In June 2022, at the XXXII Conference of the Italian Society of Parasitology, the parallels of the main endoparasitic infections of horses and donkeys were discussed. Although these 2 species are genetically different, they can be challenged by a similar range of parasites (i.e. small and large strongyles, and Parascaris spp.). Although equids can demonstrate some level of resilience to parasites, they have quite distinct helminth biodiversity, distribution and intensity among different geographical locations and breeds. Heavily infected donkeys may show fewer clinical signs than horses. Although parasite control is primarily provided to horses, we consider that there may be a risk of drug-resistance parasitic infection through passive infection in donkeys when sharing the same pasture areas. Knowing the possible lack of drug efficacy (<90 or 80%), it is advocated the use of selective treatment for both species based on fecal egg counts. Adult horses should receive treatment when the threshold exceeds 200–500 eggs per gram (EPG) of small strongyles. Moreover, considering that there are no precise indications in donkeys, a value >300 EPG may be a safe recommendation. We have highlighted the main points of the discussion including the dynamics of helminth infections between the 2 species.

Dehydration is a serious welfare issue for equines working in developing countries. Risk factors such as high ambient temperature, heavy workload and poor water availability are exacerbated by the traditional belief that provision of water to working animals will reduce their strength or cause colic and muscle cramps.

As part of the welfare assessment of 4889 working horses and donkeys during 2002/3, eight observers were trained to perform a standardised skin tent test. The prevalence of a prolonged duration of skin tenting was 50% in horses and 37% in donkeys. Two studies investigated inter-observer repeatability of skin tent test techniques, using a total of 220 horses and donkeys in India and then Egypt: measures of agreement with a ‘gold standard’ observer varied from 40 to 99%. Simplifying the test by reducing the number of possible scores for skin tent from three (immediate return of skin to normal position; delayed return up to three seconds; delayed return more than three seconds) to two (immediate return of skin to normal position; delayed return of any duration) did not improve overall repeatability of the skin tent test. Potential reasons for not achieving high levels of agreement include variations in assessment method, assessors' previous experience, subjective demarcation between score categories and biological variability.

Development agencies and animal welfare charities try to improve the health and welfare of livestock in the developing world by educating owners and providing healthcare. The impact assessment of these projects relies mainly upon input-related parameters (eg number of animals treated or educational lectures delivered). The aim of this study was to investigate whether animal-based parameters, such as scores for skin lesions, body condition and lameness, could be used to assess the impact of interventions by development agencies on working donkeys. A general checklist for integument assessment of livestock, developed and then tested on two British farms, was redefined for assessment of equine animals in West Kenya. In total, 346 donkeys were assessed over four days with a mobile clinic of the Kenyan Society for the Protection and Care of Animals, using 25 animal-based parameters. The checklist was easy to use: the parameters could be scored using visual assessment or palpation, and the procedure was completed in approximately 5 min per animal. The method was found to be acceptable for owners and animal health technicians, and no special equipment was required. Significant observations included a reduced frequency of leg lesions when head-tethering (as opposed to leg-tethering) was used, and a reduced frequency of foot lesions in regions previously visited by the charity. This animal-based method proved that the charity had made a positive impact on donkey welfare through owner education.

The well-documented versatility of donkeys (Equus africanus asinus) means that issues concerning their welfare can vary depending on how they are mainly used and their geographic location. The present study is the first assessment of donkey welfare to be systematically conducted in the Iberian Peninsula. This area is characterised by the coexistence of high levels of mechanisation and industrialisation, and human populations with low levels of formal education, still making use of donkeys in their daily work. This study aims to evaluate the main welfare problems affecting donkeys within this context. The welfare assessment was carried out in accordance with the first level of the AWIN protocol for donkeys. Only 37.6% of the animals evaluated showed all positive indicators, with no physical problem detected while the remaining 62.4% revealed at least one negative health indicator. Body Condition Score (BCS) was one of the main concerns, with around half of the donkeys exhibiting an inadequate body condition. Dental evaluation revealed similarly concerning results, with 62.8% of the animals assessed requiring treatment. Indications of hoof neglect were noted in 39.5% of the animals evaluated with a 9.84% incidence of lameness. Skin problems affected 26.7% of individuals. Only 41% of the animals displayed positive results for all behavioural indicators. Aside from BCS, the most prevalent problems were sub-optimal behaviour, pain reaction to cheek palpation, hoof disease and integument alterations. These issues should be prioritised, both as regards this specific donkey population and the education of their caretakers.

Since 2005, owners of draught and pack horses, mules and donkeys in nine districts of Uttar Pradesh, India, have received support from a UK-based charity, the Brooke. One thousand, three hundred and ninety-six village-level groups of owners and carers, responsible for 29,500 animals, were facilitated to develop their own welfare assessment protocols using a participatory learning and action process adapted from recognised good practice in human social development. Each group assessed the welfare of their animals collectively, using findings to generate action plans for improving equine health, husbandry and working practices. Welfare assessments were repeated at 1 to 3 month intervals. Competitiveness between participants to improve their animals’ welfare acted as a driver to increase the number of indicators and sensitivity of rating scales, enabling differentiation of small, incremental improvements in order to identify a ‘winner’ of each welfare assessment. Binary or three-point ‘traffic light’ (red-amber-green) scales evolved into a range of 5-, 10-, 20-point or continuous scales, then into multi-level and weighted measures to quantify the welfare improvements seen. Efforts to aggregate multi-dimensional indicators into a single ‘winning’ score led to indices describing welfare at individual animal level (‘welfare index’) and population level (‘village index’). Benefits of owner-driven monitoring include high levels of commitment and strong peer motivation or pressure to take action. Welfare monitoring and action to improve welfare are integrated within a single process carried out by the same people, in contrast to the separation of evaluation and implementation of welfare improvement seen in inspection or accreditation schemes. Challenges include aggregation of results from a variety of protocols for external analysis, reporting or certification.

Donkeys (Equus asinus) are widely used throughout Ethiopia and play essential roles in a variety of everyday and income-generating tasks for the people that use them. The challenges faced by people and their working equids vary across communities and geographic locations. This may have implications for how donkeys are perceived by the people they work for, the roles they fulfil and ultimately their welfare. Two complementary methodological approaches were used in this study to explore the socio-economic value of donkeys for their owners and the welfare of the donkeys in rural and urban Ethiopia. Using a questionnaire, donkey owners were asked about their donkeys, their attitudes and beliefs related to donkey use and ownership, and the role donkeys played in their lives. Animal-based welfare assessments were also conducted on a sample of donkeys from different locations, with the overarching aim of the study to investigate differences in use, beliefs, and donkey welfare between rural and urban locations. In both rural and urban locations, working donkeys are critical for their owners’ income-generating activity and therefore their livelihoods. The work they undertake differs substantially between locations, as does their welfare. Work in each setting presents its own challenges and these are reflected in the behaviour and physical health of the donkeys. Rural donkeys showed more apathetic behaviour, a higher ectoparasite burden and greater evidence of tethering/hobbling. Urban donkeys were more alert and had a wider range of body condition scores. The findings highlight marked differences in the role and welfare of donkeys between different areas within the same country, demonstrating the importance of understanding the context, both from the perspective of humans and working equids, prior to staging interventions intended to benefit either party.

Lectins are glycoproteins of a non-immune origin often used as histochemical reagents to study the distribution of glycoconjugates in different types of tissues. In this study, we performed a comparative cellular localization of sugar residues in bull and donkey testes using immunofluorescent lectin histochemistry. We inspected the cellular localization of the glycoconjugates within the testes using 11 biotin-labeled lectins (LCA, ConA, PNA, WGA, DBA, SBA, ECA, BPL, PTL-II, UEA-1, and PHA-E4) classified under six groups. Although the basic testicular structure in both species was similar, the cellular components showed different lectin localization patterns. The statistical analysis revealed no significant association between the intensity of labeling and different variables, including group and type of lectin and type of cell examined, at p < 0.05. However, a stronger response tended to occur in the donkey than in the bull testes (odds ratio: 1.3). These findings may be associated with the different cellular compositions of the glycoproteins and modification changes during spermatogenesis. Moreover, glycoconjugate profiling through lectin histochemistry can characterize some cell-type selective markers that will be helpful in studying bull and donkey spermatogenesis.

Donkeys have been used as working animals for transport and farm activities worldwide. Recently, in European countries, there has been an increasing interest in donkeys due to their use as pets, onotherapy or milk production. During 2014–2016, a countrywide survey was conducted to determine prevalence and risk factors of principal helminth infections in 1775 donkeys in 77 Italian farms. A questionnaire on management and parasite control practices was filled out for each farm. Faecal samples were examined using a modified McMaster technique, a centrifugation/flotation method and a sedimentation technique. Pooled coprocultures were performed for differentiation of strongylid eggs. Strongyles were the most common parasites detected (84.9%), followed by Dictyocaulus arnfieldi (6.9%), Oxyuris equi (5.8%), Parascaris spp. (3.6%), Anoplocephala spp. (1.0%), Strongyloides westeri (0.3%). Coprocultures revealed an omnipresence of cyathostomins (100%), followed by Strongylus vulgaris (31.0%), Poteriostomum spp. (25.0%), Triodontophorus spp. (9.0%), Strongylus edentatus (7.0%), Strongylus equinus (5.0%). Logistic regression analysis identified breed, co-pasture with horses, living area, herd size and number of treatments as significantly associated with strongyles. Sex, age, living area and herd size were significantly associated with Parascaris spp. Dictyocaulus arnfieldi was significantly associated with sex, grass, co-pasture with horses, living area and herd size. Strongylus vulgaris was significantly associated with living area and herd size. The mean number of anthelmintic treatments/year was 1.4; most of the donkeys (71.8%) were dewormed using an ivermectin drug. It is important to design parasite programs to specifically address both D. arnfieldi and S. vulgaris in donkeys, and this is especially important if donkeys co-graze with horses.

The lack of anthelmintic products licensed for donkeys and the rising number of small donkey milk farms in the countries of Western Europe and Italy have led to an increased interest in the study of reliable and safe plant-derived treatment alternatives. In this study, the aqueous extracts of Achillea millefolium L. (flowers), Artemisia absinthium L. (aerial parts), Centaurium erythraea Rafn. (flowers), Gentiana asclepiadea L. (rhizomes and roots), Inula helenium L. (rhizomes and roots) and Tanacetum vulgare L. (aerial parts), have been tested in vitro for their potential ovicidal and larvicidal activity against donkey nematodes. An egg-hatching assay (EHA) and larval development assay (LDA) were performed for the in vitro study, and median lethal concentration (LC-50) values for both EHA and LDA were calculated using probit analysis. All tested plant extracts showed strong anthelmintic activity against strongyle eggs and larvae at concentrations ranging between 125 and 1.95 mg/ml, except for C. erythraea, which exhibited very little or no effect at all at the tested concentrations. A strong ovicidal effect was observed in A. absinthium, with an LC-50 value of 0.486 mg/ml (95% confidence interval (CI) 0.21–1.09). Gentiana asclepiadea showed high efficacy against strongyle larvae, with an LC-50 value of 0.041 mg/ml (95% CI 0.01–0.16). The most significant (P < 0.01) anthelmintic activity was exhibited by I. helenium, with an LC-50 value of 0.041 mg/ml (95% CI 0.01–0.16) for EHA and 0.41 mg/ml (95% CI 0.27–0.62) for LDA. The results proved the anthelmintic efficacy of the tested plant extracts, highlighting the need for further research into plant bioactive molecules both in vitro and in vivo.

Information on donkey sperm bioenergetics, kinetics and oxidative status is scarce even though crucial for development of reproductive technologies and germplasm conservation. For these reasons, it is interesting to monitor sperm kinetics, bioenergetics, and oxidative status during sperm storage at +4°C and with several sperm extenders and concentrations. Donkey semen was collected from three jackasses, three times each. It was diluted with four extenders (Kenney, Equiplus, INRA96 or Hippex), set at three sperm concentrations (30, 50 or 70 × 106 spermatozoa/ml) and evaluated for its functionality after 0, 3, 24, 48 and 72 h storage at +4°C. Sperm kinetics was analyzed by Sperm Computer Analysis; sperm bioenergetics was assessed by mitochondrial membrane potential (MMP); sperm oxidative status was evaluated by lipid peroxidation (LPO), anti-LPO potential and nitroblue tetrazolium (NBT) assays. Incubation produced a progressive (P < 0.01) decline in sperm kinetics and MMP, whereas parameters related to oxidative status either increased (LPO, NBT) or decreased (anti-LPO). The anti-LPO potential was the index better related to sperm motility and kinetics. Extenders proved to be differently (P < 0.01) effective in preserving sperm kinetics, MMP, and oxidative status. The concentration of 30 × 106 spermatozoa/ml provided an optimum preservation of sperm functions. Significant correlations emerged between most parameters examined. This study identified reference criteria for storing donkey spermatozoa at +4°C. A low sperm concentration together with a proper extender are crucial requirements for optimum sperm cryopreservation efficiency. Field trials are, however, required to validate these findings, making them operational in practice.

The amount of the four caseins (αs1, αs2, β and κ-CN) in donkey milk was evaluated by Urea-PAGE analysis at pH 8.6, followed by immuno-detection with polyclonal antibodies, coupled to densitometric analysis. The results showed the percentage of each casein in decreasing order: β (54.28) > αs1 (35.59) > αs2 (7.19) > κ-CN (2.79). The mRNA quantification of donkey casein transcripts, carried out by RT-qPCR, showed that the average percentage of corresponding gene transcripts (CSN2, CSN1S1, CSN1S2 I and CSN3) was 70.85, 6.28, 14.23 and 8.65, respectively. The observed translation efficiency, assessed as percentage of single milk casein fraction out of single percentage of transcript, was 0.76, 5.66, 0.50 and 0.32, respectively. The analysis of the sequences flanking the start codon, the codon usage frequencies and the coding sequence length might explain, at least in part, the differential transcriptional and translational rate observed among the casein transcripts.

This Research Paper aimed to investigate donkey welfare in dairy husbandry systems and to identify the potential factors affecting it at animal level. In 2015, twelve dairy donkey farms (19–170 donkeys per farm, mean = 55 ± 48), distributed throughout Italy, were visited. On each farm, the Animal Welfare Indicators (AWIN) welfare assessment protocol for donkeys was used by two trained assessors to evaluate the welfare of animals for a total of 257 donkeys assessed. The protocol includes animal-based indicators that were entered in a digitalised system. Prevalence of different scores at individual, farm and category level were calculated. Farmers were asked to fill out a questionnaire including information regarding the management of donkeys and their final destination. Answers to the questionnaire were then considered as effects in the risk factor analysis whereas the scores of the animal-based indicators were considered as response variables. Most of the donkeys (80·2%) enjoyed a good nutritional status (BCS = 3). 18·7% of donkeys showed signs of hoof neglect such as overgrowth and/or incorrect trimming (Min = 0% Max = 54·5%). Belonging to a given farm or production group influenced many of the welfare indicators. The absence of pasture affected the likelihood of having skin lesions, alopecia, low BCS scores and a less positive emotional state. Lack of routine veterinary visits (P < 0·001) and having neglected hooves (P < 0·001) affected the likelihood of being thin (BCS < 3). Belonging to specific production groups, lack of access to pasture and showing an avoidance reaction to an approaching human (AD) resulted in risk factors associated with a higher prevalence of signs of hoof neglect. Our results support the idea that lack of knowledge of proper donkey care among owners was behind many welfare issues found.

Various uses of donkeys’ milk have been recently proposed for human consumption on the basis of its nutritional characteristics. Improvements in milk fatty acid profile and animal oxidative status can be induced through dietary supplementation of phenolic compounds. The study aimed to evaluate in donkeys the effects of dietary supplementation with verbascoside (VB) on: (i) the fatty acid profile and vitamins A and E contents of milk during a whole lactation, and (ii) blood biochemical parameters and markers of oxidative status of the animals. At foaling, 12 lactating jennies were subdivided into two groups (n 6): control, without VB supplement; VB, receiving a lipid-encapsulated VB supplement. Gross composition, fatty acid profile and vitamins A and E contents in milk were assessed monthly over the 6 months of lactation. Serum total cholesterol, high-density lipoproteins cholesterol and low-density lipoproteins cholesterol, tryglicerides, non-esterified fatty acid, bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase, reactive oxygen metabolites, thiobarbituric acid reactive substances (TBARs), vitamin A and vitamin E were evaluated at 8 days after foaling (D0) and then at D90, D105 and D120 of lactation. In milk, the VB supplementation decreased the saturated fatty acids (P<0.05) and increased the monounsaturated fatty acids (P<0.05), and vitamins A and E (P<0.01) values. On the serum parameters, the VB supplementation decreased total cholesterol (P<0.01), tryglicerides, bilirubin, ALT and TBARs, and increased (P<0.01) vitamin E. In conclusion, the VB dietary supplementation affects the nutritional quality of donkey’s milk with a benefit on the oxidative status and serum lipidic profile of the animals.

The study reported in this Research Communication was carried out to examine how parity and milking time affected donkey milk yield under a typical Chinese production system. Eighteen Jiangyue breed donkeys with good health condition, aged between 6 and 9 years, and with parities 3 and 4, were selected. Milk production was measured commencing from the fourth week post-foaling, with a total data collection of 170 d in milk (lactation length). Mean milk yield of the donkeys used in this study was 3·0 kg/donkey/d. Results showed milk yield decreased with days in milk from 3·3 kg/donkey/d at the start of the study, to 2·2 kg/donkey/d by the end of the 170 d. Parity 3 donkeys produced 22% more milk than parity 4 donkeys (3·3 kg/donkey/d vs 2·7 kg/donkey/d). The information provided by this study should aid producers in estimating the expected milk yields from individual donkey, and better predict milk yield over the course of a production cycle.

In this study, the first postpartum heat, termed the foal heat, characteristics and performance in female donkey (jenny) of Martina Franca are described. To this end, the follicular development of 42 jennies during foal heat was compared with that of 31 jennies at the third estrus after foaling. Estrus length (7.1±0.9 and 6.8±0.7 days), follicular development and preovulatory follicle size (43.7±3.5 and 45.1±2.5 mm) were similar between jennies during the foal heat and during the third estrus after foaling. The pregnancy rate at day 14 was significantly lower in the foal heat jennies (57.1%) than the third estrus jennies (82.3%). However, the pregnancy rate at day 14 in foal heat jennies increased significantly when the onset of foal heat was ⩾8 days after foaling (93.8%) or when the ovulation happened ⩾12 days after foaling (85.7%). The data provided in the present study suggest that the foal heat in the endangered jennies of Martina Franca could be successfully utilized to reduce the interpartum period if the onset of foal heat occurs >8 days after foaling.

Very little is known about the udder characteristics, partitioning of milk in the mammary gland and efficiency of machine milking in donkeys. This study aimed to evaluate the characteristics of the udder and teats, milk yield in relation to pulsation rates (90, 120 and 150 cycles/min), milk partitioning in the mammary gland, composition of the spontaneously removed and residual milk fractions and milking efficiency. Forty-one healthy Martina Franca jennies in the third month of lactation and routinely milked twice daily were used in three studies. Udder characteristics were evaluated by direct measurements and ultrasonographic scanning. Residual milk was obtained by milking after an oxytocin administration (40 IU i.m.). The prevalent shapes were ‘bowl’ for udders and ‘conical’ for teats. After milking the udder characteristics decreased within a range from −11·6% (udder depth) to −25·7% (diameter of teat at the base). The internal structures of the udder resulted as several pockets of ducts empting directly into the teat. The pulsation rate of 120 cycles/min improved (P<0·05) the milk yield in comparison to the 90 and 150 cycles/min, reduced the residual milk fraction, thus improved (P<0·05) milking efficiency. Residual milk composition had higher (P<0·05) fat content and somatic cell count than the spontaneously removed milk fraction. The udders revealed several pockets of ducts empting into the teat instead of a single cisternal cavity and showed a certain compliance. The use of 120 cycles/min pulsation rate improved milking efficiency.

Most donkey and local horse breeds are vulnerable to extinction as mechanization of agriculture progress throughout the world. The present study analyzed the pedigree and herd records of the donkey Asinina de Miranda breed (RAM), identifying genealogical and human factors that may affect the breed genetic diversity in the future and suggesting suitable strategies to breed preservation, early on the conservation program. The breeding rate was very low, with a ratio of foaling/live animals of 0.23 (178/760). The estimated number of founders and ancestors contributing to the reference population was 128 and 121. The number of founder herds in the reference population was 64, with an effective number of founder herds for the reference population of 7.6. The mean age of herd owners was 65.50±0.884 years, with a negative association among the herd size and owner’s age (P<0.001). In contrast, the size of the herd and the ownership of a male were both positively associated (P<0.001) with the herd number of in-born foals. Both the owners’ age and the herd location (RAM home region v. dispersal region) were negatively associated with the foaling number (P<0.001). The main identified risk factors were: low breeding rates; low number of males and their unequal contribution to the genetic pool; unequal contribution of the herds to genetic pool; and advanced age of herd owners.