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Weaning is undoubtedly one of the most crucial stages in the growth and development of all mammalian animals, including donkey foals. Weaning is a dynamic and coordinated process of the body, which is closely associated with the health, nutrition, and metabolism of the host. Many studies have shown that the intestinal microbiota and serum metabolites of mammals exhibit different changes during lactation, weaning, and postweaning. However, the alterations in serum metabolites in donkey foals before and postweaning and the correlation between serum metabolites and intestinal microbiota are largely unknown. This study is based on the fecal 16S rRNA and serum metabolomes of Dezhou donkey foals. In total, 10 samples (fecal and serum) were collected during the following three stages: before weaning (F.M.1), during weaning (F.M.3), and postweaning (F.M.6). To study the alterations in intestinal microflora, serum metabolites, and their correlation before and postweaning. We found that with the growth and weaning progress of donkey foals, the intestinal microbiota of donkey foals underwent obvious changes, and the diversity of fecal bacteria increased (Chao1 and Shannon indexes). The main intestinal microbial flora of donkey foals include Bacteroides and Firmicutes. We found many microbiota that are associated with immunity and digestion in the postweaning group, such as Verrucomicrobiales, Clostridia, Oscillospiraceae, Akkermansia, and Rikenellaceae, which can be considered microbial markers for the transition from liquid milk to solid pellet feed. Clostridia and Oscillospiraceae can produce organic acids, including butyric acid and acetic acid, which are crucial for regulating the intestinal microecological balance of donkeys. Furthermore, the metabolome showed that the serum metabolites enriched before and postweaning were mainly related to arachidonic acid metabolism and riboflavin metabolism. Riboflavin was associated with the development of the small intestine and affected the absorption of the small intestine. We also found that the changes in the gut microbiome of the foals were significantly correlated with changes in serum metabolites, including lysophosphatidylcholine (LPC; 12,0) and positively correlated with Lachnoclostridium and Roseburia. To summarize, this study provides theoretical data for the changes in the intestinal microbiome and serum metabolism during the entire weaning period of donkey foals. [ABSTRACT FROM AUTHOR]

Introduction: A better understanding of the microbiota community in donkey foals during the weaning transition is a prerequisite to optimize gut function and improve feed efficiency. The objective of the present study was to investigate the dynamic changes in fecal microbiota in donkey foals from pre-to post-weaning period. Methods: A total of 27 fecal samples of donkey foals were collected in the rectum before morning feeding at pre-weaning (30 days of age, PreW group, n = 9), durweaning (100 days of age, DurW group, n = 9) and post-weaning (170 days of age, PostW group, n = 9) period. The 16S rRNA amplicon sequencing were employed to indicate the microbial changes during the weaning period. Results: In the present study, the cessation of breastfeeding gradually and weaning onto plant-based feeds increased the microbial diversity and richness, with a higher Shannon, Ace, Chao and Sobs index in DurW and PostW than in PreW (p < 0.05). The predominant bacterial phyla in donkey foal feces were Firmicutes (>50.5%) and Bacteroidota (>29.5%), and the predominant anaerobic fungi and archaea were Neocallimastigomycota and Euryarchaeota. The cellulolytic related bacteria including phylum Firmicutes, Spirochaetota and Fibrobacterota and genus norank_f_F082, Treponema, NK4A214_group, Lachnospiraceae_AC2044_group and Streptococcus were increased from pre-to post-weaning donkey foals (p < 0.05). Meanwhile, the functions related to the fatty acid biosynthesis, carbohydrate metabolism and amino acid biosynthesis were significantly enriched in the fecal microbiome in the DurW and PostW donkeys. Furthermore, the present study provided the first direct evidence that the initial colonization and establishment of anaerobic fungi and archaea in donkey foals began prior to weaning. The relative abundance of Orpinomyces were the highest in DurW donkey foals among the three groups (p < 0.01). In terms of archaea, the abundance of Methanobrevibacter were higher in PreW than in DurW and PostW (p < 0.01), but the abundance of Methanocorpusculum were significantly increased in DurW and PostW compared to PreW donkey foals (p < 0.01). Discussion: Altogether, the current study contributes to a comprehensive understanding of the development of the microbiota community in donkey foals from pre-to post-weaning period, which may eventually result in an improvement of the digestion and feed efficiency in donkeys. [ABSTRACT FROM AUTHOR]

Simple Summary: Weaning is a stressful event as it is an abrupt separation of donkey foals from their mothers. Donkeys experience a variety of changes and stresses, such as physiological, environmental and diet challenges. The oral cavity is the gateway to total body wellness, and the oral microbiome is important in maintaining oral as well as systemic health. However, little is known on the effect of weaning on the oral microbiome of donkeys. Therefore, the present study was conducted to investigate the relationship between weaning and oral microbial changes using 16S rRNA gene sequencing. The results showed that significant changes in the oral microbiome occurred during the weaning transition period. In addition, the functions related to carbohydrate metabolic pathways were obviously enriched in the oral microbiome in postweaning donkeys compared with preweaning donkeys. The current study provides a deeper insight into the oral microbiota changes during the weaning period and helps to increase awareness of feed changes in donkey foals. As the initiation point of digestion, the oral microbiome is important in maintaining oral and systemic health. However, the composition of oral microbial communities and the influence of weaning on the oral microbiota of donkey foals remains poorly characterized. The present study used buccal swab samples to determine the changes in oral microbial communities occurring at the time of weaning. A total of 20 oral swab samples were collected from two groups: preweaning donkey foals (PreW group, n = 10) and postweaning donkey foals (PostW group, n = 10). The donkey oral microbiome was analyzed by 16S rRNA gene sequencing using Illumina MiSeq. This study is the first report of the donkey oral microbiome in association with weaning. Compared to the preweaning donkeys, the oral bacteria diversity in the postweaning donkeys was increased, with a higher Simpson index. Changes in the composition of the oral microbiota between the PreW and PostW groups were observed in the present study. At the phylum level, the relative abundance of Firmicutes and Myxococcota was significantly greater in the PostW than in the PreW group. At the genus level, the Gemella, unclassified_o__Lactobacillales, and Lactobacillus were increased in the postweaning donkeys. The donkeys' oral microbial functions were predicted using PICRUSt, and the functions related to carbohydrate metabolic pathways were significantly enriched in the oral microbiome in the PostW donkeys. In summary, the current study provides a deeper insight into the oral microbiota changes during the weaning period, and the influence of weaning together with the documented changes in diversity and composition will help us to obtain a better understanding of their long-term health impact within the oral cavities of donkey foals. However, a major limitation of the present study was that the samples were obtained from different animals in the PreW and PostW groups, which may have resulted in variability among the different individuals. Further investigation is needed to monitor the shift in oral microbes in the same individuals during the weaning period. [ABSTRACT FROM AUTHOR]

The experiment was to investigate the effect of spraying and fanning on donkey foals' serum biochemical indexes, antioxidant function and relative gene expression levels in summer, to provide reference for welfare management of donkey foals in summer. 24 healthy donkey foals with similar day-old of (51.95±2.24) d and body weight of (56.80±5.34) kg were selected and divided into four groups, with six replicates (half male and half female) in each group, and one donkey in each replicate. The test was divided into control group, spraying group, fanning group and spraying-fanning group. The experiment lasted for 28 d. The results showed that compared with control group, the donkey foals' serum activity of ALP, AST and LDH in fanning group and spraying-fanning group were extremely reduced (P<0.01), and the serum CRE content in test groups were significantly reduced (P<0.05). Compared with control group, the donkey foals' serum activity of GSH-Px in fanning group and spraying-fanning group was significantly increased (P<0.05), and the MDA content was significantly reduced (P<0.05), and the serum activity of CAT and T-SOD in test groups was significantly increased (P<0.05). Compared with control group, the relative expression levels of CAT, GSH-Px3, SOD1 and SOD2 genes in white blood cells of foals in fanning group and spraying-fanning group were significantly increased (P<0.05). The experiment indicates that spraying and fanning can alleviate the effect of high temperature environment on serum biochemical indexes of donkey foals, improve the activity of antioxidant enzymes in serum, reduce the content of MDA, and upregulate the relative expression of antioxidant related. The effect of fan treatment is relatively good. [ABSTRACT FROM AUTHOR]

Immunoglobulin type G (IgG) concentration both in jennies' colostrum and in serum of donkey foals are mostly unknown in the first 24 h after delivery. The aims of the present study were to evaluate the IgG concentrations of colostrum during the first 24 h of lactation of Amiata jennies, the absorption of colostrum and the weekly body weight gain of the donkey foals. IgG concentrations were assessed in the jennies' colostrum and in the serum of donkey foals. Colostrum was collected in 9 jennies ready after delivery, and at 6, 12, 24 h after foaling from both halves. Serum was collected at the same sampling times from 9 donkey foals. Donkey foals were weighted at birth and then weekly until the 28th days of life. Temporal changes of IgG concentrations in dam's colostrum and in donkey foal serum were analyzed by a linear regression model and a general linear model, respectively. Results showed that colostrum IgG concentration were similar between the left and the right half. Colostrum IgG concentrations decreased continuously throughout the time in all jennies by 0.0244 Log10 mg/mL per hour. Serum IgG concentrations in donkey foals at birth was significantly lower compared to other times. No correlation was found between the colostrum IgG concentrations and the average weekly body weight gain of the donkey foal. The pattern of colostrum IgG levels in jennies and serum IgG concentration in donkey foals seem to be similar to what reported for equine. However, the donkey foals seem to be less agammaglobulinemic at birth compared to the horse foal. The pattern and both serum and colostrum concentrations evaluated in the Amiata donkeys were slightly different from results reported in other donkey breeds, underlying the importance of setting references specific to breed.

Our aim was to measure the concentrations of derived reactive oxygen metabolite (d-ROMs) and biological antioxidant potential (BAP) of donkey mares and foals at delivery and to verify the protective role of the placenta against fetal oxidative stress. A total of 15 Amiata jennies with a physiological gestation length and delivery were included together with 17 foals (two twin foalings). After delivery, maternal and foal venous blood samples were collected along with blood from the artery. Circulating lactate and plasma d-ROMs and BAP were evaluated. The Wilcoxon test for paired data was applied to verify differences in d-ROMs and BAP values, while the Spearman test was used to evaluate correlations. A significantly higher d-ROMs concentration was found in jennies compared to their foals, and to the umbilical artery blood. The BAP was higher in jennies than in their foals, but no differences were observed in the umbilical artery blood. No difference was found between foals and their umbilical cord. Blood lactate was higher in foals than in their dams. Positive correlations were found between mares and umbilical cord for BAP and d-ROMs, and between mares and foals and umbilical cord for BAP. In conclusion, the placenta may be a protective factor for the fetus. As with equine foals, the antioxidant system of donkey foals does not seems to be effective at birth.

The assessment of the behavior and physiological parameters of neonatal foals is essential in the detection of early signs of illness. Modified Apgar scoring systems from human medicine exist and have been validated in foals as a guide for assessing neonatal viability after birth. This study evaluated the viability of 44 Amiata donkey foals at birth, by assessing the Apgar score and comparing the relationship between viability and various physical parameters. A total of 44 Amiata donkey foals and 27 jennies were enrolled in this study. An expert operator examined each foal within 5 min of birth. A complete physical examination was performed, along with an existing four-parameter Apgar score. The presence of the suckling reflex was evaluated. The interval time needed to acquire sternal recumbency and quadrupedal position, as well as nurse from the mare, were recorded. In addition, heart rate (HR), respiratory rate (RR), and rectal body temperature (BT) were measured. Results were expressed as median ± standard error, minimum and maximum values. The effects of the Apgar score on time to reach sternal position and quadrupedal standing, time to nurse from the mare, RR, HR, and BT were estimated along with the differences related to Apgar scoring and gender. Differences between female and male donkey foals regarding the time to acquire sternal position and quadrupedal standing, time to nurse from the mare, RR, HR, and BT were also assessed. Differences between female and male donkey foals regarding the Apgar score was evaluated using a chi-Square test. Finally, the reference values for Amiata donkeys were also calculated. Twenty/44 (45.4%) foals were colts and 24/44 (54.5%) were fillies born from 27 jennies. None of the foals showed an Apgar score lower than 6. Twenty-nine out of 44 foals showed an Apgar score of 8/8, 10/44 a score of 7/8, while 5 foals (11.3%) showed a score of 6/8. No differences between fillies and colts in relation to the Apgar score were obtained. • The assessment of neonatal foal is mandatory to notice precocious signs of illness. • The viability of Amiata donkey foals at birth was evaluated by the Apgar score. • Physical parameters were compared, and the effect Apgar score on them was assessed. • Apgar score relates to body temperature and respiratory rate, not to heart rate. [ABSTRACT FROM AUTHOR]

Abstract: Because immune transfer from jenny to donkey foal is mostly unknown, the aim of the present study was to evaluate, from 5 days before to 10 days after foaling, immunoglobulin (Ig)G, IgA, and lysozyme peripartal concentrations in serum and mammary secretions of 10 healthy, spontaneously foaling Martina Franca jennies and in serum of their mature, viable, healthy foals, in the first 10 days after birth. The results showed that, in jennies, mammary secretion of IgG levels (ranging between 16 and 75 mg/mL) and IgA (0.9–2 mg/mL), and IgG (6.8–13.5 mg/mL) and IgA (0.5–2.4 mg/mL) serum concentrations were not different along the time of study. Also, IgG concentrations in serum of foals did not show significant differences although a high level was observed at 12 hours after birth (8 mg/mL), and IgA concentrations in serum of foals did not show any significant difference, although a high level was observed at 12 hours after birth (1.2 mg/mL). Lysozyme increased significantly at Day 2 after parturition in mammary secretions of jennies (551.9 μg/mL) and at 12 hours in serum of foals (25.9 μg/mL). The study demonstrated that the pattern of passive immune transfer in donkey foals seems to be similar to that reported for the horse foal, with IgG predominating IgA in serum and mammary secretions of the jenny and also in serum of foals. The most significant early increase in foals' serum concerns lysozyme, which probably plays an important role in the innate immunity of the donkey foal in the first challenging hours after birth. [Copyright &y& Elsevier]

Abstract: A 4-day-old male Martina Franca donkey foal was evaluated for a forelimb alteration. Clinical examination and radiographs revealed the agenesia of the distal digit. Biochemical parameters were normal, and ultrasonographic evaluation did not identify any relievable organ alteration. Karyotype study revealed an abnormality on chromosome 1. The foal was discharged with a distal limb bandage in which a palmar splint was applied. A poor prognosis for the functionality of the limb was given. In endangered species, such as the Martina Franca donkey, the excessive inbreeding could result in an increase in genetic disorders. These findings shed new light on the possible pathogenesis of the digital dysgenesia. The study of the karyotype could be a useful approach to detect genetic alterations that could or could not be expressed in the animal, especially in endangered species in which a risk of an excessive inbreeding is considerable. These defects should be considered in the choice and selection of the breeders. [Copyright &y& Elsevier]

Simple Summary: The aim of the study was to measure the concentrations of derived reactive oxygen metabolite (d-ROMs) and the biological antioxidant potential (BAP) of donkey mares and foals in order to evaluate the maternal and neonatal oxidative status at delivery and to verify the protective role of the placenta against fetal oxidative stress. A total of 15 Amiata jennies and 17 foals (four foals were born from twin foaling) were included. Immediately after delivery, maternal and foal venous blood samples were drawn from the jugular vein and from one of the two umbilical arteries. Plasma samples were evaluated for d-ROMs and BAP. Lactate was also assessed from whole blood. Statistical analysis was performed. Blood lactate was higher in foals than in their dams. Our results support the hypothesis that the placenta may be a protective barrier against oxidative damage and that the antioxidant system is not well formed at birth in donkey foals. Our aim was to measure the concentrations of derived reactive oxygen metabolite (d-ROMs) and biological antioxidant potential (BAP) of donkey mares and foals at delivery and to verify the protective role of the placenta against fetal oxidative stress. A total of 15 Amiata jennies with a physiological gestation length and delivery were included together with 17 foals (two twin foalings). After delivery, maternal and foal venous blood samples were collected along with blood from the artery. Circulating lactate and plasma d-ROMs and BAP were evaluated. The Wilcoxon test for paired data was applied to verify differences in d-ROMs and BAP values, while the Spearman test was used to evaluate correlations. A significantly higher d-ROMs concentration was found in jennies compared to their foals, and to the umbilical artery blood. The BAP was higher in jennies than in their foals, but no differences were observed in the umbilical artery blood. No difference was found between foals and their umbilical cord. Blood lactate was higher in foals than in their dams. Positive correlations were found between mares and umbilical cord for BAP and d-ROMs, and between mares and foals and umbilical cord for BAP. In conclusion, the placenta may be a protective factor for the fetus. As with equine foals, the antioxidant system of donkey foals does not seems to be effective at birth. [ABSTRACT FROM AUTHOR]