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Follicular fluid is formed from the transudation of theca and granulosa cells in the growing follicular antrum. Its main function is to provide an optimal intrafollicular microenvironment to modulate oocyte maturation. The aim of this study was to determine the metabolomic profile of preovulatory follicular fluid (PFF) in jennies. For this purpose, PFF was collected from 10 follicles of five jennies in heat. Then, PFF samples were analysed by nuclear magnetic resonance (NMR) and heteronuclear single quantum correlation (2D 1H/13C HSQC). Our study revealed the presence of at least 27 metabolites in the PFF of jennies (including common amino acids, carboxylic acids, amino acid derivatives, alcohols, saccharides, fatty acids, and lactams): 3-hydroxybutyrate, acetate, alanine, betaine, citrate, creatine, creatine phosphate, creatinine, ethanol, formate, glucose, glutamine, glycerol, glycine, hippurate, isoleucine, lactate, leucine, lysine, methanol, phenylalanine, proline, pyruvate, threonine, tyrosine, valine, and τ-methylhistidine. The metabolites found here have an important role in the oocyte development and maturation, since the PFF surrounds the follicle and provides it with the needed nutrients. Our results indicate a unique metabolic profile of the jennies PFF, as it differs from those previously observed in the PFF of the mare, a phylogenetically close species that is taken as a reference for establishing reproductive biotechnology techniques in donkeys. The metabolites found here also differ from those described in the TCM-199 medium enriched with fetal bovine serum (FBS), which is the most used medium for in vitro oocyte maturation in equids. These differences would suggest that the established conditions for in vitro maturation used so far may not be suitable for donkeys. By providing the metabolic composition of jenny PFF, this study could help understand the physiology of oocyte maturation as a first step to establish in vitro reproductive techniques in this species J.C. was funded by the project “Demetra” (Dipartamente di Eccellenza 2018–2022, CUP_C46C18000530001), Italian Ministry for Education, University and Research. I.M.-R. was awarded a scholarship from the Spanish Ministry of Science, Innovation and Universities (BES-2017- 081962). I⋅Y.-O. was funded by the Secretary of Higher Education, Science, Technology and Innovation (SENESCYT), Ecuador (Scheme: Programa de Becas Internacionales de Posgrado 2019; Grant: CZ02- 000507-2019). The authors also acknowledge the support from the Ministry of Science and Innovation, Spain (Grant: AGL2017-88329-R), the Catalan Agency for Management of University and Research Grants, Regional Government of Catalonia, Spain (Grant: 2017-SGR-1229); and the Catalan Institution for Research and Advanced Studies, Spain (ICREA)

Donkey (Equus asinus) is an important livestock animal in China because of its draft and medicinal value. After a long period of natural and artificial selection, the variety and phenotype of donkeys have become abundant. We clarified the genetic and demographic characteristics of Chinese domestic donkeys and the selection pressures by analyzing 78 whole genomes from 12 breeds. According to population structure, most Chinese domestic donkeys showed a dominant ancestral type. However, the Chinese donkeys still represented a significant geographical distribution trend. In the selective sweep, gene annotation, functional enrichment, and differential expression analyses between large and small donkey groups, we identified selective signals, including NCAPG and LCORL, which are related to rapid growth and large body size. Our findings elucidate the evolutionary history and formation of different donkey breeds and provide theoretical insights into the genetic mechanism underlying breed characteristics and molecular breeding programs of donkey clades.

For thousands of years, the donkey (Equus asinus) has played an essential role in human society, underpinning the earliest forms of civilisation, facilitating critical trade networks, contributing to agricultural development, construction and mining. However, with the rise of motorised transport and agricultural machinery, the donkey was gradually turned loose in many places, and left free to roam. The emergence of freeroaming donkey populations has brought novel challenges for conservationists, land managers and animal welfarists alike. As non-native species that live and breed independently in large numbers, free-roaming donkeys appear as an ambiguous indeterminate group of wild-domestic creatures that sit uneasily with rapidly changing landscapes, societies and economies. This paper explores the status of free-roaming donkeys and the ongoing tension between the wild and the domestic, including the binary thinking underpins it and produces donkeys as non-native or ‘out of place’. Using a relational approach and paying attention to the various ways in which freeroaming donkeys are entangled and embedded within cultural historical landscapes, this paper suggests how donkeys might be re-examined in terms of their ‘entangled autonomy’, building on recent interventions in ‘wild’ animal geographies and ‘feral’ political ecologies. In doing so, it reframes the debate around the status of freeroaming donkeys and posits an argument for how they might be considered to ‘belong’ or have legitimacy within these landscapes, suggesting that more attention needs to be given to the spaces and places that donkeys create or contribute to, as well as those they disrupt and challenge.

Summary Mycobacterial infections are rare in horses, donkeys and mules. Although there are a few reports in horses, mycobacterial disease is poorly documented in the donkey. Mycobacterial infection of equine species typically affects the alimentary tract, causing granulomatous enterocolitis resulting in diarrhoea and chronic weight loss, while lymph nodes and liver may also be affected. We now document recurrent oesophageal obstruction, secondary to cranial mediastinal lymphadenitis caused by Mycobacterium avium complex (MAC). To the best of our knowledge, this is the first report of MAC infection in a donkey in the UK.

Objective As an important food therapy product with traditional Chinese medicine (TCM) applications, donkey-hide gelatin (Asini Corii Colla, ACC) has been used for thousands of years. However, till now few effective strategy had been proposed to distinguish ACC from other animal hide gelatins, especially closely related horse- and mule-hide gelatins, which was an embarrassment of ACC quality control. Methods Combined mass spectrometry and bioinformatic methods have been applied to identify and verify two ACC-specific peptides (Pep-1 and Pep-2) capable of distinguishing ACC from other closely related animal gelatins with high selectivity. Results It confirmed that these two peptides could be not only used for distinguishing ACC from highly homologous horse-hide and mule-hide gelatins as well as other animal hide gelatins. Conclusion The present study provides a simple method for species-specific peptides discovery, which can be used for assessing the quality of animal gelatin products, and ensure they are authenticable and traceable.

The objective of this study was to compare the effects of two warming protocols (three-step vs. one-step dilution) on embryo quality, post-warming embryo survival and embryo cell viability of donkey embryos vitrified by the Cryotop method. Twenty, Day 7-8, grade 1-2 donkey embryos were measured, morphologically evaluated and vitrified using the Cryotop technique. Embryos were then randomly warmed using two different warming procedures: (i) W3 (three-step dilution; n = 11): embryos were warmed in 1 M, 0.5 M and 0 M sucrose, and (ii) W1/0.5 (one-step dilution; n = 9): embryos were warmed directly in 0.5 M sucrose. After 3 and 24 h of warming, the embryos were measured and evaluated for their morphology, developmental stage and viability (Propidium Iodide-Hoechst 33,342 dyes). Although both treatments decreased embryo quality after warming (P 0.05), no significant differences (P 0.05) were observed between protocols in terms of post-warming embryo quality, diameter and embryo survival. Greater percentages of dead cells (P 0.001) were observed when embryos were warmed directly in 0.5 M sucrose (one-step dilution) when compared to the three-step protocol. The percentage of ruptured embryos was 27.3% and 0% in W3 and W1/0.5 protocols (P = 0.0893), respectively. In conclusion, warming Cryotop-vitrified donkey embryos directly in 0.5 M sucrose had no negative effects on embryo quality and post-warming embryo survival. Moreover, one-step protocol may help to prevent rupture when donkey embryos warmed directly in 0.5 M sucrose. These results observed in vitro must be verified by embryo transfer.

Many domestic donkey breeds are at risk of extinction, there is a critical urgency for genome resource banking. In the present study, we examined whether the use of Ficoll 70 added to the vitrification medium containing ethylene glycol (EG), dimethyl sulfoxide (DMSO) and sucrose improves the cryotolerance of donkey in vivo derived embryos. Day 7-8, grade 1-2 donkey embryos were measured and morphologically evaluated and then vitrified-warmed using the Cryotop technique. Before vitrification, embryos were randomly distributed into two groups: (i) VS1 (n = 14): vitrified using 15% EG + 15% DMSO + 0.5 M sucrose; and (ii) VS2 (n = 10): vitrified in the same medium supplemented also with 18% of Ficoll 70. After 24 h of warming, the embryos were measured and evaluated for their morphology, development and viability (Propidium Iodide-Hoechst 33342 dyes). Post-warming survival was numerically higher but not significantly different (P 0.05) when embryos were vitrified in VS2 (70%) compared to VS1 (57.1%). Embryo rupture was only observed in the VS1 group (21.4%, 3/14). Higher embryo diameter was observed in all groups after 24 h culture (P 0.05). No significant differences (P 0.05) were observed among treatments in terms of percentages of cell death. These results demonstrate that the addition of Ficoll 70 to the vitrification medium is not a pre-requisite for successful vitrification of donkey embryos. However, its addition seems to enhance some of the post-warming embryo quality characteristics. Since no statistically significant evidence was found, further studies should be conducted in order to confirm our findings.

As a contribution to the still open debate on the multiple african origins of the domesticated donkey, this work focuses on the wild ass of the Maghreb. It follows the recent paleontological review of the equids from Lac Karâr, a middle Pleistocene site in Northern Algeria, where most of the faunal remains were attributed to wild asses (Equus africanus Heuglin and Fitzinger, 1866). The morphometric resemblance between the equid of Lac Karâr, and Equus melkiensis Bagtach, Hadjouis and Eisenmann, 1984 which might correspond to the equid commonly referred to, as the Atlas wild ass (vernacular name) and, by descent, to Equus tabeti Arambourg, 1970, the only subspecies of ass from the early Pleistocene in Northern Africa, highlights the need for extensive biomolecular and radiometric studies on this wild and robust ass, endemic to the Maghreb. This is especially important given that progress in extracting ancient DNA from equids has been demonstrated by several recent studies and that remains of wild asses are still being uncovered in many late Pleistocene to Holocene (Neolithic) deposits in the Maghreb. The results from such studies could substantially improve the knowledge on the origin of asses. In particular, these analyses could shed light on the, as yet, undetermined clade 2 which may originate from the wild ass of the Maghreb as recently hypothesized by several authors. The archaeological and genetic data reviewed in this paper focuses on the ancient range of the Atlas wild ass mainly from sites in the northern part of the Maghreb; it contributes to current debates regarding this subspecies as one of the possible ancestors of modern asses. In addition, the paper identifies directions for further genetic researches on the extant donkey of the Maghreb.

The domestic donkey is a unique equid species with specific nutritional requirements. This article examines the importance of feeding strategies that mimic the donkey's natural environment using poor nutritional quality fibers and access to browsing materials. The relationship between nutrition and health is examined and practical approaches to the healthy and sick donkey are discussed.

The donkey evolved under harsh and arid environmental conditions, developing unique energy-efficiency traits, with an efficiency to rapidly mobilize fat in situations of increased energy demands or when food is scarce. This evolution has led to an inherent predisposition of donkeys to obesity, dyslipidemias, insulin dysregulation/metabolic syndrome, pituitary pars intermedia dysfunction, and endocrinopathic laminitis. Marked differences have been described in hormone dynamics and testing protocols for the diagnosis of these endocrine and metabolic diseases in donkeys compared with horses, underlining the necessity of a species-specific approach in order to avoid misdiagnosis, unnecessary or inadequate treatments, and additional costs.

This article provides a review of hoof anatomy and care in donkeys and mules. Hoof disease is a major cause of poor welfare and mortality globally. Problems associated with hoof disease are discussed in the context of behavior, diet, treatment, and prevention. The most common conditions encountered are discussed, including laminitis, the overgrown unbalanced hoof, white line disease, flexural deformities, and other significant issues. Differences between donkey and horse hoof anatomy are described.

Donkeys and mules often are misunderstood because of their behavioral differences compared with horses. Working with these animals requires more patience and ability to notice the slightest changes in behavior to diagnose disease. Mules and donkeys form strong bonds and trust with familiar people and other equids. Training mules and hinnies from an early age is key to modifying behavior and acceptance of unfamiliar people. Reproductive behavior is different from that of horses and requires more patience when collecting jacks. Practitioners working with mules and donkeys should take a methodological approach and get to know the animal before performing examinations.

Donkeys and mules have been critical to the development of human civilization, since being domesticated some 6000 years ago. However, they suffer from being undervalued or ignored by development agencies and animal protection nongovernmental organizations. Where they are recognized as affecting agriculture and the economy it is often because they are seen as being either invasive pests or an anachronism in the developing countries of the twenty-first century. Even in the wealthier societies of the world, donkeys suffer from ignorance about their proper management or a booming industry in health care products based on donkey skin gelatin and milk byproducts.

Donkeys are nonseasonal, polyestrous, territorial, and nonharem breeders. Although there are many similarities between horses and donkeys, there are also reproductive features that differ, from the longer cervix in the jenny to spermatogenic efficiency in the jack. Mules display reproductive cyclic activity but are rarely fertile. Frozen donkey semen has high pregnancy rates in mares, but lower rates in jennies. This article reviews key aspects of donkey and mule reproductive physiology, reproductive medicine, and assisted reproductive techniques that are useful for practitioners offering assisted reproductive techniques, and also for practitioners with the occasional client with a basic reproductive question.

Donkeys suffer from the same respiratory diseases as horses; however, owing to their nonathletic nature many conditions can present in a more advanced state before becoming clinically apparent. Anatomically, their respiratory tract is similar to the horse, with certain species-specific differences that are important to be aware of. Often donkeys do not receive the same level of routine care as horses, so many are not vaccinated against respiratory pathogens such as influenza or herpesviruses. Donkeys can act as a reservoir for certain infectious and parasitic respiratory diseases and the interpretation of diagnostic tests needs to be carried out with caution.

Research and clinical understanding of equine dentistry has progressed in recent years; however, specific knowledge about donkey dentistry is lacking. This article intends to revise unique anatomic features of the head and oral cavity of donkeys, as well as how to correctly examine, diagnose, prevent, and/or treat dental pathology, allowing for a better comprehension of oral and dental disorders affecting these animals throughout their life. It also emphasizes that these disorders need to be taken into account when assessing the welfare of donkeys.

This study compared the outcome of two new timed artificial insemination (TAI) protocols in a milk-producing donkey farm. Ninety Amiata jennies were inseminated at the moment of ovulation induction with hCG, with fresh-transported semen that had been stored at room temperature from 3 up to 6 h, with an approximate average storage time of 4 h and a half. In both protocols, on Day 0 jennies were treated with alfaprostol (PGF2α), and on Day 7 they were checked by ultrasound (US) and, if in estrus, they were treated in order to induce ovulation and were then artificially inseminated. In the slow-short TAI protocol, jennies not already inseminated were treated again with PGF2α at Day 14. On day 21 US was repeated and estrus jennies were induced to ovulate and inseminated. In the fast-long TAI protocol, US was performed once a week in jennies not already inseminated and if found in estrus, they were induced to ovulate and inseminated, while those not in estrus were treated again with PGF2α. This protocol was repeated for up to nine weeks. The rates of inseminated/treated, pregnant/inseminated and pregnant/treated jennies were 76%, 56% and 43% for the slow-short TAI protocol and 94%, 47% and 44% for the fast-long TAI protocol. The age class and the lactation status of the jennies had no significant effect on synchronization success or final pregnancy rate. This study demonstrates that it is possible to achieve reasonable pregnancy rates through simplified TAI protocols in jennies, reducing animal handling to a minimum.

In this study, probe/primers of high specificity and sensitivity were selected to analyze donkey-hide gelatin for donkey DNA and to look for horse, ox, and pig DNA as possible adulterants. The mitochondrial CO I genes in donkey, horse, and ox were selected as target sequences for design and synthesis of three pairs of specific probes and primers. In addition, eight pairs of probe/primers were obtained via literature search. Out of these eleven groups of probe/primers, those with the highest specificity and sensitivity were selected, which was fulfilled by the screening firstly with animal hide samples then the hide-glue samples. Other parameters that might affect detection specificity and efficiency-such as the amount of sampling and final concentration of primers-were also optimized. Replication tests were also conducted. The results showed that the selected probe/primers could accurately detect donkey DNA and horse, ox, and pig DNA in gelatin samples with good reproducibility. Analysis of four samples of on-market gelatin using this assay showed that two of the four samples indeed contained only donkey DNA, whereas the other two samples contained both donkey and horse DNA, indicating adulteration of these samples with horse hide. These results indicate that the TaqMan probe real-time PCR method can be used for identifying the purity of donkey DNA in gelatin samples, and can provide technical support for identifying adulterations in the gelatin market.

The diversity of the methanogenic archaea associated with the six segments of the horse and donkey hindgut (caecum, right ventral colon, left ventral colon, left dorsal colon, right dorsal colon, and rectum) was analyzed using 16S rDNA gene clone library. A total of 641 positive clones, 321 from the horse and 320 from the donkey hindgut, were examined by the RFLP, revealing 9 different ribotypes, 8 in the horse and 5 in the donkey hindgut. In both the animals Methanobacteriales (Methanobrevibacter-like sequences) and Methanomicrobiales (Methanocorpusculum-like sequences) were detected as the dominant orders followed by the uncultured Methanomassiliicoccales. The composition of the equine archaeal community was found to be dependent on the gut region. In both the two animals no Methanobrevibacter-like clones were detected in the caeca, which were instead inhabited by the Methanocorpusculum-like archeons. The Methanosarcinales were found only in distal regions of the horse hindgut.

The concentrations and distribution of polychlorinated naphthalenes (PCNs) in the whole blood of eight typical terrestrial meat animals (chicken, duck, rabbit, pig, cattle, sheep, horse and donkey) consumed daily in our life were investigated. The total concentrations (on a liquid volume basis) of PCNs were in a range from 305 to 987pg/L. Donkey blood contained the highest PCN concentrations. Mono-CNs were the dominant homolog group, accounting for 38%-71% PCNs. Apart from the mono-CNs and tri-CNs homolog groups, two hepta-CNs (mean: 9.5%) contributed most, followed by tetra-CNs (mean: 6.5%). The congeners CN1, 5/7, 24/14, 27/30, 52/60, 66/67, and 73 were the most abundant congeners or congener groups. The highest toxicity equivalencies (TEQs) were observed in cattle blood (117.4fg TEQ/L) then chicken blood (117.1fg TEQ/L). CN73 contributed 65% to total TEQs, followed by CN70 (20%) and CN66/67 (14%). The dietary intakes of PCNs were also estimated. Chicken meat, which forms the second largest component of meat product consumption in China, contributed most to the total TEQs (61%), followed by beef (27%) and pork (5.9%). The consumption of chicken might pose the highest risk from exposure to PCNs than other types of meat to populations who prefer to eat chicken meat.

The infectivity and virulence of seven Trypanosoma evansi and Trypanosoma equiperdum Venezuelan strains isolated from horses, donkeys and capybaras were compared in a mouse model up to 41 days, for parasitemia, animal weight, survival rates, packed cell volume, haemoglobin and erythrocyte count. Two T. equiperdum strains and three of the T. evansi strains resulted in 100% mice mortality, while the two T. evansi donkey strains exhibited lower infectivity and mortality. T. equiperdum strains had shorter pre-patent periods (4 days) than the T. evansi strains (4–12 days). In terms of pathogenicity, only the T. evansi horse strain and the two capybara strains produced a significant decrease of the packed cell volume, in haemoglobin concentration and in red blood cell count. In contrast, the T. evansi donkey strains did not show any changes in the hematological parameters. From the seven variables studied, only pre-patent period, day of maximum parasitemia, day of first parasitemia peak and number of parasitemia peaks were statistically significant. Weight decrease was only observed in mice infected with the T. evansi horse strain. T. equiperdum strains showed the highest mice lethality (7% survival by day 8 post-infection) with no change in the hematological parameters. The three T. evansi horse and capybara strains showed 80%, 87% and 97% survival rates, respectively by day 12 post-infection. However, by day 20 post-inoculation all the mice infected with the T. evansi horse strain died, while 53% and 27% capybara strains infected survived. Whereas by day 40 post-infection, 53 and 73% of the mice infected with the T. evansi donkey strains had survived. These results demonstrate striking infectivity and virulence differences between Venezuelan T. evansi and T. equiperdum strains in NMRI mice and open new possibilities to characterize inter and intra-species variations that may contribute to the pathogenicity of these two species.

The availability of biochemical blood data specific for the pregnant condition are essential for the correct management of both normal pregnancies and also for the prompt recognition of every abnormality. Because of the lacking knowledge about biochemical blood analysis in the donkey along the entire pregnancy, the study was designed to provide first preliminary data about the values and possible changes of blood alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), alkaline phosphatase (ALP), creatine-kinase (CK), blood urea nitrogen (BUN), creatinine (CREA), uric acid (UA), amylase (AMY), gamma-glutamyl transferase (γ-GT), triglycerides (TRI), cholesterol (CHOL), total protein (TP), albumin (ALB), glucose (GLU), phosphorus (P), calcium (Ca), occurring from the beginning of pregnancy until parturition. The study was performed on 10 Martina Franca healthy jennies with normal pregnancy course and giving birth to mature, healthy and viable foals. Blood samples were collected monthly from the 1st to the 6th month of pregnancy, then twice a month from the 6th to the 9th month and afterwards weekly until parturition. The results showed a significant slight increase of glucose and creatinine in the second quarter of pregnancy and a minor decrease of cholesterol near to parturition, while all the other parameters did not significantly change along pregnancy.

The aim of this review paper is to assess the applicability of donkey's milk to infants suffering from Cow Milk Protein Allergy (CMPA) compared to human and other available milk types. The bioactive and immune-supportive character which could be beneficial as a fortifier to the formula-fed infants is described while limitations of this type of milk are also discussed. Studies showed that human and donkey's milk have similar, overall, chemical composition as well as protein homogeneity and antigenic similarities. Several in vitro and in vivo studies showed that donkey's milk has nutraceutical and functional properties that can support immunity, alter metabolism and beneficially modify gut microbiota. Clinical studies illustrated that donkeys' milk is well tolerated (82.6%-88%) by infants. Finally, the effect that processing (i.e. thermal, non-thermal treatments, drying methods) has on donkey milk components is also discussed pointing out the need for minimally processing this type of milk.

Egg yolk (EY) has been the most widely used external cryoprotectant for donkey spermatozoa; however, it interferes with semen evaluation. Using clarified EY in equine freezing extenders has been successful, but it is difficult to apply in mountainous areas where mules are bred. Thus, the objective of this study was to evaluate the use of centrifuged EY for deep-freezing donkey spermatozoa. Twelve ejaculates were collected from four jacks of proven fertility. Seminal plasma was removed and, before freezing, samples were rediluted in four extenders, each with 7% dimethylformamide (DMF): (1) Kenney extender with 20% EY (Kenney-Y); (2) Kenney extender with 20% centrifuged EY (Kenney-CY); (3) EDTA-glucose extender with 20% EY (EDTA-Y); and (4) EDTA-glucose extender with 20% centrifuged EY (EDTA-CY). After thawing, motility, membrane function, and acrosome status were evaluated. Total and progressive motility observed in Kenney-Y and EDTA-Y was higher (P

The aim of this study was to evaluate the effect of different cooling rates on post-thaw quality of cryopreserved donkey spermatozoa. Eighteen ejaculates from six adult Andalusian donkeys (three ejaculates per donkey) were collected using an artificial vagina. Pooled semen samples (two ejaculates per pool) were divided into three aliquots, and frozen in Gent freezing extender using three different cryopreservation protocols (P): P1 (conventional slow freezing, as control): semen pre-cooled in an Equitainer for 2 h and frozen in liquid nitrogen (LN2) vapour; P2 (controlled pre-freeze cooling rate): semen pre-cooled at a controlled rate for 73 min and frozen in LN2 vapour; and P3 (rapid freezing) semen frozen immediately in LN2 vapour. After thawing at 37 °C for 30 s, semen samples were assessed for motility, morphology, acrosome and plasma membrane integrity; spermatozoa were also tested for DNA integrity. Significant (P < 0.01) differences were found between the cryopreservation protocols for all sperm parameters evaluated, except for DNA integrity. Semen samples frozen using P2 showed significantly (P < 0.01) higher values for sperm motility, morphology, sperm membrane integrity, and acrosome integrity. On the contrary, P3 reduced sperm motility (P < 0.01) and increased the percentage of spermatozoa with damaged plasma membrane (P < 0.001). In our study, we demonstrated that the sperm of Andalusian donkey is particularly sensitive to the cooling rate used before freezing. Furthermore, Andalusian donkey semen can be successfully cryopreserved using controlled cooling rates combined with freezing in LN2 vapour. Fil: Demyda-Peyrás, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico CONICET- La Plata. Instituto de Genética Veterinaria "Ing. Fernando Noel Dulout". Universidad Nacional de La Plata. Facultad de Ciencias Veterinarias. Instituto de Genética Veterinaria; Argentina. Universidad de Córdoba; España Fil: Bottrel, M.. Universidad de Córdoba; España Fil: Acha, D.. Universidad de Córdoba; España Fil: Ortiz, I.. Universidad de Córdoba; España Fil: Hidalgo, M.. Universidad de Córdoba; España Fil: Carrasco, J.J.. Centro de Selección y Reproducción Animal; España Fil: Gómez Arrones, V.. Centro de Selección y Reproducción Animal; España Fil: Gósalvez, J.. Universidad Autónoma de Madrid; España Fil: Dorado, J.. Universidad de Córdoba; España

The global equine population comprises a total of 114 million equids, which includes an estimate of 44 million donkeys (Equus asinus) and 11 million mules. Donkey populations have gradually expanded throughout the globe, particularly in developing countries. In spite of their relevance in several countries around the world, the infectious diseases affecting this group of equids are largely unknown; consequently, the available information regarding infectious diseases affecting working equids is limited. The scope of this article is to provide a brief update on the most relevant infectious diseases affecting donkeys and mules worldwide.

The aim of the study was to evaluate the effect of concentrations of egg yolk on cooling preservation and particle sizes on cryopreservation for donkey semen. Two experiments were carried out: (1) Determination of a suitable concentration of egg yolk during donkey semen cooled storage; and (2) evaluation of effects of egg yolk particles sizes on sperm quality after freezing-thawing. For experiment 1, the different concentrations (0%, 0.5%, 1.0%, 1.5%, 2.0%, and 2.5%) of egg yolk were added into the cooling extender, and the semen samples were preserved at 4°C for evaluation every 24 hours. The results demonstrated that the group of 1.0% egg yolk displayed higher total motility after 96 hours of preservation (P

There are 44 million donkeys worldwide of which 95% are working. However, only limited research programs dedicated to donkeys are carried out in the world. Indeed, donkeys have a specific nutritional status. The donkey use a selective feeding strategy; when it is foraging either poor or good feedstuffs. The ingestibility of the diverse types of forages is different. Intake rate seems to be rather slow, whereas the mean retention time of feed residues in the digestive tract is very slow. As a result, the digestion coefficients of the chemical components of the feedstuffs are high, namely for the cell walls. The digestion coefficients increase when the donkey is fed restricted compared with ad libitum, and the mean retention time slows down. So far, the volatile fatty acids (VFA) production in the large intestine is very high. The donkey takes advantage of the very high N-recycling ability. The water intake is related to the type and amount of forage which is offered and the environmental conditions. The donkey is able to manage temporary water deprivation without too high limitation of intake. And, it is able to rehydrate quickly. Energy and protein requirements are much lower than those of other equids. Mineral and vitamin requirements are not yet established. One could expect that the key ratio between some minerals and vitamins as well should be met, but we do not know if there are any specific sparing mechanisms as there is for nitrogen. For rationing, there are some key tools that are available for estimating live weight and body condition and for predicting water consumption. But the intake capacity of resting and working donkeys is poorly described. However, there is no substitution between forages and concentrate when feeding mixed diets. Unfortunately, there are neither tables of nutritive value of feeds nor recommended daily nutrients and intake allowances for donkeys. Indeed, further investigation should be carried out in the scope of large cooperation.

A cross-sectional study was conducted on 92 female donkeys. Blood samples were collected, and the following parameters were evaluated: red blood cell (RBC), white blood cell, neutrophil, lymphocyte, monocyte (MON), eosinophil (EOS) and basophil counts, hemoglobin concentration, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), and the hematocrit (HCT), alanine aminotransferase (GPT), aspartate aminotransferase, total proteins, g-glutamyl-transferase, alkaline phosphatase, creatinine, urea, and blood urea nitrogen and electrophoretic profile. Age (> 2 years > 3 [very young], > 3 years > 10 [young], and > 10 years < 17 [adult]) and lactation (early lactation [ 3 months 6 months]). Groups were independently analyzed using one-way analysis of variance or Kruskal -Wallis (post hoc test: Bonferroni's or Dunn's multiple test) tests; P was set as

Preselection of sex before conception has been one of the objectives pursued by scientists and breeders for many years. The donkey milk industry has shown a greater commercial interest, since jennies’ demand is increasing nowadays. Therefore, to get a female animal, the oocyte must be fertilized with previously sexed semen. The current technique used for sperm sexing separates spermatozoa containing the X or Y chromosome based on each cell DNA content. However, this technique exposes spermatozoa to high-pressure speed flow, DNA fluorescent stain, and UV light, factors that may affect sperm quality. The aim of this study was to test a new technique to isolate spermatozoa carrying the X chromosome by means of magnetic nanoparticles (MNPs). Results show that the MNP technique is highly effective in select X spermatozoa, without affecting several physiological sperm parameters. In conclusion, the MNP technique provides an X sperm population with similar sperm physiological state than the control sample. The new sexing technology presented here can be mainly applied to improve the donkey's milk industry. Fil: Dominguez, Esteban Mauricio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina Fil: Moreno, Ayelen. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina Fil: Ramírez Castex, Hernán. Bioteq. Reproducción y fertilidad equina; Chile Fil: Flores Bragulat, Ana Paula. Universidad Nacional de Río Cuarto. Facultad de Agronomía y Veterinaria; Argentina Fil: Ugaz, Cristian. INVESVET. Asesores en gestión veterinaria; Chile Fil: Clemente, Henry. CLEMENTE-ASSOCIATES INC.; Estados Unidos Fil: Giojalas, Laura Cecilia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Centro de Biología Celular y Molecular; Argentina Fil: Losinno, Luis. Universidad Nacional de Río Cuarto. Facultad de Agronomía y Veterinaria; Argentina

Humans have greatly benefited from their relationship with donkeys. Different roles that donkeys play in high- and low-income countries are two sides of the same coin. Their draft power is indispensable in low-income countries, and by contrast, their milk in Europe is fetching a premium. New productive niches are controversially being explored, for example, donkey meat and skin (ejiao); both are considered premium products and have increased value of donkeys in many developing regions of the world. New advances in human medicine are starting to consider possibilities of donkey serum and heart valves. Scientists and channels that they use to spread their knowledge directly influence public interest and implementation of welfare practices in domestic species, indirectly affecting their survival. The study's aim was to trace historical registry of www.sciendirect.com directory from 1896 to 2018 to assess trends followed by articles dealing with donkeys. Publications with 22 themes and 114 publications (91 JCR indexed journals) from 56 countries were included. JCR impact index was scored to study influence of publications dealing with donkeys. Chi-square test tested six variables, randomly influenced by the rest. Cramer's V measured strength of association between variables. Statistically significant differences were observed between almost every combination of variables except for year and JCR impact, year and area/topic and country of the corresponding author, and area/topic of submission. JCR impact and journal influences the area of the articles that are published. As some countries are more specialized in certain themes concerning donkey-related sciences, they are more likely to publish.

Many wild and domestic donkey breeds are now at risk of extinction, therefore there is an urgent need for genetic conservation of these breeds. The development of in vitro conditions that support oocyte maturation, fertilization, and embryo development may improve in vitro production of donkey embryos. This work was designed to identify the optimal condition for in vitro production of donkey embryos. Two experiments were conducted. In experiment 1, grade 1 and 2 cumulus-oocyte complexes were subjected to in vitro maturation in the following media: tissue culture medium 199 (TCM199), Christopher Rozenkrans 1aa medium (CR1aa), Christopher Rozenkrans 2aa medium (CR2aa), potassium simplex optimized medium (KSOMaa), centrifuged follicular fluid (CFF), or non-CFF from preovulatory follicles. In experiment 2, in vitro–matured donkey oocytes were subjected to in vitro fertilization (IVF) in Fert-TALP medium supplemented with 0%, 25%, 50%, 75%, or 100% CFF. After fertilization, the cleavage rates and embryo development were assessed across groups. The results of experiment 1 revealed that non-CFF and KSOMaa significantly increase cumulus cell expansion. The percentage of oocytes reaching the metaphase II stage nuclear maturation was significantly higher for donkey oocytes matured in non-CFF. Also, the percentage of degenerated oocytes was lower for donkey oocytes matured in vitro in non-CFF compared with those in TCM199, CR1aa, CR2aa, KSOMaa, or CFF. In experiment 2, IVF of in vitro–matured donkey oocytes using 100% CFF as IVF medium produced a significantly higher fertilization rate with respect to the number of fertilized oocytes extruding the second polar body. In conclusion, non-CFF from preovulatory follicles gave the highest cumulus cell expansion and nuclear maturation rates for in vitro–matured donkey oocytes. Hundred percent CFF was the best medium for IVF of matured donkey oocytes; however, cleavage rate and embryo development are still very low and need further investigations.

Donkey husbandry in South America is related to a social, cultural, economic, and ecological heritage. Its use is mainly based on agriculture but is also important in human feeding, production of hybrids, and transportation. Mechanization of the agricultural sector may cause concern as the decrease in a number of these animals can result in a loss of genetic variability of the species, restricting the ability of adaptation and evolution of this species. With the largest herd of these animals in South America, Brazil hosts three types of donkeys brought from Europe or Africa which are recognized as (1) Northeast (Ecotype) donkey, a class of highly rustic animals adapted to the adverse conditions of the Brazilian semiarid region; (2) Brazilian donkey, brought from Italy, with a close genetic relationship to African donkeys; and (3) Pega Donkey of Iberian origin, a special type of donkey developed mainly in the state of Minas Gerais for use in the mining industry. There are also researches performed in biotechnology for production of gate donkeys, such as artificial insemination and embryo transfer using mules and mares as recipients. In the meat business, an abattoir for donkeys has been recently built in the northeast region with capacity to slaughter 500 donkeys a day to export skin and meat to China. There are also projects to produce donkey milk but still on a small scale or at research level. All of these aspects have substantially changed the current situation of the donkey industry in South America in this new century.

International audience; In male donkeys, puberty and the related events have been poorly characterized. The aim of this study was to evaluate the age at which male donkeys reach puberty, and characterize age associated changes in testicular size, testicular blood flow, serum testosterone concentration and semen quality. Every two months, starting at 6 months and finishing at 24 months of age, five male donkeys born in May to July were subjected to B-mode ultrasound examination to assess testicular size and scrotum content and blood serum sampling for testosterone concentration. From the age of 8 months, pulsed Doppler was employed to evaluate blood flow in the testicular artery. Testosterone serum concentration was evaluated via RIA. From the age of 12 months, monthly semen collections were attempted and semen was evaluated for sperm number, motility and morphology. Onset of puberty was defined as the first ejaculate containing >= 50 x 10(6) spermatozoa with >= 10% total motility. One of the donkeys was excluded from the statistical analyses due to a hydrocele presented during the study. Testes width was affected by age (P < 0.0001) and after an initial plateau increased linearly from 10 months of age. Pulsatility and resistivity indexes were also affected by age (P < 0.01), being significantly higher at 14 months than at 24 months. Testosterone serum concentration was affected by age (P < 0.0001) and was significantly lower at 6 months (0.1 ng/ml) compared to 22-24 months (>= 0.8 ng/ml). Spermatozoa appeared in the ejaculate at a mean age of 18.7 months and puberty was attained between 19 and 20 months of age (mean: 19.5 months), between January and February. In conclusion, late spring born Amiata donkey colts reached puberty at 19-20 months of age. Puberty was accompanied by changes in testicular size, testicular blood flow and serum testosterone concentration.

While Sarcocystis parasites from the muscles of donkey and horse have been characterized as different species, similarities between the parasites in these host raises questions about this assignment (Levine and Tadros, 1980; Matuschka, 1983; Odening et al., 1995b). To resolve this, we examined the tissue cysts of Sarcocystis collected from donkeys and horses were studied by morphological and molecular methods. Morphological studies performed by light microscopy (LM) revealed that each of two types of cysts were present in samples from each host type. Under LM, villar protrusions (VP) were sometimes observed on the larger (Type I) and smaller (Type II) of these cyst types; when present, these were sometimes short and sometimes long. By electron microscopy (EM), VPs from both horse and donkey cysts were found to share similar structures, appearing to be typical of 'type 11a' VPs found on the Sarcocystis wall of Sarcocystis fayeri as described by Dubey et al., 1977. The VP of cysts in both horses and donkeys contained microtubules extending from the villar tips to the ground substance (GS). Ovoid, osmiophilic bodies (OB) were found along the length of the microtubules within the villi, but this feature was not found in all VP. To understand the phylogeny of the parasites, a portion of the coxI gene was sequenced from 22 isolated cysts (9 from donkeys and 13 from horses). Phylogenetic relationships were reconstructed from these sequences and the closest homologues available in GenBank, revealing that all of the samples, regardless of host origin or morphological appearance under LM, grouped in one clade. Ours is the first attempt to combine morphological measurements with coxI sequences in assessing such equine parasites; the results confirm a close relationship of the parasites from horse and donkey with S. fayeri. Further, the data suggest that the cysts in each host likely belong to the same species. As the first named species was Sarcocystis bertrami, we propose S. bertrami (syn. Sarcocystis fayeri) as the descriptor for this parasite of both horses and donkeys. Ultimately, this finding will only be validated by cross-transmission infection experiments that score the ability of parasite isolates from one Equus to infect the other.

In this work, the profile of the volatile compounds has been determined for the first time in fresh donkey milk and in donkey milk subjected to accelerated shelf-life test, freeze-drying and spray-drying processes. The effects of freeze-drying and spray-drying processes in donkey milk have been analyzed also on three representative whey proteins, β-lactoglobulin, α-lactalbumin and lysozyme, by determining the total amount, the enzymatic activity and the thermal stability. Considering the volatile compounds in fresh donkey milk, 2-heptanone, 1,3-bis (1,1-dimethylethyl)-benzene, nonanal, d -limonene, octanoic acid, and 1-octanol were detected. The accelerated shelf-life test revealed a constant increase of octanoic acid, 2-heptanone, and nonanal compared to fresh donkey milk (P

Diacylglycerol acyltransferase 1 (DGAT1) gene has become a potential candidate gene for milk composition and yield in cattle. A nonconservative lysine to alanine substitution (K232A) at position 10.433 and 10.434 in the eighth exon of DGAT1 gene has widely studied in cattle, goat, and sheep. This substitution conducted a considerable effect on milk fat, which was previously mapped to the centromeric end of bovine chromosome 14. Yet, no information has been found about DGAT1 gene in donkeys. In this study, the genetic variation of DGAT1 gene has been analyzed by CfrI (EaeI) restriction and DNA sequencing in 85 donkeys reared in Thrace region of Turkey. No digestion is found in the DGAT1 gene via CfrI restriction enzyme in the studied donkey populations. The DNA sequence of 437 bp of the DGAT1 gene in donkeys revealed only Alanine allele (K232A), which was reported in cattle with high milk fat content. But we found a novel polymorphism at position 10.435, from G to A, which also revealed Alanine allele. As a conclusion, CfrI digestion is not a diagnostic site in DGAT1 gene in donkeys. The DNA sequence of DGAT1 gene in donkeys is reported for the first time in this study, and this sequence is deposited to National Center for Biotechnology Information GenBank database with the accession number MK086162.

This study was carried out as part of an embryo preservation program undertaken among Hispano-Arabe (H-a) mares and Spanish (Andalusian and Zamorano-Leones) jennies, both considered to be endangered breeds. Over the course of six years, 61 donor mares and 17 donor jennies were monitored and inseminated with chilled semen from 17H-a stallions and six jackasses. A total of 199 uterine flushings (140 in mares and 59 in jennies) were carried out and the embryo recovery rate was recorded and the effect of various factors such as embryo quality and size, flushing effluent quality and individual effects were analyzed. The ovulation rate was significantly lower in donor mares (1.12) than in jennies (1.86) (p < 0.001). Embryo recovery per flush was 35.0% and 40.7% in mares and jennies respectively (p > 0.05), and it was influenced by photoperiod (p < 0.05). Individual effects were also noted for different stallions and jackasses. The embryo size was significantly affected by day of flush (p < 0.05) and embryo morphology (p < 0.001). While the effect of a range of variables is described in this study, it is important to emphasize that the endangered nature of the breeds concerned makes it harder to obtain acceptable results in an embryo transfer program, because it is difficult to select the best animals (donors, recipients and stallions), and this may play a role in the results obtained. However, it is necessary to continue the research in this field in order to improve the tools needed to preserve the pool of genetic heritage and diversity., This study was partially funded by the INIA-FEDER, project RZ2008-00025-00-00. G. Vizuete was supported by a FPU Grant Ref. AP2009-4607 from Spanish Ministry of Science and Education.

Horse meat and its products can be considered as a food with a high nutritional value. However, due to cases of economically motivated food adulteration by the intentional addition of horse meat beef products in recent years, horse meat has become a controversial issue. Consumer confidence in meat products and the meat industry has diminished, although consumers consider the differences between the food content and the label as the major issue rather than the safety and nutritional characteristics of horse meat. The elaboration of meat products from horse meat (e.g. “cecina”, dry-cured loin, salami, bressaola and pâte) is also an interesting alternative to other traditional meat products such as dry-cured pork hams, pork sausages and liver pâtes. In this review, the technological aspects, safety and storage stability of meat products elaborated from horse meat will be addressed by highlighting the nutritional and sensory aspects of these meat products. We aim to improve the existing knowledge about horse meat in the view of recent scandals.

Antimicrobial resistance results in selective colonization in animals. In the present study, 447 diarrheic foals (235 horse foals, 165 donkey foals and 47 mule foal) were selected from Lahore and Sahiwal districts of Punjab, Pakistan. Fresh fecal and blood samples from diarrheic foals were collected for isolation and confirmation of Salmonella Polymerase chain reaction. Results revealed that 50 (11.25%) foals (horse n=29, donkey n=12 and mule n=9) were positive. Fifty Salmonella enterica isolates belonging to 7 serovars S. Paratyphi B (15), S. Saintpaul (7), S. Newport (6), S. Typhimu-rium (11), S. Kottbus (4), S. Lagos (2), and S. enterica ssp salamae (5). Salmonella was common in foals that visited veterinary hospital, as compared to those in stud farms and individual foals reared in low income household. Out of the total 50 samples, 92% of isolates were resistant to three or more than three antimicrobials. The highest resistance (86%) was against Sulphamethoxazole (23.75mg) and lowest (4%) against trimethoprime (5mg). The isolates also showed resistance against Doxycycline (30mg), Oxytetracycline (30mg), Streptomycin (10mg), Neomycin (30mg), Amikacin (30mg), chloramphenicol (30mg), Ampicillin (10mg), Amoxicillin (10mg), kanamycin (30mg), Norfloxacin (10mg), Gentamicin 10mg, Cefotaxime (30mg), Ciprofloxacin (5mg) and Ceftriaxone (30mg). Blood analysis of salmonella infected foals showed That Hemoglobin, PCV and TEC were significantly higher and (while) TLC, PCV, Monocytes, Lymphocytes, Basophils, Eosinophil and Neutrophils were significantly lower than normal. Albumin were lower and BNU, Biluribin, ALT and creatinine were higher than normal values.

Despite their diffusion worldwide, many aspects of donkeys’ behaviour and cognitive abilities are still poorly understood and overshadowed by a corpus of commonplaces. The possibility of better knowing this species is of key relevance for its wellbeing and to establish a good human-donkey relationship. This review aims to investigate scientific literature on donkey behaviour and cognition, along with the behavioural tests that have been used so far to explore these topics. Searches of Web of Science (WoS) Core Collection, Scielo and Medline (through the WoS platform) and Google Scholar performed prior to March 2020 resulted in 75 records dealing with the question outlined in the present review. This research allowed us to delineate a qualitative synthesis of the studies’ characteristics, as well as a narrative summary of the topics covered by the investigated scientific literature, and to identify the behavioural tests used for donkeys. The hope is that a deeper knowledge of this species in regard to of the abovementioned aspects would allow donkeys to benefit from an improvement of human respect for their welfare.

The epidemiologic, clinical, pathologic, microbiological and immunohistochemical findings of pythiosis in equidae in northeastern Brazil are described. From January 1985 to December 2020 the Laboratory of Animal Pathology of the Federal University of Campina Grande received 1,331 tissue samples of equidae, 202 (15.17%) of which were diagnosed as pythiosis. Equidae of both sexes with ages varying from 4 months to 25 years were affected. Most animals were mixed breed (79.7%) and reared in an extensive system (73.26%). The disease occurred throughout the year but the highest incidence (70.29%) was noted after the rainy season. The clinical course was always chronic. The lesions were preferentially located on the limbs and ventral thoracoabdominal wall and characterized by nodules or tumor-like masses with ulcerations and serosanguineous discharge. The cut surface showed fistulous tracts containing kunkers. The direct examination of the kunkers and microbiological culture revealed sparsely septate and branched hyaline hyphae. Histopathology revealed a marked inflammatory infiltrate of eosinophils with multifocal well-defined areas of eosinophil necrosis and collagenolysis and intralesional negatively-stained hyphal profiles; in the donkey, a pyogranulomatous inflammatory infiltrate was noted surrounding these areas. Immunohistochemistry for Pythium insidiosum revealed strong immunolabelling of the hyphae. Pythiosis occurs endemically in equidae in northeastern Brazil, with seasonal variation in the incidence. The intralesional kunkers establishes an accurate presumptive diagnosis, but confirmation should preferably be performed through histopathology associated with immunohistochemistry, culture-based or molecular methods.

Natural ingredients as an alternative to chemical additives in the food industry are rapidly becoming widespread due to the increasing consumer concern. Lysozyme and lactoferrin are found in high amounts in donkey milk and are known to have antimicrobial effects on many microorganisms. In this study, we extracted these proteins from donkey milk with lysozyme-imprinted and lactoferrin-imprinted cryogel columns using FPLC and applied them to Kashar cheese as an antimicrobial agent by dipping into a solution. The treated cheeses were compared with control samples for their physicochemical, textural, sensory and microbiological properties. Results demonstrated that products treated with antimicrobial proteins exhibited lower microbial load, better physicochemical, textural and sensorial properties throughout the ripening periods. Results of this study highlight lysozyme and lactoferrin from donkey milk for potential usage as natural antimicrobial agents to prolong the shelf-life of products, provide consumer safety and health and obtain better quality characteristics.

Variation in donkey sperm freezability (capacity to withstand freeze-thawing) between ejaculates is a limitation for sperm cryopreservation. Seminal plasma proteins are essential for sperm function and also related to individual differences in sperm freezability. A Tandem Mass Tag (TMT) peptide labeling combine with a LC-MS/MS approach was conducted to quantitatively identify the seminal plasma proteins differentially abundant in ejaculates with optimal freezability characteristics (GFE) compared with those with suboptimal freezability characteristics (PFE). A total of 866 proteins were identified, and 99 ejaculates were in larger abundance in GFE samples. Differentially abundant proteins (DAPs) were subjected to intensive bioinformatic analysis. The majority of DAPs were involved in metabolic processes, oxidation-reduction processes and biological regulation. Results from functional protein analysis suggested that proteins functioned in oxidoreductase activity and acid phosphatase activity. This is the first report where there were analyses of the proteome of seminal plasma from donkey ejaculates with different freezability and to identify candidate proteins that could be used to explore the molecular mechanism related to donkey sperm cryotolerance. In this study, there also was elucidation of biomarkers for the early identification and selection of donkeys with optimal semen freezability.

The microorganisms in the cecum of donkeys share similar functions as those in the rumen of cattle. Transformation of the cecal microenvironment by probiotics plays an important role in the health and growth of donkeys. In order to screen out excellent donkey probiotic preparations, in this study, we isolated an antibacterial strain of Bacillus amyloliquefaciens (designated as DQB-1) from the cecum of Dezhou donkey. The strain was assessed in terms of antibacterial activity, antibacterial substance analysis, and stability. The results show that, the Bacillus amyloliquefaciens DQB-1 exhibited protease production activity and can significantly inhibit the growth of bacterial and fungal pathogens. The strongest antibacterial substance would be obtained after 24 hours of growth. The most suitable storage temperature for antibacterial extracts is -20 °C. The antibacterial substance produced by Bacillus amyloliquefaciens DQB-1 isolated from donkeys has strong antibacterial activity, protease-producing activity and good stability. Therefore, it can be developed as a probiotic preparation for preventing infectious diseases in donkey farms.

T-box transcription factor 3 (TBX3) gene encodes a transcriptional suppressor and plays an important role in embryonic development, which belongs to the T-box family. TBX3 also has been found to be associated with body size traits in horse that is a relative of donkey. Therefore, TBX3 is considered as a promising candidate gene for economic traits of donkey. This study aimed to reveal the significant variation of TBX3 gene in Dezhou donkey and explores the relationship between genotypes and body sizes. In this study, an A > G mutation was found in the intron 2 of TBX3 gene by sequencing, and three genotypes (AA, GG and AG) were identified in 380 Dezhou donkey individuals with Tm-shift method. Association analysis illustrated that there were significant differences between AA and GG genotype in body length, body height, chest depth, chest circumference, body weight, hucklebone width and rump length. Our results demonstrated that the polymorphism of TBX3 is significantly associated with body size traits, which can serve as a marker to improve donkey production performance.

Despite the extensive research on ocular ultrasonography, there is paucity of information regarding ocular biometry and color Doppler imaging of the donkey's eye. This study establishes B-scan ocular biometry and color Doppler ultrasonographic reference values in normal donkeys and describes effect of laterality, maturity and gender on ocular biometry and vasculature. Forty (80 eyes) clinically normal immature and mature donkeys of both sexes were included. Transpalpebral ocular ultrasonography was made using a 12-MHz linear transducer. Biometric ocular measurements were made including the anterior chamber depth, central lens thickness, vitreous chamber depth, axial globe length, longitudinal globe length, lens pole diameter and posterior wall thickness. Color Doppler velocimetric measurements including peak systolic velocity and end-diastolic velocity were made. Blood flow indices represented by resistive and pulsatility indices were calculated. Results revealed that laterality and gender did not represent variability factors in ocular biometry (P.05), while the animal's maturity had an influence on axial globe length (P.001), longitudinal globe length (P = 0.000) and lens pole diameter (P = .000). Laterality had no effect on velocimetric parameters and indices (P.05) while gender had significant effect on resistive index (P = .024), pulstality index (P = .000) and volume flow per area (P = .008). The state of maturity had significant effects on peak systolic velocity (P = .027) and blood volume flow per distance (P = .012), and area (P = .048). Ocular biometry and color Doppler ultrasonography provided a clinically useful tool to evaluate the donkey's eye that will be helpful in diagnosing donkeys with ocular diseases.

The aim of this study was to evaluate the impact of fibers (inulin, apple fruit, and passion fruit) on the viscosity and acceptance of donkey milk yogurt compared to cow milk yogurt. Five yogurts were developed: donkey milk control (DC), donkey milk inulin (DI), donkey milk apple fiber (DAFF), donkey milk passion fruit fiber (DPFF), and cow milk control (CC). Analyses of the physical-chemical and microbiological components were performed. Donkey milk yogurts had significantly higher LAB counts when compared to the CC treatment. The pH and color were influenced by the type of milk and fiber. The DPFF yogurt contained the best apparent viscosity (824.7 mPa s). Due to the results, the yogurts were submitted for sensory testing. The analysis evaluated the sensory attributes of the yogurts (DC, DPFF, CC, and cow milk passion fruit fiber). Passion fruit fiber provided the highest apparent viscosity. However, the addition of passion fruit fiber resulted in low acceptance.

This study aimed to evaluate the nutritional value and physicochemical characteristics of fresh donkey milk and fermented donkey milk. Furthermore, the impact of the refrigerated storage (4 °C, 21 days) on the characteristics of the fermented milk was evaluated. The fermented milk had higher acidity (lower pH values and higher titratable acidity) and presented lower lactose, mineral (calcium, phosphorous, magnesium, sodium, and zinc), and amino acid (mainly essential amino acids) contents than milk. Otherwise, it had a better fatty acid profile (lower saturated fatty acid content, and higher monounsaturated and polyunsaturated fatty acid contents). The fermented milk storage resulted in decreases in the lactose content, improvements in the amino acid profile, and maintenance of the mineral content. However, negative impacts were observed on the fatty acid profile. It can be concluded that both donkey milk and fermented milk donkey present important nutritional value, and the fermented milk could be refrigerated stored up to 21 days.

A peripheral nerve sheath tumour, with features of a benign schwannoma and arising from the endocardium of the right ventricle, was found incidentally during routine post-mortem examination of a 28-year-old gelding donkey. Macroscopically, five round to oval, white to grey and red, firm masses, firmly attached to the endocardium were identified. Microscopically, the endocardium and adjacent subendocardium were infiltrated by a variably demarcated, non-encapsulated mesenchymal neoplasm with features of a benign schwannoma, including concurrent presence of Antoni A and Antoni B areas, nuclear palisading, neoplastic cells with enlarged bizarre nuclei ('ancient change') and the formation of Verocay-like bodies. Immunohistochemically, the tumour cells were variably strongly positive for expression of S100 and glial fibrillary acidic protein. This is the first cardiac tumour reported in a donkey and is macroscopically, histologically and immunohistochemically similar to endocardial schwannoma occurring in Wistar rats.