Distribution of calcium, phosphorus, sulfur, magnesium, potassium, and sodium in major fractions of donkey milk.

The aim of this study was to evaluate the concentrations of Ca, P, S, Mg, K, and Na, and their distribution in major fractions of donkey milk (i.e., fat, casein, whey proteins, and aqueous phase). Individual milk samples were collected by mechanical milking from 16 clinically healthy lactating donkeys. Milk yield per milking was recorded and milk gross composition, casein content, and pH were determined. Whole milk samples were centrifuged to separate fat and to obtain skim milk. Skim milk samples were ultracentrifuged to separate a sedimentable casein pellet and to obtain a supernatant whey (soluble) fraction, which was then ultrafiltered to obtain the aqueous phase of donkey milk. Whole milk and the processed samples were analyzed for the aforementioned elements by inductively coupled plasma-mass spectrometry. The concentration of elements associated with fat, casein, and whey proteins was then calculated. All the Na was present in the aqueous phase. The fat fraction in donkey milk carried very little or none of the investigated elements. The majority of Ca (62.9%) and P (53.1%) was associated with casein, and the rest of these elements was mostly present in the aqueous phase. The majority of Mg was present in the aqueous phase, but a relevant part (32.6%) was associated with the casein fraction. No K was associated with casein. On a molar basis, the ratio of colloidal Ca and P to casein (mmol/g of casein) was more than double the values reported in literature for cow milk. The correlation coefficient was negative between milk pH and P in the ultracentrifuged (r = -0.81) and ultrafiltered (aqueous) fraction (r = -0.66). Milk pH correlated positively with colloidal Ca (r = 0.59) and with the ratio of colloidal Ca to casein (mmol/g of casein; r = 0.68). Colloidal Ca and P were positively correlated (r = 0.64). These data suggest that the high ratio of colloidal Ca and P to donkey casein micelles is due to a larger amount of colloidal calcium phosphate bound to casein micelles compared with literature data on cow milk. The percentage of elements associated with whey proteins was less than 5% for Ca, P, and K, but Mg reached approximately 9% of total Mg. The majority of S (63.6%) was associated with whey proteins, and only one-fourth of this element was associated with casein, indicating a higher content of sulfur-containing amino acids in donkey whey proteins than in casein.

Minor and potentially toxic trace elements in milk and blood serum of dairy donkeys.

The aim of this trial was to study the concentration of Ti, V, As, Rb, Sr, Mo, Cd, Cs, and Pb in donkey milk and blood serum. One hundred twelve individual milk and blood serum samples were collected from 16 lactating donkeys (Martina-Franca-derived population; 6 to 12 yr old; 3 to 7 parities; average live weight 205.4kg; 32 to 58 d after foaling at the beginning of the trial) during a 3-mo-long experiment. The samples were analyzed for the aforementioned elements by inductively coupled plasma-mass spectrometry. Feedstuff and drinking water were also analyzed for the investigated elements. Data were processed by ANOVA for repeated measures. Average milk concentrations (±SD) of Ti, Rb, Sr, Mo, Cs, and Pb were 77.3 (±7.7), 339.1 (±82.1), 881.7 (±270.4), 4.5 (±1.6), 0.49 (±0.09), and 3.2 (±2.7) µg/L, respectively. More than 80% of samples were below the limit of detection for V, As, and Cd in milk and for Cd, and Pb in blood serum. The lower bound calculated for milk V, As, and Cd was 0.03µg/L for the 3 elements, the upper bound was calculated at 0.23, 0.10, and 0.31µg/L and the maximum value was observed at 0.54, 0.15, and 0.51µg/L, respectively. The average milk concentrations of Ti, Rb, Sr, Mo, and Cs were 600, 458, 346, 16, and 294%, respectively, than those of blood serum. Yet, Cs concentrations were in the same order of magnitude in milk and serum. Moderate to strong positive and significant correlation coefficients were observed between milk and blood serum concentrations for Ti, Rb, Sr, and Cs. The effect of the stage of lactation was significant for all the investigated elements in milk and blood serum, but most of the elements showed only small changes or inconsistent trends, and only the concentrations of Rb and Sr showed decreasing trends both in milk and blood serum. The relationship between milk and blood serum element concentrations indicates that the mammary gland plays a role in determining the milk concentrations of Mo, Ti, Rb, Sr, Mo, and Cs. In the current experimental conditions, in agreement with the low levels in drinking water and feedstuff, donkey milk concentration of potentially toxic elements was very low and did not raise health concerns for human consumption.

Glutamate-induced epigenetic and morphological changes allow rat Müller cell dedifferentiation but not further acquisition of a photoreceptor phenotype.

Müller cells are not only the main glial cell type in the retina but also latent progenitor/stem cells, which in pathological conditions can transdifferentiate to a neuronal phenotype and regenerate the neurons lost in a mature retina. Several signal transduction pathways can induce the dedifferentiation of mature Müller cells to a progenitor-like state, including that stimulated by glutamate. However, the precise molecular mechanisms by which terminally differentiated cells are initially primed to acquire multipotency remain unclear. In the present study, we have characterized early genetic and epigenetic events that occur immediately after glutamate-induced dedifferentiation of fully differentiated Müller cells is initiated. Using Müller cell-enriched cultures from postnatal rats, we demonstrate that glutamate triggers a rapid dedifferentiation response characterized by changes in cell morphology coupled to the induction of progenitor cell marker gene expression (e.g., nestin, lin28 and sox2) within 1h. Dedifferentiation involved the activation of N-methyl-d-aspartate and type II metabotropic glutamate receptors, as well as global DNA demethylation (evident through the decrease in methyl-CpG-binding protein 2 immunoreactivity) and an increase in gadd45-ß gene expression; although, early progenitor gene expression was only partially inhibited by pharmacological impairment of DNA methylation. Importantly, the expression of Müller glia identity genes (i.e., glutamine synthetase; cellular retinaldehyde binding protein, CRALBP) is retained through the process. Dedifferentiated Müller cells held an early neuronal differentiation potential similar to that observed in retinal progenitor-enriched cultures but, contrary to the latter, dedifferentiated Müller cells failed to further differentiate into mature photoreceptor lineages. We speculate that, in spite of the initial triggering of the dedifferentiation pathways, these cells may exhibit a certain degree of epigenetic memory that precludes them from further differentiation.

Essential trace elements in milk and blood serum of lactating donkeys as affected by lactation stage and dietary supplementation with trace elements.

The aim of this trial was to study the concentration of zinc (Zn), iron (Fe), copper (Cu), manganese (Mn), selenium (Se), cobalt (Co) and iodine (I) in milk and blood serum of lactating donkeys, taking into account the effects of lactation stage and dietary supplementation with trace elements. During a 3-month period, 16 clinically healthy lactating donkeys (Martina-Franca-derived population), randomly divided into two homogeneous groups (control (CTL) and trace elements (TE)), were used to provide milk and blood samples at 2-week intervals. Donkeys in both groups had continuous access to meadow hay and were fed 2.5 kg of mixed feed daily, divided into two meals. The mixed feed for the TE group had the same ingredients as the CTL, but was supplemented with a commercial premix providing 163 mg Zn, 185 mg Fe, 36 mg Cu, 216 mg Mn, 0.67 mg Se, 2.78 mg Co and 3.20 mg I/kg mixed feed. The concentrations of Zn, Fe, Cu, Mn, Se, Co and I were measured in feeds, milk and blood serum by inductively coupled plasma-MS. Data were processed by ANOVA for repeated measures. The milk concentrations of all the investigated elements were not significantly affected by the dietary supplementation with TE. Serum concentrations of Zn, Fe, Cu Mn and Se were not affected by dietary treatment, but TE-supplemented donkeys showed significantly higher concentrations of serum Co (1.34 v. 0.69 µg/l) and I (24.42 v. 21.43 µg/l) than unsupplemented donkeys. The effect of lactation stage was significant for all the investigated elements in milk and blood serum, except for serum manganese. A clear negative trend during lactation was observed for milk Cu and Se concentrations (-38%), whereas that of Mn tended to increase. The serum Cu concentration was generally constant and that of Co tended to increase. If compared with data reported in the literature for human milk, donkey milk showed similarities for Zn, Mn, Co and I. Furthermore, this study indicated that, in the current experimental conditions, the mineral profile of donkey milk was not dependent on dietary TE supply.