Donkey Research Articles

Identifying animal-derived components in camel milk and its products by ultra-high performance liquid chromatography-tandem mass spectrometry

A method for identifying specific peptide biomarkers of animal-milk-derived components in camel milk and its products was established using proteomics. Samples were prepared by defatting, protein extraction, and trypsin hydrolysis, and proteins and peptides were identified using ultra-high performance liquid chromatography-quadrupole/electrostatic orbitrap-high resolution mass spectrometry (UHPLC-Q/Exactive-HRMS) and Protein Pilot software. Twenty two peptide biomarkers from eight species (i.e., Camelus, Bos taurus, Bubalus bubalis, Bos grunniens/Bos mutus, Capra hircus, Ovis aries, Equus asinus, Equus caballus) were identified by comparing the basic local alignment search tool (BLAST) with the Uniprot database. Verification of these marker peptides were performed quantitatively using a UHPLC-triple-quadrupole mass-spectrometry (QqQ-MS) system by multiple reaction monitoring (MRM). The pretreatment method of casein in camel milk was optimized, such as defatting, protein precipitation, and re-dissolving buffer solution. The effects of various mass-spectrometry parameters, such as atomization gas, heating- and drying-gas flow rates, and desolvation-tube (DL) and ion-source-interface temperatures on ion-response intensity were optimized. Camel milk signature peptides were detected in a mixture of milk from other seven species to ensure specificity for the selected biomarker peptides. The signature peptides of seven other species were also detected in camel milk. No mutual interference between the selected biomarker peptides of the various species was observed. Adulterated camel milk and milk powder were also quantitatively studied by adding 0, 2.5%, 5%, 10%, 25%, 50%, 75%, and 100% bovine milk or goat milk to camel milk. Similarly, the same mass proportion of bovine milk powder or goat milk powder was added to camel milk powder. A quantitative standard curve for adulteration was constructed by plotting the peak areas of characteristic cow or goat peptide segments in each mixed sample against the mass percentage of the added adulterant. The adulteration standard curves exhibited good linearity, with correlation coefficients (r2) greater than 0.99. The limits of detection and quantification (LODs and LOQs, respectively) of the method were determined as three- and ten-times the signal-to-noise ratio (S/N). The minimum adulteration LODs of bovine milk and goat milk in camel milk were determined to be 0.35% and 0.49%, respectively, and the minimum LOQs were 1.20% and 1.69%, respectively. The minimum adulteration LODs of bovine milk powder and goat milk powder in camel milk powder were determined to be 0.68% and 0.73%, respectively, and the minimum LOQs were 1.65% and 2.45%, respectively. The accuracy of the adulteration quantification method was investigated by validating the quantitative detection results for 1∶1∶1 (mass ratio) mixtures of camel milk, bovine milk, and goat milk, as well as camel-milk powder, bovine milk powder, and goat-milk powder, which revealed that this method exhibits good linearity, strong anti-interference, high sensitivity, and good repeatability for adulterated liquid-milk/solid-milk-powder samples. The adulteration results for both liquid milk and milk powder are close to the theoretical values. Finally, 11 actual commercially available samples, including five camel-milk and six camel-milk-powder samples were analyzed, which revealed that only camel signature peptides were detected in 10 samples, while camel and bovine signature peptides were both detected in one camel-milk-powder sample. The ingredient list of the latter sample revealed that it contained whole milk powder from an unidentified source; therefore, we infer that the bovine signature peptides originate from the whole milk powder. These signature peptides also demonstrate the necessity and practical significance of establishing this identification method.

Changes in the hematobiochemical, acid–base and blood gas elements as well as biomarkers of inflammation and bone metabolism in donkeys (Equus asinus) with acute bleeding

Changes in the hematobiochemical, acid–base and blood gas elements as well as biomarkers of inflammation and bone metabolism in donkeys (Equus asinus) with acute bleeding

Phylogenetic status and diet of red fox (Vulpes vulpes griffithii) inhabiting Ayubia National Park, Pakistan

Abstract The red fox (Vulpes vulpes) is a medium-sized carnivore that occurs in different regions of Pakistan, however, still lacks scientific data on its ecology and distribution. The current study investigated the phylogenetic status and diet of the red fox (V.v. griffithii) occurring in Ayubia National Park, Pakistan. Through camera trapping and molecular analysis, we confirmed the occurrence of red fox in the study area. Based on mitochondrial cytochrome B (304 bp) and limited sampling, nearly all red foxes of Ayubia National Park and surrounding Himalayan ranges fall within Holarctic maternal lineage, whereas red foxes found in plains of Pakistan are part of the basal Palearctic maternal lineage. Using 32 scats, we found that red fox diet comprises of 80% animal-based prey species (both wild and domestic) and 19% plant matter. The wild animal prey species included Cape hare (Lepus capensis) and flying squirrel (Pteromyini sp.), which constituted 17% and 15% of diet, respectively. Red foxes infrequently consumed House mouse (Mus musculus), Himalayan Palm civet (Paguma larvata) and sheep (Ovis aries), each comprising around 6% to 9% of red fox diet. The fox species also scavenged on domestic donkey opportunistically. Based on our sampling, our study suggests that the red fox (V.v. griffithii) that occurs in Ayubia National Park and across the lesser Himalayan ranges belongs to Holarctic maternal lineage. The study also highlights consumption of plant seeds by red foxes, indicating it may play an important ecological role in seed dispersal in Ayubia National Park.

Skeletal Class III phenotype: Link between animal models and human genetics: A scoping review.

This study aimed to identify evidence from animal studies examining genetic variants underlying maxillomandibular discrepancies resulting in a skeletal Class III (SCIII) malocclusion phenotype. Following the Manual for Evidence Synthesis of the JBI and the PRISMA extension for scoping reviews, a participant, concept, context question was formulated and systematic searches were executed in the PubMed, Scopus, WOS, Scielo, Open Gray, and Mednar databases. Of the 779 identified studies, 13 met the selection criteria and were included in the data extraction. The SCIII malocclusion phenotype was described as mandibular prognathism in the Danio rerio, Dicentrarchus labrax, and Equus africanus asinus models; and as maxillary deficiency in the Felis silvestris catus, Canis familiaris, Salmo trutta, and Mus musculus models. The identified genetic variants highlight the significance of BMP and TGF-β signaling. Their regulatory pathways and genetic interactions link them to cellular bone regulation events, particularly ossification regulation of postnatal cranial synchondroses. In conclusion, twenty genetic variants associated with the skeletal SCIII malocclusion phenotype were identified in animal models. Their interactions and regulatory pathways corroborate the role of these variants in bone growth, differentiation events, and ossification regulation of postnatal cranial synchondroses.

Advanced platelet-rich fibrin promotes healing of induced corneal ulcer in donkeys (Equus asinus)

Ulcerative keratitis is a common disease in horses which may cause blindness. To prevent secondary bacterial and fungal infections and promote quick re-growth of the epithelial layer, different treatment approaches have been employed. This study aimed to examine the effects of advanced platelet-rich fibrin (A-PRF) gel on the healing process of experimentally induced corneal ulcers in donkeys. Nine healthy adult donkeys were used for the study. The donkeys were divided into two groups: the control group, where no medication was applied to the corneal ulcer, and the A-PRF gel group, where A-PRF gel was applied once a day on specific days after ulcer induction. The healing process was evaluated through various examinations and analyses. The results demonstrated that the A-PRF gel group showed significant improvement in the corneal ulcer area, with epithelial and stromal regeneration. At day 35, about 60% of the A-PRF group showed negative fluorescein uptake. Additionally, fewer complications were observed during the healing process compared to the control group. In conclusion, A-PRF gel is an important and safe therapeutic option for controlling ocular surface infection and promoting corneal healing. We recommend using A-PRF gel as an alternative approach, avoiding eyelid suturing, and minimizing corneal irritation.

Transcriptome analysis reveals immune function-related mRNA expression in donkey mammary glands during four developmental stages

The low susceptibility to mastitis of female donkey (jenny) mammary glands and the strong immune properties of donkey milk are acknowledged, but little is known about the genes involved in mammary gland immunity in jennies. Herein, we used RNA-sequencing and bioinformatics analyses to explore jenny mammary gland transcriptomes and detect potential functional differentially expressed (DE) mRNAs related to immunity during four specific developmental stages: foetal (F), pubertal (P), adult parous nonlactation (N) and lactation (L). A total of 2497, 583 and 1820 DE mRNAs were identified in jenny mammary glands at F vs. P, P vs. N, and N vs. L, respectively. Gene Ontology (GO) and Kyoto Encyclopedia of Gene and Genomes (KEGG) analyses revealed numerous GO terms related to immune function, especially between F and P. Seven significantly enriched profiles were identified, among which 497 and 1261 DE mRNAs were upregulated in profiles 19 and 17. Eleven mRNAs were enriched in over 10 KEGG pathways. β-2-microglobulin (B2M), immunoglobulin heavy constant mu (IGHM), toll like receptor 2 (TLR2), toll like receptor 4 (TLR4) and myeloid differentiation factor 88 (MYD88) were mainly involved in phosphoinositide 3-kinase (PI3K)-Akt signalling, phagosome and nuclear factor kappa-B (NF-kappa B) signalling pathways. The findings provide insight into the molecular features underpinning the low prevalence of intramammary infections (i.e., mastitis) in donkeys.

Characterization of social behavior in a group of domestic donkeys (Equus asinus)

Although the popularity of domestic donkeys is increasing, the scientific literature on their behavior is still limited. This study investigated the social behavior of a group of 13 donkeys (10 females, 3 geldings, aged 3–13 years) stabled in an Animal Assisted Interventions facility in the North of Italy. Our first aim was to assess the distribution of donkeys’ intraspecific social behaviors during daytime and the second aim was to characterize affiliative and agonistic social interactions during peak activity times. Social behaviors were expressed more between 8 and 9 AM than the rest of daytime, mainly reflecting the expression of affiliative behavior. The latter consisted mostly of proximity (55.0% of all affiliative) and following (29.2%). In most cases affiliative behavior was expressed within preferential dyads, composed of either a mother and her offspring or two adult donkeys. This supports the idea that dyadic relationships are a predominant feature of the social organization of the species. However, dyadic interactions were not observed in the oldest sub-adults, suggesting that social preferences undergo a change around the time donkeys reach full growth. Agonistic behaviors were much less common than affiliative ones. The most expressed agonistic behavior was threat (51%), followed by displacement and bite (both 18.5%). They were less expressed by donkeys who were part of an affiliative pair, suggesting that limited expression of agonistic behavior is associated with an actual change in social preferences, not a mere consequence of fewer occasions of conflict over resources. While the study has limitations, the results provide insight into the social behavior of domestic donkeys which could represent a starting point for further research as well as relevant information for donkeys farming.

Molecular detection and phylogenetic analysis of Babesia caballi in domestic donkeys (Equus asinus) in Baghdad Zoo, Iraq

Molecular detection and phylogenetic analysis of Babesia caballi in domestic donkeys (Equus asinus) in Baghdad Zoo, Iraq

Listeria monocytogenes encephalitis in a donkey foal

SummaryThis 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.

PSVII-6 Effect of Summer Months on Testicular Histoarchitecture and Spermatogenesis in Dezhou Donkey: a Histological Study

Abstract This study aims to illustrate testicular histoarchitecture and progression of spermatogenesis in Dezhou donkeys during summer months. For experiment, 8 male Dezhou donkeys having body weight 260 + 20 kg and aged 2 years were analyzed using microscopy and Jonhson’s score. Based on acrosomal system, Dezhou donkey spermatogenesis comprised of XII stages. Various types of spermatogonia: A1, A2, A3, B1 and B2 were observed along with undifferentiated spermatogonia (Aund; Figure 1, 2, 3 and 4). Testicular histoarchitecture during spermatogenesis, seminiferous tubules diameter (ST), ST height (µm), Luminal tubulaR diameter (µm), Number of ST/field, Number of germ cells along with normal Jonhson’s score illustrated that stages of spermatogenesis and testicular histoarchitecture of adult Dezhou donkeys are not affected during summer months (Table 1).

Comparing methods to estimate feral burro abundance

AbstractObtaining precise and unbiased estimates of feral burro (Equus asinus) abundance in the western United States is challenging due to their cryptic pelage and the rugged terrain they inhabit. Management agencies employ helicopter‐based, simultaneous double‐observer sightability surveys (hereafter denoted as DOS) to estimate abundance of burros; but the DOS method routinely produces negatively biased estimates due to residual heterogeneity in detection probability. Consequently, testing alternative methods to improve upon current procedures is warranted. Residual heterogeneity in DOS surveys can be minimized by including radio‐collared individuals in the population. Alternatively, if distance measurements are recorded, residual heterogeneity can also be reduced via a mark‐recapture distance sampling (MRDS) approach. Aerial infrared (IR) surveys offer a safer alternative than helicopter‐based surveys because they can be flown at a higher altitude and require fewer observers in the aircraft. Further, IR surveys using a distance sampling approach have been shown to generate accurate and precise estimates of feral horse (E. caballus) populations. Accordingly, we compared results of surveys using aerial IR distance sampling, the standard DOS survey, a DOS survey incorporating detections of radio‐collared individuals, and an MRDS analysis of a feral burro population with a known minimum population size in central Utah, winter 2015–2016 and spring 2016. The minimum number of burros known alive during the winter and spring surveys were 236 and 136, respectively. The average detection probability of IR surveys was P = 0.88 (SE = 0.16) and distance models produced estimates of 127 burros (95% CIs = 99–175) for the winter survey, and 94 burros (CIs = 72–134) for the spring survey. Mean detection probability of the standard DOS surveys was P = 0.78 (SE = 0.09), and model‐generated abundance estimates were 155 burros (CIs = 133–227) in winter, and 92 burros (CIs = 79–139) in spring. Incorporating detections of radio‐collared individuals in the DOS survey resulted in a decreased detection probability (P = 0.46; SE = 0.06) and increased abundance estimates to 267 (CIs = 169–571) and 155 (CIs = 128–263) for winter and spring, respectively. Mark‐recapture distance sampling produced a mean detection probability of P = 0.48 (SE = 0.12) and resulted in estimates of 282 (CIs = 178–385) and 169 (CIs = 73–310) burros in winter and spring, respectively. Our study demonstrated that aerial IR surveys conducted using standard distance sampling can produce precise estimates of burro population sizes; however, estimates were negatively biased relative to the known population size. Small sample size limits generalization of our results, but the IR‐based distance approach did not improve upon DOS surveys. Accounting for residual heterogeneity through use of radio‐collars and mark‐recapture distance sampling eliminated the negative bias from the standard DOS survey but decreased survey precision. Managers will need to decide whether unbiased but less precise abundance estimates are preferable compared to a more precise, but biased, estimate.

Seasonal variations of the epididymis in donkeys (Equus asinus) with special reference to blood epididymal barrier.

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.

Pharmacotherapeutic monitoring of dipyrone in northeastern Brazilian donkeys (Equus asinus)

This study aimed to monitor the effects of dipyrone following multiple administrations in northeastern donkeys. Ten castrated male donkeys, aged 6.4 ± 3 years and weighing 130.6 ± 9.8 kg, were administered dipyrone (25 mg/kg IV) every 12 h, resulting in six administrations (D1 to D6) per animal. Blood samples were collected over a 72 h monitoring period. A validated UHPLC-MS/MS method was employed to determine the plasma concentrations of the 4- methylaminoantipyrine (4-MAA) and 4-aminoantipyrine (4-AA). The calculated pharmacokinetic variables of 4-MAA after D1 and D6 were, respectively: Cmax (μg/mL) = 163.60 ± 179.72 and 178.79 ± 196.94; T1/2beta (h) = 2.65 ± 0.65 and 3.37 ± 1.03; and AUC0-t (μg/mL × h) = 240.38 ± 130.87 and 373.52 ± 78.85. The same variables for 4-AA were: Cmax, (μg/mL) = 0.44 ± 0.27 and 0.90 ± 0.31, T1/2beta (h) = 14.77 ± 13.13 and 35.97 and AUC0-t (μg/mL × h) = 3.20 ± 0.43 and 27.73 ± 11.99. Concentrations of 4-MAA exceeded the minimum concentration required for 50% inhibition of cyclooxygenases 1 and 2. However, an accumulation of 4-AA, was observed. Further clinical studies are necessary to ascertain the implications of these findings on the pharmacodynamic response to dipyrone in northeastern donkeys.

Biostimulation effect of platelet-rich fibrin augmented with decellularized bovine pericardium on full-thickness cutaneous wound healing in Donkeys (Equus asinus)

AimThe current research aimed to evaluate the potential effect of adding platelet-rich fibrin (PRF) to the decellularized bovine pericardium (DBP) on the distal limb of donkeys’ full-thickness cutaneous wounds healing (Equus asinus).Materials and MethodsHealthy male donkeys (n = 12) were used in this study. Under general anesthesia, 6 cm2 full-thickness incisions were made on the middle dorsolateral surface of both forelimbs’ metacarpi. The left forelimbs were control wounds, while the right wounds were treated with PRF/DBP. Control wounds were bandaged with a standard dressing after saline irrigation and were evaluated at days 4, 7, 10, 13, 16, 19, 22, 25, and 28 post-wounding. PRF/DBP-treated wounds were dressed with a combination of PRF/DBP at the first, second, and third weeks post-wounding. Clinical and histopathological examinations of the wounds were performed to assess the healing process. Additionally, the immunohistochemical evaluation and gene expression profiles of myofibroblastic and angiogenic genes (transforming growth factor-β1, vascular endothelial growth factor-A, fibroblast growth factor 7 (FGF-7), and collagen type 3α1) were analyzed.ResultsPRF/DBP wounds had a significantly faster healing process (61.3 ± 2.6 days) than control wounds (90.3 ± 1.4 days) (p < 0.05). The immunohistochemical examination and gene expression profile revealed significant enrichment in PRF/DBP wounds compared to control wounds.ConclusionPRF/DBP dressing can be considered a natural and cost-effective biomaterial for enhancing the recovery of donkeys’ distal limb injuries.

Genomic and comparative analysis of the T cell receptor gamma locus in two Equus species.

The genus Equus is the only extant genus of the Equidae family, which belongs to Perissodactyla, an order of mammals characterized by an odd number of toes (odd-toes ungulates). Taking advantage of the latest release of the genome assembly, we studied, for the first time in two organisms belonging to the Equus genus, the horse (Equus caballus) and the donkey (Equus asinus), the T cell receptor gamma (TRG) locus encoding the gamma chain of the γδ T cell receptor. Forty-five Variable (TRGV) genes belonging to the seven IMGT-NC validated mammalian TRGV subgroups, 25 Joining (TRGJ) and 17 Constant (TRGC) genes organized in 17 V-J-(J)-C cassettes, in tandem on about 1100 Kb, characterize the horse TRG locus, making the horse TRG locus the one with the greatest extension and with a significantly higher number of genes than the orthologous loci of the other mammalian species. A clonotype analysis of an RNA-seq transcriptomic dataset derived from spleen of an adult healthy horse, using the complete set of the horse TRGJ germline gene sequences as a probe, revealed that, in addition to the most prominent V-J rearrangements within each cassette, there is a relevant proportion of trans-cassette V-J recombination, whereby the same TRGV genes can recombine with different TRGJ genes spliced to the corresponding TRGC genes. This recombinant event strongly contributes to the diversity of the γ chain repertoire. In the donkey TRG locus, 34 TRGV, 21 TRGJ and 14 TRGC genes distributed in 14 V-J-(J)-C cassettes were found in a region of approximately 860 kb. Although the donkey's TRG is smaller than that of the horse, in Equus genus, this is still the second largest locus so far found in any mammalian species. Finally, the comparative analysis highlighted differences in size and gene content between the horse and donkey TRG loci, despite belonging to the same genus, indicating a good level of diversification within Equus. These data is in agreement with the evolutionary idea of the existence of a Equus recent common ancestor in rapid evolution, for which a mutation rate between horses and donkeys is more comparable to that between species belonging to different genera rather than to species of the same genus.

Effects of Concentrate Feeding Sequence on Growth Performance, Nutrient Digestibility, VFA Production, and Fecal Microbiota of Weaned Donkeys.

In this study, effects on the growth performance, nutrient digestibility, volatile fatty acids (VFA) production, and fecal microbiota of weaned donkeys were observed using different concentrate feeding sequences. Fifteen healthy 6-month-old weaned male donkeys with a body weight of 117.13 ± 10.60 kg were randomly divided into three treatment groups, including group C1 (roughage-then-concentrate), group C2 (concentrate-then-roughage), and group C3 (total mixed ration, TMR). The experiment lasted 35 d. We measured nutrient digestion by the acid-insoluble ash method and analyzed the fecal microbiota of the weaned donkeys by high-throughput sequencing of 16s rRNA genes in the V3-V4 region. The results show that group C3 obtained the best growth performance, and the digestibility of crude protein (CP) and crude extract (EE) was significantly higher than that of group C1 (p < 0.05). Acetic acid, isobutyric acid, valeric acid, isovaleric acid, and caproic acid were notably different among all groups (p < 0.05). In addition, we observed that Firmicutes and Bacteroidetes were dominant in the fecal microbes of each group, and Firmicutes was significantly higher in group C3 (p < 0.05). At the genus level, the different genera were Treponema, Rikenellaceae-RC9-gut-group, Unidentified-F082, and Bacteroidales-RF16-group (p < 0.05). The prediction of fecal microbiota function by PICRUSt indicated that different feeding sequences had minimal impact on the function of the fecal microbiota, particularly on the high-abundance pathway. In summary, the concentrate feeding sequence changed the composition of the fecal microbe of weaned donkeys.

Feral Donkey Distribution and Ecological Impacts in a Hyper-Arid Region.

The feral donkey (Equus asinus L.) is an invasive species in Saudi Arabia and can cause severe damage to natural and cultural heritage. Over the last 30 years, feral donkeys have become a serious problem, as their abundance and geographic distribution has increased drastically. The impacts of feral donkeys are not well documented, and information about their abundance and distribution is lacking, certainly in Saudi Arabia, which hampers the implementation of effective management plans. Accordingly, we used the minimum population number approach (MPN) to determine the number of feral donkeys in this part of northwest Saudi Arabia. A total of 1135 feral donkeys were encountered in the region. The area around Khaybar harbors ~25% (n = 338) of the feral donkey population, whereas Tayma and AlGhrameel nature reserves were the least-inhabited sites (almost absent). The average population density of feral donkeys was estimated as 1.03 (0.19 SE) donkey/km2. We documented the negative ecological impact of feral donkeys on natural resources, which constituted overgrazing that resulted in habitat fragmentation and competition for resources with native species. We propose urgent actions to control the presence of feral donkeys in the region and suggest humane eradication as the most efficient and applicable to significantly reduce the negative impacts of feral donkeys.

Molecular detection of Theileria cervi in equids from México

Equine piroplasmosis is a parasitic illness caused by various protozoa of the Babesia and Theileria genera, which parasitize within red blood cells. The transmission of these pathogens occurs through certain genus of ticks, including Amblyomma, Haemaphysalis, Hyalomma, and Rhipicephalus. In recent times, an increase in the identification of new Theileria species and genotypes has been observed. This is further complicated by the presence of mixed Theileria infections in both mammals and tick vectors, particularly in regions where wildlife and livestock share habitats and vectors. Therefore, the objective of this study is to document the occurrence of Theileria cervi in a non-typical host. A total of 88 horses (Equus caballus) and 10 donkeys (Equus asinus) were sampled in three municipalities in Veracruz, Mexico. Molecular techniques were employed to identify Babesia/Theileria through the amplification of a segment of the 18S-rDNA and hsp70 genes. The phylogenetic reconstruction grouped the obtained sequences into a monophyletic cluster alongside sequences of T. cervi. This work represents the first documented occurrence of T. cervi in equids. These findings have significant implications from an epidemiological point of view. In addition, further studies are needed to determine the distribution and pathogenicity of this species for domestic animals and to develop effective control strategies.

Managing abandoned Mediterranean mountain landscapes: The effects of donkey grazing on biomass control and floral diversity in pastures

Traditional Mediterranean Mountain landscapes in Spain have suffered dramatic environmental and social changes over the last seven decades. The loss of these landscapes has had consequences for biodiversity, soil erosion, landscape quality and ecosystem services as croplands and pastures were mainly converted into forests and scrublands. Many animal breeds present in traditional land uses such as the Catalan donkey (Equus asinus var. catalana), are also at risk of extinction, but can provide environmental services while recovering traditional landscapes. In Puy de Cinca, a village in mountainous northeast Spain, we studied how grazing by Catalan donkeys reduces pasture biomass and the effects on plant diversity in pastures. We used Sentinel-2 satellite imagery to calculate the Normalized Difference Vegetation Index (NDVI), a biomass sensitive spectral index, throughout the grazing period to monitor pasture biomass and compared it to pastures without grazing. We also calculated several plant diversity indicators in pastures with and without grazing donkeys. Furthermore, we studied land use changes over the last seven decades using old (1956) and current (2018) aerial images, with forest, agricultural lands, and trails interconnecting the village being mapped to understand landscape changes. The results indicated a great increase in the forested area (348.3 ± 17.0 ha). Meanwhile, a severe decrease in cropland area (73 %) and trail length (62.5 %) was also observed. Concerning the effect of donkey grazing, biomass was lower in pastures with grazing donkeys, with NDVI values decreasing once donkeys started grazing. Nevertheless, plant diversity was higher in pastures with grazing donkeys than in abandoned pastures. This study demonstrated the capacity of low-to-moderate-intensity donkey grazing to improve plant diversity and reduce biomass in pastures. Furthermore, the study of land use changes allowed an understanding of landscape dynamics, which can help address the social and environmental recovery of the village.

Evaluation of chemical castration using intra-testicular injection of zinc gluconate into the testis of the male donkey versus surgical castration: antimullerian hormone as an endpoint marker

BackgroundChemical castration of male animals is an alternative to surgical castration for inducing azoospermia, consequent sterility. Intra-testicular injection of zinc gluconate has been used for chemical castration in several animal species. However, its application to equine species, such as donkeys, has yet to be reported. This study aimed to evaluate the use of zinc gluconate for the chemical castration of male donkeys and to compare its effectiveness relative to routine surgical castration. For this purpose, investigations of serum testosterone and anti-Müllerian hormone levels, testicular ultrasonographic echogenicity, and histopathological findings were performed.MethodsFourteen clinically healthy adult male donkeys were randomly and equally divided into two groups. The donkeys in group I (n = 7) underwent surgical castration. The donkeys in group II (n = 7) received intra-testicular zinc gluconate injections. The donkeys were kept under close clinical observation for 60 days. Abnormalities in donkey behavior and gross alterations in the external genitalia were recorded daily. Serum testosterone and anti-Müllerian hormone (AMH) levels were measured 15 days before the start of the treatment and 15, 30, 45, and 60 days after treatment. The testicles of group II donkeys were evaluated ultrasonographically. At the end of the study, the testes were removed and histologically examined.ResultsSerum testosterone levels significantly declined compared to pre-castration levels in surgically castrated donkeys (group I), but donkeys exposed to chemical castration (group II) showed a non-significant reduction in testosterone levels. Donkeys in the surgical group had considerably lower serum AMH levels. In contrast, there was a non-significant (p > 0.05) increase in AMH levels in the chemical group compared with the pre-sterilization level. In addition, ultrasonographic examination revealed that the testicular echo-density had changed, as observed by a few scattered hyperechoic regions throughout the entire testis parenchyma. The histopathological investigation confirmed the presence of necrosis of the spermatogenic epithelium, increased thickness of the basement membrane of the seminiferous tubules, marked interstitial fibrosis, and shrinkage of the seminiferous tubules. Furthermore, syncytial giant cells were present in the lumen of seminiferous tubules and were associated with Sertoli cell vacuolation. Donkeys subjected to chemical castration (group II) had orchitis, as confirmed histopathologically.ConclusionIntra-testicular injection of zinc gluconate resulted in histopathological and ultrasonographic testicular changes in adult male donkeys, which may affect their reproductive potential. However, it did not significantly alter serum testosterone or AMH levels, indicating that it cannot be used as a substitute for surgical castration in male donkeys.