A review of genetic resources and trends of omics applications in donkey research: focus on China.

Omics methodologies, such as genomics, transcriptomics, proteomics, metabolomics, lipidomics and microbiomics, have revolutionized biological research by allowing comprehensive molecular analysis in livestock animals. However, despite being widely used in various animal species, research on donkeys has been notably scarce. China, renowned for its rich history in donkey husbandry, plays a pivotal role in their conservation and utilization. China boasts 24 distinct donkey breeds, necessitating conservation efforts, especially for smaller breeds facing extinction threats. So far, omics approaches have been employed in studies of donkey milk and meat, shedding light on their composition and quality. Similarly, omics methods have been utilized to explore the molecular basis associated with donkey growth, meat production, and quality traits. Omics analysis has also unraveled the critical role of donkey microbiota in health and nutrition, with gut microbiome studies revealing associations with factors such as pregnancy, age, transportation stress, and altitude. Furthermore, omics applications have addressed donkey health issues, including infectious diseases and reproductive problems. In addition, these applications have also provided insights into the improvement of donkey reproductive efficiency research. In conclusion, omics methodologies are essential for advancing knowledge about donkeys, their genetic diversity, and their applications across various domains. However, omics research in donkeys is still in its infancy, and there is a need for continued research to enhance donkey breeding, production, and welfare in China and beyond.

Is there sufficient evidence to support the health benefits of including donkey milk in the diet?

Donkey milk has attracted attention due to its distinctive nutritional composition and potential health advantages, particularly because of its whey protein content, which includes lysozyme, α-lactalbumin, lactoferrin, and ß-lactoglobulin and vitamin C, among other components. These elements contribute to immunoregulatory, antimicrobial, antioxidant, and anti-inflammatory properties, positioning donkey milk as a possible therapeutic option. In addition, due to the low levels of caseins, the casein-to-whey protein ratio, and the ß-lactoglobulin content in donkey milk, it presents an optimal alternative for infant formula for individuals with cow's milk allergies. Moreover, research into donkey milk's potential for cancer prevention, diabetes management, and as a treatment for various diseases is ongoing, thanks to its bioactive peptides and components. Nevertheless, challenges such as its low production yield and the not fully understood mechanisms behind its potential therapeutic role necessitate more thorough investigation. This review consolidates the existing knowledge on the therapeutic possibilities of donkey milk, emphasizing its importance for human health and the need for more detailed studies to confirm its health benefits.

Effect of silver nanoparticles on donkey sperm parameters and ultrastructure.

The aim of this study was to determine the effect of silver nanoparticles (AgNPs) on donkey sperm parameters and ultrastructure. AgNPs were synthesized, purified and resuspended in the extender. Nine frozen-thawed donkey sperm samples were exposed to different concentrations of AgNPs (0, 1.25, 2.5, 5, 12.5, 25 and 50 µg/mL). Sperm parameters: total (TMOT, %) and progressive (PMOT, %) sperm motility, plasma (LIVE, %) and acrosomal membrane integrity (AIS, %), and sperm morphology (MORF, %) were evaluated immediately after AgNPs exposure (T0) and after 2 h of incubation (T2). The interaction beween AgNPs and spermatozoa was visualized by transmission electron microscopy (TEM). At T0, sperm motility and AIS were reduced (p < .05) when using concentrations ≥50 and ≥25 µg/mL, respectively. At T2, sperm motility and LIVE were significantly decreased (p < .05) in concentrations ≥25 and ≥50 µg/mL, respectively. TEM analysis revealed nanoparticle adhesion to the acrosomal region of the plasma membrane. In conclusion, AgNPs at concentrations ≥25 µg/mL impair motility, acrosome and plasma membrane integrity of donkey sperm, which may be mediated by adhesion to the acrosomal region of the sperm surface membrane.

Properties and characteristics of steam-exploded donkey bone powder and corresponding whole wheat cookies.

Steam explosion was found to be the most effective preparation method for donkey bone powder, compared with non-treatment, autoclave and steam processing. And the effect of steam-exploded donkey bone powder (SEDBP) on the quality characteristics of whole wheat cookies was evaluated. Compared with untreated powder, particle size of SEDBP was reduced by 55.60 %, while oil- and water-holding capacity, water solubility and ABTS radical-scavenging rate significantly increased by 13.94 %, 26.86 %, 298.26 % and 91.47 %, respectively. Steam explosion enhanced protein digestibility from 32.17 % to 71.43 %, increased the Ca2+ release rate from 37.47 % to 47.82 %, and increased the release of 11 amino acids during digestion. SEDBP reduced the solvent retention capacity of flour blends and improved the color, hardness and spread ratio of whole wheat cookies. Cookies with 30 % SEDBP addition had better flavor, texture and scored the highest on overall acceptability. The results will help expand animal bone applications and develop nutrition-fortified foods.

Analytical Validation of Two Point-of-Care Assays for Hematological Analysis in the Miranda Donkey.

Miranda donkeys are an endangered autochthonous breed of Portugal. The HemoCue WBC DIFF and HemoCue Hb 201 portable analyzers, developed as a simplified alternative method for total and differential WBC count and hemoglobin measurement, respectively, may be relevant tools in veterinary practice. This study aimed to validate these instruments using Miranda donkey blood samples. For the HemoCue WBC DIFF, most parameters showed acceptable intra- and inter-assay precision with coefficients of variation (CV) below 5%, except for monocytes and eosinophils with higher CVs. The HemoCue Hb 201 showed CVs of 1.98% and 4.07%. Linearity correlation coefficients (r) ranged from 0.53 to 0.99 for HemoCue WBC DIFF and 0.99 for HemoCue Hb 201. Significant levels for neutrophils, lymphocytes, monocytes, eosinophils, and Hb measurements varied. Comparisons with ProCyte Dx showed an excellent correlation for WBC (rs = 0.96), neutrophils (rs = 0.91), lymphocytes (rs = 0.94), and eosinophils (rs = 0.90) but a poor correlation for monocytes and basophils. The HemoCue Hb 201 showed a correlation of rs = 0.96 with ProCyte Dx. In conclusion, both analyzers provided reliable results and are suitable for use in Miranda's donkey breed for WBC counts and Hb measurements.

Characterization and discrimination of donkey milk lipids and volatiles across lactation stages.

The lipid and flavor in milk are key factors that affect its quality, which profiles during donkey lactation are not yet clear. In this study, the lipids and volatile compounds (VOCs) in donkey milk from stages of lactation were analyzed by using LC-MS and GC-IMS. A total of 1774 lipids were identified in donkey milk, spanning over 6 major categories and attributed to 30 subclasses. The 233 differentially expressed lipids were identified between donkey colostrum and mature milk, which participate in 20 metabolic pathways, including glycerophospholipid, linoleic acid, and sphingolipid. Additionally, 35 VOCs in donkey milk were identified, including 28.57% aldehydes, 28.57% ketones, 25.71% esters, and 8.57% alcohols. Of these VOCs, 15 were determined to be characteristic flavors in donkey milk, mainly including methyl 2-methylbutanoate, 2-pentanone, and butyl acetate. 11 significantly different VOCs were found between the groups. Acetone, 2-heptanone, and ethyl acetate-m were considered potential discriminatory markers.

First Molecular Detection and Genetic Characterization of Tetratrichomonas buttreyi and Pentatrichomonas hominis in Donkeys in Shanxi Province, China.

Two species of trichomonads, Tetratrichomonas buttreyi and Pentatrichomonas hominis, are common intestinal parasites that can impact animal health and productivity. Severe infection by these parasites can lead to diarrhea and wasting in affected animals. Notably, P. hominis is known to cause diarrhea and has the potential to be transmitted between animals and humans. Donkeys hold significant economic importance in China's agricultural sector. However, whether donkeys are infected with T. buttreyi and P. hominis remains unknown globally. To address this gap in knowledge, 815 fecal samples were collected from donkeys in three representative regions in Shanxi Province, North China. Then, the presence and genetic characteristics of T. buttreyi and P. hominis were examined using species-specific PCR primers amplifying the small subunit ribosomal RNA genes. The overall prevalence was detected to be 25.4% (207/815) for T. buttreyi and 0.7% (6/815) for P. hominis in donkeys in Shanxi Province. All obtained P. hominis sequences were identified as genotype CC1. Genetic analysis revealed that all P. hominis isolates from donkeys were clustered into the same branch with isolates detected in humans, suggesting possible zoonotic transmission. This study is the first to report the occurrence and prevalence of T. buttreyi and P. hominis in donkeys globally. These findings expand the host range of trichomonads and improve our understanding of their genetic diversity and zoonotic potential, providing essential baseline data for the prevention and control of these parasites in donkeys in the region.

The Use of Horse and Donkey Meat to Enhance the Quality of the Traditional Meat Product (Kaddid): Analysis of Physico-Chemical Traits.

The aim of this study was to evaluate the use of horse and donkey meat in the production of kaddid-a traditional dish typically not made with these meats-from a physical and chemical perspective. The results showed that both meats exhibit similar water retention during cooking, contributing to comparable tenderness and juiciness, with no significant differences in pH values, indicating similar quality (p > 0.05). However, their amino acid profiles differ: horse meat contains lower levels of glutamate (p < 0.05), methionine (p < 0.01), isoleucine (p < 0.05), and leucine (p < 0.05), but higher levels of proline (p < 0.05), histidine (p < 0.01), and lysine (p < 0.001) compared to donkey meat. Both meats provide essential amino acids. Horse meat is richer in saturated and monounsaturated fatty acids (32.44% and 39.58%, respectively), while donkey meat has a higher content of polyunsaturated fatty acids (31.51%), with a more favorable PUFA/SFA ratio, suggesting better cardiovascular health benefits. In terms of dried meat, donkey kaddid has a higher protein (17.45 g/100 g) and lower fat content (2.1 g/100 g) compared to horse kaddid (16.7 g/100 g, and 3.5 g/100 g, respectively) (p < 0.05). These findings inform consumer choices and production practices, promoting the use of horse and donkey meat for kaddid production.

Preliminary study on heart response and locomotor parameters in Donkeys (Equus asinus) during exercise using fitness tracker (Equimetre).

The welfare of donkeys remains a compelling subject for researchers, with limited literature available on the response of the donkey cardiovascular system during strenuous exercise. The study aimed to address two primary objectives. Firstly, to assess the reliability of wearable devices in detecting heart rate (HR) and ECG readings. Secondly, to determine HR, locomotor and cardiac troponin 1 (cTnI) levels in donkeys during exercise. A total of seven donkeys were outfitted with two systems for ECG measurements, namely Equimetre and the Standard base apex, to enable a comparison between the two. Additionally, fifteen apparently healthy donkeys equipped with Equimetre were divided into two groups: the race group (R), consisting of donkeys trained for racing, and the non-race group (NR), comprising donkeys used for regular riding. The results indicated a level of agreement between the two devices in intervals R-R (P = < 0.0001), S-T (P = 0.0002), Q-T(P = 0.0003), P-R (P = 0.0037), segment P-R (P = 0.0023) and HR (P = < 0.0001) at rest. This suggested that Equimetre can provide a level of accepted ECG reading in donkey. No significant difference in heart response and locomotor parameters between donkey groups, although this finding needs further studies to verify it and to understand the dynamics of donkey. This study demonstrates the feasibility of Equimetre in detection HR and present initial data of heart response and locomotor in donkeys during exercise.

Detection of equine herpesvirus antibodies in large-scale donkey farms in Liaocheng area.

BACKGROUND: Equine herpesvirus (EHV) can cause respiratory, reproductive and neurological diseases in equine animals, including donkeys. The main pathogens responsible for these diseases are EHV type 1 (EHV-1) and EHV-4. In this study, we collected serum samples from 230 donkeys on 27 large-scale donkey farms to detect EHV-1 and EHV-4 antibodies. We analyzed the presence of EHV antibodies based on region, age and season. RESULTS: Out of the 27 farms, 62.96% (17/27) tested positive for EHV. Of the 230 donkeys tested, 2.61% (6/230) were positive only for EHV-1, 5.22% (12/230) were positive only for EHV-4, and 4.78% (11/230) were positive for both EHV-1 and EHV-4. The highest percentage of positive donkeys (21.28%) was found in Dong'e County. The seropositivity rate among donkeys aged 1-4 years was significantly higher compared to the group of donkeys aged 0-1 year (p < 0.05). Additionally, the positive rate was significantly higher in fall and winter compared to spring and summer (p < 0.05). CONCLUSIONS: Altogether, our findings indicate that large-scale donkey farms in the Liaocheng area have a high prevalence of EHV antibodies. Since Liaocheng is an important donkey trading market in Shandong Province, it is crucial to consider the risk of disease transmission based on our test results. This will help in early detection and prevention of EHV outbreaks.

Analysis of miRNAs in milk of four livestock species.

BACKGROUND: Milk is essential for mammalian nutrition because it provides vital nutrients for growth and development. Milk composition, which is influenced by genetic and environmental factors, supports lactation, a complex process crucial for milk production and quality. Recent research has focused on noncoding RNAs, particularly microRNAs (miRNAs), which are present in body fluids and regulate gene expression post-transcriptionally. This study comprehensively characterizes miRNAs in milk of four livestock species, namely Bubalus bubalis, Capra hircus, Equus asinus, and Ovis aries and identifies potential target genes. RESULTS: High-throughput sequencing of milk RNA resulted in distinct read counts across species: B. bubalis (8,790,441 reads), C. hircus (12,976,275 reads), E. asinus (9,385,067 reads), and O. aries (7,295,297 reads). E. asinus had the highest RNA mapping rate (94.6%) and O. aries the lowest (84.8%). A substantially greater proportion of miRNAs over other small RNAs was observed for the donkey milk sample (7.74%) compared to buffalo (0.87%), goat (1.57%), and sheep (1.12%). Shared miRNAs, which included miR-200a, miR-200b, miR-200c, and miR-23a among others, showed varying expression levels across species, confirmed by qPCR analysis. Functional annotation of predicted miRNA target genes highlighted diverse roles, with an enrichment in functions linked to metabolism and immunity. Pathway analysis identified immune response pathways as significant, with several miRNAs targeting specific genes across species, suggesting their regulatory function in milk. CONCLUSIONS: Both conserved and species-specific miRNAs were detected in milk of the investigated species. The identified target genes of these miRNAs have important roles in neonatal development, adaptation, growth, and immune response. Furthermore, they influence milk and meat production traits in livestock.

Ultrasonographic Assessment of Testicular Biometry and Arterial Blood Flow in Pre- and Postpubertal Miranda Donkeys: Correlations With Semen Quality Parameters.

The main goals of this study were to document and compare the normal ranges of testicular haemodynamic parameters in pre- (aged 8-12 months; n = 4) and postpubertal (aged 24-259 months; n = 16) Miranda donkeys in the breeding season, and to correlate animal biometric data and testicular Doppler indices with basic semen quality parameters of sexually mature jacks. Colour and pulsed-Doppler ultrasonography were employed to assess blood flow in the left and right distal supratesticular artery (DsTA) and their marginal branches (marginal arteries-MA). Peak systolic velocity (PSV), end diastolic velocity (EDV), pulsatility index (PI) and resistive index (RI) were evaluated in both blood vessels, and TAMV (time-averaged mean velocity), TABF (total arterial blood flow) and TABF rate (TABF-R) were calculated for MA. The mean diameter of MA was greater (p < 0.05; 0.24 ± 0.05 vs. 0.19 ± 0.05 cm; mean ± SD) but TABF-R was less (p < 0.05; 0.004 ± 0.004 vs. 0.02 ± 0.01 mL/s/cm3) in sexually mature donkeys compared with prepubertal animals. Apart from RI values for the right testicle of prepubertal donkeys, PI and RI were consistently greater (p < 0.05) in DsTA compared with MA. Significant correlations were found among select biometric and haemodynamic attributes of the testes (height, width and length, TV, TTV and PSV-ST) and ejaculate characteristics (volume, sperm defects-total, head and midpiece) in sexually mature donkeys (n = 8). The present results highlight the importance of scrotal ultrasonography for the reproductive assessment of jacks and provide reference values, based on the available subpopulation of Miranda donkeys that can be used in their clinical and reproductive management and research, or conservation programmes.

Fabrication and characterization of edible Pickering emulsion stabilized by donkey myofibrillar protein.

This study aims to investigate the properties of edible Pickering emulsions (PEs) stabilized by donkey myofibrillar protein (DMP). The DMP was characterized by an atomic force microscope and interfacial tensiometer. The PEs stabilized by DMP were characterized by a confocal laser scanning microscope, rheometer, and lumisizer stability analyzer. The results showed that the DMP particles were spherical nanoparticles with an average size of 143.97 nm. The DMP could reduce the oil-water interfacial tension very well. The emulsifying activity index was below 14.06 m2/g, and the emulsion stability index was up to 93.76% when the DMP concentration was above 10 mg/mL. Increasing the concentration of DMP (5 mg/mL to 25 mg/mL) could decrease the emulsion droplet sizes and fluidity of the PEs and increase the viscoelasticity of the PEs. The increase in the oil-water ratio (1:9 to 5:5) resulted in a larger average emulsion droplet size, reduced fluidity, and enhanced viscoelasticity. The increase in DMP concentration (from 5 mg/mL to 25 mg/mL) and oil-water ratio (from 1:9 to 4:6) improved the physical stability of the PEs. These results may expand the application of DMP and provide new insight into developing edible PEs suitable for the formulation of functional foods.

Factors affecting the quality and nutritional value of donkey meat: a comprehensive review.

Donkey meat is characterized by a high content of proteins, essential amino acids, and unsaturated fatty acids and is low in fat, cholesterol, and calories. Thus, it is considered a high-quality source of meat. Based on the data from PubMed and Web of science within past 10 years, this review summarizes the factors affecting the quality of donkey meat and its nutritional value, including breed, genetics, gender, age, muscle type, feeding regimen, storage and processing conditions. Breed, gender, age, and feeding regimen mainly affect the quality of donkey meat by influencing its intramuscular fat content and carcass quality. Meanwhile, the tenderness and flavor of donkey meat depend on the muscle type, storage and processing conditions. Genetics, on the other hand, fundamentally affect donkey meat quality by influencing the polymorphism of genes. These findings provide valuable insights and guidance for producers, consumers, and decision-makers in the donkey meat industry, promoting the development of more effective marketing strategies and the improvement of meat quality, thereby enabling the expansion and progress of the entire industry.

Effect of thermal and non-thermal processing methods on the Structural and Functional Properties of Whey Protein from Donkey Milk.

This study evaluated the impact of thermal, ultrasonication, and UV treatment on the structural and functional properties of whey proteins from donkey milk (DWP). Whey proteins exhibited notable stability in non-heat-treated environments, while their structural and functional characteristics were notably impacted by excessive heat treatment. The application of high-temperature long-time thermal treatment (HTLT) resulted in a decrease in fluorescence intensity, foaming and emulsification stability, and considerable damage to the active components of the proteins. Specifically, the preservation of lysozyme activity was only 23%, and lactoferrin and immunoglobulin G exhibited a significant loss of 70% and 77%, respectively. Non-thermal treatment methods showed superior efficacy in preserving the active components in whey proteins compared with heat treatment. Ultrasonic treatment has demonstrated a notable capability in diminishing protein particle size and turbidity, and UV treatment has been observed to have the ability to oxidize internal disulfide bonds within proteins, consequently augmenting the presence of free sulfhydryl groups, which were beneficial to foaming and emulsification stability. This study not only offers a scientific basis for the processing and application of DWP but also serves as a guide to produce dairy products, aiding in the development of dairy products tailored to specific health functions.

Analgesic Effect of Dexmedetomidine-Nalbuphine Combination vs. Dexmedetomidine Alone in Donkeys Undergoing Field Castration under Total Intravenous Anesthesia.

This study evaluated the antinociceptive effect of dexmedetomidine-nalbuphine vs. dexmedetomidine alone in jacks undergoing field castration under total intravenous anesthesia. Jacks were premedicated with intravenous (IV) dexmedetomidine (5 µg/kg), either alone (Group D, n = 6) or in combination with 0.3 mg/kg nalbuphine (Group DN, n = 6). IV ketamine (1.5 mg/kg) and propofol (0.5 mg/kg) were used to induce general anesthesia, which was maintained by a continuous propofol (0.2 mg/kg/min) IV infusion. The quality of anesthesia, analgesia, and recovery were evaluated. A simple descriptive scale (SDS) was used to measure pain from the recovery time to 6 h later. The DN group exhibited improvements in analgesic and recovery quality and SDS of pain at 1-, 2-, and 3-h post-recovery. There was an apparent improvement in cardiac status, as evidenced by the enhanced heart rate and electrocardiogram findings compared to group D during surgery and recovery time. The DN group had a lower level of inflammatory cytokines, both during the surgery and shortly after recovery. Therefore, the dexmedetomidine-nalbuphine combination prior to IV anesthesia of ketamine and propofol in jacks undergoing field castration resulted in a stable surgical plane of anesthesia, improved antinociception, less pain postoperatively, and better cardiac stability.

Reproductive Disorders in Donkeys: Current Evidence and Update.

Reproductive disorders in donkeys present a significant challenge to their health and welfare, impacting their roles in agriculture, conservation, and companionship. With the development of large-scale donkey farming in recent years, reproductive disorders have become a limiting factor for the expansion of the donkey population. In general, donkeys suffer from a similar array of diseases like horses, but little is known about the specificities of donkey reproductive disorders. This review synthesizes current knowledge on the pathogenesis, distribution, presentation, diagnosis, treatment, and prognosis of a diverse array of reproductive disorders affecting donkeys. There are similar infectious and non-infectious causes for infertility and pregnancy loss in jennies compared with mares, but a difference in disease susceptibility does exist, which may be attributed to genetic influence, pathogen specificity, the environment, and reproductive management practices. Diagnostic and treatment plans need to be tailored towards the particularities of donkey reproductive disorders to increase donkey populations and to enhance the standard of care for this species. Enhancing reproductive health in donkeys not only ensures their sustainable use but also promotes their welfare and longevity in diverse human-animal interactions.

Twin Pregnancy in the Martina Franca Donkey Breed Managed by Natural Reduction and Post-Fixation Manual Crushing.

In Equids, undetected twin pregnancy represents the most important cause of abortion and is also associated with high neonatal mortality rates. Therefore, the detection and management of twin pregnancies is pivotal to allow the continuation of single pregnancies. Although pre-fixation manual crushing of one embryo is the most common management of twin pregnancies, and the impact of natural reduction has been reported in mares, very little is known about donkeys. The present study aimed to report results on the natural reduction occurrence and post-fixation manual crushing management of twin pregnancies in the Martina Franca donkey breed. METHODS: Twenty-five twin pregnancies were detected at 11-13 days after ovulation. At 16 days after ovulation, twin pregnancies were classified as unilateral or bilateral and left untreated. The occurrence of natural reduction of one embryo was assessed at 20-21 days after ovulation, and post-fixation manual crushing of one embryo was performed in those cases in which natural reduction did not occur. The pregnancy rate at 28 days after ovulation and live foal rate were recorded. RESULT: Sixteen out of 25 twin pregnancies were unilateral and nine bilateral. At 20-21 days after ovulation, the natural reduction of one embryo occurred in 87.5% of the unilateral and in none of the bilateral twin pregnancies. The remaining twin pregnancies were treated by post-fixation manual crushing of one embryo. The 28-days-after-ovulation single embryo pregnancy rate was 93.8% for unilateral twin pregnancies and 88.9% for bilateral twin pregnancies, with an overall pregnancy rate of 92%. The live foal rate was 87%. CONCLUSIONS: The success rate of natural reduction within 20-21 days after ovulation, the 28-days-after-ovulation pregnancy rate, and the live foal rate suggest that waiting for the natural reduction of one embryo and choosing the post-fixation manual crushing of one embryo could be considered as an alternative to traditional early pre-fixation manual crushing in the Martina Franca donkey breed.

Detection of Gastrointestinal Pathogens with Zoonotic Potential in Horses Used in Free-Riding Activities during a Countrywide Study in Greece.

The objectives of this study were (a) to detect zoonotic gastrointestinal pathogens in faecal samples of horses using the FilmArray® GI Panel and (b) to identify variables potentially associated with their presence. Faecal samples collected from 224 horses obtained during a countrywide study in Greece were tested by means of the BioFire® FilmArray® Gastrointestinal (GI) Panel, which uses multiplex-PCR technology for the detection of 22 pathogens. Gastrointestinal pathogens were detected in the faecal samples obtained from 97 horses (43.3%). Zoonotic pathogens were detected more frequently in samples from horses in courtyard housing (56.0%) than in samples from horses in other housing types (39.7%) (p = 0.040). The most frequently detected zoonotic pathogens were enteropathogenic Escherichia coli (19.2% of horses) and Shiga-like toxin-producing E. coli stx1/stx2 (13.8%). During multivariable analysis, two variables emerged as significant predictors for the outcome 'detection of at least one zoonotic pathogen in the faecal sample from an animal': (a) the decreasing age of horses (p = 0.0001) and (b) the presence of livestock at the same premises as the horses (p = 0.013). As a significant predictor for the outcome 'detection of two zoonotic pathogens concurrently in the faecal sample from an animal', only the season of sampling of animals (autumn) emerged as significant in the multivariable analysis (p = 0.049). The results indicated a diversity of gastrointestinal pathogens with zoonotic potential in horses and provided evidence for predictors for the infections; also, they can serve to inform horse owners and handlers regarding the possible risk of transmission of pathogens with zoonotic potential. In addition, our findings highlight the importance of continuous surveillance for zoonotic pathogens in domestic animals.

Interventional Effect of Donkey Bone Collagen Peptide Iron Chelate on Cyclophosphamide Induced Immunosuppressive Mice.

Immunodeficiency can disrupt normal physiological activity and function. In this study, donkey bone collagen peptide (DP) and its iron chelate (DPI) were evaluated their potential as immunomodulators in cyclophosphamide (Cytoxan®, CTX)-induced Balb/c mice. The femoral tissue, lymphocytes, and serum from groups of mice were subjected to hematoxylin and eosin (H&E) staining, methylthiazolyldiphenyl-tetrazolium bromide (MTT) cell proliferation assays, and enzyme-linked immunosorbent assay (ELISA), respectively. Furthermore, a non-targeted metabolomics analysis based on UPLC-MS/MS and a reverse transcription polymerase chain reaction (RT-qPCR) technology were used to explore the specific metabolic pathways of DPI regulating immunocompromise. The results showed that CTX was able to significantly reduce the proliferative activity of mouse splenic lymphocytes and led to abnormal cytokine expression. After DP and DPI interventions, bone marrow tissue damage was significantly improved. In particular, DPI showed the ability to regulate the levels of immune factors more effectively than Fe2+ and DP. Furthermore, metabolomic analysis in both positive and negative ion modes showed that DPI and DP jointly regulated the levels of 20 plasma differential metabolites, while DPI and Fe2+ jointly regulated 14, and all 3 jointly regulated 10. Fe2+ and DP regulated energy metabolism and pyrimidine metabolism pathways, respectively. In contrast, DPI mainly modulated the purine salvage pathway and the JAK/STAT signaling pathway, which are the key to immune function. Therefore, DPI shows more effective immune regulation than Fe2+ and DP alone, and has good application potential in improving immunosuppression.