In vitro evaluation of the effect of yogurt acid whey fractions on iron bioavailability.

A side effect of the raised consumption of Greek yogurt is the generation of massive amounts of yogurt acid whey (YAW). The dairy industry has tried several methods for handling these quantities, which constitute an environmental problem. Although the protein content of YAW is relatively low, given the huge amounts of produced YAW, the final protein amount in the produced YAW should not be underestimated. Taking into consideration the increased interest for bioactive peptides and the increased demand for dietary proteins, combined with protein and peptides content of YAW, efforts should be made toward reintroducing the latter in the food supply chain. In this context and in view of the prevalent dietary iron deficiency problem, the objective of the present study was the investigation of YAW fractions' effect on Fe bioavailability. With this purpose, an in vitro digest approach, following the INFOGEST protocol, was coupled with the Caco2 cell model. To evaluate whether YAW digest fractions exert positive, negative or neutral effect on Fe bioavailability, they were compared with the ones derived from milk, a well-studied food in this context. Milk and YAW showed the same effectiveness on both Fe bioavailability and the expression of relative genes (DCYTB, DMT1, FPN1, and HEPH). Focusing further on YAW fractions, by comparison with their blank digest control counterparts, it resulted that YAW 3- to 10-kDa digests fraction had a superior effect over the 0- to 3-kDa fraction on Fe-uptake, which was accompanied by a similar effect on the expression of Fe metabolism-related genes (DCYTB, FPN1, and HEPH). Finally, although the 3- to 10-kDa fraction of bovine YAW digests resulted in a nonsignificant increased Fe uptake, compared with the ovine and caprine YAW, the expression of DCYTB and FPN1 genes underlined this difference by showing a similar pattern with statistically significant higher expression of bovine compared with ovine and bovine compared with both ovine and caprine, respectively. The present study deals with the novel concept that YAW may contain factors affecting Fe bioavailability. The results show that it does not exert any negative effect and support the extensive investigation for specific peptides with positive effect as well as that YAW proteins should be further assessed on the prospect that they can be used in human nutrition.

Genomic distribution and polymorphism of G-quadruplex motifs occupying ovine promoters and enhancers.

G-quadruplexes are non-canonical DNA structures that are formed in regions with short runs of guanines. During the last decade they have gained considerable attention due to their involvement in basic cellular processes, linking them to several physiological processes and pathological conditions. Regulation of gene transcription is among the crucial roles that G-quadruplexes play in the cells. Several ways in which these structures affect transcription have been described, both negatively and positively. Recently, G-quadruplexes have been shown to be implicated in the three-dimensional rearrangement of the chromosomes that enables the interaction of enhancers and gene promoters during regulation of transcription. Sheep is a species for which almost no G-quadruplex-related studies have been conducted and thus research on this species is kept out from the progress that has been made in the G-quadruplex field. In this context, we investigated the DNA sequences with potential to form G-quadruplexes (G4-motifs) in the ovine enhancers and promoters. We describe the distribution of G4-motifs within the regulatory regions which is shown to be enriched in G4-motifs in a way similar to other mammals. Furthermore, our data suggest that G4-motifs promote promoter-enhancer interactions in sheep. The single nucleotide polymorphisms colocalizing with promoter- and enhancer-associated ovine G4-motifs constitute a considerable pool of polymorphism and given the crucial role of these specific G4-motifs on regulation of transcription, we suggest this polymorphism as an interesting target for ovine genetic studies.

Genomic landscape, polymorphism and possible LINE-associated delivery of G-quadruplex motifs in the bovine genes.

G-Quadruplex structures are non-B DNA structures that occur in regions carrying short runs of guanines. They are implicated in several biological processes including transcription, translation, replication and telomere maintenance as well as in several pathological conditions like cancer and thus have gained the attention of the scientific community. The rise of the -omics era significantly affected the G-quadruplex research and the genome-wide characterization of G-Quadruplexes has been rendered a necessary first step towards applying genomics approaches for their study. While in human and several model organisms there is a considerable number of works studying genome-wide the DNA motifs with potential to form G-quadruplexes (G4-motifs), there is a total absence of any similar studies regarding livestock animals. The objectives of the present study were to provide a detailed characterization of the bovine genic G4-motifs' distribution and properties and to suggest a possible mechanism for the delivery of G4-motifs in the genes. Our data indicate that the distribution of G4-motifs within bovine genes and the annotation of said genes to Gene Ontology terms are similar to what is already shown for other organisms. By investigating their structural characteristics and polymorphism, it is obvious that the overall stability of the putative quadruplex structures is in line with the current notion in the G4 field. Similarly to human, the bovine G4-motifs are overrepresented in specific LINE repeat elements, the L1_BTs in the case of cattle. We highlight the potential role of these elements as vehicles for delivery of G4-motifs in the introns of the bovine genes. Lastly, it seems that a basis exists for connecting traits of agricultural importance to the genetic variation of G4-motifs, thus, the value of cattle as an interesting new model organism for G4-related genetic studies might be worth investigating.

Factors affecting public transport performance due to the COVID-19 outbreak: A worldwide analysis.

In this paper we investigate the public transport trip frequency variations, as well as the reasons that led to the shift away from public transport means, due to the COVID-19 pandemic. We studied relevant data from the Moovit platform, and we compared operational and trip frequency characteristics of public transport systems before and after the outbreak of the pandemic in 87 cities worldwide. On average, waiting times at public transport stops/stations increased while trip distances decreased, apparently due to the mobility restriction and social distancing measures implemented in 2020. Most of the Moovit users who said that they abandoned public transport in 2020 were found in Italy and Greece. We developed linear regression analysis models to investigate (among the 35 variables examined in the study) the relationship between public transport abandonment rates and socioeconomic factors, quality of service characteristics, and indicators of pandemic's spread. Empirical findings show that public transport dropout rates are positively correlated with the COVID-19 death toll figures, the cleanliness of public transport vehicles and facilities, as well as with the income inequality (GINI) index of the population, and thus reconfirm previous research findings. In addition, the waiting time at stops/stations and the number of transfers required for commute trips appeared to be the most critical public transport trip segments, which significantly determine the discontinuation of public transport use under pandemic circumstances. Our research findings indicate specific aspects of public transport services, which require tailored adjustments in order to recover ridership in the post-pandemic period.

Schizochytrium sp. Dietary supplementation modify Toll-like receptor 4 (TLR4) transcriptional regulation in monocytes and neutrophils of dairy goats.

Animals protect themselves against pathogens or abiotic factors by innate or adaptive mechanisms. Long-chain polyunsaturated fatty acids (ω3) of microalgae modify both human and mice' immune systems resulting in a beneficial balance between pro-inflammatory and anti-inflammatory pathways. However, scarce information exists on their impact on lactating animals' immunity. The objective of this study was to investigate the impact of dietary inclusion of Schizochytrium sp. (rich in docosapentaenoic and docosahexaenoic acid), on the expression of several genes involved in the innate immunity of goats. Twenty-four dairy goats were divided into four homogeneous sub-groups (n = 6). All goats were fed individually with alfalfa hay and concentrate. The concentrate of the control group (CON) had no microalgae while those of the treated groups were supplemented daily with 20 (ALG20), 40 (ALG40), and 60 (ALG60) g Schizochytrium sp. Monocytes and neutrophils were isolated from goats' blood in the 20th, 40th, and 60th days from the beginning of the experimental period. The relative transcript levels of TLR4, MYD88, MAPK, IRF3, IFNG, and pro-inflammatory cytokines (IL1B, IL2, IL8, TNF), and chemokines (CCL5 and CXCL16) were decreased in monocytes of microalgae treated goats compared to the CON. In contrast, MAPK and IL1B relative transcript levels were increased in neutrophils of ALG40 and ALG60 groups. In conclusion, the supplementation of goats' diet with 20 g Schizochytrium sp. resulted in a downregulation of the pro-inflammatory transcriptions, and following further research could be considered as a sustainable alternative strategy to improve immune function.

DNA G-quadruplexes: functional significance in plant and farm animal science.

G-quadruplexes (G4s) are non-canonical structures that can be formed in DNA and RNA sequences which carry four short runs of guanines. They are distributed in the whole genome but are enriched in gene promoter regions, gene UTRs and chromosome telomeres. The whole array of their functional roles is not fully explored yet but there is solid evidence supporting their implication in a number of processes like regulation of transcription, replication and telomere organization, among others. During the last decade, there is an increased research interest for G4s that has resulted in a better understanding of their role in several physiological and pathological conditions. On the other hand, these structures are poorly studied in plant species and animals of agricultural interest. Here, we summarize the current methods that are used for studying G4s, we review the studies concerning plants and farm animals and we discuss the advantages of a more thorough inclusion of G4s research in the agricultural sciences.

Set-style yoghurts made from goat milk bases fortified with whey protein concentrates.

This research paper addresses the hypothesis that the fortification of goat milk base with whey protein concentrate (WPC) could affect both the textural and the biofunctional properties of set-style yoghurt. The effect of fortification of goat milk base with two different WPCs on thermophilic bacteria counts, proteolysis, physical and biofunctional properties of set-style yoghurts was studied at specific sampling points throughout a 4-week storage period. Fortification and storage did not influence thermophilic counts. Physical properties were affected significantly (P < 0.05) by the composition of the protein and the mineral fraction of the WPC but not by the storage. ACE-inhibitory activity was moderate in accordance to low lactobacilli counts and lack of proteolysis. DPPH-radical scavenging activity, Fe2+-chelating activity and superoxide scavenging activity were high. At 28 d an anti-inflammatory effect was observed, which was not affected by WPC addition.

The effect of extended milking intervals (24, 48 and 72 h) on milk yield, milk composition, mammary physiology and welfare traits in dairy ewes.

The majority of the work today regarding the effects of extended milking intervals has focused on dairy cattle and only to a limited extent on dairy goats and sheep. The aim of this study was to investigate the effect of different non-milking intervals on milk yield and composition, mammary physiology and welfare indices in dairy ewes. Thirty-six multiparous ewes in late lactation were allocated to one of four groups of nine and subjected to 24, 48 or 72 h of non-milking or normal milking interval (12 h) (group A, B, C and D, respectively). Data showed that there were no significant differences in milk yield among the experimental groups during the third day after re-milking. Furthermore, no significant differences in milk lactose, protein and fat concentration among the experimental groups were observed after 7, 14 and 21 d of re-milking, respectively. Non-milking for 72 h resulted in a temporary increase of sodium concentration, Na + /K + ratio and enzymatic activities of plasmin (PL), plasminogen (PG) and plasminogen activator (PA) in milk. However, these parameters had similar values among the experimental groups on day 5 after re-milking. The concentration of lactose in blood was also significantly increased as a result of the 72 h non-milking interval and returned to its initial levels the second day after re-milking. These data, taken together, suggest that early involution events that occurred as an effect of non-milking were fully reversible within a short period of time. Finally, no significant signs of welfare impairment were observed in ewes due to extended milking intervals. In conclusion, non-milking up to 72 h had no negative effects on milk yield and composition, mammary physiology and welfare parameters in dairy ewes.

Leucocyte expression of genes implicated in the plasminogen activation cascade is modulated by yoghurt peptides.

The urokinase-plasminogen activator (u-PA), its receptor (u-PAR) and the inhibitors of u-PA (PAI-1 and PAI-2) provide a multi-molecular system in leucocytes that exerts pleiotropic functions influencing the development of inflammatory and immune responses. The objective of the present study was to examine the ability of water soluble extracts (WSE) obtained from traditional Greek yoghurt made from bovine or ovine milk to modulate the expression of u-PA, u-PAR, PAI-1 and PAI-2 in ovine monocytes and neutrophils. WSE were obtained from 8 commercial traditional type Greek yoghurts made from ovine or bovine milk. WSE upregulated the expression of all 4 u-PA related genes in monocytes but the upregulation was much higher in the PAI-1 (10-fold) than in u-PA and u-PAR (3-4 fold) thus, shifting the system towards inhibition. In line with this observation, WSE reduced total and membrane-bound u-PA activity in monocytes. In neutrophils, WSE caused small (50-60%) but significant (P < 0·05) reductions in expression of u-PAR and PAI-2 but had no effect on expression of u-PA, PAI-1 and on total cell-associated and membrane-bound u-PA activity. WSE from yoghurts made from bovine or ovine milk were essentially equally effective in affecting the u-PA system except for the u-PAR gene in ovine neutrophils that was affected (reduced) by the ovine and not the bovine WSE. In conclusion, peptides present in WSE modulated the expression of u-PA related genes but the effect was much more prominent in monocytes than in neutrophils.

The influence of dietary supplementation with oyster mushroom waste on laying hens' performance, egg quality and immune parameters.

The objective of the present study was to evaluate the effects of dietary supplementation with oyster mushroom (Pleurotus ostreatus) waste (OMW) on performance, egg quality, fatty acid (FA) profile and oxidative stability, serum and yolk cholesterol and immune parameters of laying hens. Two hundred fifty-six laying hens were allocated into 4 treatment groups, with eight replicate cages, and were fed for 28 d either a control diet, or diets supplemented with OMW at 1, 2 or 4 g per 100 g feed (P1, P2 and P4 experimental groups, respectively). No significant effects were detected on the performance and egg quality (P > 0.05) except from a modest decrease in the intensity of orange yolk color from hens fed with OMW, as indicated by the reduced DSM YolkFan score and color parameter a* (redness) (P < 0.05), and the tendency for increased color parameter L* (lightness) (P < 0.1). Serum total cholesterol and high-density lipoprotein content were decreased in OMW experimental groups compared with the control group (P, P-linear < 0.05) whereas yolk cholesterol content was unaffected (P > 0.05). The ratio of heterophils to lymphocytes were not influenced by dietary treatment (P > 0.05), whereas T helper lymphocyte (Th) percentage was increased in OMW supplemented groups in comparison with control (P < 0.05, P-quadratic < 0.05). The yolk FA profile was beneficially affected, as shown by the linear increase in polyunsaturated FA and a linear decrease in saturated FA in OMW supplemented groups of hens (P-linear < 0.05), with most notable effects observed in the P4 group. Furthermore, oxidative stability, that was expressed as malondialdehyde content, of both fresh and stored egg yolks for up to 90 d, was significantly improved in OMW supplemented groups (P < 0.05). The beneficial effects of OMW on yolk oxidative stability and FA profile, without adverse effects on performance or egg quality, shows that this byproduct may be successfully employed in laying hens' diets, in a circular economy scheme, with benefits not only for the consumers and farmers but for the environment as well.

Impact of Spent Mushroom Substrate Combined with Hydroponic Leafy Vegetable Roots on Pleurotus citrinopileatus Productivity and Fruit Bodies Biological Properties.

Agricultural activities produce large quantities of organic byproducts and waste rich in lignocellulosic materials, which are not sufficiently utilized. In this study, alternative agricultural waste products, namely, spent mushroom substrate (SMS) from the cultivation of edible Pleurotus ostreatus mushrooms and the roots of leafy vegetables from hydroponic cultivation (HRL), were evaluated for their potential to be used as substrates for the cultivation of Pleurotus citrinopileatus and their effects on the quality, the nutritional value, the chemical properties (lipid, protein, carbohydrate, ash, fatty acid and carbohydrate composition) and the bioactive content (total phenolic compounds and antioxidant activity) of produced mushrooms. SMS and HRL (in different ratios with and without additives) and wheat straw with additives (WS-control) were used. During incubation, the linear growth rate of the mycelium (Kr, mm/day) was measured and used for screening. Mushroom cultivation took place in bags, where several characteristics were examined: earliness (duration between the day of substrate inoculation and the day of first harvest) and biological efficiency (B.E. %, the ratio of the weight of fresh mushrooms produced per dry weight of the substrate × 100). Furthermore, this study aimed to investigate the effect of the protein extract (PE) and carbohydrate extract (CE) of P. citrinopileatus after in vitro digestion (fraction less than 3kDa: PE-DP-3; digestate fraction: CE-D, respectively) on the expression of antioxidant-related genes in the THP-1 cell line. The results showed that mushrooms grown on SMS 50%-HRL 40% had the fastest growth (6.1 mm/d) and the highest protein and lipid contents (34.7% d.w.; 5.1% d.w.). The highest B.E. (73.5%), total carbohydrate (65.7%) and total phenolic compound (60.2 mg GAE/g d.w.) values were recorded on the control substrate. Antioxidant activity was observed in all extracts; the total flavonoid content was low in the samples, and the maximum total triterpene value was detected in SMS 80%-HRL 20% (9.8 mg UA/g d.w.). In all mushrooms, linoleic acid (C18:2) was the main fatty acid (above 60%), and fructose was the dominant individual saccharide. In the investigation of the regulation pathway, NFE2L2 gene expression was upregulated only in the SMS 60%-HRL 40% intervention during incubation with CE-D samples. Additionally, the transcription levels of antioxidant-related genes, SOD1, CAT, HMOX1 and GSR, were increased in the SMS 60-30% intervention. Compared to WS, the alternative substrates are observed to trigger a pathway concerning CE that may resist oxidative stress. This study supports the utilization of agricultural byproducts through sustainable and environmentally friendly practices while simultaneously producing high-value-added products such as mushrooms. Therefore, alternative substrates, particularly those containing HRL, could serve as natural sources of antioxidant potential.

Antioxidant Activity of Sweet Whey Derived from Bovine, Ovine and Caprine Milk Obtained from Various Small-Scale Cheese Plants in Greece before and after In Vitro Simulated Gastrointestinal Digestion.

Whey-derived peptides have been associated with different biological properties, but most peptides are usually further hydrolyzed during the digestive process. In the present study, the antioxidant capacity of 48 samples of sweet whey (SW) derived from cheeses obtained from small-scale cheese plants made with bovine, ovine, caprine or a mixture of ovine/caprine milk was assessed using both cell-free and cell-based assays. SW digestates (SW-Ds) and a fraction (<3 kDa; SW-D-P3) thereof were obtained after in vitro digestion and subsequent ultrafiltration. Antioxidant properties using four different assays were evaluated before and after digestion. Our data showed higher values (p < 0.05) for ORAC, ABTS, FRAP and P-FRAP after in vitro digestion (SW-Ds and SW-D-P3) when compared with the corresponding values before digestion. In the non-digested SW, ORAC values were higher (p < 0.05) for the bovine SW compared with all the other samples. In contrast, the ABTS assay indicated a higher antioxidant activity for the ovine SW both before digestion and for SW-D-P3 compared with the bovine SW. The fraction SW-D-P3 of the ovine SW, using HT29 cells and H2O2 as an oxidizing agent, increased (p < 0.05) the cellular antioxidant activity. Furthermore, the same fraction of the ovine/caprine mixed SW increased, through the NF-κB pathway, the expression of SOD1 and CAT, genes implicated in the oxidative response in macrophage-like THP-1 cells. These findings indicate that SW, and particularly bovine and ovine SW, could be a candidate source for physical antioxidants in human and animal nutrition.

Effect of Milk Origin and Seasonality of Yogurt Acid Whey on Antioxidant Activity before and after In Vitro Gastrointestinal Digestion.

Yogurt acid whey (YAW) is a by-product of Greek strained yogurt production. The disposal of YAW constitutes an environmental problem, and given the increasing demand of Greek yogurt worldwide, its handling is a challenge. However, whey-derived peptides, resulting from microbial fermentation as well as those resulting from further hydrolysis during the digestion process, have been linked to enhanced biological activities. In this study, the antioxidant capacity of 33 samples of YAW obtained from Greek dairy companies of bovine, ovine or caprine origin was investigated using both cell-free and cell-based assays. The YAW samples, their in vitro digestion products (YAW-Ds) and a fraction of the digests (less than 3 kDa; YAW-D-P3) were assessed using four biochemical assays, namely ORAC, ABTS, FRAP and P-FRAP. Our data revealed a higher antioxidant capacity for digested samples compared with undigested samples, with all four methods. ORAC values after in vitro digestion were higher for the ovine samples compared to their bovine (YAW-D and YAW-D-P3) and caprine (YAW-D-P3) counterparts. Furthermore, the YAW-D-P3 fraction derived from samples collected in the summer months exhibited higher ORAC values when compared to the respective fraction from the winter months' samples. The cellular antioxidant activity of ovine YAW-D-P3 was improved in H2O2-treated HT29 cells compared to the control H2O2-treated cells. However, YAW-D-P3 could not trigger either the pathways involving the transcription factors NF-κB or NFE2L2 or the gene expression of SOD1, CAT and HMOX1 in LPS-challenged THP-1-derived macrophages. These results suggest that YAW, and particularly YAW from ovine origin, could be used as a natural source for its antioxidant potential in human and animal nutrition.

The Effects of Yoghurt Acid Whey Marination on Quality Parameters of Pork and Chicken Meat.

Large amounts of yoghurt acid whey (YAW) are annually generated as a result of Greek-style yoghurt production, which poses a great hazard to the environment. In terms of sustainability, YAW application in the meat industry appears as a great alternative since meat marination with natural solutions is a practice that continuously gains ground due to its positive effects on meat sensory attributes. The aim of the present study was to determine the quality characteristics and oxidative status of pork and chicken meat after their marination in yoghurt acid whey. Forty samples per meat type were randomly assigned into five groups: CON, without YAW marination; YAW1 and YAW3, in which meat was marinated at 4 °C and a pH of 4.5 for 15 and 10 h, respectively; or YAW2 and YAW4, in which meat was handled as in the YAW1 and YAW3 group, respectively, while hesperidin at 2 g/L was also incorporated into the marinade. As shown, meat shear force values were decreased in pork but not in chicken meat samples. Meat pH values were also generally decreased, while lightness was increased in raw but not in cooked meat samples as an effect of marination. Moreover, meat oxidative stability was improved to a greater extent in chicken than in pork meat. In order to find the ideal marination period for pork meat, we further immersed it into YAW for 5 h. However, this treatment affected neither meat tenderness and the other quality properties nor meat oxidation rates. In general, hesperidin addition did not have an additional or side effect on the quality traits of pork and chicken meat. As it can be concluded, pork meat marination in YAW for 10-15 h improves tenderness, but 5 h of marination does not. On the other hand, the tenderness of chicken meat was not affected, but its oxidative stability was greatly improved after remaining in the YAW marinade for 10-15 h.

The Effect of Milking Frequency, Breed, and Stage of Lactation on the Milk Fat Globule Size and Fatty Acid Composition in Sheep's Milk.

This study examined the effects of milking frequency, breed, and stage of lactation on the milk fat globules (MFG) size and fatty acids (FA) composition of sheep milk. Milk from Karagouniko (n = 13) and Chios (n = 13) ewes was sampled postpartum on the 93rd, 101st, 108th, 121st, 156th, and 188th days of lactation. On the 108th day, the ewes were divided randomly into two milking groups: Once daily at 06:00 a.m. or twice daily at 06:00 a.m. and 16:00 p.m. Morphometric characteristics of MFG and FA composition were determined for each sample. Once versus twice daily milking had no effect on MFG dimensions, which tended to vary according to breed (smaller MFG were secreted from Chios with p = 0.065), while the stage of lactation had a significant effect (p < 0.001). FA composition differed significantly according to the stage of lactation and breed. The FA profile of the Karagouniko breed showed higher concentrations of short-chain FA. The milk samples from late lactation were characterized by higher concentrations of mono-unsaturated FA (MUFA) compared to early and mid-lactation. Moreover, correlations were found between the average diameter of MFG and FA concentrations, where the size of MFG was positively correlated with saturated FA (SFA) and negatively correlated with MUFA.

Dietary Inclusion Level Effects of Yoghurt Acid Whey Powder on Performance, Digestibility of Nutrients and Meat Quality of Broilers.

In recent years, the increasing demand for Greek strained yoghurt produced massive amounts of acid whey, which constitutes a major environmental pollutant. Whether yoghurt acid whey can be included in poultry diets is not known. The purpose of this study was to evaluate the effects of four dietary levels of yoghurt acid whey powder (YAWP) on the growth performance, nutrient digestibility, meat quality traits and oxidative stability. A total of 300 male 1-day-old Ross 308 broilers were assigned into four groups with five replicates of 15 broilers each: control-fed basal diet with no YAWP addition (WO) or basal diet supplemented with YAWP at 25 g/kg of diet (WA), 50 g/kg of diet (WB), or 100 g/kg of diet (WC). At the starter period, body weight and body weight gain were reduced after WB and WC treatments compared to the WO treatment. Breast meat oxidative stability was improved during refrigerated storage for 1 and 3 d in all YAWP treatments compared to control, while the WA treatment showed an improved oxidative stability after 6 and 9 d. The results suggest that YAWP inclusion at 25 g/kg of diet did not impair performance and extended the meat shelf life by reducing lipid oxidation rates.

Soya protein hydrolysates modify the expression of various pro-inflammatory genes induced by fatty acids in ovine phagocytes.

The objective of the present study was to test the hypothesis that fatty acids are the circulating mediators acting in a pro-inflammatory manner towards activated circulating ovine monocyte/macrophages and neutrophils. Furthermore, whether soya protein hydrolysates (SPH) inhibit the fatty acid-induced increase in the production of pro-inflammatory responses by ovine phagocytes was tested in vitro. All the fatty acids tested (myristic, palmitic, palmitoleic, stearic and oleic) increased (P<0·01; C18>C16>C14) membrane-bound urokinase plasminogen activator (u-PA) and u-PA free binding sites in cell membranes of activated ovine blood monocytes/macrophages, but only the C18 fatty acids (stearic, oleic) were effective towards blood neutrophils. The C18 fatty acids up-regulated (P<0·05) the gene expression of u-PA, u-PA receptor, intercellular adhesion molecule 1 and inducible NO synthase (in monocytes) but not that of cyclo-oxygenase-2, integrin α X and plasminogen activator inhibitor types 1 and 2 by ovine phagocytes. SPH blocked completely or partially all C18 fatty acid-induced changes in the expression of various pro-inflammatory genes. In conclusion, fatty acids selectively 'activate' ovine phagocytes, suggesting that these cells 'sense' metabolic signals derived from adipocytes. Soya protein peptides inhibit all changes in gene expression induced by fatty acids in ovine phagocytes in vitro. This constitutes a novel mechanism of action.

The Effects of Replacing Soybean Meal with Rapeseed Meal, Cottonseed Cake, and Fava Beans on the Milk Yield and Quality Traits in Milking Ewes.

The replacement of soybean meal (SBM) from intensively reared dairy sheep diets has emerged as a significant challenge for sustainable production. However, the effects of this replacement on milk production have not been sufficiently elucidated. The objective of this study was to prospectively assess the effects of replacing SBM with a mixture of alternative protein sources on the milk yield (MY) and the milk quality traits (MQT) in intensively reared dairy sheep. A total of 112 multiparous, purebred milking ewes of the Chios and Frizarta breeds, from two intensive dairy sheep farms, were involved in the study, postweaning, and were assigned to either the control (CR) or the experimental ration (ER) group. In the ER, 3/4 of the SBM was replaced by a mixture of rapeseed meal, cottonseed cake, and fava beans, producing a ration of a similar nutritional value. MY, MQT, and body condition scores were recorded for each individual ewe monthly for a period of 4 months during lactation. The experimental ration was associated with beneficial effects on daily and 100-day fat yields and on the electrical conductivity of milk as an improved udder health status indicator, with no adverse effects on any of the rest of the studied milk production traits.

Effect of growth factors and lactogenic hormones on expression of plasminogen activator-related genes and cell proliferation in a bovine mammary epithelial cell line.

There is conflicting evidence in the literature as to whether up-regulation of urokinase plasminogen activator (u-PA) expression is related to bovine mammary epithelial cell growth. The role of u-PA receptor (u-PAR) and that of the plasminogen activator inhibitors type 1 and type 2 (PAI-1 and PAI-2) in bovine mammary epithelial cell proliferation is not known. The effect of growth factors and various hormones known to affect mammary function on expression of u-PA, u-PAR, PAI-1, PAI-2 and cell proliferation using the BME-UV1 bovine mammary epithelial cell line was examined. Cell proliferation was measured using the MTT assay and direct cell enumeration. Results showed that both IGF-1 and EGF increased cell proliferation but EGF was a more potent mitogen than IGF-1. Furthermore, IGF-1 increased by 2-fold expression of both u-PA and u-PAR while EGF increased by 3·8-fold the expression of only u-PAR. Both growth factors had no effect on expression of PAI-1 and PAI-2. In a manner consistent with changes in gene expression, EGF and to a lesser extent IGF-1 up-regulated total cell associated, membrane-bound and secreted u-PA activity. Thus, a strong correlation exists between u-PAR gene expression along with the activity of u-PA present on cell membranes and cell proliferation. Dexamethasone, prolactin and surprisingly insulin had no effect on cell proliferation. Dexamethasone alone and when combined with insulin or prolactin up-regulated gene expression of both PAI- and PAI-2 but not that of u-PA and u-PAR. Decreased total cell-associated, membrane-bound and secreted u-PA activity was detected in cells cultured in the presence of dexamethasone when combined with insulin or prolactin. However no such effect was observed in the presence of dexamethasone alone. Thus, dexamethasone acting synergistically with prolactin or insulin inhibits the activation of the plasmin-plasminogen system but this inhibition is not correlated with any changes in cell proliferation.

A Note on the Effects of Yoghurt Acid Whey Marination on the Tenderness and Oxidative Stability of Different Meat Types.

The aim of this preliminary study was to examine the effects of yoghurt acid whey (YAW) marination on quality parameters and the oxidative stability of pork, lamb, rabbit and chicken meat. Twenty-four samples per meat type were randomly allocated to one of four groups: CON, without any treatment; YAW1 and YAW2, where samples were marinated for 20 h at 4 °C at a pH of 5 or 4.5, respectively; and YAW3, where samples were treated as in the YAW2 group except hesperidin was also added at the level of 2 g/L. As indicated, meat tenderness was improved as a result of YAW marination, apart from the chicken samples. In general, values of pH, redness and yellowness were decreased after immersion in YAW both in raw and cooked samples. However, lightness was increased in the raw meat samples as a result of YAW marination, though this effect was not observed in the cooked meat samples with the exception of chicken meat. Chroma values were higher in controls compared to YAW-treated groups in raw pork and lamb meat, while no significant differences regarding chroma were found among groups in cooked lamb and rabbit meat. Hue angle values were greater in YAW-treated groups compared to controls in raw samples, whereas no significant differences among groups were indicated in cooked meat. Meat oxidation rates were not affected by treatment with YAW and the hesperidin addition, which improved the oxidative stability of lamb and chicken meat. Thus, YAW marination could be recommended as a novel strategy that improves meat tenderness without negative effects on the other quality characteristics.