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Metabolomics can allow for the comprehensive identification of metabolites within biological systems, at given time points, in physiological and pathological conditions. In the last few years, metabolomic analysis has gained popularity both in human and in veterinary medicine, showing great potential for novel applications in clinical activity. The aim of applying metabolomics in clinical practice is understanding the mechanisms underlying pathological conditions and the influence of certain stimuli (i.e., drugs, nutrition, exercise) on body systems, in the attempt of identifying biomarkers that can help in the diagnosis of diseases. Proton Nuclear Magnetic Resonance spectroscopy (1H-NMR) is well tailored to be used as an analytical platform for metabolites' detection at the base of metabolomics studies, due to minimal sample preparation and high reproducibility. In this mini-review article, the scientific production of NMR metabolomic applications to equine medicine is examined. The research works are very different in methodology and difficult to compare. Studies are mainly focused on exercise, reproduction, and nutrition, other than respiratory and musculoskeletal diseases. The available information on this topic is still scant, but a greater collection of data could allow researchers to define new reliable markers to be used in clinical practice for diagnostic and therapeutical purposes. [ABSTRACT FROM AUTHOR]

Summary Background Objectives Study design Methods Results Main limitation Conclusion Long‐toe, underrun heel conformation is a term referring to a long toe, collapsed frog and atrophied heels in equids, including donkeys. However, data on the radiological evaluation and classification of the long‐toe, underrun heel conformation in the donkey are relatively limited.The present study aimed to determine the relationship between the dorsal hoof wall angle and the radiological angular and linear features of the distal phalanx in long‐toe, underrun heel conformation in Egypt Baladi donkeys.Post‐mortem collected feet.Forty‐seven front hooves with long‐toe, underrun heels from cadavers were radiographically examined. Lateromedial radiometric parameters included dorsal hoof wall angle, heel angle, distal phalanx angle, distal phalanx solar border angle, coronet angle, axis of the distal phalanx angle, proximal palmar cortex angle, distal phalanx to ground distance, distal phalanx to frog distance, distal phalanx to toe distance, foot length and hoof balance%. Long‐toe, underrun heel cadaver feet were classified into four groups according to the dorsal hoof wall angle (DHWA).The DHWA of the first group was less than 58.25°–54° (<58.25°:54°), the second group was less than 54°–52° (<54°:52°), the third group was less than 52°–47° (<52°:47°), and the fourth group was less than 47° (<47°). From the first to the fourth groups DHWA, heel angle (HA), distal phalanx solar border angle (P3BA), distal phalanx angle (P3A), axis of the distal phalanx angle (LP3A), proximal palmar cortex angle (PPCA), distal phalanx to frog distance (P3F) and foot length (FL) were decreased, while the coronet angle (CA), distal phalanx to toe distance (P3T) and hoof balance (HB) percentages were increased. Statistical analysis revealed strong and moderate correlations between DHWA and HA, P3A, P3BA, LP3A, PPCA, P3G, P3F and P3T.Cadaver feet were collected from different yards and body sizes, the collection of radiographic parameters without trimming and the lack of a record of the work history of these donkeys.This study will assist veterinarians in evaluating the long‐toe, underrun heel conformation during hoof trimming. [ABSTRACT FROM AUTHOR]

Helminths are widespread infectious agents, causing severe economic and performance losses in all animals, especially donkeys. Strongylus (S.) vulgaris and Fasciola (F.) hepatica are abundantly present in the equines. Their control using herbal is being necessitated because of the resistance of parasites towards routinely used chemicals. In this study, 60 positive animals were divided equally into 4 groups, each having three replicates named A, B, C, and D. Donkeys in groups A, B, and C received 1200, 1000, and 800 mg/kg mixture of 4 plants of family Apiaceae while the group D was kept as non-medicated control. The medication of the animals was done for 4 weeks. Fecal, hematological, and hepato-renal function-related serum samples were collected on days 0, 14, and 28 of the experiment to estimate the effects of herbal mixture on fecal egg count reduction, red blood cell counts, white blood cells, hemoglobin, packed cell volume, alanine aminotransferase, aspartate transferase, serum proteins, serum albumins, urea, and creatinine. The donkeys were weighed at the start and end of the trial to estimate the effect of the herbal mixture on weight gain. The results suggested that total helminth, S. vulgaris, and F. hepatica egg counts were significantly (p<0.05) less than the control group. Herbal mixture significantly (p<0.05) improved the red blood cells, packed cell volume, hemoglobin, total proteins, albumins, and weight gains of the donkeys treated with 1200 mg/kg herbal mixture. The results suggested that the herbal mixture from the plants of the family Apiaceae was effective in controlling the helminths and improving hematological and serological parameters. [ABSTRACT FROM AUTHOR]

The donkey's extraordinary capacity to endure substantial loads over long distances while maintaining equilibrium suggests a distinctive cerebellar architecture specialized in balance regulation. Consequently, our study aims to investigate the intricate histophysiology of the donkey's cerebellum using advanced ultrastructural and immunohistochemical methodologies to comprehend the mechanisms that govern this exceptional ability. This study represents the pioneering investigation to comprehensively describe the ultrastructure and immunohistochemistry within the donkey cerebellum. Five adult donkeys' cerebella were utilized for the study, employing stains such as hematoxylin, eosin, and toluidine blue to facilitate a comprehensive histological examination. For immunohistochemical investigation, synaptophysin (SP), calretinin, and glial fibrillary acidic protein were used and evaluated by the Image J software. Furthermore, a double immunofluorescence staining of SP and neuron‐specific enolase (NSE) was performed to highlight the co‐localization of these markers and explore their potential contribution to synaptic function within the donkey cerebellum. This investigation aims to understand their possible roles in regulating neuronal activity and synaptic connectivity. We observed co‐expression of SP and NSE in the donkey cerebellum, which emphasizes the crucial role of efficient energy utilization for motor coordination and balance, highlighting the interdependence of synaptic function and energy metabolism. The Purkinje cells were situated in the intermediate zone of the cerebellum cortex, known as the Purkinje cell layer. Characteristically, the Purkinje cell's bodies exhibited a distinct pear‐like shape. The cross‐section area of the Purkinje cells was 107.7 ± 0.2 µm2, and the Purkinje cell nucleus was 95.7 ± 0.1 µm2. The length and diameter of the Purkinje cells were 36.4 × 23.4 µm. By scanning electron microscopy, the body of the Purkinje cell looked like a triangular or oval with a meandrous outer surface. The dendrites appeared to have small spines. The Purkinje cells' cytoplasm was rich with mitochondria, rough endoplasmic reticulum, ribosomes, Golgi apparatus, multivesicular bodies, and lysosomes. Purkinje cell dendrites were discovered in the molecular layer, resembling trees. This study sheds light on the anatomical and cellular characteristics underlying the donkey's exceptional balance‐maintaining abilities. Highlights: The donkey's remarkable ability to endure substantial loads over long distances while maintaining equilibrium suggests the presence of a specialized cerebellar architecture dedicated to balance regulation.The objective of this study is to investigate the intricate histophysiology of the donkey's cerebellum using advanced ultrastructural and immunohistochemical methodologies, aiming to comprehend the mechanisms governing this exceptional balancing ability.This study pioneers the comprehensive description of the ultrastructure and immunohistochemistry within the donkey cerebellum, representing a groundbreaking investigation in the field.Five adult donkeys' cerebella were utilized in this study, employing staining techniques such as hematoxylin, eosin, and toluidine blue to facilitate a comprehensive histological examination.Immunohistochemical investigation involved synaptophysin, calretinin, and glial fibrillary acidic protein, evaluated using Image J software, providing valuable insights into their potential roles in regulating neuronal activity and synaptic connectivity.Double immunofluorescence staining of synaptophysin and neuron‐specific enolase highlighted their co‐localization and potential contribution to synaptic function within the donkey cerebellum.Scanning electron microscopy revealed that Purkinje cells had a triangular or oval shape with a meandering outer surface, and the dendrites appeared to have small spines.Transmission electron microscopy showed that the cytoplasm of Purkinje cells contained various organelles, including mitochondria, rough endoplasmic reticulum, ribosomes, Golgi apparatus, multivesicular bodies, and lysosomes.In conclusion, this study sheds light on the anatomical and cellular characteristics underlying the donkey's exceptional balance‐maintaining abilities. [ABSTRACT FROM AUTHOR]

Most of the northern hemisphere donkey breeds are faced with the risk of extinction, thus donkey reproduction is considered an emerging branch of theriogenology, and the management of artificial insemination and induction of ovulation is a crucial point in setting up preservation protocols. For four consecutive cycles, six jennies' ovarian activity was routinely monitored; an ultrasound examination was performed daily from the evidence of a follicle diameter =27 mm until ovulation. Upon reaching a follicular diameter =32 ± 2 mm (Hour 0), oestrous jennies were treated alternatively with 0.1 mg triptorelin acetate, sc, (TRIP), 0.4 mg/sc of buserelin acetate (BUS) or saline, sc, (CTRL) and examined ultrasonographically at Hours 14, 24, 38, 42, 48, 62 and every 24 h until ovulation. Induction of ovulation was considered successful if ovulation occurred from 24 to 48 h after treatment. 11/12 cycles resulted in ovulation for TRIP and 12/12 for BUS and CTRL groups, respectively. Mean ± SD ovulation time after treatment was 37.3 ± 8.3, 47.1 ± 21.0 and 66.8 ± 25.9 h for BUS, TRIP and CTRL, respectively (p = .0032). Ovulation rates between h24 and h48 were 10/12 (83.3%) for both TRIP/BUS and 2/12 (16.7%) for CTRL, respectively (p = .003). Buserelin and triptorelin-treated jennies had a 25 times higher probability to ovulate between Hours 24 and 48 than controls (p = .003), while there were no jenny and cycle effects on the ovulatory rate. The results of this study show how triptorelin successfully induced ovulation in jennies, like other GnRH analogues previously evaluated. [ABSTRACT FROM AUTHOR]

Enrofloxacin is a broad-spectrum antimicrobial agent, but the study of its pharmacokinetics/pharmacodynamics (PKs/PDs) in donkeys is rarely reported. The present study aimed to investigate the pharmacokinetics of enrofloxacin administered intragastrically, and to study the pharmacokinetics of enrofloxacin and its metabolite ciprofloxacin in plasma, urine, and feces, and the PK/PD parameters were investigated to provide a rationale for enrofloxacin treatment in donkeys. A total of five healthy donkeys were selected for intragastric administration of 7.5 mg·kg−1 BW of enrofloxacin by gavage, and blood, urine, and fecal samples were collected. The results showed that the elimination half-life of plasma enrofloxacin was 11.40 ± 6.40 h, Tmax was 0.55 ± 0.12 h, Cmax was 2.46 ± 0.14 mg·L−1, AUC0–∞ was 10.30 ± 3.37 mg·L−1·h, and mean residence time (MRT) was 7.88 ± 1.26 h. The Tmax of plasma ciprofloxacin was 0.52 ± 0.08 h, Cmax was 0.14 ± 0.03 mg·L−1, and AUC0–∞ was 0.24 ± 0.16 mg·L−1·h. Urinary Cmax was 38.18 ± 8.56 mg·L−1 for enrofloxacin and 15.94 ± 4.15 mg·L−1 for ciprofloxacin. The total enrofloxacin and ciprofloxacin recovered amount in urine was 7.09 ± 2.55% of the dose for 144 h after dosing. The total enrofloxacin and ciprofloxacin recovered amount in feces was 25.73 ± 10.34% of the dose for 144 h after dosing. PK/PD parameters were also examined in this study, based on published MICs. In conclusion, 7.5 mg/kg BW of enrofloxacin administered intragastrically to donkeys was rapidly absorbed, widely distributed, and slowly eliminated in their bodies, and was predicted to be effective against bacteria with MICs < 0.25 mg·L−1. [ABSTRACT FROM AUTHOR]

The involvement of Helicobacter-like agents in EGGD is not clear. Much evidence supports the presence of Helicobacter in the horse's stomach, but it is not so clear about the donkey. The present study was conducted to evaluate the presence of Helicobacter in the stomach of donkeys and assess the best method for its identification and its possible participation in EGGD and serum antioxidant status. Gastric juice sampling and biopsy from NMP and PA were performed by gastroscopic method from 12 donkeys. Histopathology and RUT were used to identify Helicobacter. TAC, THIOL, NO, and DPPH were measured to assess the antioxidant status of serum. Helicobacter was detected only by the RUT method in one donkey (8.33%), but it is unlikely that this infection was effective in causing EGGD. The RUT results for the three samples of gastric juice, NMP, and PA were not different. Histopathology of NMP and PA did not show Helicobacter infection. Helicobacter is present in the stomach of a donkey but does not change the antioxidant status of serum. The RUT is more efficient than H&E histopathology for the assessment of H. pylori in the equine stomach, and the RUT of gastric juice is preferable to tissue samples because it is easy, fast, and non-invasive. In conclusion, further studies using more accurate methods are recommended to evaluate the effect of Helicobacter in the gastric diseases of donkeys and other equine. [ABSTRACT FROM AUTHOR]

The microanatomy of the donkey eye is important to understand because pathological disorders affecting them are relatively common. The current study aimed to document the cellular components of donkey's retinae using light and electron microscopic studies. Ten donkey retinae were dissected and processed for semi-thin sections and electron microscopic studies. The photoreceptor layer was made up of the outer and inner segments of rods and cones. The outer segments were filled with invaginations of cell membranes that form stacks of membranous disks. Shed discs of photoreceptor outer segments could be seen in the photoreceptor layer as well as near the Müller cells. The inner segments of cones were conical in shape, while those of rods were slim rod-shaped. Both were filled with long thin mitochondria and free ribosomes. Three rows of photoreceptor cell nuclei made up the outer nuclear layer. The rod nuclei had more electron-dense chromatin than those of the cones. There were two rows of cell nuclei in the inner nuclear layer that represent the following four cell classes: horizontal cells, bipolar cells, amacrine cells, and Müller cells. Bipolar cells constitute the bulk of the inner nuclear layer. They were elongated in shape and had thick branched dendrites. Amacrine cells were in the inner face of the INL. It could be observed within IPL and known as displaced amacrine cells. Muller glial cells were irregular elongated in shape with many cytoplasmic processes. They were distributed in the INL among the bipolar cells. Müller cells were observed in the inner plexiform layer, the ganglion cell layer, and the nerve fiber layer. In conclusion, this study characterized the detailed cytological organization and ultrastructure of the healthy donkey retina. which maintains the fundamental laminar architecture characteristic of other mammalian retinas, and consists of 10 distinguishable layers. When compared to previously described retinal morphologies in domestic species, some distinctive characters were observed in donkey retinal cells. [ABSTRACT FROM AUTHOR]

Background: Millions of working equids provide socio-economic support for many low-income communities worldwide. With the prevalence of harness-related wounds reported as higher than 60%, this study aims to describe the equipment used by working equids in three locations of the Ethiopian national regional state of Oromia (Fiche, Bishoftu and Shashamene), and the attitudes and practices of equid owners, users and harness makers regarding work equipment. This mixed-methods study consists of cross-sectional surveying of working equids used for taxi or transport of goods or water, as well as cart-driver questionnaires and focus groups (FG) with working equid stakeholders. Activities conducted with FG included participatory ranking of equipment attributes and equipment drawing exercises. Indicators of equipment design and assembly, as well as cart-driver attitudes and practices were described quantitatively. Associations between equipment characteristics and species, work-type and cart-driver indicators were investigated through univariable logistic regression models, whereas focus group discussions were transcribed and analysed thematically. Results: In total, 368 working equid surveys and cart-driver questionnaires were completed and 87 participants took part in nine FG. Equipment composition and characteristics differed considerably from ideal animal draught and harnessing principles described in the literature, with none of the observed harnesses adhering to all principles and thus not considered fully adequate. Various harness compositions were used, with only saddles and breast collars present in all. Donkey equipment had fewer components than that of horses, such as swingle trees (OR 0.02; 95% CI 0.01–0.06; p < 0.001) or girths (OR 0.09; 95%CI 0.02–0.4; p = 0.002). Horse equipment was more likely to have functional elements such as breeching (OR 7.8; 95% CI 2.9–20.9; p < 0.001). Of all equipment attributes, FG participants ranked cost, design and impact on animal wellbeing as having the highest importance. Thematic analysis identified motivations and priorities regarding equipment; awareness and knowledge of design and function; barriers to using ideal equipment; and consequences of equipment design and practices as key themes. Conclusions: Various weaknesses of design, assembly and use of equid work equipment were identified. Promoting behavioural change through awareness and training could lead to a sustainable improvement of work equipment quality, access, and prevention of equipment-related problems. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate carcass and meat quality traits in Masri (n = 14) and North African (n = 14) male donkeys, raised in a semi-intensive breeding system, grazing on mountainous areas, with supplementation of 1 kg of barley per day per animal, slaughtered at 5 (±0.48) years old. Compared to Masri, the North African population exhibited higher (p < 0.05) final body weights (181.7 ± 2.1 and 212.5 ± 7.6 kg) and cold carcass weights (101.7 ± 1.3 and 116.2 ± 4.5 kg), but lower (p < 0.05) cold dressing percentages (56.0 ± 0.4 and 54.6 ± 0.4%). Meat quality analyses showed higher (p < 0.05) cooking loss values in Masri meat (43.9 ± 0.8 vs. 39.9 ± 1.2%). Among the meat color parameters, the hue value was higher (p < 0.05) in North African samples (0.42 ± 0.01 vs. 0.39 ± 0.1). Meat from North African donkeys had higher (p < 0.05) dry matter, fat, and protein contents. Meat amino acid analysis revealed abundant levels of lysine, leucine, and methionine, in both populations. Donkey meat from both populations presented a high polyunsaturated fatty acids content, resulting in polyunsaturated fatty acids/saturated fatty acids and omega 6/omega 3 ratios for all breeds close to the recommended values for human health. Atherogenic and thrombogenic indices were also suggested to have positive effects on consumers’ artery health. The characteristics of donkey meat present intriguing nutritional aspects compared to ruminant meat, and its production should be encouraged in the rural development of inner-Mediterranean areas. [ABSTRACT FROM AUTHOR]

Objectives: The aim of the present study was to examine donkey sperm quality after intratesticular injection of hypertonic mannitol (HM) and saline (HS). Methods: Randomly assigned to five treatment groups were 15 adult male donkeys: (1) Control group (no treatment), (2) Surgery group (surgical castration for testosterone control), (3) NS group (normal saline intratesticular injection), (4) HS group (hypertonic saline), and (5) HM group. We injected 20 mL per testicle. We took 5 mL blood from all donkeys before injection. Castration was performed under general anesthesia 60 days later. Samples included blood and testicular tissue. Total motility (TM), progressive motility (PM), movementy features, DNA damage, morphology, viability, and plasma membrane functionality were evaluated. Hormone analyses, histomorphometric studies and oxidative stress indices including total antioxidant capacity (TAC), glutathione peroxidase (GPx), glutathione (GSH), superoxide dismutase (SOD), malondialdehyde (MDA), and NADP+/NADPH were evaluated. Apoptosis, pyroptosis-related Bax, Caspase-1, GSDMD, and Bcl-2 expression were also assessed. Results: In HS and HM groups, testosterone, epididymal sperm count, motility, viability, and plasma membrane functionality dropped while sperm DNA damage increased. HS and HM groups had significantly lower histomorphometric parameters, TAC, GPx, SOD, GSH, and Bcl-2 gene expression. MDA, NADP+/NADPH, Bax, Caspase-1, and GSDMD gene expression were substantially higher in the HS and HM groups than in the control group. Conclusions: Toxic effects of hypertonic saline and mannitol on reproductive parameters were seen following, hence, they might be considered as a good chemical sterilizing treatment in donkeys. [ABSTRACT FROM AUTHOR]

Simple Summary: Equids often live and work below welfare standards till exhaustion and die painfully. Euthanasia for humane reasons should be considered in some situations. Scientific evidence on the most common diseases of working equids is still limited. This study aimed to describe the population and disorders of the equids that arrived at the Egypt Equine Aid clinic (Cairo, Egypt) from 2019 to 2022. Secondly, it aimed to find any possible association between physical parameters at admission and outcome (i.e., discharged, euthanasia, or dead). The results of this study showed that working equids suffered mainly from wounds, orthopaedic problems, colics, and infectious diseases. In addition, in the univariable ordinal regression, some easy-to-assess physical parameters, namely the abnormal colour of the mucous membrane, highly increased capillary refill time, and increased heart rate, were associated with the non-survival outcome (i.e., euthanasia or dead). In conclusion, owner education and timely medical assistance in case of need remain the central focus to ensure a life worth living for working equids. This study to our best knowledge represents the first one to describe the disease prevalence of the working equid population around the Cairo area and to identify critical points to improve their quality of life. Working equids are often used to exhaustion, living and dying in conditions below minimal welfare standards. Due to their poor welfare status, euthanasia should be considered in certain conditions. The study aimed to describe the population and the disease frequency of the working equids admitted at an equine clinic in Cairo (i.e., Egypt Equine Aid (EEA)) from 2019 to 2022 and identify possible associations between physical parameters at admission and the outcome. Records of 1360 equids admitted at EEA were reviewed. The majority of the admitted equids were horses (65.6%), followed by donkeys (33%), in particular stallions (68.7%), from 1 to 15 years old (74.8%). Hospitalisation was mainly due to wounds (28.9%), orthopaedic problems (27.4%), colic (8.5%), or infectious diseases (7.4%). The majority of the equids were discharged, but 5.1% died on their own, without human intervention, and 23% were euthanised. Text mining revealed the anamnesis's most frequent words were 'accident', 'lameness', and 'wound'. In addition, owners sometimes reported using inappropriate remedies (e.g., firing) before hospitalisation. Multivariable ordinal regression analysis performed between physical parameters and the outcome (ordered based on severity: discharged, euthanasia, and dead) revealed that sex (male vs. female: OR = 1.33; p < 0.05), colour of the mucous membrane (pathological vs. physiological: OR = 1.72; p < 0.01), and capillary refill time (pathological vs. physiological: OR = 1.42; p = 0.02) increased the likelihood of a non-survival outcome. In conclusion, early euthanasia should be considered for these equids, to minimise prolonged suffering. Moreover, owners' education is recommended to guarantee minimal welfare standards to the working equids. [ABSTRACT FROM AUTHOR]

Summary: This is the first known case report of encephalitic listeriosis in a donkey (Equus asinus). A 5‐day‐old female donkey foal presented for fever, progressive weakness and loss of suckle reflex. Upon arrival, its vital parameters were normal, but the foal was collapsed. It had horizontal nystagmus, front limb paddling, minimal response to stimuli, prolapse of the left third eyelid and curling the neck to the left when in sternal recumbency. Samples collected for diagnostic evaluation included blood and cerebral spinal fluid. The cerebrospinal fluid (CSF) showed a pleocytosis consisting mainly of nondegenerative neutrophils and a normal protein concentration. Bacteria were not seen on Gram stain. After 24 h of treatment with intravenous fluids, anti‐inflammatories, antibiotics, nasogastric feedings of the dam's milk and supplemental nasal oxygen, little improvement was observed and the owners elected to euthanise the foal. Post‐mortem examination revealed multifocal, irregularly distributed hepatic necrosis and multifocal lymphohistiocytic encephalitis. Listeria monocytogenes was isolated from a section of brainstem. [ABSTRACT FROM AUTHOR]

Various ophthalmic disorders (conjunctivitis, corneal wounds, keratitis) have been reported in donkeys. There are no studies on the effect of medetomidine or tramadol on Schirmer tear test (STT) readings in donkeys. This prospective study investigated changes in STT readings in 24 clinically healthy donkeys (Equus asinus) (14 geldings and 10 mares) treated with commonly used doses of medetomidine hydrochloride and tramadol hydrochloride as mono- or combined therapy. Analgesia, sedation, ataxia, and STT readings were measured before treatment (baseline) and at different periods after administration (5-120 min) of the specific drug in each group. Tramadol monotherapy induced a mild analgesic effect (score 1) at 10 min post-administration. All treated donkeys exhibited mild to moderate ataxia. Medetomidine alone or in combination with tramadol induced a significant decrease (P<0.05) in the STT readings in both right and left eyes at 5, 15, 30, and 60 min relative to baseline, and the lowest values were observed 60 min after drug administration in both groups. Intravenous administration of medetomidine alone or in combination with tramadol induced a significant reduction in STT readings in clinically healthy donkeys. Therefore, in donkeys, the ocular surface treated with these sedatives should be carefully examined and adequately covered by an artificial tear solution or ophthalmic gel. [ABSTRACT FROM AUTHOR]

Abstract Animals adjust their habitat use patterns in response to changes in their physiological needs and environmental conditions. Understanding the mechanisms underlying resource selection and space use across time and space reveals effects of the environment on animals' decisions. We explored the effects of habitat availability and heterogeneity on the seasonal and annual space use and resource selection of two free‐roaming feral burro (Equus asinus) populations in the United States within distinct climate and habitat conditions: the Sonoran Desert and the Colorado Plateau. As an introduced yet protected species in the United States, understanding burros' interactions with habitat elements is important for their conservation and management, as well as the conservation of sympatric wildlife. We used GPS locations of female burros (72 animals across both study areas) to delineate annual and seasonal ranges and resource selection patterns. We evaluated effects of mean and CV of habitat covariates, including forage, distance to water, and topography, representing availability and heterogeneity of resources, on seasonal and annual range size of burros. Moreover, we explored how burro seasonal and annual resource selection patterns were affected by availability and heterogeneity of resources. In the Sonoran Desert study area, burros had smaller seasonal and annual ranges and constant resource selection patterns across a year, likely due to a freshwater lake in the area, making water a nonlimiting resource. Human presence was the greatest factor affecting range size and resource selection in the Sonoran Desert, where burros selected for areas near roads and human recreation. In the Colorado Plateau study area, where resources were more seasonal, we found larger range sizes and fluctuating resource selection patterns compared to the Sonoran Desert population. Spatial variation in forage, water, and topography significantly affected range size of burros inhabiting the Colorado Plateau study area. Productive habitats with available water support smaller ranges and a more consistent pattern of resource selection. Our results highlight the positive effect of habitat heterogeneity and the negative effect of habitat productivity on range size of animals. Our findings contribute to an improved understanding of habitat requirements for free‐roaming burros that currently live under various climate and habitat conditions globally.

This study investigated the description and feasibility of a surgical procedure for enucleation–exenteration of the equine eye via the supraorbital fossa. A preliminary study was performed on both eyes of four cadaveric heads of native breed donkeys (Equus asinus) to describe the surgical anatomy and demonstrate a new supraorbital enucleation surgical approach. For the clinical study, eight donkeys were admitted for unilateral enucleation. All procedures were performed in a lateral recumbent position under the influence of inhalation anesthesia in combination with a retrobulbar nerve block. A semi-circular incision was made in the skin and fascia of the supraorbital fossa to gain access into the orbital cavity, after which the periorbital fat was dissected and removed. Bleeding was controlled by electrocautery and large blood vessels were ligated, then the eyeball was dissected sharply and freed from its bony attachment. The procedure was successfully accomplished in all clinical cases, and no significant complications occurred during or throughout the postoperative follow-up period. The initial results suggest the feasibility and safety of the supraorbital enucleation technique described in this study for equine eye enucleation. This new technique seems promising due to its feasibility, safety, and positive outcomes observed in both cadaveric and clinical studies.

Abstract Background Lymphoma is a common neoplasm in horses but is reported much less commonly in donkeys. In this case report, we describe the macroscopic, microscopic and immunohistochemical features of a multicentric lymphoma with intestinal and bone marrow involvement. Case presentation A geriatric female donkey with history of chronic lameness was found dead. Post-mortem examination revealed advanced emaciation, periodontal disease, left front foot laminitis and multiple, soft, white to yellow tan intestinal transmural masses, up to 12 cm in diameter. Cytology suggested a round cell intestinal neoplasm. The femur of the left hint limb was double the size of the normal contralateral, with suspected neoplastic infiltration and replacement of bone marrow and bone. Histologically we diagnosed a lymphoma in the intestine and left femur. Immunohistochemically, the neoplastic cells showed CD3 immunolabelling, supporting a diagnosis of a multicentric T-cell lymphoma. Conclusions To the authors’ knowledge, this is the first time multicentric lymphoma is diagnosed in donkeys. Further studies of the genetic background, clinical, laboratory, histopathologic, and immunohistochemical, as well as the pathogenesis of lymphoma, is needed to better understand the uniquely low frequency of this neoplasm in donkeys.

The surging demand for donkey milk has led to a stronger focus on donkey husbandry and reproductive management, particularly in the peripartum period. Techniques to predict parturition are needed to avoid losses due to dystocias, which are frequently related to unattended foaling. In mares, pH and electrolyte analyses of pre-colostrum are used to predict parturition with a good accuracy. The purpose of this study was to evaluate the pH of the mammary secretion using both a pH metre and test strips and the electrolytes (Na+, K + and Ca2+) in the mammary gland secretions of 9 jennies during the 5 days preceding parturition, to understand if the trends of these cations were comparable to those of the mare and to identify possible markers/thresholds of impending parturition. Ca2+ concentrations increased in the days before delivery from day −2 (p

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.

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 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.

Background: Lymphoma is a common neoplasm in horses but is reported much less commonly in donkeys. In this case report, we describe the macroscopic, microscopic and immunohistochemical features of a multicentric lymphoma with intestinal and bone marrow involvement. Case presentation: A geriatric female donkey with history of chronic lameness was found dead. Post-mortem examination revealed advanced emaciation, periodontal disease, left front foot laminitis and multiple, soft, white to yellow tan intestinal transmural masses, up to 12 cm in diameter. Cytology suggested a round cell intestinal neoplasm. The femur of the left hint limb was double the size of the normal contralateral, with suspected neoplastic infiltration and replacement of bone marrow and bone. Histologically we diagnosed a lymphoma in the intestine and left femur. Immunohistochemically, the neoplastic cells showed CD3 immunolabelling, supporting a diagnosis of a multicentric T-cell lymphoma. Conclusions: To the authors' knowledge, this is the first time multicentric lymphoma is diagnosed in donkeys. Further studies of the genetic background, clinical, laboratory, histopathologic, and immunohistochemical, as well as the pathogenesis of lymphoma, is needed to better understand the uniquely low frequency of this neoplasm in donkeys. [ABSTRACT FROM AUTHOR]

The epididymis, a key component of the male reproductive system, controls spermatozoa's maturation, fertility, and storage. The objective of this study is to evaluate the histological, ultrastructural, and immunohistochemical variations in the epididymis of donkeys that occur throughout the year. During the breeding season (spring) and nonbreeding seasons (summer, autumn, and winter), 20 epididymis were collected from adult, clinically healthy donkeys. Compared to non‐breeding seasons, the epididymal duct displayed a more active lining epithelium and more sperm in the lumen during the breeding season. The epithelial height is the lowest and the lumen is the widest during the breeding season. Furthermore, the epididymal epithelium in the tail region exhibits undulations with polyps‐like projections. The epididymal epithelium is composed mainly of the principal, basal, and dark cells. Tight junction between adjacent principal cells is more obvious in the breeding season as compared to the non‐breeding seasons. However, intraepithelial lymphocytes, phagocytic, and other immune cells are more frequent in non‐breeding seasons. β‐catenin, which is a component of the adherent junctions between adjacent PCs, exhibits more immunoreactivity during the spring. On the other hand, iNOS, an indicator of oxidative stress, reacts positively during the summer. Additionally, during non‐breeding seasons, autophagy was detected within the epididymal epithelium which may be linked to stress adaptation. In conclusion, our findings suggest that the histological and ultrastructural characteristics of the epididymal epithelium are more active during spring compared to other seasons of the year. Research Highlights: The study aimed to evaluate the histological, ultrastructural, and immunohistochemical variations in the blood epididymal barrier (BEB) and epididymal epithelium of donkeys that occur throughout the year.In comparison to non‐breeding seasons, the epididymal duct displayed a more active lining epithelium and more sperm in the lumen during the breeding season.The epithelial height is the lowest and the lumen is the widest during the breeding season.The epididymal epithelium in the tail region exhibits undulations with polyps‐like projections that increase the surface area.β‐catenin, which is a component of the adherent junctions between adjacent PCs, exhibits more immunoreactivity during the spring. On the other hand, iNOS, an indicator of oxidative stress, reacts positively during the summer. [ABSTRACT FROM AUTHOR]

Florfenicol (FF) is a commonly used antibacterial agent in animals. We investigated the pharmacokinetics of FF and its metabolite florfenicol amine (FFA) in donkeys. Donkeys were administered FF (30 mg/kg bodyweight, p.o.). Pharmacokinetic parameters were calculated using a non-compartmental model. The FF (FFA) pharmacokinetics parameters were characterized by along elimination half-life (t1/2 kz) of 5.92 h (15.95 h), plasma peak concentration (Cmax) of 0.13 μg/mL (0.08 μg/mL), and the time taken to reach Cmax (Tmax) of 0.68 h (0.72 h). The area under plasma concentration-time curve and mean residence time of FF (FFA) in plasma were 1.31 μg·mL-1·h (0.47 μg·mL-1·h) and 10.37 h (18.40 h), respectively. The t1/2 kz of FF and FFA in urine was 21.93 and 40.26 h, and the maximum excretion rate was 10.56 and 4.03 μg/h reached at 25.60 and 32.20 h, respectively. The respective values in feces were 0.02 and 0.01 μg·h-1 reached at 33.40 h. The amount of FF and FFA recovered in feces was 0.52 and 0.22 μg, respectively. In conclusion, FF (FFA) is rapidly absorbed and slowly eliminated after a single oral administration to donkeys. Compared to FF, FFA was more slowly eliminated. FF (FFA) is mostly excreted through urine. [ABSTRACT FROM AUTHOR]

The application of embryo recovery and transfer technology in the donkey industry is far lower than that of horses and cattle. Sometimes the recovered embryos could not be transferred in time, which required embryo cryopreservation. The embryo cryopreservation technology is more conducive to the preservation and transportation of recovered embryos with excellent genetic traits. However, this technique for donkey embryos is not efficient and needs further optimization. The objective of this study was to evaluate the effect of different manual puncture methods on the viability and pregnancy rates of vitrified-thawed donkey embryos. A total of 117 donkey embryos were recovered on day 7–8 post-ovulation and were divided into four groups by random assortment. There were 28 embryos without puncture or cryopreservation (Control). 58 embryos were manually punctured using a 29G needle (VG, n = 29) or microneedle with a sharp tip of <10 μm (VM, n = 29), then vitrified in 15% EG + 15% DMSO + 0.5 M sucrose. Another 31 embryos were punctured with a microneedle and vitrified with 10% EG + 10% DMSO +0.5 M sucrose +2 mol/L proline (VMP). Both fresh embryos and vitrified-thawed embryos were incubated for 3 h (38.5 °C, 5% CO 2 in air) before embryo transfer. The results showed that the embryo recovery rates on day 7.5 and 8 were higher than day 7 (P < 0.05). After incubation, dead cells were assessed and the percentages of dead cells in VM and VMP were lower than that in VG (P < 0.05), although both were higher than those in Control (P < 0.05). The pregnancy rates after 23 days post transfer were assessed and the results showed that the pregnancy rate in VG (8.0%) was lower than that in Control (41.7%), VM (24.0%) and VMP (29.6%) (P < 0.05). No pregnancies resulted from the 10 embryos with diameters ≤650 μm in VG, which lower than either VM (33.3%) or VMP (38.9%) (P < 0.05). While, there was no difference in pregnancy rates among all vitrification groups when embryos were >650 μm in diameter (P > 0.05). In conclusion, the embryo recovery rate on day 7 after ovulation was relatively low, and it was more appropriate to extend it to day 8. Microneedle puncture could reduce embryo damage and achieve a higher pregnancy rate compared with 29G needles. Proline has the potential to improve donkey embryo cryopreservation. • Donkey embryos could be vitrified successfully after puncture with the needle under stereomicroscope. • The vitrified-thawed embryos showed higher viability and pregnancy rates when puncture using a microneedle. • Proline has potential to improve donkey embryo cryopreservation. [ABSTRACT FROM AUTHOR]

Simple Summary: In recent years, with the expansion of animal husbandry, the competition for grain between its use as feedstuffs for animals and as foodstuffs for humans has become increasingly prominent. Traditional feeds such as corn, soybean, and alfalfa are no longer sufficient to meet the growing needs of the livestock industry. Consequently, it is imperative to develop novel, unconventional feedstuffs. Broussonetia papyrifera (BP) emerges as a promising substitute, being a woody feedstuff with wide environmental adaptability, high yield, and high protein content. The purpose of this experiment was to study the apparent digestibility and the effects of Broussonetia papyrifera (BP) branch/leaf powder supplementation on growth performance and serum indicators in donkeys. The results showed that the apparent digestibility of dry matter (DM), crude protein (CP), crude fiber (CF), neutral detergent fiber (NDF), acidic detergent fiber (ADF), and digestible energy content (DE) of BP branch/leaf powder were 51.88%, 67.27%, 64.86%, 49.59%, 54.73%, 40.87%, and 6.37 MJ/kg, respectively. The average daily gain (ADG) in the 20% group was significantly higher than in the 0% and 30% groups. The serum albumin (ALB) levels in the 0% and 10% groups were significantly higher than those in the 20% and 30% groups, while the serum globulin (GLB) content in the 10% group was significantly lower than in the other groups. The 20% group showed decreased serum triglyceride (TG) levels compared to the other groups. Both the 20% and 30% groups exhibited lower total cholesterol (TC) levels and increased alanine aminotransferase (ALT) compared to the 0% and 10% groups and higher serum lactate dehydrogenase (LDH) levels than the 10% group. The 30% group had higher serum immunoglobulin A (IgA) levels than the other groups, while all three BP branch/leaf powder groups had lower serum tumor necrosis factor (TNF-α) levels than the 0% group. There was a gradual increase in serum total antioxidant capacity (T-AOC) with the increasing amount of BP branch/leaf powder added. In conclusion, the optimal supplemental proportion of BP branch/leaf powder in the diet is 20%. Furthermore, BP branch/leaf powder can improve growth performance, serum immune indices, and antioxidant capacity in Dezhou donkeys. [ABSTRACT FROM AUTHOR]

Zearalenone (ZEN) is a mycotoxin produced by various Fusarium strains, that is present in food and feed raw materials worldwide, causing toxicity effects in animals and humans. This research aimed to explore the toxicokinetics of ZEN on female Dezhou donkeys following a single oral exposure dosage of 2 mg/kg BW (body weight). The sample collection of donkeys plasma was carried out at 0, 5, 10, 15, 20, 30, 45, 60, 90 min, 2 h, 2.5 h, 3 h, 3.5 h, 4 h, 4.5 h, 6 h, 9 h, 12 h, 24 h, 48 h, 72 h, 96 h and 120 h via intravenous catheter, and fecal and urinary samples were severally collected at 0 h and every 6 h until 120 h. The concentrations of ZEN, α-zearalenol (α-ZOL), β-zearalenol (β-ZOL), α-zearalanol (α-ZAL), β-zearalanol (β-ZAL), zearalanone (ZAN) in plasma, urine, and feces were detected by UPLC-MS/MS. Only ZEN was detected in plasma, and the maximum was 15.34 ± 5.12 µg/L occurred at 0.48 h after gavage. The total plasma clearance (Cl) of ZEN was 95.20 ± 8.01 L·kg·BW−1·h−1. In addition, the volume of distribution (Vd) was up to 216.17 ± 58.71 L/kg. The percentage of total ZEN (ZEN plus the main metabolites) excretion in feces and urine was 2.49% and 2.10%, respectively. In summary, ZEN was fast absorbed and relatively slowly excreted in female donkeys during 120 h after a single gavage, indicating a trend of wider tissue distribution and longer tissue persistence. [ABSTRACT FROM AUTHOR]

Enterocytozoon bieneusi and Blastocystis may cause diarrhea in humans and various animals. However, little information is available regarding the prevalence and genetic diversity of E. bieneusi and Blastocystis in donkeys. To fill this gap, we molecularly assessed E. bieneusi and Blastocystis in fecal samples from donkeys (n = 815) in Shanxi Province, north China. The overall prevalence of E. bieneusi and Blastocystis in donkeys was 8.1% and 0.2%, respectively. Region and age were risk factors associated with E. bieneusi infection in donkeys. Three internal transcribed spacer (ITS) genotypes of E. bieneusi were identified in the current study, including two previously described genotypes (D and Henan-IV) and one novel genotype (named SXD1). Of which, genotype D was found to be the most prevalent. Phylogenetic analysis demonstrated that the three genotypes belonged to group 1, implying a potential of zoonotic transmission. Multilocus sequence typing showed that 19, 15, 13, and 22 types were identified at the loci MS1, MS3, MS4, and MS7, respectively, forming six multilocus genotypes (MLGs) distributed in the genotype D. One Blastocystis subtype (ST33) was identified, which has previously been reported only in horses. This is the first molecular-based description of E. bieneusi and Blastocystis infections in donkeys in Shanxi Province, north China, contributing to a better understanding of transmission dynamics and molecular epidemiological characteristics of the two intestinal protozoa. [ABSTRACT FROM AUTHOR]

Equines' ability in racing and riding as well as gaitedness have influenced the human civilization. Aim of this study was to identify and characterize the novel polymorphisms or SNPs in DMRT3 gene in Indian horse and donkey breeds. In this study, the DMRT3 gene was sequenced and characterized in 72 Indian horses' and 33 Indian donkeys' samples. One SNP (A > C) at 878 was found in studied horses while identical SNPs (A > C) at two different nucleotide positions i.e., 878 and 942 in DMRT3 gene (chromosome 23) were observed in studied Indian donkey breeds. Horses and donkeys both have a non-synonymous mutation (A > C) at nucleotide 878 (codon 61) that converts a Stop codon (TAG > TCG) to coding codon Serine, whereas donkeys have a synonymous mutation at nucleotide 942 (codon 82) that converts Serine (TCA > TCC) into Serine. A phylogenetic tree indicated that the DMRT3 gene was equally distributed among the equine breeds. Most of the donkey breeds have been shown high levels of genetic diversity while horse breeds and Halari donkey showed the least genetic diversity. Mutation in DMRT3 has a major impact on gaitedness in horses and is presented at a high frequency in gaited breeds and in horses breed for harness racing. [ABSTRACT FROM AUTHOR]

Simple Summary: Giardia duodenalis is a common zoonotic intestinal protozoan and causes significant economic losses and potential risks to public health. China is among the several major producers of donkeys worldwide, with approximately 2.6 million donkeys being reared in 2019, but studies on G. duodenalis infection in donkeys were limited. Thus, in order to better understand the molecular epidemiology of G. duodenalis in donkeys in China, 815 fecal samples were collected from donkeys in three representative geographical locations in Shanxi Province, North China, to determine the presence and genotypes of G. duodenalis based on three established loci (tpi, bg, and gdh). A total of 137 out of 815 fecal samples were detected as G. duodenalis-positive with an overall prevalence of 16.81%. Among the detected assemblages A, B, and E, the most predominant assemblage was B in the three study areas. Three MLGs (MLG-novel-1 to 3) were identified with multilocus genotypes (MLGs) analysis and validated by a phylogenetic tree. This is the first report of G. duodenalis infection in donkeys in Shanxi Province, which not only enhances our understanding of the genetic diversity of G. duodenalis in donkeys in China but also provides essential baseline data for the prevention and control of G. duodenalis infection donkeys in the study areas. Giardia duodenalis is a ubiquitous flagellated protozoan, causing significant economic losses to animal husbandry and posing threats to public health. China ranks the world's sixth largest major producer of donkeys, rearing approximately 2.6 million donkeys in 2019, but limited investigation of G. duodenalis prevalence has been conducted in the past, and it is yet to be known whether donkeys in Shanxi Province are infected with G. duodenalis. In the present study, a total of 815 fecal samples collected from donkeys in representative regions of Shanxi Province, North China, were examined for G. duodenalis using nested PCR. Then, the assemblages and multilocus genotypes (MLGs) were examined based on three established loci: namely, β-giardin (bg), triosephosphate isomerase (tpi), and glutamate dehydrogenase (gdh). The overall prevalence of G. duodenalis in donkeys in Shanxi Province was 16.81% (137/815). The region was identified as the main risk factor for the observed difference in G. duodenalis prevalence in donkeys among the three study areas (χ2 = 21.611, p < 0.001). Assemblages A, E, and B were identified, with the latter as the predominant assemblage. Three MLGs (MLG-novel-1 to 3) were formed based on sequence variation among the three loci. The present study reveals the presence of G. duodenalis in donkeys in Shanxi Province, North China, for the first time, which not only enriches the data on the distribution of G. duodenalis in donkeys in China but also provides useful baseline data for planning control strategies against G. duodenalis infection in the sampled areas. [ABSTRACT FROM AUTHOR]

Domestic donkeys (Equus asinus) have been maintained mainly for service purposes in the past. Nowadays, there is an increasing interest in donkey milk and meat production in several countries, including China. Donkey meat is highly consumed because of its nutritional value and unique flavor. However, genomic studies on donkey meat are limited. Therefore, in this study, we aimed to examine the molecular difference of longissimus dorsi muscles of donkey, cow, and goat. RNA sequencing and Proteome sequencing technology were used to analyze the transcriptome and proteome of the longissimus dorsi muscle of donkey, cow, and goat. A total of 1338 and 1780 differentially expressed genes (DEGs) were identified in donkey meat compared with that in cow and goat meat, respectively. Most of the DEGs were involved in biological processes, including small GTPase-mediated signal transduction, protein ubiquitination, protein glycosylation, and MAP kinase tyrosine/serine/threonine phosphatase activity. Additionally, 764 and 1024 differentially expressed proteins (DEPs) were identified in cow vs. donkey, and goat vs. donkey, respectively; these DEPs were mainly involved in metabolism. Genetic variation and regulatory factors can combine as a database to provide more valuable molecular information for further analysis. [ABSTRACT FROM AUTHOR]

Long non-coding RNAs (lncRNAs) play a critical role in the development of muscles. However, the role of lncRNAs in regulating skeletal muscle development has not been studied systematically in the donkey. In this study, we performed the RNA sequencing for different stages of muscles in donkeys, and investigate their expression profile, which showed that 3215 mRNAs (p-adjust <0.05) and 471 lncRNAs (p-value <0.05) were significantly differently expressed (DE) verified by RT-qPCR. GO and KEGG enrichment analysis indicated that DE genes and target genes of DE lncRNAs were associated with muscle development in the donkey. We also found these four target genes (DCN, ITM2A, MUSTN1, ARRDC2) involved in skeletal muscle growth and development. Combined with transcriptome data, network, and RT-qPCR results showed that four co-expression networks of DCN and lnc-008278, ITM2A and lnc_017247, MUSTN1 and lnc_030153, and ARRDC2 and lnc_033914, which may play an important role in the formation and development of muscle in the donkey. [ABSTRACT FROM AUTHOR]

The surging demand for donkey milk has led to a stronger focus on donkey husbandry and reproductive management, particularly in the peripartum period. Techniques to predict parturition are needed to avoid losses due to dystocias, which are frequently related to unattended foaling. In mares, pH and electrolyte analyses of pre-colostrum are used to predict parturition with a good accuracy. The purpose of this study was to evaluate the pH of the mammary secretion using both a pH metre and test strips and the electrolytes (Na+, K + and Ca2+) in the mammary gland secretions of 9 jennies during the 5 days preceding parturition, to understand if the trends of these cations were comparable to those of the mare and to identify possible markers/thresholds of impending parturition. Ca2+ concentrations increased in the days before delivery from day −2 (p < 0.01) and 78% of the jennies foaled within 48 h of reaching the threshold of 20 mmol/L. Na + and K + levels remained constant as parturition approached. The pH decreased in the days before foaling, starting from day −1 (p < 0.05). However, it was not possible to deduce certain indications on the date of delivery based on electrolyte concentrations, nor on pH and at the current state of knowledge foaling alert systems are the only methods for ensuring prompt assistance during parturition. The ability to predict parturition for providing adequate assistance is essential in a donkey farm. pH and electrolytes of mammary gland secretions are used in mares to predict foaling, but no reference values are available for jennies. In our work, neither electrolyte nor pH evaluations proved to be reliable to predict with an adequate level of confidence the impeding parturition. [ABSTRACT FROM AUTHOR]

The process of domestication of animals allowed for a change in the way of human life. Farm animals provided easy and relatively constant access to milk, meat, leather and wool. Farm animals are also increasingly used as pets and companions. Unfortunately, the appearance of animals in the human environment has also provided contact with new allergens. Allergy to fur animals has become a significant problem in the practice of an allergist. At present, it is a relatively common form of allergy and affects both adults and children. In the second part of the review of fur allergens, approved by the WHO/IUIS Subcommittee on allergen nomenclature, the allergens of rabbit, domestic cattle, horse, donkey, pig, goat and sheep were characterized. [ABSTRACT FROM AUTHOR]

Abstract Background Various anti‐parasitic drugs are used to control donkey parasitic diseases. The abuse of donkey drugs leads to the disposition of residues in the edible parts of treated donkeys. Objectives The aim of this study was to (1) analyse the pharmacokinetics of ABZSO to serve as reference for the dosage regimen in donkey; and (2) calculate the withdrawal times of the ABZSO in the tissue of the donkey. Methods The concentrations of ABZSO and its metabolites in plasma and tissues were determined using high‐performance liquid chromatography with an ultraviolet detector. Pharmacokinetic analysis was performed by the programme 3p97. Results The plasma concentrations of ABZSO and ABZSO2 concentration–time data in donkey conformed to the absorption one‐compartment open model. The t1/2ke of ABZSO was 0.67 h, whereas the t1/2ke was 12.93 h; the Cmax and the Tp were calculated as 0.58 μg mL−1 and 3.01 h. The Vd/F of ABZSO was estimated to be 10.92 L kg−1; the area under the curve (AUC) was 12.81 μg mL−1 h. The Cmax and AUC values of ABZSO were higher than those of ABZSO2; however, t1/2Ke and Vd/F were lower. Other pharmacokinetics parameters were similar between the two metabolites. Conclusions The results revealed that ABZSO2 was the main metabolite of ABZSO in donkey plasma. The concentrations of ABZSO and its chief metabolite (ABZSO2) were detected in liver, kidney, skin and muscle; however, ABZ‐SO2NH2 was only detected in liver and kidney. The results also revealed that the depletion of ABZSO and its metabolite in donkey was longer, especially in skin.

In June 2022, at the XXXII Conference of the Italian Society of Parasitology, the parallels of the main endoparasitic infections of horses and donkeys were discussed. Although these 2 species are genetically different, they can be challenged by a similar range of parasites (i.e. small and large strongyles, and Parascaris spp.). Although equids can demonstrate some level of resilience to parasites, they have quite distinct helminth biodiversity, distribution and intensity among different geographical locations and breeds. Heavily infected donkeys may show fewer clinical signs than horses. Although parasite control is primarily provided to horses, we consider that there may be a risk of drug-resistance parasitic infection through passive infection in donkeys when sharing the same pasture areas. Knowing the possible lack of drug efficacy (300 EPG may be a safe recommendation. We have highlighted the main points of the discussion including the dynamics of helminth infections between the 2 species.

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 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.

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.

The reproductive performance of horse sperm and donkey sperm has been reported to differ. Sperm proteins play a crucial role in sperm viability and fertility. Although differences between species are known, no prior study has investigated disparities in the sperm proteome between horses and donkeys. Therefore, this study characterized and compared the sperm proteomes of horses and donkeys using 4D-DIA mass spectrometry technology. We identified 3436 proteins in horse sperm and 3404 proteins in donkey sperm. Of these, 3363 proteins were expressed in both horse and donkey sperm, with 73 proteins being specifically expressed in horse sperm, and 41 in donkey sperm. According to data analysis, donkeys exhibited a greater percentage of motility and progressive movement in straight-line sperm than horses, as well as lower percentages of static and slow sperm than horses. Joint analysis of the results from the horse and donkey sperm proteomes and their CEROS II-read parameters demonstrated a possible association between sperm proteins and their sperm viability patterns. These findings suggest that there are discrepancies in the expression levels and protein compositions of horse and donkey sperm and that certain specific proteins may be responsible for the differences in performance between these two species.

Previous hematologic and serum biochemistry reference interval (RI) values have been established for donkeys in various geographic regions, life-stages, or for specific donkey breeds. The last extensive investigation establishing RIs for adult donkeys in the United States (U.S.) was published over three decades ago. We aimed to establish updated robust RIs using a reference population of apparently healthy adult donkeys from across the U.S. Standard sized (n = 102), miniature (n = 17), and mammoth (n = 1) donkeys from four different states were enrolled, with 20% of the study population including donkeys captured directly from the wild in Death Valley National Park, CA. RIs were established in accordance with the American Society for Veterinary Clinical Pathology and Clinical Laboratory Standards Institute guidelines. The findings will assist practitioners with the interpretation of their complete blood count and biochemistry panel results in U.S. donkeys. This study also highlights a comparison of results for some important analytes in U.S. donkeys compared to U.S. horses and previously established donkey RIs.

ABSTRACT: β-lactoglobulin I (β-LG I) is one of the most important whey proteins in donkey milk. However, to our knowledge, there has been no study focusing on the full nucleotide sequences of this gene (BLG I). Current investigation of donkey BLG I gene is very limited with only 2 variants (A and B) characterized so far at the protein level. Recently, a new β-LG I variant, with a significantly higher mass (+1,915 Da) than known variants has been detected. In this study, we report the whole nucleotide sequence of the BLG I gene from 2 donkeys, whose milk samples are characterized by the β-LG I SDS-PAGE band with a normal electrophoretic mobility (18,514.25 Da, β-LG I B1 form) the first, and by the presence of a unique β-LG I band with a higher electrophoretic mobility (20,428.5 Da, β-LG I D form) the latter. A high genetic variability was found all over the 2 sequenced BLG I alleles. In particular, 16 polymorphic sites were found in introns, one in the 5′ flanking region, 3 SNPs in the 5′ untranslated region and one SNP in the coding region (g.1871G > A) located at the 40th nucleotide of exon 2 and responsible for the AA substitutions p.Asp28 > Asn in the mature protein. Two SNPs (g.920–922CAC > TGT and g.1871G/A) were genotyped in 93 donkeys of 2 Italian breeds (60 Ragusana and 33 Amiatina, respectively) and the overall frequencies of g.920–922CAC and g.1871A were 0.3065 and 0.043, respectively. Only the rare allele g.1871A was observed to be associated with the slower migrating β-LG I. Considering this genetic diversity and those found in the database, it was possible to deduce at least 5 different alleles (BLG I A, B, B1, C, D) responsible for 4 potential β-LG I translations. Among these alleles, B1 and D are those characterized in the present research, with the D allele of real novel identification. Haplotype data analysis suggests an evolutionary pathway of donkey BLG I gene and a possible phylogenetic map is proposed. Analyses of mRNA secondary structure showed relevant changes in the structures, as consequence of the g.1871G > A polymorphism, that might be responsible for the recognition of an alternative initiation site providing an additional signal peptide. The extension of 19 AA sequence to the mature protein, corresponding to the canonical signal peptide with an additional alanine residue, is sufficient to provide the observed molecular weight of the slower migrating β-LG I encoded by the BLG I D allele.

Paslahepevirus balayani (HEV) is an important emerging zoonotic virus in Europe. Although domestic pigs and wild boar are the main reservoirs of this pathogen, susceptibility to this virus has been confirmed in a growing number of animal species, including equines. However, their role in the epidemiology of this virus remains poorly understood. Our aim was to assess HEV circulation and identify potential risk factors associated with exposure in equid species in different European countries. A total of 596 equines, including 496 horses, 63 donkeys and 37 mules/hinnies bred in four European countries (Spain, Italy, United Kingdom and Ireland) were sampled. Thirty-three animals (5.5%; 95%CI: 3.7–7.4) had anti-HEV antibodies. Seropositivity was found in 4.6% of horses, 11.1% of donkeys and 8.1% of mules/hinnies tested. By country, 6.3%, 5.4%, 5.0% and 4.0% of the equines sampled in Spain, Italy, United Kingdom and Ireland, respectively, were seropositive, respectively. Statistical analysis showed that “species” and “drinking water from ponds and streams” were potential risk factors associated with HEV seropositivity in equines in Europe. HEV RNA was not detected in any (0.0%; 95%CI: 0.0–1.8) of the 202 equines tested. Our results provide evidence of a low, spatially homogeneous and widespread viral circulation that is not equal across species in equid populations in the European countries analyzed and indicate that these species appear to play a limited role in the epidemiology of this virus. Further studies are required to elucidate the differences in seroprevalence between donkeys, mules/hinnies and horses and to determine the risk of zoonotic transmission of this pathogen from equid species.

Introduction: Streptococcus equi subspecies equi (S. equi) is the causative agent of strangles, which is one of the most common and highly contagious respiratory infectious illnesses in horses. Streptococcus equi (S. equi) is a horsespecific pathogen that originated from the closely related zoonotic pathogen Streptococcus equi subspecies zooepidemicus (S. zooepidemicus). Despite decades of research, the movement of genetic material across host-restricted diseases remains a mystery. Methods: Three S. equi donkey isolates (HTP133, HTP232, and HT1112) were recently isolated from a strangles epidemic on donkey farms in China's Xinjiang Province. In this study, we performed a comprehensive comparative analysis of these isolates using whole genome sequencing and compared them to the published genomic sequences of equine strain S. equi 4047 to uncover evidence of genetic events that shaped the evolution of these donkey S. equi isolates' genomes. Results: Whole genome sequencing indicated that both strains were closely related, with comparable gene compositions and a high rate of shared core genomes (1788-2004). Our comparative genomic study indicated that the genome structure is substantially conserved across three donkey strains; however, there are several rearrangements and inversions when compared to the horse isolate S. equi 4047. The virulence factors conveyed by genomic islands and prophages, in particular, played a key role in shaping the pathogenic capacity and genetic diversity of these S. equi strains. Furthermore, we discovered that the HT133 isolate had a strong colonization ability and increased motility; the HT1112 isolates had a significantly higher ability for antimicrobial resistance and biofilm formation, and the HT232 isolate gained pathogenic specialization by acquiring a bacteriophage encoding hyaluronate lyase. Discussion: In summary, our findings show that genetic exchange across S. equi strains influences the development of the donkey S. equi genome, offering important genetic insights for future epidemiological studies of S. equi infection. [ABSTRACT FROM AUTHOR]

Background: Donkeys with clinical signs of pituitary pars intermedia dysfunction are treated with oral pergolide mesylate despite the lack of species‐specific pharmacokinetic data. Objective: To evaluate the pharmacokinetics of intragastric and oral pergolide mesylate in healthy donkeys (Equus asinus). Study design: Pharmacokinetic study. Methods: Six healthy donkeys were administered pergolide mesylate (Prascend®) at 2 μg/kg bodyweight (bwt) intragastrically once, then once daily per os (PO) for 5 days. Blood samples were collected at 0, 10, 20, 30 and 45 min and 1, 1.5, 2, 3, 4, 6, 8, 12, 24, 36 and 48 h after the single intragastric dose, once daily immediately before the PO dose, and then again at the above times after Day 5 of once daily oral dosing. Plasma pergolide concentration was quantified via ultra‐performance liquid chromatography–tandem mass spectrometry. Pergolide concentration versus time data after the first and last doses were analysed based on noncompartmental pharmacokinetics using commercial software. Paired t‐tests were used to compare single and multiple doses (p < 0.05). In a follow‐up study, a single oral dose was then administered to two donkeys followed by concurrent blood sampling from the jugular and cephalic veins to evaluate the effect of route of administration on pergolide pharmacokinetics. Results: Cmax, Tmax AUC, and t½λz differed significantly (p ≤ 0.03) after single and multiple doses, with significantly lower Cmax (0.16 ± 0.16 ng/ml) and t½λz (9.74 ± 1.35 h) after intragastric dosing on Day 1 than after 5 days of oral dosing (3.74 ± 2.26 ng/ml and 16.35 ± 5.21 h, respectively). Pergolide plasma concentrations were higher in jugular vein samples compared to cephalic vein samples after a single oral dose. Main limitations: Small sample size; varied administration routes. Conclusions: Pergolide mesylate (dosed at 2 μg/kg bwt) is bioavailable in donkeys after intragastric and PO administration. Differences in pharmacokinetics were noted after multiple doses, related to different routes of administration and sublingual absorption of pergolide. [ABSTRACT FROM AUTHOR]

Background: Morphine is the prototypical μ‐opioid receptor agonist used to provide analgesia in veterinary species. Its effects are well‐described in horses but not donkeys. Objectives: To determine the antinociceptive effects of two doses of morphine in donkeys. To describe preliminary pharmacokinetic parameters of morphine in donkeys. Study design: In vivo experiment. Methods: Eight adult castrated male donkeys were given intravenous (IV) 0.9% saline, morphine 0.1 mg/kg bwt (LDM), or morphine 0.5 mg/kg bwt (HDM) in a randomised order with a minimum 1‐week washout period. Mechanical nociceptive thresholds (MNTs) were determined by a blinded investigator pre‐injection and 15, 30, 45, 60, 90, 120, 150, 180, 210, 240, 300, and 360 min post‐injection. Venous blood samples were collected pre‐injection and 2, 5, 10, 15, 30, 45, 60, 90, and 120 min post‐injection. Data were analysed using Friedman's test with Dunn's post hoc test for multiple comparisons. Pharmacokinetic parameters were calculated for the HDM treatment. Results: Baseline MNT was [median (interquartile range)] 8.9 (7.1–10.3) N and did not differ between treatments. Peak MNTs occurred at 60 min for both LDM (16.2 N) and HDM (25.0 N) treatments. MNTs after HDM treatment were higher than saline (p < 0.04) at 15, 60, 90, 120, 150, 180, 240, and 300 min post‐injection. MNTs after LDM treatment were higher than baseline (p < 0.05) at 45 and 60 min post‐injection. Terminal half‐life for HDM was (mean ± SD) 51.0 ± 10.7 min, the volume of distribution at steady‐state 2.07 ± 0.33 L/min and clearance 49.2 ± 4.16 ml * min/kg using noncompartmental analysis. The concentration of morphine‐3‐glucuronide (M3G) was higher than morphine‐6‐glucuronide (M6G) at all sampled time points. Main limitations: Short duration of plasma sampling for pharmacokinetic analysis; lack of objective measure of gastrointestinal function. Conclusions: The HDM treatment provided mechanical antinociception in donkeys with no significant adverse effects. [ABSTRACT FROM AUTHOR]

This paper represents the fundamental report of the survey of genome-wide changes of four Chinese indigenous donkey breeds, Dezhou (DZ), Guangling (GL), North China (NC), and Shandong Little donkey (SDL), and the findings will prove usefully for identification of biomarkers that perhaps predict or characterize the growth and coat color patterns. Three genomic regions in CYP3A12, TUBGCP5, and GSTA1 genes, were identified as putative selective sweeps in all researched donkey populations. The loci of candidate genes that may have contributed to the phenotypes in body size (ACSL4, MSI2, ADRA1B, and CDKL5) and coat color patterns (KITLG and TBX3) in donkey populations would be found in underlying strong selection signatures when compared between large and small donkey types, and between different coat colors. The results of the phylogenetic analysis, FST, and principal component analysis (PCA) supported that each population cannot clearly deviate from each other, showing no obvious population structure. We can conclude from the population history that the formation processes between DZS and NC, GL, and SDL are completely different. The genetic variants discovered here provide a rich resource to help identify potential genomic markers and their associated molecular mechanisms that impact economically important traits for Chinese donkey breeding programs. [ABSTRACT FROM AUTHOR]

Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) is one of the most common pathogens of endometritis in intensive donkey farms. In order to investigate the molecular epidemiological characteristics of S. zooepidemicus isolates associated with endometritis in intensive donkey breeding farms in China, isolates (n=40) from 8 small and medium-sized breeding farms were selected for multilocus sequence typing analysis. Results revealed that 4 sequence types (STs) were identified, ST360, ST380, ST413, and a novel genotype ST-novel, and these were genetically distant from each other, which suggested that S. zooepidemicus strains caused endometritis in intensive donkey farms possessed high genetic diversity and were lack of genotype specificity. The result of present study contributed to the understanding of the molecular epidemiological characteristics of S. zooepidemicus isolates. [ABSTRACT FROM AUTHOR]

The experiment was to study the effect of different roughages on cecal fermentation parameters in vitro. A single factor completely randomized experimental design was used to compare in vitro cecal fermentation parameters of alfalfa, oat grass, whole plant corn silage, Leymus chinensis, corn straw and cereal grass. Cereal grass was used as control group, and different coarse fodder were used as treatment group. The results showed that the gas production of alfalfa was the highest, and that of cereal grass was lower. The concentration of ammonium nitrogen was the highest in alfalfa and the lowest in corn straw. Mycoprotein concentration was the highest in whole plant corn silage and the lowest in cereal grass. The acetic acid content of alfalfa and oat grass was higher, while that of corn straw and cereal grass was lower. Propionic acid content of whole plant corn silage and oat grass was higher, cereal grass was the lowest. The content of butyric acid, isobutyric acid, isovaleric acid, valeric acid and total volatile fatty acid were the highest in oat grass, while the content of corn straw and millet grass were lower. The multiple combination effect value of roughage was shown as follow: alfalfa > oat grass > whole plant corn silage > Leymus chinensis > corn straw > cereal grass. The study indaicates that the in vitro cecal fermentation of alfalfa, oat grass, whole plant corn silage and Leymus chinensis are greater, and that of corn straw and cereal grass are lower. [ABSTRACT FROM AUTHOR]