Giovanni Martemucci

Immunological properties of donkey's milk: its potential use in the prevention of atherosclerosis.

Donkeys milk is the best substitute of human milk for its content in lactose, proteins, minerals, and omega-3 fatty acids. Here, we have evaluated the effects of colostrum and milk from donkeys (Martina Franca breed) on the function of human peripheral blood mononuclear cells (PBMCs) at different intervals from lactation. Colostrum induced more IgA responses, while milk induced predominantly more IgG responses. Both milk and colostrum induced expression of CD25 and CD69 on PBMCs. The ability to induce release of interleukins (IL) (IL-12, IL-1 beta and IL-10) and tumor necrosis factor-alpha was confined only to milk, while colostrum was devoid of this capacity. Finally, both colostrum and milk induced nitric oxide (NO) release from PBMCs but milk exhibited a greater capacity than colostrum in NO generation. Taken together, these immunological activities exerted by both colostrum and milk from donkeys may be useful in the treatment of human immune-related diseases. In particular, NO induction by donkeys milk may be very useful in the prevention of atherosclerosis, being a strong vasodilator and an effective antimicrobial agent since pathogens and/or their products may play a proatherogenic role.

Fat content, energy value and fatty acid profile of donkey milk during lactation and implications for human nutrition

Background and aimsMilk contains numerous nutrients. The content of n-3 fatty acids, the n-6/n-3 ratio, and short- and medium-chain fatty acids may promote positive health effects. In Western societies, cow’s milk fat is perceived as a risk factor for health because it is a source of a high fraction of saturated fatty acids. Recently, there has been increasing interest in donkey’s milk. In this work, the fat and energetic value and acidic composition of donkey’s milk, with reference to human nutrition, and their variations during lactation, were investigated. We also discuss the implications of the acidic profile of donkey’s milk on human nutrition.MethodsIndividual milk samples from lactating jennies were collected 15, 30, 45, 60, 90, 120, 150, 180 and 210days after foaling, for the analysis of fat, proteins and lactose, which was achieved using an infrared milk analyser, and fatty acids composition by gas chromatography.ResultsThe donkey’s milk was characterised by low fat and energetic (1719.2kJ·kg-1) values, a high polyunsaturated fatty acids (PUFA) content of mainly α-linolenic acid (ALA) and linoleic acid (LA), a low n-6 to n-3 FA ratio or LA/ALA ratio, and advantageous values of atherogenic and thrombogenic indices. Among the minor PUFA, docosahesaenoic (DHA), eicosapentanoic (EPA), and arachidonic (AA) acids were present in very small amounts (<1%). In addition, the AA/EPA ratio was low (0.18). The fat and energetic values decreased (P < 0.01) during lactation.The fatty acid patterns were affected by the lactation stage and showed a decrease (P < 0.01) in saturated fatty acids content and an increase (P < 0.01) in the unsaturated fatty acids content. The n-6 to n-3 ratio and the LA/ALA ratio were approximately 2:1, with values <1 during the last period of lactation, suggesting the more optimal use of milk during this period.ConclusionsThe high level of unsaturated/saturated fatty acids and PUFA-n3 content and the low n-6/n-3 ratio suggest the use of donkey’s milk as a functional food for human nutrition and its potential utilisation for infant nutrition as well as adult diets, particular for the elderly.

Donkey and Goat Milk Intake and Modulation of the Human Aged Immune Response

In a group of 14 healthy aged subjects, donkey and goat milk was administered respectively, for a period of one month. Cytokine profile [interleukin (IL)-12, IL-10, IL-1beta, IL-8, IL-6 and Tumor Necrosis Factor (TNF)-alpha] was assessed before and after milk intake by means of a cytometric bead array test. Data demonstrated that IL-12 was undetectable, while IL-10, IL-1beta and TNF-alpha were released in very low amounts. Quite interestingly, IL-8 was increased by donkey milk administration, while same cytokine was dramatically decreased following goat milk intake. Same pattern of response was noted with IL-6 even if levels of these cytokine were lower than those detectable in the case of IL-8. Taken together, these findings indicate that administration of donkey milk in the aged host is able to upregulate the immune response, while goat milk seems to reduce the exaggerated acute phase response in elderly.

Ability of Goat Milk to Modulate Healthy Human Peripheral Blood Lymphomonocyte and Polymorphonuclear Cell Function: In vitro Effects and Clinical Implications

The in vitro effects of goats milk from different sources (Jonica, Saanen, and Priska breeds plus a commercial preparation) on healthy human peripheral blood mononuclear cells (PBMCs) were evaluated in terms of nitric oxide (NO) and cytokine release. According to the incubation time (24 h or 48 h) used all milks could induce release of NO from monocytes. In this context, however, in the presence of a commercial milk preparation inhibition of lypopolysaccharide (LPS)-induce NO generation was evident. Also polymorphonuclear cells stimulated with the various milks released detectable amounts of NO. In the case of Priska milk inhibition of LPS-mediated NO generation was observed. Despite a broad array of cytokines tested [Interleukin (IL)-4, IL-5, IL-6, IL-10, IL-12, IL-13, IL-17, Tumor Necrosis Factor (TNF)-alpha, Transforming Growth Factor-beta and Granulocyte Colony Stimulating Factor] only IL-10, TNF-alpha, and IL-6 were released by PBMCs upon stimulation with various milks. Taken together, these data indicate that goats milk for its capacity to produce NO may exert a cardioprotective and anti-atherogenic effect in consumers. Moreover, induction of proinflammatory (TNF-alpha and IL-6) and anti-inflammatory (IL-10) cytokines suggests the ability of this milk to maintain immune homeostasis in the immunocompromised host (e.g., aged people).

Major whey proteins in donkey's milk: effect of season and lactation stage. Implications for potential dietary interventions in human diseases

According to current literature, donkey’s milk has been suggested as a hypoallergenic substitute in children affected by cow’s milk protein allergy as well as a promising nutraceutical for aged people. However, the biologically active components of donkey’s milk have not yet completely elucidated. In this framework this study is aimed at measuring α-lactalbumin (α-LA), β-lactoglobulin (β-LG), and lysozyme (LYS), the principal whey proteins in donkey’s milk, in relation to lactation stage and production season. Analysis were performed by reversed-phase high-performance liquid chromatography. α-LA, β-LG, and LYS resulted to be affected by lactation stage (P < 0.01) and production season (P < 0.01). Overall, the protein content was higher (0.01 > P < 0.05) during the first four lactation’s months and decreased until the month 8. The β-LG was the major protein (1.75 mg mL−1 as mean; peak 2.24 ± 0.09 mg mL−1), while the α-LA had a mean concentration of 1.32 mg mL−1 and peaked at month 1 (1.57 ± 0.09 mg mL−1) and LYS (0.66 mg mL−1 as mean) showed the highest value equal to 0.76 ± 0.03 mg mL−1. The highest (P < 0.01) concentration of all proteins was recorded at spring (α-LA: 1.69 mL−1; β-LG: 2.07 mL−1; LYS: 0.76 mL−1).

Yield and quality of milk and udder health in Martina Franca ass: effects of daily interval and time of machine milking

Abstract Twenty asses of Martina Franca breed, machine milked twice a day, were used to assess the influence of milking interval (3-h, 5-h, and 8-h; N=5) and time (700, 1200 and 1900) on milk yield and udder health. Individual milk samples were taken to determine fat, protein and lactose content. Sensory analysis profile was also assessed. Milk’s total bacterial count (TBC), somatic cell content (SCC) and udder’s skin temperature were considered to assess udder health. Milk yield increases by 28.4% (P<0.01) with a milking interval from 3-h to 8-h and is higher (P<0.01) at morning milking. The maximum milk yield per milking corresponds to 700 milking (1416.9 mL) thus indicating a circadian rhythm in milk secretion processes. Milking intervals of 5 and 8 hours cause a decrease (P<0.01) in milk fat and lactose content. The 8-h interval leads to an increase (P<0.01) in SCC but without any significance for the health udder. No alterations about CBT, clinical evaluation and temperature of udder were observed. Milk organoleptic characteristics were better in the 3-h interval milking.

Effects of Verbascoside Administration on the Blood Parameters and Oxidative Status in Jennies and Their Suckling Foals: Potential Improvement of Milk for Human Use

BACKGROUND Oxidative damage of tissues and cellular components is a primary or secondary cause of many human diseases and is associated with the welfare and productivity of farm animals. Natural antioxidants have gained attention for the prevention of oxidative damage-related diseases. AIM OF THE STUDY To determine the effects of dietary supplementation with a natural polyphenol (verbascoside, VB) on the serum lipid profile, the hepatic functionality and oxidative status of jennies and their suckling foals. RESULTS Supplementation with VB over 30 days decreased in jennies the serum levels of total cholesterol, LDL cholesterol, triglycerides, bilirubin, AST and ALT, and it increased the HDL cholesterol. As markers of the oxidative status, a decrease of ROMs and TBARs, and an increase in vitamin E levels were observed. Interestingly, the suckling foals showed the same trends in the blood parameters and oxidative status. CONCLUSIONS Supplementation with VB influenced the lipidic and hepatic profiles, and oxidative status of jennies and the suckling foals, and may represent a potentially novel strategy for improving the functional properties of donkeys milk for human diet and for improving the welfare of young animals.

PrP genotype frequencies and risk evaluation for scrapie in dairy sheep breeds from southern Italy

Concerns regarding scrapie in sheep breeding have increased in the last few decades. The present study was carried out in dairy sheep breeds from southern Italy. In order to find breeding animals resistant to scrapie, the PrP genes of 1,205 animals from entire flocks of dairy native Apulian Leccese and Altamurana breeds, and Sicilian Comisana breed, were analysed for polymorphisms at codons 136, 154, and 171 related to scrapie resistance/susceptibility. The Altamurana breed was considered as two populations (Alt-Cav and Alt-Cra-Zoe), based on presumed cross-breeding. A total of five alleles and ten different genotypes were found. The ARQ allele was predominant for all breeds followed by ARR, the most resistant allele to scrapie, which was highly prevalent in Comisana (50%) and in native Alt-Cav (42.4%). The VRQ allele, associated with the highest susceptibility to scrapie, was detected at not negligeable levels in allocthonous Comisana (3.5%), at a low frequency (0.2%) in native Leccese and Alt-Cra-Zoe, while it was absent in Alt-Cav. The frequencies of PrP genotypes with a very low susceptibility risk to scrapie (R1) was higher in Comisana and Alt-Cav. The most susceptible genotype, ARQ/VRQ, was found only in Comisana. Within the Altamurana breed, there were notable differences between Alt-Cav and Alt-Cra-Zoe sheep. The Alt-Cav was characterised by the absence of VRQ and AHQ alleles and by the higher frequency of the ARR/ARR genotype (18.7%). Breeding programs, mainly in endangered breeds such as Altamurana, should be conducted gradually, combining resistance to scrapie, maintenance of genetic variability, and production.

Post-thaw survival and acrosome integrity of spermatozoa of Leccese rams frozen in different seasons with a milk-egg yolk extender

Abstract The influence of the period of semen collection on post-thawing survival, motility and acrosome integrity of spermatozoa in Leccese rams was studied throughout an entire year. The year was divided into the seasons: winter and spring (first semester) and summer and autumn (second semester). Semen from 5 adult rams was collected every two weeks by artificial vagina and frozen according to a freezing system based on milk-lactose egg yolk to constitute semen doses of 400 x 106 spermatozoa. At thawing, survival and acrosomal status of cells were assessed and the motility of the sperm and their kinetic rating (scale 0 to 5 score) were determined at thawing (0 h), and after 1 and 3 h incubation (37° C). Semen collected during the first semester (winter – spring) of the year showed the highest (P<0.01) proportion of post-thaw live spermatozoa, with the maximum value in winter (P<0.01), and the best acrosomal status of spermatozoa, considered as both total proportion of spermatozoa with acrosome break down and spermatozoa without acrosomes. Acrosome integrity was positively correlated (r = 0.32; P<0.01) with post-thaw sperm viability. Motility of spermatozoa at thawing was not influenced by the period of semen collection. However, after 3 h incubation sperm motility was higher (P<0.01) during the first semester of the year, without a difference between winter and spring. A marked individual ram effect was found on freezability of semen. The results provide evidence that the period of semen collection can influence freezability of spermatozoa in Leccese rams. The best characteristics of spermatozoa were observed during the first semester of the year, corresponding to the sexual hypoactivity season for this breed.

Use of purified FSH and LH for embryo production, cryopreservation by conventional freezing or vitrification and transfer of embryos in dairy ewes

Abstract Three experiments were carried out with the aim of evaluating the efficiency of techniques of in vivo production, storage and transfer of embryos in dairy sheep. Experiment I - For embryo production, thirty-one ewes were synchronized with FGA (vaginal sponges, 40 mg, 9 d) and PGF2α (ICI; 50 µg, 7th d), and subdivided into three groups corresponding to the following superovulatory treatments over 3 days with purified gonadotrophic preparations: A) control, FSH/LH ratio = 1 (250 IU p-FSH : 250 UI p-LH); B) FSH/LH ratio = 2 (250 IU p-FSH : 125 IU p-LH) and daily FSH/LH ratio of 3.4 – 1.7 – 0.8 in the 3 days of treatment, respectively; C) FSH/LH ratio = 2 (250 IU p-FSH : 125 IU p-LH) and daily FSH/LH ratio of 5.0 – 1.0 – 0.3. On the 7th day after oestrus and mating, ovarian response and embryo production were evaluated. Experiment II – Three freezing methods were evaluated based upon post-thaw embryo quality: CF) conventional slow freezing by 1.5 M ethylene glycol (EG); V-1) one-step vitrification based on exposure of the embryos to one solution (EG 7.15 M + ficoll 2.5 mM); V-3) vitrification in three steps, corresponding to three solutions at increasing concentration of glycerol (GLY) and EG (GLY 1.4 M; GLY 3.4 M + EG 1.4 M; GLY 4.6 M + EG 3.4 M). V-1) and V-3) frozen embryos were directly plunged in liquid nitrogen. At thawing, embryo viability was evaluated on the basis of morphological features. Experiment III – For embryo transfer, a total of 26 recipient ewes were synchronized with donors. On the 7th d from oestrus, 11 recipient ewes received fresh embryos (Group FE – control) and 15 recipients received vitrified-thawed embryos (Group VTE). Each recipient received 2 embryos. Superovulatory treatment B) significantly advanced the onset of oestrus compared to the control (27.3 vs 34.7 h; P<0.05). Ovulation rate did not differ among the groups (6.5 to 10.8). Transferable embryos in Group B) (7.2) resulted similar to Group A) (5.3) and significantly (P<0.05) different when compared to Group C) (3.2). V3-method resulted in the highest (P<0.01) transferable embryos (74.5%) compared to CF- and V1-methods. After transfer, in FE and VTE recipient ewes were comparable in fertility rates (72.7 vs 73.3%; P>0.05) and embryo survival (63.6 vs 56.7%; P>0.05). In conclusion, the results demonstrated that treatments B) and C) did not improve superovulatory response compared to A); for embryo cryopreservation the V3 method can successfully be used for embryo transfer in ewes.