscle were collected for histological evaluation at the same time points. RESULTS: Physical and organoleptic assessments revealed excellent specimen quality (mean scores higher than 4 on a 5-point scale) in most cases. In some specimens, lower scores (3) were assigned to the range of mouth opening, particularly on D0 and D10. A reduced the range of mouth opening may be a limiting factor in teaching activities involving structures located in the oral cavity. CONCLUSIONS: The excellent physical, histologic, and organoleptic characteristics of the specimens in this sample support their usability in teaching within the time frame considered. Appropriate physical and organoleptic characteristics (color, texture, odor, and flexibility) of the specimens in this study support the use of the method described for preparation of reusable anatomical specimens.

Predicted genetic burden and frequency of phenotype-associated variants in the horse.

Disease-causing variants have been identified for less than 20% of suspected equine genetic diseases. Whole genome sequencing (WGS) allows rapid identification of rare disease causal variants. However, interpreting the clinical variant consequence is confounded by the number of predicted deleterious variants that healthy individuals carry (predicted genetic burden). Estimation of the predicted genetic burden and baseline frequencies of known deleterious or phenotype associated variants within and across the major horse breeds have not been performed. We used WGS of 605 horses across 48 breeds to identify 32,818,945 variants, demonstrate a high predicted genetic burden (median 730 variants/horse, interquartile range: 613-829), show breed differences in predicted genetic burden across 12 target breeds, and estimate the high frequencies of some previously reported disease variants. This large-scale variant catalog for a major and highly athletic domestic animal species will enhance its ability to serve as a model for human phenotypes and improves our ability to discover the bases for important equine phenotypes.

Green treasures: Investigating the biodiversity potential of equine yards through the presence and quality of landscape features in the Netherlands.

At a time of mounting ecological crises and biodiversity loss, there is an urgent need for nature-based solutions. Equestrian properties cover a considerable proportion of the European rural and peri-urban landscape and provide much potential for integrating ecosystem services, such as the inclusion of small landscape features. The aim of this study was to investigate the presence and quality of landscape features (LF) to help determine how the equine sector can contribute to the agro-ecological transition. Using a citizen science approach, 87 commercial and 420 private yard owners reported the type, frequency and geometric dimension of LFs and additional biodiversity enhancing features. A hierarchical multivariate regression was used to determine how equine property characteristics explain variation in the Percentage Property Coverage (PPC) of LFs. The model explained 47% of the variation of PPC. The variables that explained significant variation in PPC included Yard size, Number of LFs, Tree rows, Fruit orchard, Wild hedges, Flowering strips, Buffer strips, Embankments and Cluttered corners. Commercial yards are significantly larger with significantly more horses and on average only 9% (±13.87%) of the property was covered by LFs whilst private yards had significantly more coverage of LFs with on average 12% (±14.77%). These findings highlight the substantial yet untapped potential of equine yards in fostering biodiversity, suggesting that the equine sector could play an important role in the agro-ecological transition. To encourage more biodiverse-inclusive yard designs, tailored strategies should consider the diverse factors influencing equine yard design, including existing knowledge, client demands, financial considerations, and equine health and welfare.

Integrated analysis of gut metabolome, microbiome, and exfoliome data in an equine model of intestinal injury.

BACKGROUND: The equine gastrointestinal (GI) microbiome has been described in the context of various diseases. The observed changes, however, have not been linked to host function and therefore it remains unclear how specific changes in the microbiome alter cellular and molecular pathways within the GI tract. Further, non-invasive techniques to examine the host gene expression profile of the GI mucosa have been described in horses but not evaluated in response to interventions. Therefore, the objectives of our study were to (1) profile gene expression and metabolomic changes in an equine model of non-steroidal anti-inflammatory drug (NSAID)-induced intestinal inflammation and (2) apply computational data integration methods to examine host-microbiota interactions. METHODS: Twenty horses were randomly assigned to 1 of 2 groups (n = 10): control (placebo paste) or NSAID (phenylbutazone 4.4 mg/kg orally once daily for 9 days). Fecal samples were collected on days 0 and 10 and analyzed with respect to microbiota (16S rDNA gene sequencing), metabolomic (untargeted metabolites), and host exfoliated cell transcriptomic (exfoliome) changes. Data were analyzed and integrated using a variety of computational techniques, and underlying regulatory mechanisms were inferred from features that were commonly identified by all computational approaches. RESULTS: Phenylbutazone induced alterations in the microbiota, metabolome, and host transcriptome. Data integration identified correlation of specific bacterial genera with expression of several genes and metabolites that were linked to oxidative stress. Concomitant microbiota and metabolite changes resulted in the initiation of endoplasmic reticulum stress and unfolded protein response within the intestinal mucosa. CONCLUSIONS: Results of integrative analysis identified an important role for oxidative stress, and subsequent cell signaling responses, in a large animal model of GI inflammation. The computational approaches for combining non-invasive platforms for unbiased assessment of host GI responses (e.g., exfoliomics) with metabolomic and microbiota changes have broad application for the field of gastroenterology. Video Abstract.

Immunogenic profile of a plant-produced nonavalent African horse sickness viral protein 2 (VP2) vaccine in IFNAR-/- mice.

A safe, highly immunogenic multivalent vaccine to protect against all nine serotypes of African horse sickness virus (AHSV), will revolutionise the AHS vaccine industry in endemic countries and beyond. Plant-produced AHS virus-like particles (VLPs) and soluble viral protein 2 (VP2) vaccine candidates were developed that have the potential to protect against all nine serotypes but can equally well be formulated as mono- and bi-valent formulations for localised outbreaks of specific serotypes. In the first interferon α/ß receptor knock-out (IFNAR-/-) mice trial conducted, a nine-serotype (nonavalent) vaccine administered as two pentavalent (5 µg per serotype) vaccines (VLP/VP2 combination or exclusively VP2), were directly compared to the commercially available AHS live attenuated vaccine. In a follow up trial, mice were vaccinated with an adjuvanted nine-serotype multivalent VP2 vaccine in a prime boost strategy and resulted in the desired neutralising antibody titres of 1:320, previously demonstrated to confer protective immunity in IFNAR-/- mice. In addition, the plant-produced VP2 vaccine performed favourably when compared to the commercial vaccine. Here we provide compelling data for a nonavalent VP2-based vaccine candidate, with the VP2 from each serotype being antigenically distinguishable based on LC-MS/MS and ELISA data. This is the first preclinical trial demonstrating the ability of an adjuvanted nonavalent cocktail of soluble, plant-expressed AHS VP2 proteins administered in a prime-boost strategy eliciting high antibody titres against all 9 AHSV serotypes. Furthermore, elevated T helper cells 2 (Th2) and Th1, indicative of humoral and cell-mediated memory T cell immune responses, respectively, were detected in mouse serum collected 14 days after the multivalent prime-boost vaccination. Both Th2 and Th1 may play a role to confer protective immunity. These preclinical immunogenicity studies paved the way to test the safety and protective efficacy of the plant-produced nonavalent VP2 vaccine candidate in the target animals, horses.

Expression pattern of germ cell markers in cryptorchid stallion testes.

Cryptorchidism affects spermatogenesis and testis development, often resulting in stallion subfertility/infertility. This study aims to identify the specific germ cells impacted by cryptorchism in stallions. In a previous study, we found that PGP9.5 and VASA are molecular markers expressed in different germ cells within stallions. Herein, we assessed the heat stress-induced response of spermatogonial stem cells (SSCs) in the seminiferous tubules (ST) of cryptorchid stallion testes (CST) and normal stallion testes (NST). This goal was accomplished by comparing PGP9.5 and VASA expression patterns through reverse transcription quantitative PCR and immunofluorescence assays. We also compared the cross-sectional ST area between groups. Six post-pubertal Thoroughbred unilateral cryptorchid stallions were used. The relative abundance of the mRNA transcripts of PGP9.5 and VASA was significantly upregulated in the NST group than in the CST group. Additionally, the cross-sectional ST area and localization of PGP9.5 and VASA in germ cells were significantly higher in the NST group than in the CST group. Regarding Leydig cells, PGP9.5 staining was observed in both groups. Spermatogonia, primary spermatocytes and secondary spermatocytes were immunostained with VASA in the NST group, while immunostaining was only observed in spermatogonia in the CST group. These results indicate long-term exposure to heat stress conditions, such as cryptorchidism, directly impacts germ cell proliferation and differentiation, leading to impaired spermatogenesis and compromised fertility in stallions.

Diet modulates strongyle infection and microbiota in the large intestine of horses.

The use of anthelminthic drugs has several drawbacks, including the selection of resistant parasite strains. Alternative avenues to mitigate the negative effects of helminth infection involve dietary interventions that might affect resistance and/or tolerance by improving host immunity, modulating the microbiota, or exerting direct anthelmintic effects. The aim of this study was to assess the impact of diet on strongyle infection in horses, specifically through immune-mediated, microbiota-mediated, or direct anthelmintic effects. Horses that were naturally infected with strongyles were fed either a high-fiber or high-starch diet, supplemented with either polyphenol-rich pellets (dehydrated sainfoin) or control pellets (sunflower and hay). When horses were fed a high-starch diet, they excreted more strongyle eggs. Adding sainfoin in the high-starch diet reduced egg excretion. Additionally, sainfoin decreased larval motility whatever the diet. Moreover, the high-starch diet led to a lower fecal bacterial diversity, structural differences in fecal microbiota, lower fecal pH, lower blood acetate, and lower hematocrit compared to the high-fiber diet. Circulating levels of Th1 and Th2 cytokines, lipopolysaccharides, procalcitonin, and white blood cells proportions did not differ between diets. Overall, this study highlights the role of dietary manipulations as an alternative strategy to mitigate the effect of helminth infection and suggests that, in addition to the direct effects, changes in the intestinal ecosystem are the possible underlying mechanism.

Bacteriological evaluation and advanced SYBR-green multiplex real-time PCR assay for detection of minced meat adulteration.

Background: Minced meat is a valuable source of nutrients, but it is vulnerable to contamination by microorganisms commonly present in the environment. In addition, there is a risk of adulteration with cheaper meat sources, which can be harmful to consumers. Aim: It is crucial to identify meat adulteration with distinct microbiological analysis for legal, economic, religious, and public health purposes. Methods: A total of 100 minced meat samples were collected from several markets in Sharkia Governorate, Egypt. These samples were then subjected to bacteriological testing and an advanced multiplex PCR method. This method enables the detection of bovine, equine, porcine, and dog species in meat samples with just one step. Results: The adulterated samples had a higher total bacterial count and pH values compared to pure bovine meat. These differences in bacterial count and pH values were statistically significant, with p-values of 0.843 (log10) and 0.233, respectively. The frequency of Escherichia coli occurrence was 13%, and the O111 serotype was predominant in the adulterated samples. Listeria monocytogenes and Staphylococcus aureus were isolated with prevalence rates of 3% and 29%, respectively. Besides, the SYBR-green multiplex real-time PCR assay used in this study detected adulteration with dog, equine, and porcine meats in the examined samples at rates of 9%, 5%, and 4%, respectively. Conclusion: This method provides a sensitive and specific approach to detect issues related to well-being and safety.