Effects of the addition of non-fibre carbohydrates with different rumen degradation rates in dairy cow high-forage diets using the Rumen Simulation Technique.

Nutrient synchronisation of protein and carbohydrates is a promising practice to improve ruminal nutrient utilisation. However, dietary sources supplying these nutrients can vary in ruminal nutrient availability due to differing degradation rates, therefore potentially affecting utilisation of nitrogen (N). The effects of the addition of non-fibre carbohydrates (NFCs) with different rumen degradation rates in high-forage diets on ruminal fermentation, efficiency and microbial flow were investigated in vitro using the Rumen Simulation Technique (RUSITEC). Four diets were tested: control with 100% ryegrass silage (GRS) and substitution of 20% on a DM basis of ryegrass silage with corn grain (CORN), processed corn (OZ) or sucrose (SUC). The four diets were assigned to 16 vessels in two sets of RUSITEC apparatuses in a randomised block design over a 17 d experimental trial; 10 d consisted of adaptation and 7 d for sample collection. Rumen fluid was collected from four rumen-cannulated dry Holstein-Friesian dairy cows and was treated without mixing. Then, rumen fluid from each cow was used to inoculate four vessels, and diet treatments were randomly allocated to each one. This was repeated for all cows resulting in 16 vessels. The inclusion of SUC in ryegrass silage diets improved DM and organic matter digestibility. The only diet to significantly lower ammonia-N concentration compared with GRS was SUC. The outflows of non-ammonia-N, microbial-N, and efficiency of microbial protein synthesis were not affected by diet type. However, the efficiency of nitrogen utilisation was improved by SUC compared with GRS. This indicates that the inclusion of an energy source with a high rumen degradation rate in high-forage diets improves rumen fermentation, digestibility, and N utilisation. Specifically, this effect was observed for the more readily available energy source, SUC, compared with the more slowly degradable NFC sources, CORN and OZ.

The amino acid profile of Camelina sativa seeds correlates with the strongest immune response in dairy ewes.

Camelina sativa is an alternative protein source (with a specific amino acid profile) rich also in bioactive compounds (such as polyunsaturated fatty acids and antioxidants), which have immunomodulatory properties. This study aimed to assess the impact of the dietary inclusion level of Camelina seeds, on the expression levels of key genes involved in ewes' innate immunity. Forty-eight dairy ewes were assigned into four homogenous groups of 12 animals that were fed individually with alfalfa hay, wheat straw, and concentrate. The concentrate of the control group (CON) had no Camelina seeds, while in the treated groups, Camelina seeds (CSs) were incorporated at 6 (CS6), 11 (CS11), and 16% (CS16) in the concentrates, respectively, as partial substitution of both soybean meal and maize grain. The relative transcript levels of the immune-related genes were determined using a real-time PCR platform. The relative transcript levels of toll-interleukin receptor-domain-containing adapter-inducing interferon-ß, tumour necrosis factor receptor-associated factor 3, Interferon regulatory factor 5, and Mitogen-activated protein kinase were upregulated in monocytes of the CS11-fed ewes. Furthermore, in the CS6-fed ewes, the relative transcript levels of Interleukin-1 beta (IL1B) were upregulated in monocytes compared to the CON, while those of IL1B, Interleukin-8, and Interleukin-10 were upregulated in neutrophils compared to the CON and the CS11-fed ewes. The highest inclusion level of CS (CS16) did not have a negative impact on ewes' innate immunity. The response of monocytes on dietary amino acid (mainly threonine, tyrosine, serine, and lysine) changes related to Camelina inclusion is different from that of neutrophils. The observed responses need to be further investigated.

Effects of spent coffee grounds on production traits, haematological parameters, and antioxidant activity of blood and milk in dairy goats.

Spent coffee ground (SCG) is a byproduct of coffee beverage preparation and a potential source of carbohydrate, protein, and phenolic compounds for livestock feeding. In this study, the effects of SCG supplementation in the diet of lactating goats on milk production traits and health status were studied. The antioxidant status of blood and milk was also evaluated. Twenty-four Saanen goats were fed a total mixed ration containing commercial concentrate, soybean, and haylage; they were divided into three groups: control diet (CON), SCG50 (50 g/d SCG), and SCG100 (100 g/d SCG). The experiment lasted 6 weeks. Linear and quadratic contrasts were used to evaluate the effects of the byproduct doses. SCG supplementation did not affect milk production, but influenced some milk fatty acids. SCG supplementation increased the contents of C18:1, cis-9, trans-11 C18:2, odd and branched-chain fatty acids, and total conjugated linoleic acid. Most of the haematological and biochemical parameters were within the physiological range for goats. The basophil, eosinophil, and glucose contents were quadratically affected by SCG, whereas platelet count increased linearly with the SCG dose. The SCG supplementation had a positive effect on the blood antioxidant status, as evidenced by an increase in ferric reducing antioxidant power and a decrease in malondialdehyde. The SCG supplementation had no effect on the milk antioxidant status. The results show that SCG (up to 100 g/d) did not negatively affect milk production and health status in goats. However, quadratic effects on some antioxidant and biochemical parameters suggest that further investigations are necessary, especially with regard to the optimisation of the supplement dose.

Evaluation of nitrogen excretion equations for ryegrass pasture-fed dairy cows.

Accurate and precise estimates of nitrogen (N) excretion in faeces and urine of dairy cattle may provide direct tools to improve N management and thus, to mitigate environmental pollution from dairy production. Empirical equations of N excretion have been evaluated for indoor dairy cattle but there is no evaluation for cows fed high proportions of fresh forage. Therefore, the objective of the current study was to evaluate N excretion equations with a unique data set of zero-grazing experiments. Through literature searches, 89 predictive equations were identified from 13 studies. An independent data set was developed from seven zero-grazing experiments with, in total, 55 dairy Holstein-Friesian cows. Models' performance was evaluated with statistics derived from a mixed-effect model and a simple regression analysis model. Squared sample correlation coefficients were used as indicators of precision and based on either the best linear unbiased predictions (R2BLUP) or model-predicted estimates (R2MDP) derived from the mixed model and simple regression analysis, respectively. The slope (ß0), the intercept (ß1) and the root mean square prediction error (RMSPEm%) were calculated with the mixed-effect model and used to assess accuracy. The root mean square prediction error (RMSPEsr%) and the decomposition of the mean square prediction error were calculated with the simple regression analysis and were used to estimate the error due to central tendency (mean bias), regression (systematic bias), and random variation. Concordance correlation coefficient (CCC) were also calculated with the simple regression analysis model and were used to simultaneously assess accuracy and precision. Considering both analysis models, results suggested that urinary N excretion (UN; R2MDP = 0.76, R2BLUP = 0.89, RMSPEm% = 17.2, CCC = 0.82), total manure N excretion (ManN; R2MDP = 0.83, R2BLUP = 0.90, RMSPEm% = 11.0, CCC = 0.84) and N apparently digested (NAD; R2MDP = 0.97, R2BLUP = 0.97, RMSPEm% = 5.3, CCC = 0.95) were closely related to N intake. Milk N secretion was better predicted using milk yield as a single independent variable (MilkN; R2MDP = 0.77, R2BLUP = 0.97, RMSPEm% = 6.0, CCC = 0.74). Additionally, DM intake was a good predictor of UN and ManN and dietary CP concentration of UN and ManN. Consequently, results suggest that several evaluated empirical equations can be used to make accurate and precise predictions concerning N excretion from dairy cows being fed on fresh forage.

The food for feed concept. Performance of broilers fed hotel food residues.

1. In recent times the use of food waste in animal diets has gained considerable attention because of the increasing demand to cover the needs of human population and the high prices of conventional, arable based, animal feeds.2. The aim of the present study was to investigate the effect of adding dried human food waste to the diet of meat-type chickens (broilers). Two hundred, one-day-old broilers were divided into two treatment groups, with 10 replicate pens containing 10 birds per pen. The duration of the study was 42 days. In the control (C), the diet did not contain any food waste, whereas in the second treatment (T) food waste residues from hotels made up 15% of the diet. Diets had similar crude protein and metabolisable energy content.3. Feed intake and body weight were recorded in order to calculate weight gain and feed conversion ratio (FCR). Carcase and breast muscle yield, the weight of selected internal organs and the level of selected biochemical and haematological parameters were determined. Quality of breast muscle meat was assessed.4. Broilers fed the control treatment consumed more feed and gained more weight compared to broilers fed waste; however, the FCR was similar. No major differences were seen for internal organ weights and haematological parameters, although some differences were observed in colour traits and shear force of meat. It was concluded that there is a potential for use of food waste in broiler diets.

The effect of dietary Chlorella vulgaris inclusion on goat's milk chemical composition, fatty acids profile and enzymes activities related to oxidation.

The impact of dietary supplementation with microalgae on goat's milk chemical composition, fatty acids (FA) profile and enzymes activities related to antioxidant mechanism has not been well documented. Thus, this study aimed to investigate the effects of dietary inclusion of Chlorella vulgaris on the following: (i) milk yield, chemical composition and FA profile, (ii) the activities of superoxide dismutase (SOD), catalase (CAT), glutathione reductase (GR), glutathione transferase (GST) and glutathione peroxidase (GSH-Px) in blood plasma and (iii) the activities of SOD, GR and lactoperoxidase (LPO) in milk of goats. Furthermore, the oxidative stress indicators for measuring total antioxidant and free radical scavenging activity [ferric reducing ability of plasma (FRAP) and 2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays] and oxidative stress biomarkers [malondialdehyde (MDA) and protein carbonyls (PC)] were also determined in blood plasma and milk of the animals. For this purpose, 16 cross-bred goats were divided into two homogenous groups. Each goat of both groups was fed individually with alfalfa hay and concentrates separately. The concentrates of the control group (Control) had no microalgae, while those of the Chlorella group were supplemented with 10 g lyophilized Chlorella vulgaris/kg concentrates (Chlorella). Thus, the average intake was 5.15 g Chlorella vulgaris/kg DM. The results showed that the dietary inclusion of Chlorella vulgaris had not noticeable impact on goat's milk yield, chemical composition and FA profile. Significantly higher SOD (by 10.31%) and CAT (by 18.66%) activities in the blood plasma of goats fed with Chlorella vulgaris compared with the control were found. Moreover, the dietary supplementation with Chlorella vulgaris caused a significant increase in SOD (by 68.84%) activity and a reduction in PC (by 24.07%) content in goat's milk. In conclusion, the Chlorella vulgaris inclusion in goat's diets improved the antioxidant status of both animals and milk.

Effect of under- and overfeeding on sheep and goat milk and plasma enzymes activities related to oxidation.

Twenty-four dairy sheep and goats, respectively, were assigned each to three homogenous subgroups per animal species and fed the same diet in quantities which met 70% (underfeeding), 100% (control) and 130% (overfeeding) of their energy and crude protein requirements. The results showed that the underfed sheep in comparison with the control had significantly lower glutathione reductase (GR), superoxide dismutase (SOD), catalase and glutathione peroxidase (GPX) activities and total antioxidant capacity (measured with Ferric Reducing Ability of Plasma [FRAP] assay) in their blood plasma. A significant increase in the glutathione transferase (GST) and GPX activities, malondialdehyde content and total antioxidant capacity (measured with 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) [ABTS] assay) in the blood plasma of underfed goats compared with controls was observed, while the opposite happened for the GR and SOD activities. The underfeeding in both animal species caused a significant increase in the protein carbonyls (PC) content of their blood plasma. The overfeeding, compared with the control, caused a significant decline in the GPX activity and total antioxidant capacity (measured with FRAP) in the blood plasma of sheep while the opposite happened for the GPX and GST activities in the case of goats. The overfed animals, of both species, compared with the respective controls, had higher PC content in their blood plasma. The feeding level had no noticeable impact on the antioxidants' enzymes activities of milk in both animal species. Moreover, the underfeeding in the blood plasma and the overfeeding in milk of both animal species resulted into a significant increase in the PC content. Finally, only in sheep milk, the underfeeding, compared with the respective control, and overfeeding reduced significantly the total antioxidant capacity (measured with ABTS). The feeding level caused oxidative stress in both organism and milk but the response was different in animal species and needs further investigation.

Effect of soya bean and fish oil inclusion in diets on milk and plasma enzymes from sheep and goat related to oxidation.

This study investigated the effects of dietary inclusion of soya bean oil combined with fish oil (SFO) on the activities of a) superoxide dismutase (SOD), glutathione reductase (GR), catalase (CAT) and glutathione transferase (GST) in blood plasma and b) SOD, GR, CAT and lactoperoxidase (LPO) in the milk of sheep and goats. Furthermore, the oxidative stress indicators for measuring total antioxidant activity and free radical scavenging activity [ferric reducing ability of plasma (FRAP) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays] and oxidative stress biomarkers [malondialdehyde (MDA) and protein carbonyl (PC)] were also determined in the blood plasma and milk of the animals. For this purpose, twelve dairy sheep and twelve dairy goats were assigned each to two homogenous subgroups. Treatments in both animal species involved a control diet without added oil and a diet supplemented with 5% soya bean oil and 1% fish oil. The results showed that the inclusion of SFO in the diets of sheep and goats increased significantly the activities of CAT and GR in their blood plasma. The same effect was observed for the activities of GST and FRAP in the blood plasma of goats. Moreover, the fact that the goats had significantly higher average daily PUFA intake (3.62 g/kg BW0.75 ) compared to sheep (2.51 g/kg BW0.75 ) resulted in an enhancement in the MDA content in their plasma. A significant increase in CAT activity in the milk in both animal species fed with SFO diets was also found. Finally, due to the higher apparent transfer rate of n-3 FA from the diet to the milk in sheep, the PC concentrations were found to be enhanced in their plasma and milk. In conclusion, the impact of dietary SFO supplementation on the oxidative status of body and/or on the milk of small ruminants depends not only on the daily PUFA intake, but also on the amount of n-3 FA that reach their milk.

The effect of dietary supplementation with rumen-protected methionine alone or in combination with rumen-protected choline and betaine on sheep milk and antioxidant capacity.

This study investigated the effects of dietary inclusion of rumen-protected methionine alone or in combination with rumen-protected choline and betaine on: (i) milk yield, chemical composition and fatty acids (FA) profile and (ii) blood plasma glutathione transferase (GST) activity of periparturient ewes. Furthermore, the oxidative stress indicators for measuring total antioxidant and free radical scavenging activity [ferric reducing ability of plasma (FRAP) and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays] were also determined in plasma and milk of ewes. Thus, 45 ewes were divided into three equal groups. Each animal of the control group fed daily with a basal diet. The same diet was offered also in each animal of the other two groups. However, the concentrate fed to M group was supplemented with 2.5 g/kg rumen-protected methionine, while the concentrate fed to MCB group with 5 g/kg of a commercial product which contained a combination of methionine, choline and betaine, all three in rumen-protected form. The results showed that the M diet, compared with the control, increased significantly the ewe's milk fat and the total solids content. Likewise, a tendency for higher milk fat and total solids content in ewes fed the MCB diet was also observed. Both M and MCB diets had not noticeable impact on ewes milk FA profile. Significantly higher FRAP values in the blood plasma of ewes fed the MCB and in the milk of ewes fed with the M diet compared with the control were found. Additionally, significantly higher GST activity in the blood plasma of ewes fed the M diet, compared with the control, was observed. Moreover, a significant increase (by 20%) and a tendency for increase (by 16.72%) in the growth rate of lambs nursing ewes fed with M and MCB diets, respectively, compared to controls, were found.

The effect of dietary Chlorella vulgaris supplementation on micro-organism community, enzyme activities and fatty acid profile in the rumen liquid of goats.

Microalgae might be considered as an alternative source of fat and/or protein for ruminant's diets. However, changes in populations of ruminal micro-organisms associated with biohydrogenation process, methane and ammonia production in response to microalgae dietary supplementation have not been well characterized. Thus, 16 cross-bred goats were divided into two groups. Each goat of both groups was fed individually with alfalfa hay and concentrates separately. The concentrates of the control group had no microalgae while those of the treated group were supplemented with 10 g lyophilized Chlorella vulgaris/kg concentrate (chlor). On the 30th experimental day, samples of rumen fluid were collected for microbial DNA extraction, fatty acid profile and enzyme activity analyses. The results showed that the chlor diet compared with the control increased significantly the populations of Methanosphaera stadtmanae, Methanobrevibacter ruminantium and Methanogens bacteria and protozoa in the rumen of goats. A significant reduction in the cellulase activity and in the abundance of Ruminococcus albus, and a significant increase in the protease activity and in the abundance of Clostridium sticklandii in the rumen liquid of goats fed with the chlor diet, compared with the control, were found. Chlorella vulgaris supplementation promoted the formation of trans C18:1 , trans-11 C18:1 and monounsaturated fatty acids (MUFA), while the proportions of C18:0 and long-chain fatty acids (LCFA) reduced significantly in the rumen liquid of goats. This shift in ruminal biohydrogenation pathway was accompanied by a significant increase in Butyrivibrio fibrisolvens trans C18:1 -producing bacteria. In conclusion, the supplementation of diets with microalgae needs further investigation because it enhances the populations of methane-producing bacteria and protozoa.

The role of bentonite binders in single or concomitant mycotoxin contamination of chicken diets.

Concomitant presence of mycotoxins is more likely to appear than a single mycotoxicosis since many mycotoxigenic fungi grow and produce their toxic metabolites under similar conditions. The present study was designed to evaluate the efficacy of 4 mycotoxin binders to protect meat-type chickens against single and concomitant administration in the feed of two mycotoxins, namely aflatoxin B1 (AFB1) and ochratoxin A (OTA) both at concentration of 0.1 mg/kg. A total of 440 as hatched, d-old, Ross 308 broilers were reared for 42 d. There were 11 dietary treatments. Chickens were fed on either an uncontaminated basal diet, basal diet and AFB1, basal with concomitant presence of AFB1 and OTA, basal diet and three binders A, B and C (1%) with or without AFB1 or basal diet and binder D (0.5%) with or without concomitant presence of AFB1 and OTA. Performance, carcass yield and several biochemical parameters were examined. Mycotoxin concentration in liver and breast muscle samples was determined. Broiler performance under concomitant mycotoxin contamination was poorer than that under single mycotoxicosis. Mycotoxin presence increased relative heart weight compared to that of broilers fed on uncontaminated diets. Only OTA and not AFB1 was detected and only in the liver. OTA concentration was four-fold lower in broilers fed on a diet with binder compared to those fed on contaminated diets without binder. In conclusion, the study revealed that binder composition and presence or not of multiple toxins may be important factors for optimum broiler performance under mycotoxicosis.

The effect of long-term under- and overfeeding on the expression of six major milk proteins' genes in the mammary tissue of goats.

Milk protein synthesis in the mammary gland involves expression of six major milk proteins' genes whose nutritional regulation remains poorly defined. In this study, the effect of long-term under- and overfeeding on the expression of as1-casein: CSN1S1, as2-casein: CSN1S2, ß-casein: CSN2, κ-casein: CSN3, α-lactalbumin: LALBA and ß-lactoglobulin: BLG gene in goat mammary tissue (MT) was examined. Twenty-four lactating dairy goat, at 90-98 days in milk, were divided into three homogenous subgroups and fed the same ration, for 60 days, in quantities which met 70% (underfeeding), 100% (control) and 130% (overfeeding) of their energy and crude protein requirements. The results showed a significant decrease in mRNA of CSN1S2, CSN2, CSN3 and LALBA genes in the MT of underfed goats compared with the overfed and on the CSN1S1 and BLG gene expressions in the MT of underfed goats compared with the respective control and overfed. CSN2 was the most abundant transcript in goat MT relative to the other milk proteins' genes. Significantly positive correlations were observed between the mRNA levels of caseins' and BLG genes with the milk yield. Moreover, a significant correlation was found between the mRNA levels of CSN1S2 with the milk protein, lactose content and lactose yield and also between the LALBA gene expression with the lactose content and lactose yield respectively. In conclusion, the feeding level and consequently the nutrients availability affected the milk lactose content, protein and lactose yield as well as the milk volume by altering the CSN1S1, CSN1S2, CSN2, CSN3, LALBA and BLG gene expression involved in their metabolic pathways.

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.

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.

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.