E. Tsiplakou

Pesticides residues in milks and feedstuff of farm animals drawn from Greece

The objective of this study was to investigate if milk from dairy sheep and goats, fed mainly with supplementary feed during the winter months, was contaminated with pesticides residues. Tauwo hundred milk samples from sheep and goats were collected from 10 farms of each animals. The sheep and goats farms were selected from those which represent common conventional production and feeding systems in Greece. Milk and feed samples (alfalfa hay, wheat straw, shrubs, pasture and concentrates) were taken from each farm to analyze for pesticides residues. The results showed that the summation operatorendosulfan was the main pesticide residue which was detected in all the concentrates samples at a mean concentration of 5.36 mgkg(-1), which is much higher from the maximum residue level (MRL). In addition, the summation operatorendosulfan was also detected in all the alfalfa hay samples but at a mean concentration of 0.10 mgkg(-1) which is lower than the MRL. The mean concentrations of endosulfan alpha and beta were 2.82 and 2.39 mgkg(-1) in the concentrates samples and 0.08 and 0.02 mgkg(-1) respectively in alfalfa hay samples. In the wheat straw, shrubs and pasture samples no pesticides residues were detected. No pesticide residues were also detected in milk samples of sheep and goats. Thus, this milk from the farms sampled presents no human health risks as far as the contaminants analyzed concerned.

Differences in sheep and goats milk fatty acid profile between conventional and organic farming systems

The objective of this study was to investigate whether there is a difference in chemical composition and particularly in fatty acid (FA) profile, with emphasis on cis-9, trans-11 CLA, of milk obtained from conventional and organic dairy sheep and goats farms under the farming conditions practiced in Greece. Four dairy sheep and four dairy goat farms, representing common conventional production systems and another four dairy sheep and four dairy goat farms, organically certified, representing organic production and feeding systems were selected from all over Greece. One hundred and sixty two individual milk samples were collected from those farms in January-February 2009, about three months after parturition. The milk samples were analyzed for their main chemical constituents and their FA profile. The results showed that the production system affected milk chemical composition: in particular fat content was lower in the organic sheep and goats milk compared with the corresponding conventional. Milk from organic sheep had higher content in MUFA, PUFA, alpha-LNA, cis-9, trans-11 CLA, and omega-3 FA, whereas in milk from organic goats alpha-LNA and omega-3 FA content was higher than that in conventional one. These differences are, mainly, attributed to different feeding practices used by the two production systems. The results of this study show that the organic milk produced under the farming conditions practiced in Greece has higher nutritional value, due to its FA profile, compared with the respective conventional milk.

The effect of dietary inclusion of olive tree leaves and grape marc on the content of conjugated linoleic acid and vaccenic acid in the milk of dairy sheep and goats.

Olive tree leaves (OTL) and grape marc (GM) are by-products with high linolenic (LNA) and linoleic (LA) acid content, respectively, which can be used as dietary ingredients to increase the cis-9 trans-11 conjugated linoleic acid (CLA) content of milk fat in sheep and goats. An experiment was conducted with 16 Friesian ewes and 16 Alpine goats to study the effect of OTL and GM inclusion in sheep and goat diets on their milk fatty acid profile, with emphasis on cis-9, trans-11 CLA and vaccenic acid (VA). Ewes and the goats were fed the control (C) diets from parturition to the 90 days in milk (DIM) and then both groups were divided into two sub-groups (treatments). The control groups of both species continued to be fed the C diets, whereas the treated groups were gradually switched over a 2-week period (DIM=91-105) from the C diets to that of treatment 1, which contained air-dried OTL. These OTL diets were fed ad libitum for 1 month (DIM=106-135). After that period, the same treated groups, after 2 weeks of gradual adaptation (DIM=136-150), were switched to treatment 2, which contained air-dried GM. The GM diets were fed ad libitum for 1 month (151-180 DIM). Concentrations of polyunsaturated fatty acids (PUFA) and mono-unsaturated fatty acids increased significantly in milk fat of sheep fed OTL v. C. For goats, only the PUFA in milk fat was increased by feeding OTL compared with C. Relative to C, GM increased significantly the concentration of PUFA only in milk fat of sheep. OTL and GM diets increased the cis-9, trans-11 CLA and VA content in milk fat, compared with C, only in sheep. GM caused a sharp increase in 18:0 only in sheep milk fat, while the OTL diet increased significantly the 18:0 in milk fat of goats. GM and OTL diets also had opposite effects on the 18:1/18:0 ratio of sheep milk fat. In conclusion, OTL and GM, when included in sheep diets altered the milk fatty acid profile with a pronounced increase in cis-9, trans-11 CLA and VA contents. The results show that the response of sheep and goats to OTL and GM diets was different, suggesting a species difference that needs further investigation.

Sheep and goats differences in CLA and fatty acids milk fat content in relation with mRNA stearoyl-CoA desaturase and lipogenic genes expression in their mammary gland

An experiment was conducted with 12 lactating dairy ewes and 12 goats with the objective to determine whether, under the same dietary treatments, the differences in their fatty acid (FA) profile with emphasis on cis-9 trans-11 CLA milk fat content, are reflected in the transcript levels of genes involved in FA and cis-9, trans-11 CLA biosynthesis. The animals were fed with two diets (A, B) in different days of milk (DIM) due to the different milk yield, body weight etc, in order to have the same food intake and to avoid dietary effects. Diet A was fed to the animals on a group basis as it is traditionally used in practice, while diet B was chosen to avoid individual feed intake variation which is usually observed in group feeding. The results showed that there are significantly lower mRNA levels of acetyl-CoA carboxylase (ACC) in sheep mammary gland compared with those of goats, independently from the diet fed. The same trend was observed with the mRNA level of FA synthase (FAS), but the results were significant only for diet A. The mRNA level of lipoprotein lipase (LPL) in the mammary gland did not differ between sheep and goats fed with diet A. In addition, the concentration of cis-9 trans-11 CLA content was significantly higher in sheep milk fat compared with those of goats. This is in accordance with the significant higher levels on mRNA of stearoyl-CoA desaturase (SCD) which were observed in their mammary adipocytes of sheep compared with those of goats, independently of the fed diet (A or B). In conclusion, these findings demonstrate that the differences between sheep and goats, concerning cis-9, trans-11 CLA and FA milk fat content, under the same dietary treatments could be explained in part by the differences in mRNA of SCD and lipogenic genes in their mammary gland.

Determination of mycotoxins in feedstuffs and ruminant's milk using an easy and simple LC-MS/MS multiresidue method.

Mycotoxin contamination is a common problem on feedstuffs, that can be formed on crops in the field, during harvest, storage, processing or feeding. The scope of the current study was to investigate the levels of Aflatoxin B1, Aflatoxin B2, Aflatoxin G1, Aflatoxin G2, Diacetoxyscirpenol, Ochratoxin A, Toxin HT-2, Toxin T-2 and zearalenone in a variety of feedstuffs (maize silage, alfalfa hay, cottonseed cake, corn grain and concentrates) fed to ruminants and the possible contamination of milk though consumption. For this purpose an easy and simple multiresidue LC-MS/MS method without any clean-up step was developed and successfully validated in feed and milk matrices. The LOQ of the method was set at 10 μg/kg for all analytes and 0.05 μg/kg for Aflatoxin M1 and Ochratoxin A in milk. The results showed that 7 cottonseed cake samples, out of 13 were contaminated with Aflatoxin B1 at a level higher than the maximum levels as set by EU Regulations and with Toxin T-2 with values ranging from 8 to 562 μg/kg. Nine maize silages and 6 alfalfa hay samples were contaminated with Aflatoxin G2 at levels higher than the maximum tolerance limit. No mycotoxins or their metabolites were found above the LOQ in any of the analyzed milk samples.

Changes in milk and plasma fatty acid profile in response to fish and soybean oil supplementation in dairy sheep

An effective strategy for enhancing the bioactive fatty acids (FA) in sheep milk could be dietary supplementation with a moderate level of a combination of soybean oil with fish oil (SFO) without negative effects on milk yield and its chemical composition. Thus, the objective of this study was to determine the effects of a moderate forage diet supplementation with SFO on milk chemical composition and FA profile, as well as on plasma FA. Twelve dairy sheep were assigned to two homogenous sub-groups. Treatments involved a control diet without added oil, and a diet supplemented with 23.6 g soybean oil and 4.7 g fish oil per kg dry matter (DM) of the total ration. The results showed that SFO diet had no effect on milk yield and chemical composition. In blood plasma the concentrations of trans-11 C(18:2) (VA), C(18:2n-6), C(20:5n-3) (EPA) and C(22:6n-3) (DHA) were significantly higher while those of C(14:0), C(16:0) and C(18:0) were lower in sheep fed with SFO diet compared with control. The SFO supplementation of sheep diet increased the concentrations of VA, cis-9, trans-11 C(18:2) CLA, trans-10, cis-12, C(18:2) CLA, EPA, DHA, monounsaturated FA (MUFA), polyusaturated fatty acids (PUFA) and n-3 FA and decreased those of short chain FA (SCFA), medium chain FA (MCFA), the saturated/unsaturated ratio and the atherogenicity index value in milk compared with the control. In conclussion, the SFO supplementation at the above levels in a sheep diet, with moderate forage to concentrate ratio, improved the milk FA profile from human health standpoint without negative effects on its chemical composition.

The effect of long term under- and over-feeding of sheep on milk and plasma fatty acid profiles and on insulin and leptin concentrations.

Since sheeps milk is mainly used for cheese making and milk chemical composition and fatty acids (FA) profile affect cheese yield and quality, the objective of this study was to determine the effects of different feeding levels on milk chemical composition and FA profile, as well as on plasma FA profile, and on insulin and leptin concentrations. Twenty-four sheep were assigned to three homogeneous sub-groups. Throughout the experimental period each group was fed the same diet but in quantities which met 70% (under-feeding), 100% (control) and 130% (over-feeding) of their respective energy and crude protein requirements. The results showed that the underfed sheep had higher milk fat content compared with overfed. In blood plasma the concentrations of C18:0 and C18:1 in the underfed sheep were significantly higher compared with control and overfed sheep. The concentrations of leptin and insulin were significantly higher in overfed compared with underfed sheep. Underfeeding reduced the concentrations of short chain FA (SCFA) and medium chain FA (MCFA) and increased that of C18:0 and mono unsaturated FA (MUFA) in sheep milk fat compared with controls and overfed. The concentrations of C18:0, long chain FA (LCFA) and monounsaturated FA (MUFA) in milk were significantly higher and those of SCFA, MCFA and saturated FA (SFA) significantly lower in the underfed compared with the overfed sheep. In conclusion, long term under- and over-feeding affected the sheep milk chemical composition and FA profile which consequently has an impact on milk products yield (cheese and yogurt) and quality (human health).

The effect of long term under- and over-feeding on the expression of genes related to glucose metabolism in mammary tissue of sheep

Glucose utilisation for lactose synthesis in the mammary gland involves expression of a large number of genes whose nutritional regulation remains poorly defined. In this study, the effect of long term under- and over-feeding on the expression of genes [glucose transporter 1: GLUT1, glucose transporter 3: GLUT3, Sodium glucose contransporter 1: SGLT1, two isoforms of β- (1,4) galactosyltransferase: β- (1,4) GAT1, β- (1,4) GAT3 and α-lactalbumin: LALBA] related to glucose metabolism in sheep mammary tissue (MT) was examined. Twenty-four lactating dairy sheep were divided into three homogenous sub-groups and fed the same ration in quantities which met 70% (underfeeding), 100% (control) and 130% (overfeeding) of their energy and crude protein requirements. The results showed a significant reduction on mRNA of GLUT1 and LALBA gene in the MT of underfed sheep, compared with the respective controls and overfed and a significant reduction on mRNA level of SGLT1 and β- (1,4) GAT1 in the MT of underfed sheep, compared with the overfed ones. A significant increase in the GLUT3 mRNA accumulation in the MT of both under- and over- fed sheep was found. Additionally, a trend of reduction on β- (1,4) GAT3 mRNA level in the MT of the underfed sheep, compared with the overfed, was observed. A close positive relationship was obtained between the mRNA transcripts accumulation of GLUT1, SGLT1, β- (1,4) GAT1 and LALBA gene with the milk lactose content and milk lactose yield respectively. In conclusion, feeding level and consequently nutrient availability, may affect glucose uptake and utilisation in sheep MT by altering the GLUT1, GLUT3, SGLT1, β- (1,4) GAT1 and LALBA gene expression involved in their metabolic pathways.

Changes in essential oil content and composition of Origanum vulgare spp. hirtum during storage as a whole plant or after grinding and mixing with a concentrate ruminant diet

Essential oils (EO) from aromatic plants (AP) are used in ruminants’ diets due to several biological activities. The inclusion of AP in ruminants’ diets can be done as dried material, as they are harvested once a year. However, storage is one of the main factors that affect the EO content and chemical composition. Thus, the objective of this study was to determine the effect of storage, in the shade under ambient conditions, on the EO content and chemical composition (a) of Origanum vulgare spp. hirtum as a whole plant (OP) and/or (b) as the powdered form after mixing with the concentrate diet. The results showed that storage of the whole OP in the shade for six months has a very low EO loss (5.8%), while grinding and mixing OP with concentrates causes different losses, which depend on the OP/concentrate ratio and the storage period.

Effects of soyabean meal- or whey-based diets on lipid metabolism in weaned piglets.

The present study aimed to test the hypothesis that dietary protein source influences lipid metabolism-related parameters weaned piglets. The effects of soyabean meal (SB) and whey proteins (WP) on gene expression of several genes involved in the lipogenic process in liver, visceral (VAT) and subcutaneous (SAT) adipose tissues, plasma insulin concentration and fatty acid (FA) profile were investigated in 18 weaned piglets. Weaned piglets were fed one of two diets containing either SB or WP as the main protein source. Following a 10-h fasting period, plasma insulin concentration and FA profile were assessed at 56 and 72 days of age, whereas gene expression in liver, VAT and SAT was assessed at 72 days of age. Plasma insulin concentration was not affected by diet, although it was 40% lower in SB fed pigs. The SB pigs had lower 14:0 (p < 0.01) and higher 18:3n-3 (p < 0.001) levels in plasma in comparison with WP pigs. However, these changes were attributed to background differences in the dietary FA profile and not to a direct protein source effect. Gene expression of sterol regulatory element-binding protein 1 (SREBP-1) in liver and VAT were lower (p < 0.01 and p < 0.05, respectively) in SB compared to WP fed piglets, but no differences occurred in SAT. No changes were observed in sterol regulatory element-binding protein 2, liver X receptor, peroxisome proliferator-activated receptors α and γ and plasminogen activator inhibitor 1 mRNA levels, either in liver or in adipose tissues. In conclusion, dietary protein source, accompanied likely by side alterations in the dietary composition, affects lipid metabolism in pigs through the downregulation of SREBP-1, which is a crucial determinant of lipogenic process.